bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization and cross‐species amplification of microsatellite loci in the plant pathogenic fungus Leptosphaeria maculans
Authors: HAYDEN HELEN L. , WILSON LEANNE M. , COZIJNSEN ANTON J. , HOWLETT BARBARA J. 
Published: August 2004

AbstractSeven polymorphic microsatellite markers suitable for population genetic studies and genetic mapping were developed for Leptosphaeria maculans, a fungal pathogen of canola (Brassica napus). Polymorphism was evaluated using 14 isolates from diverse geographical locations. Each locus had either two or three alleles. Cross‐species amplification was observed for almost all loci in L. biglobosa ‘brassicae’ and L. maculans ‘lepidii’.
Microsatellite markers in coast redwood (Sequoia sempervirens)
Authors: BRUNO D. , BRINEGAR C. 
Published: August 2004

AbstractFive polymorphic microsatellite markers are described for coast redwood, a hexaploid conifer. Dinucleotide (three), trinucleotide (one) and tetranucleotide (one) repeat loci were isolated from a genomic plasmid library and amplified in a test population of 25 second‐growth redwoods. There were three to 11 alleles per locus and observed heterozygosities ranged from 0.00 to 0.80. The nonheterozygous locus (a CTTA repeat) is probably of chloroplast origin and may have potential as a marker for paternal inheritance.
Characterization and isolation of DNA microsatellite primers in the spiny dogfish (Squalus acanthias)
Authors: McCAULEY LINDA , GOECKER COURTNEY , PARKER PAUL , RUDOLPH TOM , GOETZ FREDERICK , GERLACH GABRIELE 
Published: August 2004

AbstractThe North Atlantic spiny dogfish (Squalus acanthias) population has been declining since the 1980s. Proper management of this population is essential as dogfish are prone to rapid collapse based on the unique biology of this species. We characterized eight microsatellite loci for spiny dogfish to use in determining the genetic structure of this species along the Atlantic coast of the USA.
Characterization of six microsatellite primers for the grey fox (Urocyon cinereoargenteus)
Authors: WESTON JULIE L. , SCHABLE NANCY A. , GLENN TRAVIS C. 
Published: August 2004

AbstractWe describe polymerase chain reaction (PCR) primers and conditions to amplify one dinucleotide and five tetranucleotide microsatellite DNA loci isolated from the grey fox (Urocyon cinereoargenteus). The PCR primers were tested on nine to 12 individuals collected from the Department of Energy's Savannah River site (Aiken, SC, USA). The grey fox microsatellite primers developed had three to 10 alleles per locus that yielded observed heterozygosities ranging from 0.222 to 0.889.
Long‐Term Results of Endovascular Exclusion of Extracranial Internal Carotid Artery Aneurysms and Dissecting Aneurysm
Authors: BERGERON Patrice , KHANOYAN Patrick , MEUNIER Jean‐Pierre , GRAZIANI Jean‐Noël , GAY Joel 
Published: August 2004

Objectives: Surgery remains the standard option to treat symptomatic or complicated aneurysms of the extracranial internal carotid arteries (EICA). When located more distally to the EICA, surgery appears to be very invasive and disabling. Endovascular treatment of high aneurysmal EICA has been poorly reported. We report our experience in this particular field.
Methods: We treated five EICA endovascularly, using covered stents and stentgrafts in four patients, two males and two females. One male was treated bilaterally. The average age was 59.2 years (39–80). Three patients were symptomatic (two transient ischemic attack and one stroke). Patients were followed by duplex scan, CT scan, or angio MR.
Results: Protecting devices were used in two cases. No in‐hospital complication was observed. During follow‐up (3.6 ± 1.3 years), no adverse event was observed and all devices remained patent at duplex scan and angiography. One early endoleak was observed and treated with covered stent extension. No sign of in‐stent stenosis was observed. All the aneurysmal sacs thrombosed.
Conclusion: Covered stents and stentgrafts allow a less invasive approach to treat highly located internal carotid aneurysms. Larger series are needed to assess the role of covered stents in treating aneurysmal EICA as first choice.
UK patients with primary Sjögren's syndrome are at increased risk from clinical depression
Authors: Stevenson H. A. , Jones M. E. , Rostron J. L. , Longman L. P. , Field E. A. 
Published: August 2004

Objective:  This study was undertaken to assess the presence and degree of anxiety and depression in a group of UK patients with primary Sjögren's syndrome (1°SS).Design:  Cross‐sectional.Setting:  Department of Oral Medicine, Liverpool University Dental Hospital.Subjects:  Eighty adult patients; 40 diagnosed with 1°SS according to the revised European Criteria and 40 age/gender‐matched controls with no history of chronic illness.Intervention:  Hospital Anxiety and Depression Scale (HADS), a self‐administered questionnaire designed to evaluate the presence and degree of anxiety and depression in a clinical setting.Main outcome measures:  Age, gender, Hospital Anxiety and Depression Scale (HADS).Results:  Forty patients with 1°SS and 40/age/gender‐matched controls completed the HADS. Scores for anxiety in both the 1°SS and control groups showed no statistically significant difference. Patients with 1°SS had statistically significant higher, mean HADS scores for depression than the controls. There was an increased prevalence of ‘definite’ clinical depression in the 1°SS group.Conclusion:  Patients with 1°SS appear to be at increased risk from clinical depression. Early recognition and appropriate intervention is therefore essential to reduce the negative impact of depression on the patient's quality of life and outcome of their disease.
Association between functional dental state and dietary intake of Chinese vegetarian old age home residents
Authors: Kwok Timothy , Yu Charles N.F. , Hui Hing W. , Kwan Mandy , Chan Veronica 
Published: August 2004

Objective:  To examine the association between dental functional status and dietary intakes of Chinese vegetarian old age home residents.Subjects and methods:  A cross‐sectional survey was performed in a group of 76 older vegetarian Chinese women living in an old age home. Oral examination was performed by a dental surgeon. Sixty‐eight of them had a 24‐hour food record by direct observation, analysed by a Chinese food composition table. Nutritional status was measured by Body Mass Index (BMI), and the functional status by modified Barthel Index (BI).Results:  Forty‐two subjects (55.3%) were edentulous; 59 subjects (77.6%) had chewing difficulties; 35 subjects (46%) had poor dental functional status defined by having five or less functional teeth unit (FTU). When compared with those older people with better dental functional status, the poor dental functional status group was more likely to have chewing difficulties, tolerate soft diet only, and be functionally dependent (BI < 15/20). Poor dental functional status was associated with lower mean daily fibre intake, but not with intakes of macronutrients and micronutrients, after adjustment by BI categories.Conclusion:  Poor dental functional status is associated with impaired chewing and lower fibre intake in Chinese vegetarian old age home residents, but it is not associated with reduced intakes of macronutrients or micronutrients.
Phospholipid Scramblase 1 Potentiates the Antiviral Activity of Interferon
Authors: Dong Beihua , Zhou Quansheng , Zhao Ji , Zhou Aimin , Harty Ronald N. , Bose Santanu , Banerjee Amiya , Slee Roger , Guenther Jeanna , Williams Bryan R. G. , Wiedmer Therese , Sims Peter J. , Silverman Robert H. 
Published: August 2004

ABSTRACTPhospholipid scramblase 1 (PLSCR1) is an interferon (IFN)- and growth factor-inducible, calcium-binding protein that either inserts into the plasma membrane or binds DNA in the nucleus depending on its state of palmyitoylation. In certain hematopoietic cells, PLSCR1 is required for normal maturation and terminal differentiation from progenitor cells as regulated by select growth factors, where it promotes recruitment and activation of Src kinases. PLSCR1 is a substrate of Src (and Abl) kinases, and transcription of the PLSCR1 gene is regulated by the same growth factor receptor pathways in which PLSCR1 potentiates afferent signaling. The marked transcriptional upregulation of PLSCR1 by IFNs led us to explore whether PLSCR1 plays an analogous role in cellular responses to IFN, with specific focus on antiviral activities. Accordingly, human cells in which PLSCR1 expression was decreased with short interfering RNA were rendered relatively insensitive to the antiviral activity of IFNs, resulting in higher titers of vesicular stomatitis virus (VSV) and encephalomyocarditis virus. Similarly, VSV replicated to higher titers in mousePLSCR1−/−embryonic fibroblasts than in identical cells transduced to express PLSCR1. PLSCR1 inhibited accumulation of primary VSV transcripts, similar to the effects of IFN against VSV. The antiviral effect of PLSCR1 correlated with increased expression of a subset of IFN-stimulated genes (ISGs), including ISG15, ISG54, p56, and guanylate binding proteins. Our results suggest that PLSCR1, which is itself an ISG-encoded protein, provides a mechanism for amplifying and enhancing the IFN response through increased expression of a select subset of potent antiviral genes.
Palindrome Regeneration by Template Strand-Switching Mechanism at the Origin of DNA Replication of Porcine Circovirus via the Rolling-Circle Melting-Pot Replication Model
Authors: Cheung Andrew K. 
Published: August 2004

ABSTRACT
Palindromic sequences (inverted repeats) flanking the origin of DNA replication with the potential of forming single-stranded stem-loop cruciform structures have been reported to be essential for replication of the circular genomes of many prokaryotic and eukaryotic systems. In this study, mutant genomes of porcine circovirus with deletions in the origin-flanking palindrome and incapable of forming any cruciform structures invariably yielded progeny viruses containing longer and more stable palindromes. These results suggest that origin-flanking palindromes are essential for termination but not for initiation of DNA replication. Detection of template strand switching in the middle of an inverted repeat strand among the progeny viruses demonstrated that both the minus genome and a corresponding palindromic strand served as templates simultaneously during DNA biosynthesis and supports the recently proposed rolling-circle “melting-pot” replication model. The genome configuration presented by this model, a four-stranded tertiary structure, provides insights into the mechanisms of DNA replication, inverted repeat correction (or conversion), and illegitimate recombination of any circular DNA molecule with an origin-flanking palindrome.
Inhibition of Hepatitis C Virus-Like Particle Binding to Target Cells by Antiviral Antibodies in Acute and Chronic Hepatitis C
Authors: Steinmann Daniel , Barth Heidi , Gissler Bettina , Schürmann Peter , Adah Mohammed I. , Gerlach J. Tilman , Pape Gerd R. , Depla Erik , Jacobs Dirk , Maertens Geert , Patel Arvind H. , Inchauspé Geneviève , Liang T. Jake , Blum Hubert E. , Baumert Thomas F. 
Published: August 2004

ABSTRACTHepatitis C virus (HCV) is a leading cause of chronic viral hepatitis worldwide. The study of antibody-mediated virus neutralization has been hampered by the lack of an efficient and high-throughput cell culture system for the study of virus neutralization. The HCV structural proteins have been shown to assemble into noninfectious HCV-like particles (HCV-LPs). Similar to serum-derived virions, HCV-LPs bind and enter human hepatocytes and hepatoma cell lines. In this study, we developed an HCV-LP-based model system for a systematic functional analysis of antiviral antibodies from patients with acute or chronic hepatitis C. We demonstrate that cellular HCV-LP binding was specifically inhibited by antiviral antibodies from patients with acute or chronic hepatitis C in a dose-dependent manner. Using a library of homologous overlapping envelope peptides covering the entire HCV envelope, we identified an epitope in the N-terminal E2 region (SQKIQLVNTNGSWHI; amino acid positions 408 to 422) as one target of human antiviral antibodies inhibiting cellular particle binding. Using a large panel of serum samples from patients with acute and chronic hepatitis C, we demonstrated that the presence of antibodies with inhibition of binding activity was not associated with viral clearance. In conclusion, antibody-mediated inhibition of cellular HCV-LP binding represents a convenient system for the functional characterization of human anti-HCV antibodies, allowing the mapping of envelope neutralization epitopes targeted by naturally occurring antiviral antibodies.
Global Effects of Human Papillomavirus Type 18 E6/E7 in an Organotypic Keratinocyte Culture System
Authors: Garner-Hamrick Peggy A. , Fostel J. M. , Chien Wei-Ming , Banerjee N. Sanjib , Chow Louise T. , Broker Thomas R. , Fisher Chris 
Published: August 2004

ABSTRACT
The effects of human papillomavirus type 18 (HPV-18) E6 and E7 proteins on global patterns of host gene expression in primary human keratinocytes grown in organotypic raft culture system were assessed. Primary human keratinocytes were infected with retroviruses that express the wild-type HPV-18 E6 and E7 genes from the native differentiation-dependent HPV enhancer-promoter. Total RNA was isolated from raft cultures and used to generate probes for querying Affymetrix U95A microarrays, which contain >12,500 human gene sequences. Quadruplicate arrays of each E6/E7-transduced and empty vector-transduced samples were analyzed by 16 pairwise comparisons. Transcripts altered in ≥12 comparisons were selected for further analysis. With this approach, HPV-18 E6/E7 expression significantly altered the expression of 1,381 genes. A large increase in transcripts associated with DNA and RNA metabolism was observed, with major increases noted for transcription factors, splicing factors, and DNA replication elements, among others. Multiple genes associated with protein translation were downregulated. In addition, major alterations were found in transcripts associated with the cell cycle and cell differentiation. Our study provides a systematic description of transcript changes brought about by HPV-18 E6/E7 in a physiologically relevant model and should furnish a solid source of information to guide future studies.
Subcellular Localization and Calcium and pH Requirements for Proteolytic Processing of the Hendra Virus Fusion Protein
Authors: Pager Cara Theresia , Wurth Mark Allen , Dutch Rebecca Ellis 
Published: August 2004

ABSTRACT

Proteolytic cleavage of the Hendra virus fusion (F) protein results in the formation of disulfide-linked F
1
and F
2
subunits, with cleavage occurring after residue K
109
in the sequence GDVK↓L. This unusual cleavage site and efficient propagation of Hendra virus in a furin-deficient cell line indicate that the Hendra F protein is not cleaved by furin, the protease responsible for proteolytic activation of many viral fusion proteins. To identify the subcellular site of Hendra F processing, Vero cells transfected with pCAGGS-Hendra F or pCAGGS-SV5 F were metabolically labeled and chased in the absence and presence of inhibitors of exocytosis. The addition of carbonyl-cyanide-3-chlorophenylhydrazone, monensin, brefeldin A, or NaF-AlCl
3
or incubation of cells at 20°C all inhibited processing of the Hendra F protein, suggesting that cleavage of Hendra F occurs either in secretory vesicles budding from the
trans
-Golgi network or at the cell surface. In contrast to proteolytic cleavage of the simian virus 5 (SV5) F protein by the Ca
2+
-dependent protease furin, proteolytic cleavage of the Hendra F protein was not significantly inhibited by decreases in Ca
2+
levels following incubation with EGTA or A23187. However, in the presence of weak amines and H
+
V-ATPase inhibitors, known to raise intracellular pH, cleavage of Hendra F protein was inhibited while processing of the SV5 F protein was not significantly affected. The subcellular location, sensitivity to pH changes, and decreased Ca
2+
requirement suggest that the protease responsible for cleavage of Hendra F protein differs from proteases previously shown to be involved in the processing of other viral glycoproteins.
Glycosylation of Immunodominant Linear Epitopes in the Carboxy-Terminal Region of the Caprine Arthritis-Encephalitis Virus Surface Envelope Enhances Vaccine-Induced Type-Specific and Cross-Reactive Neutralizing Antibody Responses
Authors: Trujillo J. D. , Kumpula-McWhirter N. M. , Hötzel K. J. , Gonzalez M. , Cheevers W. P. 
Published: August 2004

ABSTRACTThis study evaluated type-specific and cross-reactive neutralizing antibodies induced by immunization with modified surface glycoproteins (SU) of the 63 isolate of caprine arthritis-encephalitis lentivirus (CAEV-63). Epitope mapping of sera from CAEV-infected goats localized immunodominant linear epitopes in the carboxy terminus of SU. Two modified SU (SU-M and SU-T) and wild-type CAEV-63 SU (SU-W) were produced in vaccinia virus and utilized to evaluate the effects of glycosylation or the deletion of immunodominant linear epitopes on neutralizing antibody responses induced by immunization. SU-M contained two N-linked glycosylation sites inserted into the target epitopes by R539S and E542N mutations. SU-T was truncated at 518A, upstream from the target epitopes, by introduction of termination codons at 519Y and 521Y. Six yearling Saanen goats were immunized subcutaneously with 30 μg of SU-W, SU-M, or SU-T in Quil A adjuvant and boosted at 3, 7, and 16 weeks. SU antibody titers determined by indirect enzyme-linked immunosorbent assay demonstrated anamnestic responses after each boost. Wild-type and modified SU-induced type-specific CAEV-63 neutralizing antibodies and cross-reactive neutralizing antibodies against CAEV-Co, a virus isolate closely related to CAEV-63, and CAEV-1g5, an isolate geographically distinct from CAEV-63, were determined. Immunization with SU-T resulted in altered recognition of SU linear epitopes and a 2.8- to 4.6-fold decrease in neutralizing antibody titers against CAEV-63, CAEV-Co, and CAEV-1g5 compared to titers of SU-W-immunized goats. In contrast, immunization with SU-M resulted in reduced recognition of glycosylated epitopes and a 2.4- to 2.7-fold increase in neutralizing antibody titers compared to titers of SU-W-immunized goats. Thus, the glycosylation of linear immunodominant nonneutralization epitopes, but not epitope deletion, is an effective strategy to enhance neutralizing antibody responses by immunization.
Unique Features of Hepatitis C Virus Capsid Formation Revealed by De Novo Cell-Free Assembly
Authors: Klein Kevin C. , Polyak Stephen J. , Lingappa Jaisri R. 
Published: August 2004

ABSTRACTThe assembly of hepatitis C virus (HCV) is poorly understood, largely due to the lack of mammalian cell culture systems that are easily manipulated and produce high titers of virus. This problem is highlighted by the inability of the recently established HCV replicon systems to support HCV capsid assembly despite high levels of structural protein synthesis. Here we demonstrate that up to 80% of HCV core protein synthesized de novo in cell-free systems containing rabbit reticulocyte lysate or wheat germ extracts assembles into HCV capsids. This contrasts with standard primate cell culture systems, in which almost no core assembles into capsids. Cell-free HCV capsids, which have a sedimentation value of ≈100S, have a buoyant density (1.28 g/ml) on cesium chloride similar to that of HCV capsids from other systems. Capsids produced in cell-free systems are also indistinguishable from capsids isolated from HCV-infected patient serum when analyzed by transmission electron microscopy. Using these cell-free systems, we show that HCV capsid assembly is independent of signal sequence cleavage, is dependent on the N terminus but not the C terminus of HCV core, proceeds at very low nascent chain concentrations, is independent of intact membrane surfaces, and is partially inhibited by cultured liver cell lysates. By allowing reproducible and quantitative assessment of viral and cellular requirements for capsid formation, these cell-free systems make a mechanistic dissection of HCV capsid assembly possible.
Evidence of Unique Genotypes of
Beak and Feather Disease Virus
in Southern Africa
Authors: Heath Livio , Martin Darren P. , Warburton Louise , Perrin Mike , Horsfield William , Kingsley Chris , Rybicki Edward P. , Williamson Anna-Lise 
Published: August 2004

ABSTRACT

Psittacine beak and feather disease (PBFD), caused by
Beak and feather disease virus
(BFDV), is the most significant infectious disease in psittacines. PBFD is thought to have originated in Australia but is now found worldwide; in Africa, it threatens the survival of the indigenous endangered Cape parrot and the vulnerable black-cheeked lovebird. We investigated the genetic diversity of putative BFDVs from southern Africa. Feathers and heparinized blood samples were collected from 27 birds representing 9 psittacine species, all showing clinical signs of PBFD. DNA extracted from these samples was used for PCR amplification of the putative BFDV coat protein (CP) gene. The nucleotide sequences of the CP genes of 19 unique BFDV isolates were determined and compared with the 24 previously described sequences of BFDV isolates from Australasia and America. Phylogenetic analysis revealed eight BFDV lineages, with the southern African isolates representing at least three distinctly unique genotypes; 10 complete genome sequences were determined, representing at least one of every distinct lineage. The nucleotide diversity of the southern African isolates was calculated to be 6.4% and is comparable to that found in Australia and New Zealand. BFDVs in southern Africa have, however, diverged substantially from viruses found in other parts of the world, as the average distance between the southern African isolates and BFDV isolates from Australia ranged from 8.3 to 10.8%. In addition to point mutations, recombination was found to contribute substantially to the level of genetic variation among BFDVs, with evidence of recombination in all but one of the genomes analyzed.
Blocking of the Alpha Interferon-Induced Jak-Stat Signaling Pathway by Japanese Encephalitis Virus Infection
Authors: Lin Ren-Jye , Liao Ching-Len , Lin Elong , Lin Yi-Ling 
Published: August 2004

ABSTRACTThe induction of alpha/beta interferon (IFN-α/β) is a powerful host defense mechanism against viral infection, and many viruses have evolved strategies to overcome the antiviral effects of IFN. In this study, we found that IFN-α had only some degree of antiviral activity against Japanese encephalitis virus (JEV) infection, in contrast to another flavivirus, dengue virus serotype 2, which was highly sensitive to IFN-α in the cultured cell system. JEV infection appeared to render cells resistant to IFN-α since the IFN-α-induced luciferase reporter activity driven by the IFN-stimulated response element (ISRE) was gradually reduced as the JEV infection progressed. Since the biological activities of IFNs are triggered by the Janus kinase (Jak) signal transducer and activation of transcription (Stat) signaling cascade, we then studied the activation of Jak-Stat pathway in the virus-infected cells. The IFN-α-stimulated tyrosine phosphorylation of Stat1, Stat2, and Stat3 was suppressed by JEV in a virus replication and de novo protein synthesis-dependent manner. Furthermore, JEV infection blocked the tyrosine phosphorylation of IFN receptor-associated Jak kinase, Tyk2, without affecting the expression of IFN-α/β receptor on the cell surface. Consequently, expression of several IFN-stimulated genes in response to IFN-α stimulation was also reduced in the JEV-infected cells. Overall, our findings suggest that JEV counteracts the effect of IFN-α/β by blocking Tyk2 activation, thereby resulting in inhibition of Jak-Stat signaling pathway.