bluuebunny/crossref_metadata_2025 huggingface dataset:
Chemical Dependency and Standardized Nursing Language
Authors: Scherb Cindy A. , Cox‐Kolek Marlae
Published: August 2004

BACKGROUND. Chemical dependency (CD) documentation and nursing documentation within a hospital have differed in many ways. The organization's direction toward a multidisciplinary patient record using NANDA, NIC, and NOC standardized nursing languages (SNL) was a challenge for the CD counselors. The use of SNLs caused some discord with the counseling staff as to how a care plan could be individualized with the use of standardized language.

MAIN CONTENT POINTS. The CD counselors needed to resolve some issues before the change in documentation could proceed. The words “standardized language” provoked thoughts of “cookbook” care, so the counselors needed to reconcile the use of SNLs and the ability to develop individualized treatment plans. The counselors were initially uncertain about using nursing languages to describe their practice and the minimal counseling terms within the languages. These issues were overcome by studying the languages and seeing that they did address the practice of the CD counselors even though nursing developed the SNLs. Implementation included education about NANDA, NIC, and NOC. The staff needed time to accept the need for standardized languages to enhance interdisciplinary documentation and to remove the attitude barrier of “we've always done it this way.” The next step was to create and revise care plans and documentation screens.

CONCLUSIONS. The practice of providing care to patients in CD treatment is enhanced by using NANDA, NIC, and NOC because of the ability to develop a true interdisciplinary patient record, retain individualization of patient plans of care, use the outcomes to substantiate discharge status and recommendations, and assess effectiveness through use of outcome data. Pilot outcomes data indicate that CD counselors and nursing exhibit statistically significant results related to coping, decision making, knowledge: substance use control, leisure participation, psychosocial adjustment, life change, substance addiction consequences, self‐esteem, and spiritual well‐being. Future research efforts should include monitoring patient outcomes over the continuum of care. Most patients participate in an aftercare program following inpatient or outpatient treatment. It would be beneficial to follow outcomes throughout the entire course of a patient's treatment.
Spatial Representation in the Entorhinal Cortex
Authors: Fyhn Marianne , Molden Sturla , Witter Menno P. , Moser Edvard I. , Moser May-Britt
Published: August 2004

As the interface between hippocampus and neocortex, the entorhinal cortex is likely to play a pivotal role in memory. To determine how information is represented in this area, we measured spatial modulation of neural activity in layers of medial entorhinal cortex projecting to the hippocampus. Close to the postrhinal-entorhinal border, entorhinal neurons had stable and discrete multipeaked place fields, predicting the rat's location as accurately as place cells in the hippocampus. Precise positional modulation was not observed more ventromedially in the entorhinal cortex or upstream in the postrhinal cortex, suggesting that sensory input is transformed into durable allocentric spatial representations internally in the dorsocaudal medial entorhinal cortex.
Powering Fuel Cells with CO via Aqueous Polyoxometalates and Gold Catalysts
Authors: Kim Won Bae , Voitl T. , Rodriguez-Rivera G. J. , Dumesic J. A.
Published: August 2004

Electricity was produced by catalytic oxidation of carbon monoxide (CO) by using gold catalysts at room temperature. The observed rates are faster than conventional processes operating at 500 kelvin or higher for the conversion of CO with water to produce hydrogen and carbon dioxide through the water-gas shift (WGS). By eliminating the WGS reaction, we remove the need to transport and vaporize liquid water in the production of energy for portable applications. This process can use CO-containing gas streams from the catalytic reforming of hydrocarbons to produce an aqueous solution of reduced polyoxometalate compounds that can be used to generate power. The reduced polyoxometalate can be reoxidized in fuel cells that contain simple carbon anodes.
Direct Measurement of Light Waves
Authors: Goulielmakis E. , Uiberacker M. , Kienberger R. , Baltuska A. , Yakovlev V. , Scrinzi A. , Westerwalbesloh Th. , Kleineberg U. , Heinzmann U. , Drescher M. , Krausz F.
Published: August 2004

The electromagnetic field of visible light performs ∼10
15
oscillations per second. Although many instruments are sensitive to the amplitude and frequency (or wavelength) of these oscillations, they cannot access the light field itself. We directly observed how the field built up and disappeared in a short, few-cycle pulse of visible laser light by probing the variation of the field strength with a 250-attosecond electron burst. Our apparatus allows complete characterization of few-cycle waves of visible, ultraviolet, and/or infrared light, thereby providing the possibility for controlled and reproducible synthesis of ultrabroadband light waveforms.
Transparent, Conductive Carbon Nanotube Films
Authors: Wu Zhuangchun , Chen Zhihong , Du Xu , Logan Jonathan M. , Sippel Jennifer , Nikolou Maria , Kamaras Katalin , Reynolds John R. , Tanner David B. , Hebard Arthur F. , Rinzler Andrew G.
Published: August 2004

We describe a simple process for the fabrication of ultrathin, transparent, optically homogeneous, electrically conducting films of pure single-walled carbon nanotubes and the transfer of those films to various substrates. For equivalent sheet resistance, the films exhibit optical transmittance comparable to that of commercial indium tin oxide in the visible spectrum, but far superior transmittance in the technologically relevant 2- to 5-micrometer infrared spectral band. These characteristics indicate broad applicability of the films for electrical coupling in photonic devices. In an example application, the films are used to construct an electric field–activated optical modulator, which constitutes an optical analog to the nanotube-based field effect transistor.
Plasminogen Is a Critical Host Pathogenicity Factor for Group A Streptococcal Infection
Authors: Sun Hongmin , Ringdahl Ulrika , Homeister Jonathon W. , Fay William P. , Engleberg N. Cary , Yang Angela Y. , Rozek Laura S. , Wang Xixi , Sjöbring Ulf , Ginsburg David
Published: August 2004

Group A streptococci, a common human pathogen, secrete streptokinase, which activates the host's blood clot–dissolving protein, plasminogen. Streptokinase is highly specific for human plasminogen, exhibiting little or no activity against other mammalian species, including mouse. Here, a transgene expressing human plasminogen markedly increased mortality in mice infected with streptococci, and this susceptibility was dependent on bacterial streptokinase expression. Thus, streptokinase is a key pathogenicity factor and the primary determinant of host species specificity for group A streptococcal infection. In addition, local fibrin clot formation may be implicated in host defense against microbial pathogens.
The Melaminophenyl Arsenicals Melarsoprol and Melarsen Oxide Interfere with Thiamine Metabolism in the Fission Yeast
Schizosaccharomyces pombe
Authors: Schweingruber M. Ernst
Published: August 2004

The melaminophenyl arsenical melarsoprol is the main drug used against late-stage sleeping sickness caused by
Trypanosoma brucei
subspecies. Its active metabolite in the human body is melarsen oxide. Here, it is shown that this metabolite inhibits growth of the fission yeast
Schizosaccharomyces pombe
and that its toxicity can be abolished efficiently by thiamine (vitamin B
1
), thiamine analogues, and the pyrimidine moiety of the thiamine molecule. Uptake of melarsen oxide is mediated by a membrane protein (car1p), which is involved in the uptake of thiamine and its pyrimidine moiety. Melarsoprol is taken up by cells in a thiamine- and car1p-dependent manner but is not toxic to cells.
Rotavirus Enterotoxin NSP4 Binds to the Extracellular Matrix Proteins Laminin-β3 and Fibronectin
Authors: Boshuizen J. A. , Rossen J. W. A. , Sitaram C. K. , Kimenai F. F. P. , Simons-Oosterhuis Y. , Laffeber C. , Büller H. A. , Einerhand A. W. C.
Published: August 2004

Rotavirus is the most important cause of viral gastroenteritis and dehydrating diarrhea in young children. Rotavirus nonstructural protein 4 (NSP4) is an enterotoxin that was identified as an important agent in symptomatic rotavirus infection. To identify cellular proteins that interact with NSP4, a two-hybrid technique with
Saccharomyces cerevisiae
was used. NSP4 cDNA, derived from the human rotavirus strain Wa, was cloned into the yeast shuttle vector pGBKT7. An intestinal cDNA library derived from Caco-2 cells cloned into the yeast shuttle vector pGAD10 was screened for proteins that interact with NSP4. Protein interactions were confirmed in vivo by coimmunoprecipitation and immunohistochemical colocalization. After two-hybrid library screening, we repeatedly isolated cDNAs encoding the extracellular matrix (ECM) protein laminin-β3 (amino acids [aa] 274 to 878) and a cDNA encoding the ECM protein fibronectin (aa 1755 to 1884). Using deletion mutants of NSP4, we mapped the region of interaction with the ECM proteins between aa 87 and 145. Deletion analysis of laminin-β3 indicated that the region comprising aa 726 to 875 of laminin-β3 interacts with NSP4. Interaction of NSP4 with either laminin-β3 or fibronectin was confirmed by coimmunoprecipitation. NSP4 was present in infected enterocytes and in the basement membrane (BM) of infected neonatal mice and colocalized with laminin-β3, indicating a physiological interaction. In conclusion, two-hybrid screening with NSP4 yielded two potential target proteins, laminin-β3 and fibronectin, interacting with the enterotoxin NSP4. The release of NSP4 from the basal side of infected epithelial cells and the subsequent binding to ECM proteins localized at the BM may signify a new mechanism by which rotavirus disease is established.
Consequences of Cytotoxic T-Lymphocyte Escape: Common Escape Mutations in Simian Immunodeficiency Virus Are Poorly Recognized in Naïve Hosts
Authors: Friedrich Thomas C. , McDermott Adrian B. , Reynolds Matthew R. , Piaskowski Shari , Fuenger Sarah , de Souza Ivna P. , Rudersdorf Richard , Cullen Candice , Yant Levi J. , Vojnov Lara , Stephany Jason , Martin Sarah , O'Connor David H. , Wilson Nancy , Watkins David I.
Published: August 2004

Cytotoxic T lymphocytes (CTL) are associated with control of immunodeficiency virus infection but also select for variants that escape immune recognition. Declining frequencies of epitope-specific CTL frequencies have been correlated with viral escape in individual hosts. However, escape mutations may give rise to new epitopes that could be recognized by CTL expressing appropriate T-cell receptors and thus still be immunogenic when escape variants are passed to individuals expressing the appropriate major histocompatibility complex class I molecules. To determine whether peptide ligands that have been altered through escape can be immunogenic in new hosts, we challenged naïve, immunocompetent macaques with a molecularly cloned simian immunodeficiency virus (SIV) bearing common escape mutations in three immunodominant CTL epitopes. Responses to the altered peptides were barely detectable in fresh samples at any time after infection. Surprisingly, CTL specific for two of three escaped epitopes could be expanded by in vitro stimulation with synthetic peptides. Our results suggest that some escape variant epitopes evolving in infected individuals do not efficiently stimulate new populations of CTL, either in that individual or upon passage to new hosts. Nevertheless, escape variation may not completely abolish an epitope's immunogenicity. Moreover, since the mutant epitope sequences did not revert to wild type during the study period, it is possible that low-frequency CTL exerted enough selective pressure to preserve epitope mutations in viruses replicating in vivo.
A Long-Distance Translocatable Phloem Protein from Cucumber Forms a Ribonucleoprotein Complex In Vivo with
Hop Stunt Viroid
RNA
Authors: Gómez Gustavo , Pallás Vicente
Published: August 2004

Viroids are highly structured plant pathogenic RNAs that do not code for any protein, and thus, their long-distance movement within the plant must be mediated by direct interaction with cellular factors, the nature of which is presently unknown. In addition to this type of RNAs, recent evidence indicates that endogenous RNAs move through the phloem acting as macromolecular signals involved in plant defense and development. The form in which these RNA molecules are transported to distal parts of the plant is unclear. Viroids can be a good model system to try to identify translocatable proteins that could assist the vascular movement of RNA molecules. Here, we demonstrate by use of immunoprecipitation experiments, that the phloem protein 2 from cucumber (CsPP2) is able to interact in vivo with a viroid RNA. Intergeneric graft assays revealed that both the CsPP2 and the
Hop stunt viroid
RNA were translocated to the scion. The translocated viroid is symptomatic in the nonhost scion, indicating that the translocated RNA is functional. The
CsPP2
gene was cloned and sequenced. The analysis of its primary structure revealed the existence of a potential double-spaced-RNA-binding motif, previously identified in a set of proteins that bind to highly structured RNAs, which could explain its RNA-binding properties. The possible involvement of this phloem protein in assisting the long-distance movement of the viroid RNA within the plant is discussed.
Generation of Influenza A Virus NS2 (NEP) Mutants with an Altered Nuclear Export Signal Sequence
Authors: Iwatsuki-Horimoto Kiyoko , Horimoto Taisuke , Fujii Yutaka , Kawaoka Yoshihiro
Published: August 2004

The NS2 (NEP) protein of influenza A virus contains a highly conserved nuclear export signal (NES) motif in its amino-terminal region (
12
ILMRMSKMQL
21
, A/WSN/33), which is thought to be required for nuclear export of viral ribonucleoprotein complexes (vRNPs) mediated by a cellular export factor, CRM1. However, simultaneous replacement of three hydrophobic residues in the NES with alanine does not affect NS2 (NEP) binding to CRM1, although the virus with these mutations is not viable. To determine the extent of sequence conservation required by the NS2 (NEP) NES for its export function during viral replication, we randomly introduced mutations by degenerative mutagenesis into the region of NS cDNA encoding the NS2 (NEP) NES and then attempted to generate mutant viruses containing these alterations by reverse genetics. Sequence analysis of the recovered viruses showed that although some of the mutants possessed amino acids other than those conserved in the NES, hydrophobicity within this motif was maintained. Nuclear export of vRNPs representing all of the mutant viruses was completely inhibited in the presence of a CRM1 inhibitor, leptomycin B, as was the transport of wild-type virus, indicating that the CRM1-mediated pathway is responsible for the nuclear export of both wild-type and mutant vRNPs. The vRNPs of some of the mutant viruses were exported in a delayed manner, resulting in limited viral growth in cell culture and in mice. These results suggest that the NES motif may be an attractive target for the introduction of attenuating mutations in the production of live vaccine viruses.
Introduction of a Temperature-Sensitive Phenotype into Influenza A/WSN/33 Virus by Altering the Basic Amino Acid Domain of Influenza Virus Matrix Protein
Authors: Liu Teresa , Ye Zhiping
Published: August 2004

Our previous studies with influenza A viruses indicated that the association of M1 with viral RNA and nucleoprotein (NP) is required for the efficient formation of helical ribonucleoprotein (RNP) and for the nuclear export of RNPs. RNA-binding domains of M1 map to the following two independent regions: a zinc finger motif at amino acid positions 148 to 162 and a series of basic amino acids (RKLKR) at amino acid positions 101 to 105. Altering the zinc finger motif of M1 reduces viral growth slightly. A substitution of Ser for Arg at either position 101 or position 105 of the RKLKR domain partially reduces the nuclear export of RNP and viral replication. To further understand the role of the zinc finger motif and the RKLKR domain in viral assembly and replication, we introduced multiple mutations by using reverse genetics to modify these regions of the M gene of influenza virus A/WSN/33. Of multiple mutants analyzed, a double mutant, R101S-R105S, of RKLKR resulted in a temperature-sensitive phenotype. The R101S-R105S double mutant had a greatly reduced ratio of M1 to NP in viral particles and a weaker binding of M1 to RNPs. These results suggest that mutations can be introduced into the RKLKR domain to control viral replication.
Intranasal Immunization with Inactivated Influenza Virus Enhances Immune Responses to Coadministered Simian-Human Immunodeficiency Virus-Like Particle Antigens
Authors: Kang Sang-Moo , Guo Lizheng , Yao Qizhi , Skountzou Ioanna , Compans Richard W.
Published: August 2004

Intranasal immunization with inactivated influenza virus vaccine can provide protective immunity, whereas many other antigens are less effective when used for mucosal immunization. To determine whether influenza virus could enhance immune responses to an antigen coadministered to a mucosal surface, we studied the intranasal immunization of mice with a mixture of simian-human immunodeficiency virus (SHIV) virus-like particles (VLPs) and inactivated influenza virus. Compared to mice immunized with SHIV VLPs alone, mice coimmunized with SHIV VLPs and inactivated influenza virus showed significant increases in serum immunoglobulin G (IgG) and mucosal IgA antibodies specific to the human immunodeficiency virus envelope protein, neutralizing activities, numbers of gamma interferon- and interleukin 4-secreting lymphocytes, and cytotoxic-T-lymphocyte activities. The levels of enhancement of immune response by coimmunization with inactivated influenza virus were equivalent to those induced by inclusion of immunostimulatory CpG oligodeoxynucleotides (CpG DNA). We also observed that SHIV VLPs bind to influenza virus virions, forming mixed aggregates. These results indicate that inactivated influenza virus can play a role as a mucosal adjuvant to coadministered antigens.
Ubiquitous Activation of the Nipah Virus Fusion Protein Does Not Require a Basic Amino Acid at the Cleavage Site
Authors: Moll Markus , Diederich Sandra , Klenk Hans-Dieter , Czub Markus , Maisner Andrea
Published: August 2004

Nipah virus (NiV), a highly pathogenic paramyxovirus, causes a systemic infection in vivo and is able to replicate in cultured cells of many species and organs. Such pantropic paramyxoviruses generally encode fusion (F) proteins with multibasic cleavage sites activated by furin or other ubiquitous intracellular host cell proteases. In contrast, NiV has an F protein with a single arginine (R
109
) at the cleavage site, as is the case with paramyxoviruses that are activated by trypsin-like proteases only present in specific cells or tissues and therefore only cause localized infections. Unlike these viruses, cleavage of the NiV F protein is ubiquitous and does not require the addition of exogenous proteases in cell culture. To determine the importance of the amino acid sequence at the NiV F protein cleavage site for ubiquitous activation, we generated NiV F proteins with mutations around R
109
. Surprisingly, neither the exchange of amino acids upstream of R
109
nor replacement of the basic residue itself interfered with F cleavage. Thus, R
109
is not essential for F cleavage and activation. Our data demonstrate that NiV F-protein activation depends on a novel type of proteolytic cleavage that has not yet been described for any other paramyxovirus F protein. NiV F activation is mediated by a ubiquitous protease that requires neither a monobasic nor a multibasic cleavage site and therefore differs from the furin- or trypsin-like proteases known to activate other ortho- and paramyxovirus fusion proteins.
Two African Swine Fever Virus Proteins Derived from a Common Precursor Exhibit Different Nucleocytoplasmic Transport Activities
Authors: Eulálio A. , Nunes-Correia I. , Carvalho A. L. , Faro C. , Citovsky V. , Simões S. , Pedroso de Lima M. C.
Published: August 2004

African swine fever virus (ASFV), a large icosahedral deoxyvirus, is the causative agent of an economically relevant hemorrhagic disease that affects domestic pigs. The major purpose of the present study was to investigate the nuclear transport activities of the ASFV p37 and p14 proteins, which result from the proteolytic processing of a common precursor. Experiments were performed by using yeast-based nucleocytoplasmic transport assays and by analysis of the subcellular localization of different green fluorescent and Myc fusion proteins in mammalian cells. The results obtained both in yeast and mammalian cells clearly demonstrated that ASFV p14 protein is imported into the nucleus but not exported to the cytoplasm. The ability of p37 protein to be exported from the nucleus to the cytoplasm of both yeast and mammalian cells was also demonstrated, and the results clearly indicate that p37 nuclear export is dependent on the interaction of the protein with the CRM-1 receptor. In addition, p37 was shown to exhibit nuclear import activity in mammalian cells. The p37 protein nuclear import and export abilities described here constitute the first report of a nucleocytoplasmic shuttling protein encoded by the ASFV genome. Overall, the overlapping results obtained for green fluorescent protein fusions and Myc-tagged proteins undoubtedly demonstrate that ASFV p37 and p14 proteins exhibit nucleocytoplasmic transport activities. These findings are significant for understanding the role these proteins play in the replication cycle of ASFV.
Recombinant Interleukin-7 Induces Proliferation of Naive Macaque CD4+and CD8+T Cells In Vivo
Authors: Moniuszko Marcin , Fry Terry , Tsai Wen-Po , Morre Michel , Assouline Brigitte , Cortez Pierre , Lewis Mark G. , Cairns Scott , Mackall Crystal , Franchini Genoveffa
Published: August 2004

Interleukin-7 (IL-7) regulates T-cell homeostasis, and its availability is augmented in lymphopenic hosts. Naive CD8+T cells transferred to lymphopenic mice acquire a memory-like phenotype, raising the possibility that IL-7 is the biological mediator of this effect. Here, we provide direct evidence that IL-7 induces the acquisition of memory-cell markers not only in CD8+T cells but also in CD4+T-cell subsets in immune-competent Indian rhesus macaques. The increase of these memory-like populations was dependent on the dose of the cytokine, and these cells were found in the blood as well as secondary lymphoid organs. Memory-like CD4+and CD8+T cells acquired the ability to secrete tumor necrosis factor alpha and, to a lesser extent, gamma interferon following stimulation with a cognate antigen. The phenotypic change observed in naive T cells was promptly reversed after discontinuation of IL-7. Importantly, IL-7 induced cycling of both CD4+and CD8+central memory and effector memory T cells, demonstrating its contribution to the maintenance of the entire T-cell pool. Thus, IL-7 may be of benefit in the treatment of iatrogenic or virus-induced T-cell depletion.
Human Cytomegalovirus UL76 Encodes a Novel Virion-Associated Protein That Is Able To Inhibit Viral Replication
Authors: Wang Shang-Kwei , Duh Chang-Yih , Wu Cheng-Wen
Published: August 2004

The human cytomegalovirus (HCMV) UL76 gene encodes a highly conserved herpesvirus protein, pUL76, which is able to modulate gene expression in either activation or repression. In this study, two specific transcripts were found to contain the reading frame of UL76, one a 4.5-kb and the other a 5.5-kb tricistronic mRNA encoding the UL76, UL77, and UL78 open reading frames. Both transcripts were expressed with true late kinetics, as revealed by data showing inhibition of production in the presence of phosphonoformic acid. Immediately after viral infection, pUL76 was found in the nuclear fraction and was detected in cells in the presence of the protein synthesis inhibitor cycloheximide. Subsequent virus particle purification and Western blot analysis revealed that two forms of pUL76 are associated within mature virions. The high-molecular-mass protein (H-pUL76) was verified as originating from a free form of pUL76 by cross-linking with an unknown protein(s). By performing a biochemical fractionation experiment with purified virions, we provide evidence that pUL76 and H-pUL76 are associated with the detergent-soluble (envelope) and -insoluble (tegument/capsid) fractions, respectively. Both results were consistent with the images exhibited by immunoelectron microscopy, which showed that the distribution of gold particles labeled by the anti-pUL76 antibody juxtaposed the compartments of the envelope and the tegument/capsid of the virion. Evidence indicated that expression of pUL76 at the immediate-early phase of the viral replication cycle leads to the inhibition of HCMV production. The viral constituent pUL76, with a dominant-negative effect on replication, may provide a novel mechanism for HCMV's resumption of latency.
Capsid Region Involved in Hepatitis A Virus Binding to Glycophorin A of the Erythrocyte Membrane
Authors: Sánchez Glòria , Aragonès Lluís , Costafreda M. Isabel , Ribes Enric , Bosch Albert , Pintó Rosa M.
Published: August 2004

Hepatitis A virus (HAV) has previously been reported to agglutinate human red blood cells at acidic pHs. Treatment of erythrocytes with different enzymes and chemical reagents indicated that HAV attachment is mediated through an interaction with sialylglycoproteins. HAV hemagglutination could be blocked by incubating the virus with glycophorin A, indicating that this sialylglycoprotein is the erythrocyte receptor. The number of receptors used was estimated to be around 500 per cell. At the same time, HAV-induced hemagglutination could also be blocked by either monoclonal antibody H7C27 or an anti-VP3(102-121) ascitic fluid, indicating that lysine 221 of VP1 and the surrounding VP3 residues lining the capsid pit are involved in HAV binding to erythrocytes.
A Consecutive Priming-Boosting Vaccination of Mice with Simian Immunodeficiency Virus (SIV)gag/polDNA and Recombinant Vaccinia Virus Strain DIs Elicits Effective Anti-SIV Immunity
Authors: Someya Kenji , Xin Ke-Qin , Matsuo Kazuhiro , Okuda Kenji , Yamamoto Naoki , Honda Mitsuo
Published: August 2004

To evaluate immunity induced by a novel DNA prime-boost regimen, we constructed a DNA plasmid encoding thegagandpolgenes from simian immunodeficiency virus (SIV) (SIVgag/polDNA), in addition to a replication-deficient vaccinia virus strain DIs recombinant expressing SIVgagandpolgenes (rDIsSIVgag/pol). In mice, priming with SIVgag/polDNA, followed by rDIsSIVgag/polinduced an SIV-specific lymphoproliferative response that was mediated by a CD4+-T-lymphocyte subset. Immunization with either vaccine alone was insufficient to induce high levels of proliferation or Th1 responses in the animals. The prime-boost regimen also induced SIV Gag-specific cellular responses based on gamma interferon secretion, as well as cytotoxic-T-lymphocyte responses. Thus, the regimen of DNA priming and recombinant DIs boosting induced Th1-type cell-mediated immunity, which was associated with resistance to viral challenge with wild-type vaccinia virus expressing SIVgag/pol, suggesting that this new regimen may hold promise as a safe and effective vaccine against human immunodeficiency virus type 1.
High Levels of Human Immunodeficiency Virus Infection of CD8 Lymphocytes Expressing CD4 In Vivo
Authors: Cochrane Alexandra , Imlach Stuart , Leen Clifford , Scott Gordon , Kennedy Dermot , Simmonds Peter
Published: August 2004

Human immunodeficiency virus (HIV)-infected CD8 lymphocytes have been reported in vivo, but the mechanism of infection remains unclear. Experiments using the thy/hu mouse model support export of intrathymically infected CD8 precursors, while recent in vitro data suggest that mature CD8 lymphocytes upregulate CD4 upon activation (generating a CD8brightCD4dimphenotype) and are susceptible to HIV infection. To determine whether these mechanisms operate in vivo and to assess their relative importance in the generation of circulating HIV-infected CD8 lymphocytes, we quantified HIV long terminal repeat (LTR) DNA in CD8+CD4−and CD8brightCD4dimlymphocytes isolated from HIV-infected individuals by fluorescence-activated cell sorting. HIV infection of CD8 lymphocytes was demonstrated in 17 of 19 subjects, with a significant inverse relationship between level of infection and CD4 lymphocyte count (R= −0.73;P< 0.001). The level of HIV infection of CD8brightCD4dimlymphocytes was significantly higher (median, 1,730 HIV LTR copies/106cells;n= 9) than that of CD8+CD4−lymphocytes (undetectable in seven of nine individuals;P< 0.01) and approached that of CD4 lymphocytes from the same individuals (median, 3,660 HIV LTR copies/106cells). CD8brightCD4dimlymphocytes represented 0.8 to 3.3% of total CD8 lymphocytes and were most prevalent in the memory subset. Thus, HIV-infected CD8 lymphocytes commonly circulate in HIV-infected individuals and are generated through infection of activated CD8 lymphocytes rather than through export of intrathymically infected precursors. The high level of infection of CD8brightCD4dimlymphocytes could have a direct role in the decline in CD8 lymphocyte function that accompanies HIV disease progression.
Rheumatoid Factors Induce Signaling from B Cells, Leading to Epstein-Barr Virus and B-Cell Activation
Authors: Yang Lixin , Hakoda Masayuki , Iwabuchi Kazuya , Takeda Tsuyoshi , Koike Takao , Kamatani Naoyuki , Takada Kenzo
Published: August 2004

B-cell antigen receptor signaling is initiated upon binding of the antigen to membrane-bound immunoblobulin (Ig), and the anti-Ig antibody (Ab) mimics this signaling. In B cells latently infected with Epstein-Barr virus (EBV), the same signals induce virus activation. We examine here whether rheumatoid factors (RFs), autoantibodies directed against the Fc portion of IgG, induce EBV and B-cell activation. As a source of RFs, RF-producing lymphoblastoid cell line (LCL) clones were isolated from peripheral blood mononuclear cells (PBMC) and synovial cells from patients with rheumatoid arthritis (RA) by EBV transformation. Burkitt's lymphoma-derived Akata cells, which are highly responsive to EBV activation by anti-Ig Abs, were used for the assay of EBV activation. Akata cells expressed IgG3 as membrane-bound Ig. RFs from a synovium-derived LCL were directed to IgG3 and induced EBV activation in 16 to 18% of Akata cells, whereas RFs from another synovium-derived LCL were directed to IgG1 and did not induce EBV activation. Pretreatment of RFs with the purified Fc fragment of human IgG completely abolished EBV activation. Furthermore, B-cell activation was assessed by incorporation of [3H]thymidine. RFs from synovium-derived LCLs efficiently induced B-cell activation, and the addition of CD40 ligand had a synergistic effect. On the other hand, RFs from PBMC-derived LCLs were polyreactive, had a lower affinity to IgG, and did not induce EBV and B-cell activation. The present findings imply a possible role for RFs as EBV and B-cell activators.
Poly(ADP-Ribose) Polymerase 1 Binds to Kaposi's Sarcoma-Associated Herpesvirus (KSHV) Terminal Repeat Sequence and Modulates KSHV Replication in Latency
Authors: Ohsaki Eriko , Ueda Keiji , Sakakibara Shuhei , Do Eunju , Yada Kaori , Yamanishi Koichi
Published: August 2004

During latency, Kaposi's sarcoma-associated herpesvirus (KSHV) is thought to replicate once and to be partitioned in synchrony with the cell cycle of the host. In this replication cycle, the KSHV terminal repeat (TR) sequence functions as a replication origin, assisted by the latency-associated nuclear antigen (LANA). Thus, TR seems to function as a
cis
element for the replication and partitioning of the KSHV genome. Viral replication and partitioning are also likely to require cellular factors that interact with TR in either a LANA-dependent or -independent manner. Here, we sought to identify factors that associate with TR by using a TR DNA column and found that poly(ADP-ribose) polymerase 1 (PARP1) and known replication factors, including ORC2, CDC6, and Mcm7, bound to TR. PARP1 bound directly to a specific region within TR independent of LANA, and LANA was poly(ADP-ribosyl)ated by PARP1. Drugs such as hydroxyurea and niacinamide, which raise or lower PARP activity, respectively, affected the virus copy number in infected cells. Thus, the poly(ADP-ribosyl)ation status of LANA appears to affect the replication and/or maintenance of the viral genome. Drugs that specifically up-regulate PARP activity may lead to the disappearance of latent KSHV.
Activation of the Tumor-Specific Death Effector Apoptin and Its Kinase by an N-Terminal Determinant of Simian Virus 40 Large T Antigen
Authors: Zhang Ying-Hui , Kooistra Klaas , Pietersen Alexandra , Rohn Jennifer L. , Noteborn Mathieu H. M.
Published: August 2004

Apoptin, a viral death protein derived from chicken anemia virus, displays a number of tumor-specific behaviors. In particular, apoptin is phosphorylated, translocates to the nucleus, and induces apoptosis specifically in tumor or transformed cells, whereas it is nonphosphorylated and remains primarily inactive in the cytoplasm of nontransformed normal cells. Here, we show that in normal cells apoptin can also be activated by the transient transforming signals conferred by ectopically expressed simian virus 40 (SV40) large T antigen (LT), which rapidly induces apoptin's phosphorylation, nuclear accumulation, and the ability to induce apoptosis. Further analyses with mutants of LT showed that the minimum domain capable of inducing all three of apoptin's tumor-specific properties resided in the N-terminal J domain, a sequence which is largely shared by SV40 small t antigen (st). Interestingly, the J domain in st, which lacks its own nuclear localization signal (NLS), required nuclear localization to activate apoptin. These results reveal the existence of a cellular pathway shared by conditions of transient transformation and the stable cancerous or precancerous state, and they support a model whereby a transient transforming signal confers on apoptin both the upstream activity of phosphorylation and the downstream activity of nuclear accumulation and apoptosis induction. Such a pathway may reflect a general lesion contributing to human cancers.
Identification and Characterization of Severe Acute Respiratory Syndrome Coronavirus Replicase Proteins
Authors: Prentice Erik , McAuliffe Josephine , Lu Xiaotao , Subbarao Kanta , Denison Mark R.
Published: August 2004

The severe acute respiratory syndrome coronavirus (SARS-CoV) encodes proteins required for RNA transcription and genome replication as large polyproteins that are proteolytically processed by virus-encoded proteinases to produce mature replicase proteins. In this report, we generated antibodies against SARS-CoV predicted replicase protein and used the antibodies to identify and characterize 12 of the 16 predicted mature replicase proteins (nsp1, nsp2, nsp3, nsp4, nsp5, nsp8, nsp9, nsp12, nsp13, nsp14, nsp15, and nsp16) in SARS-CoV-infected Vero cells. Immunoblot analysis of infected-cell lysates identified proteins of the predicted sizes. Immunofluorescence microscopy detected similar patterns of punctate perinuclear and distributed cytoplasmic foci with all replicase antibodies and as early as 6 h postinfection. Dual-labeling studies demonstrated colocalization of replicase protein nsp8 with nsp2 and nsp3 in cytoplasmic complexes and also with LC3, a protein marker for autophagic vacuoles. Antibodies directed against mouse hepatitis virus (MHV) virions and against the putative RNA-dependent RNA polymerase (Pol) detected SARS-CoV nucleocapsid and nsp12 (Pol), respectively, in SARS-CoV-infected Vero cells. These results confirm the predicted protein processing pattern for mature SARS-CoV replicase proteins, demonstrate localization of replicase proteins to cytoplasmic complexes containing markers for autophagosome membranes, and suggest conservation of protein epitopes in the replicase and nucleocapsid of SARS-CoV and the group II coronavirus, MHV. Further, the results demonstrate the ability of replicase antibodies to detect SARS-CoV-infected cells as early as 6 h postinfection and thus represent important tools for studies of SARS-CoV replication, inhibition, and diagnosis.
A Single Amino Acid Substitution in the Envelope Protein of Chimeric Yellow Fever-Dengue 1 Vaccine Virus Reduces Neurovirulence for Suckling Mice and Viremia/Viscerotropism for Monkeys
Authors: Guirakhoo F. , Zhang Z. , Myers G. , Johnson B. W. , Pugachev K. , Nichols R. , Brown N. , Levenbook I. , Draper K. , Cyrek S. , Lang J. , Fournier C. , Barrere B. , Delagrave S. , Monath T. P.
Published: August 2004

A chimeric yellow fever-dengue 1 (ChimeriVax-DEN1) virus was produced by the transfection of Vero cells with chimeric in vitro RNA transcripts. The cell culture supernatant was subjected to plaque purification for the identification of a vaccine candidate without mutations. Of 10 plaque-purified clones, 1 containing no mutation (clone J) was selected for production of the vaccine virus. During subsequent cell culture passaging of this clone for vaccine production, a single amino acid substitution (K to R) occurred in the envelope (E) protein at residue 204 (E204) (F. Guirakhoo, K. Pugachev, Z. Zhang, G. Myers, I. Levenbook, K. Draper, J. Lang, S. Ocran, F. Mitchell, M. Parsons, N. Brown, S. Brandler, C. Fournier, B. Barrere, F. Rizvi, A. Travassos, R. Nichols, D. Trent, and T. Monath, J. Virol.
78:
4761-4775, 2004). The same mutation was observed in another clone (clone E). This mutation attenuated the virus in 4-day-old suckling mice inoculated by the intracerebral (i.c.) route and led to reduced viremia in monkeys inoculated by the subcutaneous or i.c. route. The histopathology scores of lesions in the brain tissue of monkeys inoculated with either the E204K or E204R virus were reduced compared to those for monkeys inoculated with the reference virus, a commercial yellow fever 17D vaccine (YF-VAX). Both viruses grew to significantly lower titers than YF-VAX in HepG2, a human hepatoma cell line. After intrathoracic inoculation into mosquitoes, both viruses grew to a similar level as YF-VAX, which was significantly lower than that of their wild-type DEN1 parent virus. A comparison of the E-protein structures of nonmutant and mutant viruses suggested the appearance of new intramolecular bonds between residues 204R, 261H, and 257E in the mutant virus. These changes may be responsible for virus attenuation through a change in the pH threshold for virus envelope fusion with the host cell membrane.
Minors’ tobacco possession law violations and intentions to smoke: implications for tobacco control
Authors: Gottlieb N H , Loukas A , Corrao M , McAlister A , Snell C , Huang P P
Published: August 2004

Objectives:To test: (1) whether citation under the Minors in Possession (MIP) law, vicarious citation (knowing someone who was cited), and threat of driving licence suspension are associated with decreased intentions to smoke next year; and (2) whether the policy is differentially enforced.

Subjects:28 249 white, Hispanic, and African American students in grades 6–12 (11–18 years old) participated in the study.

Method:The 86 item anonymous Texas Youth Tobacco Survey was completed by students attending 37 schools in 14 east and central Texas communities.

Results:Hierarchical linear modelling showed that MIP citation was unrelated to the future smoking intentions of most youth. However, there was a negative association between citation and smoking intentions for ever daily smoking youth at four schools. Threat of licence suspension was associated with a lower likelihood of future smoking intentions among ever daily smoking youth and vicarious citation did not deter youth from future smoking. African American and Hispanic youth had a higher probability of being cited than their peers.

Conclusions:Threat of driving licence suspension has the intended effect upon youth who are/were committed smokers and MIP citation has the intended effect upon committed smokers at only four schools. However, differential enforcement of the law based on ethnicity may be occurring. Before drawing firm conclusions, current findings must be replicated with longitudinal data to determine the consequences of citation on subsequent tobacco use.
Short term patterns of early smoking acquisition
Authors: Wellman R J , DiFranza J R , Savageau J A , Dussault G F
Published: August 2004

Objective:To describe short term patterns of smoking acquisition exhibited by adolescent smokers.

Design:Interview records from the prospective development and assessment of nicotine dependence in youth study were examined retrospectively. Interviews were conducted three times per year over 30 months.

Subjects:164 students in grades 7–9 (ages 12–15 years, 86 girls, 78 boys) who had used cigarettes at least twice.

Main outcome measures:A continuous timeline of smoking activity, beginning with the subject’s first use of tobacco and continuing through follow up, was translated into six patterns—abstinent, sporadic, occasional, daily, escalating, and intermittent. Outcome measures were the proportion of subjects starting/ending in each pattern, and the number of transitions per subject between patterns.

Results:There was a general but discontinuous progression from infrequent to more frequent use, with many interspersed periods of not smoking. Escalation to daily smoking was common after the development of dependence symptoms, but was rare among those who did not have symptoms. After the appearance of symptoms, both transitions to heavier daily smoking and attempts at cessation increased.

Conclusions:Movement to heavier, more frequent smoking is generally unidirectional, although many youths attempt to quit one or more times. The appearance of any symptom of dependence altered the subsequent pattern of smoking behaviour. Future investigators might consider using more frequent data points and a continuous timeline to track smoking behaviour.
Prevalence and correlates of internet cigarette purchasing among adult smokers in New Jersey
Authors: Hrywna M , Delnevo C D , Staniewska D
Published: August 2004

Objective:To examine the prevalence and correlates of internet cigarette purchasing among adult smokers.

Design:Analysis of internet purchasing in data from a population based telephone survey of New Jersey households. Logistic regression was used to determine factors associated with internet cigarette purchasing, adjusting for year, demographic, and smoking behaviour variables.

Participants:3447 current cigarette smokers pooled from three cross sectional surveys conducted in 2000, 2001, and 2002.

Main outcome measures:Ever purchasing tobacco and usually buying cigarettes via the internet.

Results:Among all current cigarette smokers, ever having purchased tobacco via the internet increased from 1.1% in 2000 to 6.7% in 2002 and usually buying cigarettes via the internet increased from 0.8% in 2000 to 3.1% in 2002. Among current cigarette smokers with internet access, ever having purchased tobacco via the internet was higher among those who reported smoking 31 or more cigarettes per day (adjusted odds ratio (OR) 3.9, 95% confidence interval (CI) 1.5 to 10.2) and those without a past year quit attempt (adjusted OR 1.8, 95% CI 1.1 to 3.0). Usually purchasing cigarettes via the internet was higher among those aged 45–64 years (adjusted OR 4.4, 95% CI 1.1 to 17.1) and who reported having their first cigarette ⩽ 30 minutes after waking (adjusted OR 3.3, 95% CI 1.2 to 9.2).

Conclusions:Although higher prices are known to reduce the demand for cigarettes, internet cigarette purchasing is likely to weaken this effect, particularly among heavy, more dependent smokers who are less interested in quitting.
Public health foundations and the tobacco industry: lessons from Minnesota
Authors: Ibrahim J K , Tsoukalas T H , Glantz S A
Published: August 2004

Objective:To investigate whether private foundations can be created in a way that will insulate them from attacks by the tobacco industry, using the Minnesota Partnership for Action Against Tobacco (MPAAT) as a case study.

Design:Information was collected from internal tobacco industry documents, court documents, newspapers, and interviews with health advocates and elected officials.

Results:The creation of MPAAT as an independent foundation did not insulate it from attacks by tobacco industry allies. During 2001–2002, MPAAT was repeatedly attacked by Attorney General Mike Hatch and major media, using standard tobacco industry rhetoric. This strategy of attack and demands for information were reminiscent of previous attacks on Minnesota’s Plan for Nonsmoking and Health and the American Stop Smoking Intervention Study (ASSIST). MPAAT was ultimately forced to restructure its programme to abandon effective community norm change interventions around smoke-free policies and replace them with less effective individual cessation interventions. Neither MPAAT nor other health advocates mounted an effective public response to these attacks, instead relying on the insider strategy of responding in court.

Conclusion:It is not possible to avoid attacks by the tobacco industry or its political allies. Like programmes administered by government agencies, tobacco control foundations must be prepared for these attacks, including a proactive plan to educate the public about the principles of community based tobacco control. Public health advocates also need to be willing to take prompt action to defend these programmes and hold public officials who attack tobacco control programmes accountable for their actions.
Fine structure of Rossipollis reticulatus from the Upper Cretaceous of southern Sweden
Authors: Skarby Annie , Morbelli Marta A , Rowley John R
Published: August 2004

Dispersed monosulcate pollen identified as Rossipollis reticulatus Ross and similar pollen in two pollen sacs from a single anther were recovered from Upper Cretaceous fluvial sediments in southern Sweden. In most pollen grains the wall consists of a foot layer, an inner solid reticulum, crowned by columellae, which appear as an infratectal reticulum, and a tectum with irregularly spaced fine perforations. An additional papillate innermost layer, interpreted as a transitory endexin, is present in some pollen grains. A monocotyledonous origin is indicated by the configuration of exine and aperture along with the presence of papillate endexine structures in grains considered to have aborted at earlier stages of development.Key words: Cretaceous, fossil pollen, endexine, monocots.