bluuebunny/crossref_metadata_2025_split huggingface dataset:
Changes in hepatic gene expression in dogs with experimentally induced nutritional iron deficiency
Authors: Fry Michael M. , Kirk Claudia A. , Liggett Jason L. , Daniel Gregory B. , Baek Seung Joon , Gouffon Julia S. , Chimakurthy Pradeep M. , Rekapalli Bhanu 
Published: February 2009

Background and Objective: We investigated hepatic gene expression in dogs with experimentally induced nutritional iron deficiency (ID). Our hypothesis was that ID would result in decreased hepcidin gene expression, and possibly in altered expression of other genes associated with iron metabolism.Methods: Liver biopsies were collected from each of 3 dogs before induction of ID, at the point of maximal ID, and after resolution of ID. Using Affymetrix microarray technology and analytical tools specifically designed for microarray data, we identified genes that had at least a 2‐fold change in expression in response to ID. Four genes were selected for further analysis by reverse transcriptase PCR (RT‐PCR).Results: Dogs with ID had markedly decreased expression of the hepcidin gene (mean decrease of 40‐fold for one probe and >100‐fold for another probe) and increased expression of the transferrin receptor gene (mean increase of >7‐fold). There was also mildly decreased expression of the “similar to calreticulin” gene and a gene of unknown function. Results of RT‐PCR analysis were consistent with microarray findings.Conclusion: Changes in hepcidin and transferrin receptor gene expression were consistent with the known biology of iron metabolism. The decrease in expression of a gene identified as “similar to calreticulin,” while not statistically significant, was consistent with the findings of other investigators that suggest iron plays a role in calreticulin expression.
Canine complete blood counts: a comparison of four in‐office instruments with the ADVIA 120 and manual differential counts
Authors: Welles Elizabeth G. , Hall Amy S. , Carpenter D. Mark 
Published: February 2009

Background: With more use of bench‐top in‐office hematology analyzers, the accuracy of reported values is increasingly important. Instruments use varied methods for cell counting and differentiation, and blood smears may not always be examined.Objective: The purpose of this study was to compare canine CBC results using 4 bench‐top instruments (Hemavet 950, Heska CBC‐Diff, IDEXX LaserCyte, and IDEXX VetAutoread) with ADVIA 120 and manual leukocyte counts.Methods: EDTA‐anticoagulated canine blood samples (n=100) were analyzed on each instrument. Manual differentials were based on 100‐cell counts. Linear regression, difference plots, paired t‐tests, and estimation of diagnostic equivalence were used to analyze results.Results: Correlations of HCT, WBC, and platelet counts were very good to excellent between all in‐office instruments and the ADVIA 120, but results varied in accuracy (comparability). Hemavet 950 and Heska CBC‐Diff results compared best with ADVIA results and manual leukocyte differentials. HCT and platelet counts on the IDEXX VetAutoread compared well with those from the ADVIA. Except for neutrophil counts, leukocyte differentials from all instruments compared poorly with ADVIA and manual counts. Reticulocyte counts on the LaserCyte and VetAutoread compared poorly with those from the ADVIA.Conclusions: The Hemavet 950 and Heska CBC‐Diff performed best of the 4 analyzers we compared. HCT, WBC, and platelet counts on the LaserCyte had minimally sufficient comparability for diagnostic use. Except for neutrophils (granulocytes), leukocyte differential counts were unreliable on all in‐office analyzers. Instruments with a 5‐part leukocyte differential provided no added benefit over a 3‐part differential. Assessment of erythrocyte regeneration on the LaserCyte and VetAutoread was unreliable compared with the ADVIA 120.
Serum acute phase protein concentrations in dogs with hyperadrenocorticism with and without concurrent inflammatory conditions
Authors: Caldin Marco , Tasca Silvia , Carli Erika , Bianchini Silvia , Furlanello Tommaso , Martinez‐Subiela Silvia , Cerón Jose J 
Published: February 2009

Background:Acute phase proteins (APPs) are promising markers of inflammation in dogs, because they are more sensitive than WBC counts in detecting clinical and subclinical inflammation. Endogenous corticosteroids can mask an acute phase response and make it more difficult to identify underlying inflammatory disease.Objective:The purpose of this study was to evaluate the acute phase protein response in dogs with spontaneous hyperadrenocorticism (HAC) with and without concurrent inflammatory conditions.Methods:Serum concentrations of C‐reactive protein (CRP), haptoglobin (Hp), fibrinogen, and albumin were measured in 44 healthy adult dogs and 39 dogs with HAC; the HAC group was further divided into dogs with and without concurrent infection/inflammation. A fourth group of dogs with severe sepsis and without HAC was compared with the dogs with HAC and severe sepsis.Results:Dogs with uncomplicated HAC had significantly higher Hp and fibrinogen concentrations compared with healthy control dogs (P<.001). Dogs with HAC and severe inflammatory disease also had significantly higher CRP and lower albumin concentrations than control dogs and dogs with HAC without concurrent inflammation. Dogs with sepsis but without HAC had significantly higher CRP concentrations than dogs with HAC and sepsis.Conclusions:Dogs with HAC had increases in the moderate APPs (Hp and fibrinogen), and no significant changes in CRP and albumin compared with healthy dogs. Although concurrent HAC appeared to blunt the CRP response in dogs with sepsis, increased serum CRP concentration in dogs with HAC is likely indicative of severe concurrent inflammation.
Platelet aggregation responses in clinically healthy adult llamas
Authors: Gilbert Rosanne M. , Bird Karyn E. , Kutzler Michelle A. 
Published: February 2009

Background:Limited information exists regarding hemostasis in camelids despite the importance of platelet function testing in the accurate identification of platelet disorders. As further importation of llamas to North America is restricted, variability in breeding stock will continue to decrease, potentially leading to an increase in heritable bleeding disorders.Objective:The objective of this study was to measure platelet aggregation responses in clinically healthy llamas and provide baseline data to which abnormal platelet function may be compared in the future.Methods:Blood samples were collected from 39 healthy adult llamas, citrated, and centrifuged to produce platelet‐rich plasma (PRP). Within 4 hours of the blood draw, 20 μL of each agonist reagent were added to 180 μL of PRP. Final concentrations of agonists were 2 × 10−5 M ADP, 0.19 mg collagen/mL PRP, 1 × 10−4 M epinephrine, and 500 μg arachidonic acid/mL PRP.Results:Llama platelets were most responsive to ADP and collagen, with a maximum percent aggregation (mean±SD) of 71.3±18.6% and 55.8±19% and aggregation rates of 9.5±3.9 and 6.7±3.7 cm/min, respectively. Llama platelet aggregation in response to epinephrine and arachidonic acid was minimal to absent.Conclusions:This study is the first of its kind to establish baseline values for platelet aggregation in healthy adult llamas.
Evidence of Prior Exposure to Human Bocavirus as Determined by a Retrospective Serological Study of 404 Serum Samples from Adults in the United States
Authors: Cecchini Sylvain , Negrete Alejandro , Virag Tamas , Graham Barney S. , Cohen Jeffrey I. , Kotin Robert M. 
Published: February 2009

ABSTRACT

Recently, molecular screening for pathogenic agents has identified a partial genome of a novel parvovirus, called human bocavirus (HBoV). The presence of this newly described parvovirus correlated with upper and lower respiratory tract infections in children. Lower respiratory tract infections are a leading cause of hospital admission in children, and the etiological agent has not been identified in up to 39% of these cases. Using baculovirus expression vectors (BEVs) and an insect cell system, we produced virus-like particles (VLPs) of HBoV. The engineered BEVs express the HBoV capsid proteins stoichiometrically from a single open reading frame. Three capsid proteins assemble into the VLP rather than two proteins predicted from the HBoV genome sequence. The denatured capsid proteins VP1, VP2, and VP3 resolve on silver-stained sodium dodecyl sulfate-polyacrylamide gels as three bands with apparent molecular masses of 72 kDa, 68 kDa, and 62 kDa, respectively. VP2 apparently initiates at a GCT codon (alanine) 273 nucleotides downstream from the VP1 start site and 114 nucleotides upstream from the VP3 initiation site. We characterized the stable capsids using physical, biochemical, and serological techniques. We found that the density of the VLP is 1.32 g/cm
3
and is consistent with an icosahedral symmetry with approximately a 25-nm diameter. Rabbit antiserum against the capsid of HBoV, which did not cross-react with adeno-associated virus type 2, was used to develop enzyme-linked immunosorbent assays (ELISAs) for anti-HBoV antibodies in human serum. Using ELISA, we tested 404 human serum samples and established a range of antibody titers in a large U.S. adult population sample.
Genotyping of Clinical Varicella-Zoster Virus Isolates Collected in China
Authors: Liu Jingjing , Wang Mingli , Gan Lin , Yang Sen , Chen Jason 
Published: February 2009

ABSTRACT
Varicella-zoster virus (VZV) is genetically stable; and various schemes for the genotyping of VZV based on restriction fragment length polymorphisms (RFLPs), PCR, and sequencing have been developed. At least three major genotypes have been recognized among VZV isolates or clinical samples from different locations around the world; however, few data were available for viral isolates from China. In the current study, a collection of 19 VZV isolates from patients with zoster or varicella in the middle eastern region of China was examined for genetic variations. RFLP analysis of DNA fragments of open reading frames (ORFs) 38, 54, and 62 showed that all 19 isolates were PstI and BglI positive and SmaI negative, and this may represent the major restriction pattern of wild-type VZV strains in China. Further analysis of the R5 variable-repeat region in those strains revealed that 9 (47.4%) were type R5A, while the remaining 10 strains (52.6%) were type R5B. On the basis of the sequencing data for ORFs 1, 21, 22, and 54, all 19 Chinese strains could be grouped into genotype J or J1. A novel in-frame 3-nucleotide insertion (CGG) in ORF1 was found in 4 (21%) of the 19 isolates. Additionally three new nucleotide substitutions were detected in two of the isolates. A varicella isolate from the United States, strain MLS, was included in this study as a control for American wild-type VZV, and was found to be type M1, which represents one of the minor genotypes in North America.
Development and Validation of a Microsphere-Based Luminex Assay for Rapid Identification of Clinically Relevant Aspergilli
Authors: Etienne Kizee A. , Kano Rui , Balajee S. Arunmozhi 
Published: February 2009

ABSTRACT

A Luminex-based assay for the rapid identification of
Aspergillus
species was designed, optimized, and validated with 131 clinical isolates of
Aspergillus fumigatus
,
A. flavus
,
A. niger
,
A. terreus
,
A. ustus
, and
A. versicolor
. The six species-specific probes were directed toward the internal transcribed spacer 1 (ITS-1) region and tested in a multiplex format with results generated within 6 h. Species identifications generated by the
Aspergillus
Luminex assay were 100% concordant with results from comparative sequence analyses of the ITS-1 region and showed excellent specificity. The
Aspergillus
Luminex assay is a rapid, relatively simple method that may prove to be a useful diagnostic tool for rapid
Aspergillus
identification in clinical laboratory settings.
Suitability of Simple Human Immunodeficiency Virus Rapid Tests in Clinical Trials in Community-Based Clinic Settings
Authors: Everett Dean B. , Baisley Kathy , Changalucha John , Vallely Andrew , Watson-Jones Deborah , Cook Claire , Knight Louise , Ross David A. , Mugeye Kokugonza , McCormack Sheena , Lacey Charles J. , Jentsch Ute , Hayes Richard J. 
Published: February 2009

ABSTRACT
The suitability and accuracy of using simple human immunodeficiency virus (HIV) rapid (SR) tests in community-based clinics in northwest Tanzania were determined to assess eligibility for participation in clinical trials. The HIV rapid and ELISA test results for 789 women aged 16 to 54 who were screened for two clinical trials of HIV prevention were compared. Women were offered voluntary HIV counseling and testing (VCT) at screening; those who accepted were tested with the Abbott Determine and Trinity Biotech Capillus SR tests in parallel. The results were confirmed by two parallel HIV enzyme-linked immunosorbent assay (ELISA) tests (Abbott Murex HIV Ag/Ab combination and Vironostika Uniform II HIV Ag/Ab) to determine eligibility. Positive samples for any of the four assays were confirmed by a line immunoassay and p24 testing. The parallel SR tests had high concordance (96.2%) with the parallel ELISA algorithm. The sensitivities of the SR tests were 98.6% for Capillus (95% confidence interval [CI], 95.1 to 99.8%), 99.3% for Determine (95% CI, 96.2 to 100%), and 98.6% for the parallel SR (95% CI, 95.1 to 99.8%). The specificities were 99.7% for Capillus (95% CI, 98.9 to 100%), 99.7% for Determine (95% CI, 98.9 to 100%), and 100% for the parallel SR (95% CI, 99.4 to 100%). SR tests are suitable for use in community-based clinical research settings to assess eligibility both for trial participation and for the provision of on-site VCT services.
Evaluation of a Rapid Fluorescent Staining Method for Detection of Mycobacteria in Clinical Specimens
Authors: Hendry Cindy , Dionne Kim , Hedgepeth Annie , Carroll Karen , Parrish Nicole 
Published: February 2009

ABSTRACT
Rapid detection of mycobacterial disease is essential. Using multiple specimen types and concentrations of mycobacteria, we compared two commercial auramine O stains. The more rapid stain permitted consistent acid-fast bacillus quantitation and exhibited less debris staining, and the staining procedure required less time (∼2 min) to perform. These results suggest that the rapid stain may be more cost-effective and efficient for use in clinical laboratories.
Suppression of Tetherin-Restricting Activity upon Human Immunodeficiency Virus Type 1 Particle Release Correlates with Localization of Vpu in the
trans
-Golgi Network
Authors: Dubé Mathieu , Roy Bibhuti Bhusan , Guiot-Guillain Pierre , Mercier Johanne , Binette Julie , Leung Grace , Cohen Éric A. 
Published: February 2009

ABSTRACT

Vpu promotes the efficient release of human immunodeficiency virus type 1 (HIV-1) by overcoming the activity of tetherin, a host cell restriction factor that retains assembled virions at the cell surface. In this study, we analyzed the intracellular localization and trafficking of subtype B Vpu in HIV-1-producing human cells. We found that mutations of conserved positively charged residues (R30 and K31) within the putative overlapping tyrosine- and dileucine-based sorting motifs of the Vpu hinge region affected both the accumulation of the protein in the
trans
-Golgi network (TGN) and its efficient delivery to late endosomal degradative compartments. A functional characterization of this mutant revealed that the mislocalization of Vpu from the TGN correlated with an attenuation of HIV-1 release. Interestingly, clathrin light chain small interfering RNA-directed disruption of Vpu trafficking from the TGN to the endosomal system slightly stimulated Vpu-mediated HIV-1 release and completely restored the activity of the Vpu R30A,K31A mutant. An analysis of the C-terminal deletion mutants of Vpu identified an additional determinant in the second helical structure of the protein, which regulated TGN retention/localization, and further revealed the functional importance of Vpu localization in the TGN. Finally, we show that a large fraction of Vpu colocalizes with tetherin in the TGN and provide evidence that the degree of Vpu colocalization with tetherin in the TGN is important for efficient HIV-1 release. Taken together, our results reveal that Vpu traffics between the TGN and the endosomal system and suggest that the proper distribution of Vpu in the TGN is critical to overcome the restricting activity of tetherin on HIV-1 release.
CD4 T Cells Mediate Killing during Persistent Gammaherpesvirus 68 Infection
Authors: Stuller Kathleen A. , Flaño Emilio 
Published: February 2009

ABSTRACT
CD4 T cells are critical for the control of gammaherpesvirus persistence, but their direct effector mechanisms of virus control in vivo are still poorly understood. In this study, we use murine gammaherpesvirus 68 (γHV68) in in vitro and in vivo cytotoxicity assays to show CD4-dependent killing of γHV68-loaded cells in mice persistently infected with γHV68. Our results underscore the cytotoxic capacity of CD4 T cells during γHV68 persistence.
Reduced Ribavirin Antiviral Efficacy via Nucleoside Transporter-Mediated Drug Resistance
Authors: Ibarra Kristie D. , Pfeiffer Julie K. 
Published: February 2009

ABSTRACTTreatment for hepatitis C virus infection currently consists of pegylated interferon and ribavirin (RBV), a nucleoside analog. Although RBV clearly plays a role in aiding the treatment response, its antiviral mechanism is unclear. Regardless of the specific mechanism of RBV, we hypothesize that differences in levels of cellular uptake of RBV may affect antiviral efficacy and treatment success and that cells may become RBV resistant through reduced uptake. We monitored RBV uptake in various cell lines and determined the effect of uptake capacity on viral replication. RBV-resistant cells demonstrated reduced RBV uptake and increased growth of a model RNA virus, poliovirus, in the presence of RBV. Overexpression of equilibrative nucleoside transporter 1 (ENT1) or concentrative nucleoside transporter 3 (CNT3) increased RBV uptake in RBV-sensitive cell lines and restored the uptake defect in most RBV-resistant cell lines. However, CNT3 is not expressed in Huh-7 liver cells, and inhibition of concentrative transport did not affect RBV uptake. Blocking equilibrative transport using the inhibitor nitrobenzylmercaptopurine riboside recapitulated the RBV-resistant phenotype in RBV-sensitive cell lines, with a reduction in RBV uptake and increased poliovirus growth. Taken together, these results indicate that RBV uptake is restricted primarily to ENT1 in the cell lines examined. Interestingly, some RBV-resistant cell lines may compensate for reduced ENT1-mediated nucleoside uptake by increasing the activity of an alternative nucleoside transporter, ENT2. It is possible that RBV uptake affects the antiviral treatment response, either through natural differences in patients or through acquired resistance.
Protein Array Identification of Substrates of the Epstein-Barr Virus Protein Kinase BGLF4
Authors: Zhu Jian , Liao Gangling , Shan Liang , Zhang Jun , Chen Mei-Ru , Hayward Gary S. , Hayward S. Diane , Desai Prashant , Zhu Heng 
Published: February 2009

ABSTRACT

A conserved family of herpesvirus protein kinases plays a crucial role in herpesvirus DNA replication and virion production. However, despite the fact that these kinases are potential therapeutic targets, no systematic studies have been performed to identify their substrates. We generated an Epstein-Barr virus (EBV) protein array to evaluate the targets of the EBV protein kinase BGLF4. Multiple proteins involved in EBV lytic DNA replication and virion assembly were identified as previously unrecognized substrates for BGLF4, illustrating the broad role played by this protein kinase. Approximately half of the BGLF4 targets were also in vitro substrates for the cellular kinase CDK1/cyclin B. Unexpectedly, EBNA1 was identified as a substrate and binding partner of BGLF4. EBNA1 is essential for replication and maintenance of the episomal EBV genome during latency. BGLF4 did not prevent EBNA1 binding to sites in the EBV latency origin of replication,
oriP
. Rather, we found that BGLF4 was recruited by EBNA1 to
oriP
in cells transfected with an
oriP
vector and BGLF4 and in lytically induced EBV-positive Akata cells. In cells transfected with an
oriP
vector, the presence of BGLF4 led to more rapid loss of the episomal DNA, and this was dependent on BGLF4 kinase activity. Similarly, expression of doxycycline-inducible BGLF4 in Akata cells led to a reduction in episomal EBV genomes. We propose that BGLF4 contributes to effective EBV lytic cycle progression, not only through phosphorylation of EBV lytic DNA replication and virion proteins, but also by interfering with the EBNA1 replication function.
Role of Conserved Histidine Residues in the Low-pH Dependence of the Semliki Forest Virus Fusion Protein
Authors: Qin Zhao-ling , Zheng Yan , Kielian Margaret 
Published: February 2009

ABSTRACT
A wide variety of enveloped viruses infects cells by taking advantage of the low pH in the endocytic pathway to trigger virus-membrane fusion. For alphaviruses such as Semliki Forest virus (SFV), acidic pH initiates a series of conformational changes in the heterodimeric virus envelope proteins E1 and E2. Low pH dissociates the E2/E1 dimer, releasing the membrane fusion protein E1. E1 inserts into the target membrane and refolds to a trimeric hairpin conformation, thus driving the fusion reaction. The means by which E1 senses and responds to low pH is unclear, and protonation of conserved E1 histidine residues has been proposed as a possible mechanism. We tested the role of four conserved histidines by mutagenesis of the wild-type (wt) SFV infectious clone to create virus mutants with E1 H3A, H125A, H331A, and H331A/H333A mutations. The H125A, H331A, and H331A/H333A mutants had growth properties similar to those of wt SFV and showed modest change or no change in the pH dependence of virus-membrane fusion. By contrast, the E1 H3A mutation produced impaired virus growth and a markedly more acidic pH requirement for virus-membrane fusion. The dissociation of the H3A heterodimer and the membrane insertion of the mutant E1 protein were comparable to those of the wt in efficiency and pH dependence. However, the formation of the H3A homotrimer required a much lower pH and showed reduced efficiency. Together, these results and the location of H3 suggest that this residue acts to regulate the low-pH-dependent refolding of E1 during membrane fusion.