bluuebunny/crossref_metadata_2025_split huggingface dataset:
Five hundred and twenty‐eight microsatellite markers for ecological genomic investigations using Daphnia
Authors: COLBOURNE J. K. , ROBISON B. , BOGART K. , LYNCH M. 
Published: August 2004

AbstractUntil recently, an enormous effort was needed to apply genomic tools to ecological investigations, especially when striving to uncover the functional mechanisms of phenotypic plasticity and the genetic basis of evolutionary adaptation within natural populations. This present study aimed to develop a genomic resource for an organism ideally suited for functional ecology and evolutionary research. Over 760 unique DNA fragments containing microsatellite loci were isolated and characterized from Daphnia to provide more than 500 molecular markers for constructing a genetic map and for localizing chromosomal regions containing genes of ecological importance via quantitative trait locus analyses. Although primarily developed to genotype members of the Daphnia pulex species complex, a significant fraction of these markers is potentially valuable for population genetics and recombination mapping of distantly related species. Over 60% of markers tested in cross‐specific amplifications are possibly conserved within the subgenus Daphnia, whereas 48 and 18% of tested primers are found to amplify subgenus Hyalodaphnia and subgenus Ctenodaphnia DNA, which represents ∼140 and 200 million years of evolutionary preservation.
Characterization of microsatellite loci in the dwarf eelgrass Zostera noltii (Zosteraceae) and cross‐reactivity with Z. japonica
Authors: COYER J. A. , REUSCH T. B. H. , STAM W. T. , SERRÃO E. A. , PEARSON G. , PROCACCINI G. , OLSEN J. L. 
Published: August 2004

Abstract Zostera noltii is an important species of eelgrass occurring along European, north African, Mediterranean, Black Sea and Azov Sea coasts. Nine microsatellite loci were developed and no linkage disequilibrium was observed. Cross‐amplification was observed for all loci (polymorphic) in Z. japonica; only four loci amplified (monomorphic) in Z. marina.
Cutting Balloon for In‐Stent Restenosis:
Authors: RAHEL BRAIM M. , SUTTORP MAARTEN JAN , BERG JURRIËN M. TEN , BAL EGBERT T. , ERNST SJEF M.P.G. , RENSING BENNO J. , KELDER JOHANNES C. , PLOKKER H.W. THIJS 
Published: August 2004

Introduction: Conventional percutaneous coronary intervention for the treatment of in‐stent restenosis (ISR) has shown a high rate of ISR (30–55%). Considering the need for both extrusion of hyperplastic intima and additional stent expansion, a cutting balloon might be more effective for the treatment of ISR.
Methods: We prospectively assessed the immediate and 8‐month outcome of balloon angioplasty using the Barath Cutting Balloon in 100 consecutive patients (mean age: 60.5 ± 10.8 years, 71% male).
Results: In 73 lesions (73%), a good result was reached with the cutting balloon only. In 21 lesions (21%) postdilatation and in 6 lesions (6%) predilatation with a conventional balloon was necessary. The mean inflation pressure was 8.7 ± 2.0 (range: 6.0–18.0) atm. Before the procedure the mean minimal luminal diameter (MLD) was 0.95 ± 0.45 mm. Quantitative coronary analysis showed a mean diameter stenosis of 65%± 16%. Immediately after the procedure the mean MLD was 2.42 ± 0.54 mm with a mean diameter stenosis of 19%± 13%. Two patients died during the follow‐up period (1 stroke, 1 nonvascular). At 8‐month follow‐up 26 patients (26%) reported to have anginal complaints CCS class II–IV of whom 16 (16%) needed target lesion revascularization.
Conclusion: Treatment of ISR using the Barath Cutting Balloon can be performed safely with good immediate results and a relatively low need for repeated target lesion revascularization at 8‐month follow‐up.
Conflict Behavior in Clinical Couples: Interpersonal Perceptions and Stable Outcomes
Authors: EPSTEIN NATHAN B. , SANTA‐BARBARA JACK 
Published: August 2004

The purpose of the present study was to examine the relation between a couple's perceptions of each other while engaged in a conflict situation, and their management of the conflict itself.One‐hundred‐eighty couples undergoing family therapy interacted in a mixed‐motive game that served as the standard conflict situation. Each couple's interactions were classified into one of four categories, depending upon how they managed this conflict. The interpersonal perceptions of each group were then examined, and several differences were found. Couples who resolved the conflict in a cooperative manner perceived each other as cooperative and themselves expressed more appeasing intentions than any other group. Couples who managed the conflict in a mutually destructive manner perceived each other as competitive and themselves expressed the highest level of exploitative and defensive intentions. Couples who developed either a dominant‐submissive relationship, or those who waivered between cooperating and competing and failed to attain any stable solution to the conflict, also differed from each of the other groups.
Sisterhood‐Brotherhood is Powerful: Sibling Sub‐Systems and Family Therapy
Authors: BANK STEPHEN , KAHN MICHAEL D. 
Published: August 2004

Sibling interaction is an often overlooked aspect of family functioning. Individual development and many family behavior patterns may be attributed to autonomous activities within the sibling sub‐system.A number of phenomena in which siblings have profound influence upon one another are explored. Siblings collude and align with each other, at times help each other resist the powerful vertical influences of parents. Other sibling systems serve to enmesh the youngsters even more with parents. Important sib‐behavior patterns include: the death or departure of siblings; the interplay between the sibling sub‐system and the parenting system; and the roles that “well” siblings play vis‐à‐vis their “sick” siblings during family crises. The sibling relationship is seen as a life‐long process, highly influential throughout the life cycle.Understanding of sibling sub‐system structure and dynamics can lead to more flexible therapeutic interventions. Direct work with siblings provides the therapist with more options and greater leverage in producing change for all siblings, as well as for other family members.
Replication and Cytopathic Effect of Oncolytic Vesicular Stomatitis Virus in Hypoxic Tumor Cells In Vitro and In Vivo
Authors: Connor John H. , Naczki Christine , Koumenis Costas , Lyles Douglas S. 
Published: August 2004

ABSTRACT
Tumor hypoxia presents an obstacle to the effectiveness of most antitumor therapies, including treatment with oncolytic viruses. In particular, an oncolytic virus must be resistant to the inhibition of DNA, RNA, and protein synthesis that occurs during hypoxic stress. Here we show that vesicular stomatitis virus (VSV), an oncolytic RNA virus, is capable of replication under hypoxic conditions. In cells undergoing hypoxic stress, VSV infection produced larger amounts of mRNA than under normoxic conditions. However, translation of these mRNAs was reduced at earlier times postinfection in hypoxia-adapted cells than in normoxic cells. At later times postinfection, VSV overcame a hypoxia-associated increase in α subunit of eukaryotic initiation factor 2 (eIF-2α) phosphorylation and initial suppression of viral protein synthesis in hypoxic cells to produce large amounts of viral protein. VSV infection caused the dephosphorylation of the translation initiation factor eIF-4E and inhibited host translation similarly under both normoxic and hypoxic conditions. VSV produced progeny virus to similar levels in hypoxic and normoxic cells and showed the ability to expand from an initial infection of 1% of hypoxic cells to spread through an entire population. In all cases, virus infection induced classical cytopathic effects and apoptotic cell death. When VSV was used to treat tumors established in nude mice, we found VSV replication in hypoxic areas of these tumors. This occurred whether the virus was administered intratumorally or intravenously. These results show for the first time that VSV has an inherent capacity for infecting and killing hypoxic cancer cells. This ability could represent a critical advantage over existing therapies in treating established tumors.
Characterization of a Highly Pathogenic Molecular Clone of Feline Immunodeficiency Virus Clade C
Authors: de Rozières Sohela , Mathiason Candace K. , Rolston Matthew R. , Chatterji Udayan , Hoover Edward A. , Elder John H. 
Published: August 2004

ABSTRACT

We have derived and characterized a highly pathogenic molecular isolate of feline immunodeficiency virus subtype C (FIV-C) CABCpady00C. Clone FIV-C36 was obtained by lambda cloning from cats that developed severe immunodeficiency disease when infected with CABCpady00C (Abbotsford, British Columbia, Canada). Clone FIV-C36 Env is 96% identical to the noninfectious FIV-C isolate sequence deposited in GenBank (FIV-Cgb; GenBank accession number
AF474246
) (A. Harmache et al.) but is much more divergent in Env when compared to the subgroup A clones Petaluma (34TF10) and FIV-PPR (76 and 78% divergence, respectively). Clone FIV-C36 was able to infect freshly isolated feline peripheral blood mononuclear cells and primary T-cell lines but failed to productively infect CrFK cells, as is typical of FIV field isolates. Two-week-old specific-pathogen-free cats infected with FIV-C36 tissue culture supernatant became PCR positive and developed severe acute immunodeficiency disease similar to that caused by the uncloned CABCpady00C parent. At 4 to 5 weeks postinfection (PI), 3 of 4 animals developed CD4
+
-T-cell depletion, fever, weight loss, diarrhea, and opportunistic infections, including ulcerative stomatitis and tonsillitis associated with abundant bacterial growth, pneumonia, and pyelonephritis, requiring euthanasia. Histopathology confirmed severe thymic and systemic lymphoid depletion. Interestingly, the dam also became infected with a high viral load at 5 weeks PI of the kittens and developed a similar disease syndrome, requiring euthanasia at 11 weeks PI of the kittens. This constitutes the first report of a replication-competent, infectious, and pathogenic molecular clone of FIV-C. Clone FIV-C36 will facilitate dissection of the pathogenic determinants of FIV.
Measles Virus (MV) Hemagglutinin: Evidence that Attachment Sites for MV Receptors SLAM and CD46 Overlap on the Globular Head
Authors: Massé Nicolas , Ainouze Michelle , Néel Benjamin , Wild T. Fabian , Buckland Robin , Langedijk Johannes P. M. 
Published: August 2004

ABSTRACT

Measles virus hemagglutinin (MVH) residues potentially responsible for attachment to the wild-type (wt) MV receptor SLAM (CD150) have been identified and localized on the MVH globular head by reference to a revised hypothetical structural model for MVH (
www.pepscan.nl/downloads/measlesH.pdb
). We show that the mutation of five charged MVH residues which are conserved among morbillivirus H proteins has major effects on both SLAM downregulation and SLAM-dependent fusion. In the three-dimensional surface representation of the structural model, three of these residues (D505, D507, and R533) align the rim on one side of the cavity on the top surface of the MVH globular head and form the basis of a single continuous site that overlaps with the 546-548-549 CD46 binding site. We show that the overlapping sites fall within the footprint of an anti-MVH monoclonal antibody that neutralizes both wt and laboratory-vaccine MV strains and whose epitope contains R533. Our study does not exclude the possibility that Y481 binds CD46 directly but suggests that the N481Y mutation of wt MVH could influence, at a distance, the conformation of the overlapping sites so that affinity to CD46 increases. The relevance of these results to present concepts of MV receptor usage is discussed, and an explanation is proposed as to why morbillivirus attachment proteins are H, whereas those from the other paramyxoviruses are HN (hemagglutinin-neuraminidase).
trans
-Complementation Allows Recovery of Human Respiratory Syncytial Viruses That Are Infectious but Deficient in Cell-to-Cell Transmission
Authors: Oomens A. G. P. , Wertz Gail W. 
Published: August 2004

ABSTRACT

Human respiratory syncytial virus (HRSV) expresses three transmembrane glycoproteins: small hydrophobic protein SH, attachment protein G, and fusion protein F. The genes encoding SH and G can be deleted from the HRSV genome and infectious virus recovered. In contrast, HRSVs lacking the F gene or a functional replacement thereof have not been reported. To generate a system with which to study the roles of the viral transmembrane glycoproteins, including F, in the HRSV life cycle, we generated a cell line expressing a heterologous viral glycoprotein for complementation of glycoprotein function in
trans
. We previously demonstrated that the baculovirus GP64 protein or a chimeric form of GP64 carrying the 12 C-terminal amino acids of the HRSV F protein (GP
64/F
) can efficiently mediate HRSV infectivity and improve its stability, when expressed from an engineered HRSV genome. Here, we report the development of a stably transfected Vero cell line (Vbac) constitutively expressing the GP
64/F
protein. From the Vbac cell line, viruses that lacked the SH and F open reading frames (ORFs) or the SH, G, and F ORFs could be recovered from cDNAs. These viruses, designated RSΔ
sh,f
and RSΔ
sh,g,f
, respectively, had place-keeper ORFs inserted in place of the deleted ORFs to maintain authentic transcription levels. In the Vbac cell line, RSΔ
sh,f
and RSΔ
sh,g,f
could be amplified to near wild-type-level titers, and the resulting viruses were infectious to Vero and HEp-2 cells. After entry into Vero or HEp-2 cells, however, neither virus RSΔ
sh,f
nor virus RSΔ
sh,g,f
was able to spread in the infected cultures. Growth analyses showed that infectious virions were not produced in Vero or HEp-2 cells infected with RSΔ
sh,f
and RSΔ
sh,g,f
. Combined, these data provide direct evidence that HRSV F is an essential viral protein required for cell-to-cell transmission and demonstrate that complementation with the GP64 protein in
trans
constitutes a powerful tool for the study of the role of individual HRSV transmembrane glycoproteins in virus assembly, morphogenesis, and pathogenesis. In addition, the ability to generate infectious but nonspreading viruses may provide an alternative approach for the development of safe and stable HRSV vaccine candidates.
Adjuvant Activities of Novel Cytokines, Interleukin-23 (IL-23) and IL-27, for Induction of Hepatitis C Virus-Specific Cytotoxic T Lymphocytes in HLA-A*0201 Transgenic Mice
Authors: Matsui Masanori , Moriya Osamu , Belladonna Maria Laura , Kamiya Sadahiro , Lemonnier François A. , Yoshimoto Takayuki , Akatsuka Toshitaka 
Published: August 2004

ABSTRACTSearching the sequence databases has revealed two novel cytokines: interleukin-23 (IL-23) and IL-27. These cytokines are quite similar to, but clearly distinct from IL-12 in their structures and T-cell stimulatory fashions. In contrast to IL-12, however, little is known about the roles of IL-23 and IL-27 in the immune regulation. Previously, we evaluated the prime-boost immunization consisting of priming and the first boosting with the hepatitis C virus (HCV)-core expression plasmid, followed by a second boosting with recombinant adenovirus expressing HCV core for induction of HCV core-specific cytotoxic T lymphocytes (CTLs) in BALB/c mice. The present study demonstrates that HCV-specific CTL induction was greatly enhanced by coinoculation of an IL-12 expression plasmid in the prime-boost immunization, indicating the potent adjuvant activity of IL-12. We investigated whether similar adjuvant effects could be exerted by either IL-23 or IL-27 in a prime-boost immunization with HLA-A*0201 transgenic mice. Coadministration of either an IL-23 or an IL-27 expression plasmid, as well as an IL-12 expression plasmid, in a prime-boost immunization enhanced induction of HCV-specific CTLs and led to dramatic increases in the numbers of gamma interferon (IFN-γ)-producing, HCV-specific CD8+cells. Further, preinjections of IL-12, IL-23, or IL-27 expression plasmids before immunization resulted in great increases in the number of IFN-γ-producing, HCV-specific CD8+cells in response to immunization with recombinant adenovirus. These data revealed that both IL-23 and IL-27, as well as IL-12, are potent adjuvants for epitope-specific CTL induction. The two novel cytokines might offer new prophylactic and therapeutic strategies against infectious pathogens such as HCV.
Analysis of Human Immunodeficiency Virus Type 1 Transcriptional Elongation in Resting CD4+T Cells In Vivo
Authors: Lassen Kara G. , Bailey Justin R. , Siliciano Robert F. 
Published: August 2004

ABSTRACTA stable latent reservoir for human immunodeficiency virus type 1 (HIV-1) in resting memory CD4+T cells presents a barrier to eradication of the infection even in patients on highly active antiretroviral therapy. Potential mechanisms for latency include inaccessibility of the integrated viral genome, absence of key host transcription factors, premature termination of HIV-1 RNAs, and abnormal splicing patterns. To differentiate among these mechanisms, we isolated extremely pure populations of resting CD4+T cells from patients on highly active antiretroviral therapy. These cells did not produce virus but retained the capacity to do so if appropriately stimulated. Products of HIV-1 transcription were examined in purified resting CD4+T cells. Although short, prematurely terminated HIV-1 transcripts have been suggested as a marker for latently infected cells, the production of short transcripts had not been previously demonstrated in purified populations of resting CD4+T cells. By separating RNA into polyadenylated and nonpolyadenylated fractions, we showed that resting CD4+T cells from patients on highly active antiretroviral therapy produce abortive transcripts that lack a poly(A) tail and that terminate prior to nucleotide 181. Short transcripts dominated the pool of total HIV-1 transcripts in resting CD4+T cells. Processive, polyadenylated HIV-1 mRNAs were also present at a low level. Both unspliced and multiply spliced forms were found. Taken together, these results show that the nonproductive nature of the infection in resting CD4+T cells from patients on highly active antiretroviral therapy is not due to absolute blocks at the level of either transcriptional initiation or elongation but rather relative inefficiencies at multiple steps.
Conformation- and Fusion-Defective Mutations in the Hypothetical Phospholipid-Binding and Fusion Peptides of Viral Hemorrhagic Septicemia Salmonid Rhabdovirus Protein G
Authors: Rocha A. , Ruiz S. , Tafalla C. , Coll J. M. 
Published: August 2004

ABSTRACT
Fourteen single and two double point mutants in the highly conserved region (positions 56 to 159) of the G gene of viral hemorrhagic septicaemia virus (VHSV), a salmonid rhabdovirus, were selected and obtained in plasmids by site-directed mutagenesis. Fish cell monolayers transfected with the mutant plasmids were then assayed for protein G (pG) expression, conformation-dependent monoclonal antibody (MAb) reactivity, and cell-cell fusion. Some mutations located in the phospholipid-binding p2 peptide (positions 82 to 110; mutants P86A, A96E, G98A, and R107A) abolished both MAb recognition and fusion activity, while others (P79A, L85S, and R103A) abolished MAb recognition but retained fusion at similar or lower pHs compared to those for the wild type. Phospholipid-binding assays of p2-derived synthetic peptides suggested that phosphatidylserine binding was not affected by the mutations studied. On the other hand, three (P79A, L85S, and T135E) of the four mutants retaining fusion activity mapped around two locations showing amino acid variation in 22 VHSV isolates and in neutralizing MAb-resistant mutants described previously. Mutations located in the hypothetical fusion peptide (positions 142 to 159; mutants F147K, P148K, and W154K) abolished both MAb recognition and fusion activity. The existence of mutants with altered conformation and defective fusion in both p2 and fusion peptides provides further evidence in favor of the participation of these and adjacent regions in some of the steps of the VHSV fusion processes, as suggested by previous studies. In addition, because the studied region induced strong immunological responses in trout, some of the mutants described here might be used to design attenuated VHSV vaccines.
Identification of
cis
Sequences Required for Lytic DNA Replication and Packaging of Murine Gammaherpesvirus 68
Authors: Deng Hongyu , Chu Julia T. , Park No-Hee , Sun Ren 
Published: August 2004

ABSTRACT

Human gammaherpesviruses are associated with lymphomas and other malignancies. Murine gammaherpesvirus 68 (MHV-68) infection of mice has emerged as a model for understanding gammaherpesvirus pathogenesis in vivo. In contrast to human gammaherpesviruses, MHV-68 replicates in permissive cell lines in a robust manner, presenting an efficient model to study the basic mechanisms for DNA replication and recombination processes. In addition, MHV-68 also infects a broad range of cells of different tissue types and from different host species, and the viral genome persists as an episome in infected cells. These features make MHV-68 an attractive system on which to build gene delivery vectors. We have therefore undertaken a study to identify the
cis
elements required for MHV-68 genome replication and packaging. Here we report that an 8.4-kb MHV-68 genomic fragment between ORF66 and ORF73 conferred on the plasmid the ability to replicate; replication required the presence of either de novo viral infection or viral reactivation from latency. We further mapped the origin of lytic replication (
ori
Lyt) to a 1.25-kb region. Moreover, we demonstrated that the terminal repeat of the viral genome is sufficient for packaging of the replicated
ori
Lyt plasmid into mature viral particles. Functional identification of the MHV-68
ori
Lyt and packaging signal has laid a foundation for investigating the mechanisms controlling gammaherpesvirus DNA replication during the viral lytic phase and will also serve as a base on which to design gene delivery vectors.
The PA Subunit Is Required for Efficient Nuclear Accumulation of the PB1 Subunit of the Influenza A Virus RNA Polymerase Complex
Authors: Fodor Ervin , Smith Matt 
Published: August 2004

ABSTRACT
The RNA genome of influenza virus is transcribed and replicated by the viral RNA polymerase complex in the cell nucleus. We have generated green fluorescent protein (GFP)-tagged polymerase subunits to study the assembly of the polymerase complex. Our results show that individually expressed polymerase basic protein 1 (PB1) and polymerase acidic protein (PA) subunits were distributed in both the cytoplasm and the nucleus, while the polymerase basic protein 2 (PB2) subunit accumulated in the nucleus. Although it has been reported that PB1 alone accumulates in the nucleus, we demonstrate that PB1 requires the coexpression of PA for efficient nuclear accumulation. Our results support a model which proposes that PB1 and PA are transported into the nucleus as a complex.