bluuebunny/crossref_metadata_2025_split huggingface dataset:
Point‐of‐care method for total white cell count: an evaluation of the HemoCue WBC device
Authors: OSEI‐BIMPONG A. , JURY C. , McLEAN R. , LEWIS S. M. 
Published: August 2008

SummaryPoint‐of‐care testing (POCT) is becoming an important adjunct to haematology laboratory practice. An important component of the blood count is the total white cell count (WBC). Previously, this required laborious microscopic cell counting, but it can now be performed by means of automation; however, in many under‐resourced countries, costly automated counters are only available in very few central hospitals. Moreover, neither method is practical in most POCT situations. The HemoCue WBC has been developed as a simplified alternative method, consisting of a reagent pre‐loaded disposable cuvette together with basic image analysis technology. This report describes an assessment of its utility. The WBC of 500 routine blood samples from the hospital were tested in parallel by the HemoCue WBC and by a reference analyser to assess accuracy and utility of the former. The tests included precision, linearity, type of blood sample and anticoagulant and potential interfering substances in blood specimens. In the tests for accuracy, 192 of the 200 showed percentage difference from the NEQAS reference of <10% whilst the remaining eight samples differed by <12%, thus meeting the requirements of Clinical laboratory improvement amendments (CLIA)‐88 regulations. Of the samples tested with potential interfering substances only those with >2% normoblasts or reticulocytosis showed significant differences from the reference measurements. The HemoCue WBC is reliable for WBC counts within the analytical range of 0.4–30.0 × 109/l, except in samples where there are significant numbers of normoblasts or reticulocytes. It is simple to use and provides a valuable advance in the facilities available for POCT in haematology.
Impact of
Trichomonas vaginalis
Transcription-Mediated Amplification-Based Analyte-Specific-Reagent Testing in a Metropolitan Setting of High Sexually Transmitted Disease Prevalence
Authors: Munson Erik , Napierala Maureen , Olson Robin , Endes Tina , Block Timothy , Hryciuk Jeanne E. , Schell Ronald F. 
Published: August 2008

ABSTRACT

Trichomoniasis is a significant sexually transmitted disease (STD) in the spectrum of public health and primary care because of its association with agents such as human immunodeficiency virus and
Neisseria gonorrhoeae
. However, its true significance may be underestimated due to diagnostic modalities that exhibit poor sensitivity. A total of 1,086 genital specimens from two urban emergency departments, a suburban urgent-care facility, and a metropolitan outpatient physician group were subjected to transcription-mediated amplification-based
Trichomonas vaginalis
analyte-specific-reagent (ASR) testing (Gen-Probe, Inc.). The rate of positive molecular ASR results (14.5%) doubled that of direct saline preparation (7.0%;
P
< 0.0002). Analogous increases were observed at one emergency department and within the outpatient physician group (
P
< 0.0002). No significant increase in the rate of positive molecular ASR results was observed from the facilities that encountered a lower frequency of black/African American patients. While positive
T. vaginalis
findings via direct saline preparation did not have a significant association with concomitant
Chlamydia trachomatis
or
N. gonorrhoeae
infection overall, a positive
T. vaginalis
ASR result was a better predictor of concomitant
C. trachomatis
or
N. gonorrhoeae
infection (odds ratios of 2.34 and 4.46, respectively;
P
< 0.0001). The increased rate of positive
T. vaginalis
ASR results was observed in both point-of-care (
P
= 0.02 versus direct saline preparation) and laboratory (
P
= 0.003) testing. Highly sensitive
T. vaginalis
molecular ASR not only transcends issues of specimen integrity and microscopic acumen but also has an increased ability to predict the likelihood of additional STDs in defined populations.
Rapid Method for Sensitive Screening of Oligosaccharide Epitopes in the Lipooligosaccharide from
Campylobacter jejuni
Strains Isolated from Guillain-Barré Syndrome and Miller Fisher Syndrome Patients
Authors: Dzieciatkowska Monika , Liu Xin , Heikema Astrid P. , Houliston R. Scott , van Belkum Alex , Schweda Elke K. H. , Gilbert Michel , Richards James C. , Li Jianjun 
Published: August 2008

ABSTRACT

Campylobacter jejuni
lipooligosaccharide (LOS) can trigger Guillain-Barré syndrome (GBS) due to its similarity to human gangliosides. Rapid and accurate structural elucidation of the LOS glycan of a strain isolated from a GBS patient could help physicians determine the spectrum of anti-ganglioside antibodies likely to be found and therefore provide valuable assistance in establishing an appropriate course of treatment. The ability of implemented mass spectrometry-based approaches in a clinical setting has been limited by the laborious and time-consuming nature of the protocols, typically 3 to 4 days, used to prepare LOS. In order to improve the analytical throughput, microwave-assisted enzymatic digestion was investigated. In this study, the bacterial cells were suspended in 50 μl of 20 mM ammonium acetate buffer containing DNase and RNase and treated by direct microwave irradiation for 3 min. Then, proteinase K was added and the samples were again microwaved. The intact LOS samples were analyzed using electrophoresis-assisted open-tubular liquid chromatography-mass spectrometry. The reliability of the rapid, high-throughput technique was demonstrated through analysis of LOS glycans from 73
C. jejuni
strains. The structure was elucidated using material from a single colony. The total time for sample preparation and MS analysis is less than 60 min.
Quantification of DNA in Plasma by an Automated Real-Time PCR Assay (Cytomegalovirus PCR Kit) for Surveillance of Active Cytomegalovirus Infection and Guidance of Preemptive Therapy for Allogeneic Hematopoietic Stem Cell Transplant Recipients
Authors: Gimeno Concepción , Solano Carlos , Latorre José C. , Hernández-Boluda Juan C. , Clari María A. , Remigia María J. , Furió Santiago , Calabuig Marisa , Tormo Nuria , Navarro David 
Published: August 2008

ABSTRACT

The performance of a plasma real-time PCR (cytomegalovirus [CMV] PCR kit; Abbott Diagnostics) was compared with that of the antigenemia assay for the surveillance of active CMV infection in 42 allogeneic hematopoietic stem cell transplantation (Allo-SCT) recipients. A total of 1,156 samples were analyzed by the two assays. Concordance between the two assays was 82.2%. Plasma DNA levels correlated with the number of pp65-positive cells, particularly prior to the initiation of preemptive therapy. Fifty-seven episodes of active CMV infection were detected in 37 patients: 18 were defined solely by the PCR assay and four were defined on the basis of the antigenemia assay. Either a cutoff of 288 CMV DNA copies/ml or a 2.42-log
10
increase of DNAemia levels between two consecutive PCR positive samples was an optimal value to discriminate between patients requiring preemptive therapy and those not requiring therapy on the basis of the antigenemia results. The real-time PCR assay allowed an earlier diagnosis of active CMV infection and was a more reliable marker of successful clearance of CMV from the blood. Analysis of the kinetics of DNAemia levels at a median of 7 days posttreatment allowed the prediction of the response to CMV therapy. Two patients developed CMV colitis. The PCR assay tested positive both before the onset of symptoms and during the disease period. The plasma real-time PCR from Abbott is more suitable than the antigenemia assay for monitoring active CMV infection in Allo-SCT recipients and may be used for guiding preemptive therapy in this clinical setting.
Detection of Resistance to Macrolides in Thermotolerant
Campylobacter
Species by Fluorescence In Situ Hybridization
Authors: Haas Michaela , Essig Andreas , Bartelt Edda , Poppert Sven 
Published: August 2008

ABSTRACT

The resistance of enteritis-causing
Campylobacter
strains to erythromycin is an emerging problem. We therefore evaluated fluorescence in situ hybridization (FISH) for the rapid detection of resistance using 74 campylobacter isolates. FISH showed specificity and sensitivity of 100% for the detection of high-level resistance.
Double-Stranded RNA Adenosine Deaminases Enhance Expression of Human Immunodeficiency Virus Type 1 Proteins
Authors: Phuphuakrat Angsana , Kraiwong Romchat , Boonarkart Chompunuch , Lauhakirti Darat , Lee Tun-Hou , Auewarakul Prasert 
Published: August 2008

ABSTRACT

ADARs (adenosine deaminases that act on double-stranded RNA) are RNA editing enzymes that catalyze a change from adenosine to inosine, which is then recognized as guanosine by translational machinery. We demonstrate here that overexpression of ADARs but not of an ADAR mutant lacking editing activity could upregulate human immunodeficiency virus type 1 (HIV-1) structural protein expression and viral production. Knockdown of ADAR1 by RNA silencing inhibited HIV-1 production. Viral RNA harvested from transfected ADAR1-knocked-down cells showed a decrease in the level of unspliced RNA transcripts. Overexpression of ADAR1 induced editing at a specific site in the
env
gene, and a mutant with the edited sequence was expressed more efficiently than the wild-type viral genome. These data suggested the role of ADAR in modulation of HIV-1 replication. Our data demonstrate a novel mechanism in which HIV-1 employs host RNA modification machinery for posttranscriptional regulation of viral protein expression.
Differences in Virulence of Marine and Freshwater Isolates of Viral Hemorrhagic Septicemia Virus In Vivo Correlate with In Vitro Ability To Infect Gill Epithelial Cells and Macrophages of Rainbow Trout (
Oncorhynchus mykiss
)
Authors: Brudeseth Bjørn E. , Skall Helle F. , Evensen Øystein 
Published: August 2008

ABSTRACT

Two strains of viral hemorrhagic septicemia virus (VHSV) with known different virulence characteristics in vivo were studied (by a time course approach) for their abilities to infect and translocate across a primary culture of gill epithelial cells (GEC) of rainbow trout (RBT;
Oncorhynchus mykiss
). The strains included one low-virulence marine VHSV (ma-VHSV) strain, ma-1p8, and a highly pathogenic freshwater VHSV (fw-VHSV) strain, fw-DK-3592B. Infectivities toward trout head kidney macrophages were also studied (by a time course method), and differences in in vivo virulence were reconfirmed, the aim being to determine any correlation between in vivo virulence and in vitro infectivity. The in vitro studies showed that the fw-VHSV isolate infected and caused a cytotoxic effect in monolayers of GEC (demonstrating virulence) at an early time point (2 h postinoculation) and that the same virus strain had translocated over a confluent, polarized GEC layer by 2 h postinoculation. The marine isolate did not infect monolayers of GEC, and delayed translocation across polarized GEC was seen by 48 h postinoculation. Primary cultures of head kidney macrophages were also infected with fw-VHSV, with a maximum of 9.5% virus-positive cells by 3 days postinfection, while for the ma-VHSV strain, only 0.5% of the macrophages were positive after 3 days of culture. In vivo studies showed that the fw-VHSV strain was highly virulent for RBT fry and caused high mortality, with classical features of viral hemorrhagic septicemia. The ma-VHSV showed a very low level of virulence (only one pool of samples from the dead fish was VHSV positive). This study has shown that the differences in virulence between marine and freshwater strains of VHSV following the in vivo infection of RBT correlate with in vitro abilities to infect primary cultures of GEC and head kidney macrophages of the same species.
The M, E, and N Structural Proteins of the Severe Acute Respiratory Syndrome Coronavirus Are Required for Efficient Assembly, Trafficking, and Release of Virus-Like Particles
Authors: Siu Y. L. , Teoh K. T. , Lo J. , Chan C. M. , Kien F. , Escriou N. , Tsao S. W. , Nicholls J. M. , Altmeyer R. , Peiris J. S. M. , Bruzzone R. , Nal B. 
Published: August 2008

ABSTRACT
The production of virus-like particles (VLPs) constitutes a relevant and safe model to study molecular determinants of virion egress. The minimal requirement for the assembly of VLPs for the coronavirus responsible for severe acute respiratory syndrome in humans (SARS-CoV) is still controversial. Recent studies have shown that SARS-CoV VLP formation depends on either M and E proteins or M and N proteins. Here we show that both E and N proteins must be coexpressed with M protein for the efficient production and release of VLPs by transfected Vero E6 cells. This suggests that the mechanism of SARS-CoV assembly differs from that of other studied coronaviruses, which only require M and E proteins for VLP formation. When coexpressed, the native envelope trimeric S glycoprotein is incorporated onto VLPs. Interestingly, when a fluorescent protein tag is added to the C-terminal end of N or S protein, but not M protein, the chimeric viral proteins can be assembled within VLPs and allow visualization of VLP production and trafficking in living cells by state-of-the-art imaging technologies. Fluorescent VLPs will be used further to investigate the role of cellular machineries during SARS-CoV egress.
Evidence for Protection against Chronic Hepatitis C Virus Infection in Chimpanzees by Immunization with Replicating Recombinant Vaccinia Virus
Authors: Youn Jin-Won , Hu Yu-Wen , Tricoche Nancy , Pfahler Wolfram , Shata Mohamed Tarek , Dreux Marlene , Cosset François-Loic , Folgori Antonella , Lee Dong-Hun , Brotman Betsy , Prince Alfred M. 
Published: August 2008

ABSTRACTGiven the failures of nonreplicating vaccines against chronic hepatitis C virus (HCV) infection, we hypothesized that a replicating viral vector may provide protective immunity. Four chimpanzees were immunized transdermally twice with recombinant vaccinia viruses (rVV) expressing HCV genes. After challenge with 24 50% chimpanzee infective doses of homologous HCV, the two control animals that had received only the parental VV developed chronic HCV infection. All four immunized animals resolved HCV infection. The difference in the rate of chronicity between the immunized and the control animals was close to statistical significance (P= 0.067). Immunized animals developed vigorous gamma interferon enzyme-linked immunospot responses and moderate proliferative responses. To investigate cross-genotype protection, the immunized recovered chimpanzees were challenged with a pool of six major HCV genotypes. During the acute phase after the multigenotype challenge, all animals had high-titer viremia in which genotype 4 dominated (87%), followed by genotype 5 (13%). However, after fluctuating low-level viremia, the viremia finally turned negative or persisted at very low levels. This study suggests the potential efficacy of replicating recombinant vaccinia virus-based immunization against chronic HCV infection.
Role of cdk9 in the Optimization of Expression of the Genes Regulated by ICP22 of Herpes Simplex Virus 1
Authors: Durand Lizette Olga , Roizman Bernard 
Published: August 2008

ABSTRACTICP22 is a multifunctional herpes simplex virus 1 (HSV-1) regulatory protein that regulates the accumulation of a subset of late (γ2) proteins exemplified by UL38, UL41, and US11. ICP22 binds the cyclin-dependent kinase 9 (cdk9) but not cdk7, and this complex in conjunction with viral protein kinases phosphorylates the carboxyl terminus of RNA polymerase II (Pol II) in vitro. The primary function of cdk9 and its partners, the cyclin T variants, is in the elongation of RNA transcripts, although functions related to the initiation and processing of transcripts have also been reported. We report two series of experiments designed to probe the role of cdk9 in infected cells. In the first, infected cells were treated with 5,6-dichloro-1-β-d-ribofuranosylbenzimidazole (DRB), a specific inhibitor of cdk9. In cells treated with DRB, the major effect was in the accumulation of viral RNAs and proteins regulated by ICP22. The accumulation of α, β, or γ proteins not regulated by ICP22 was not affected by the drug. The results obtained with DRB were duplicated in cells transfected with small interfering RNA (siRNA) targeting cdk9 mRNAs. Interestingly, DRB and siRNA reduced the levels of ICP22 but not those of other α gene products. In addition, cdk9 and ICP22 appeared to colocalize with RNA Pol II in wild-type-virus-infected cells but not in ΔUL13-infected cells. We conclude that cdk9 plays a critical role in the optimization of expression of genes regulated by ICP22 and that one function of cdk9 in HSV-1-infected cells may be to bring ICP22 into the RNA Pol II transcriptional complex.
A paleomagnetic test for northward tectonic transport of the eastern Selwyn Basin, northern Canadian Cordillera: data from the mid-Cretaceous Ragged Pluton
Authors: Symons D. T.A. , Kawasaki K. , Hart C. J.R. , Harris M. J. 
Published: August 2008

Ragged Pluton is a member of the Tungsten Plutonic Suite in the eastern Selwyn Basin. This circular massive 97 Ma granitic pluton is ∼5 km in diameter. It intrudes deformed and metamorphosed clastic strata of the Neoproterozoic to Lower Cambrian Vampire Formation. Thermal and alternating field step demagnetization data for 216 specimens from 21 plutonic sites isolated a stable characteristic remanent magnetization (ChRM) direction at 20 sites of declination D = 325.9°, inclination I = 80.4° (α95 = 2.7°, k = 142). Thermal demagnetization and saturation remanence analysis show that the ChRM is carried by both single-domain pyrrhotite and magnetite. Ragged Pluton’s paleopole at 73.9°N latitude, 191.4°E longitude (dp = 5.0°, dm = 5.2°) is concordant with its co-eval 97 Ma North American reference paleopole, showing at 95% confidence that the pluton has not been significantly rotated or translated relative to North America. This paleomagnetic result favours tectonic models in which the eastern Selwyn Basin is autochthonous, or nearly so, since the mid Cretaceous.
The ratio of ±KTS splice variants of the Wilms’ tumour suppressor protein WT1 mRNA is determined by an intronic enhancer
Authors: Yang Cheng , Romaniuk Paul J. 
Published: August 2008

Alternative splicing of the primary transcript of the Wilms’ tumour suppressor gene WT1 involving 2 overlapping 5′ splice sites at the end of exon 9 results in the insertion of 2 amino acids (KTS) between zinc fingers 3 and 4 of the protein. The presence or absence of these 3 amino acids has consequences for DNA binding affinity, protein–protein interactions, and subnuclear localization. Disruption of the characteristic +KTS to –KTS ratio of mRNA isoforms as a result of mutations in the +KTS splice site results in Frasier syndrome. Mutational analysis of a WT1 minigene construct was carried out to search for sequences that regulate the ±KTS alternative splicing event. A strong pyrimidine-rich intronic enhancer that increases the use of the +KTS splice site was identified. Cross-linking experiments with nuclear extracts demonstrated that this enhancer specifically binds a protein with a molecular mass of 42 ± 2 kDa. One candidate for this trans-factor is the splicing regulator TIA-1, which binds to the pyrimidine-rich enhancer in the primary transcript from the minigene construct and increases the ±KTS splicing ratio. The observation that TIA-1 and WT1 are both involved in apoptosis supports our proposal for a functional link between these proteins.
Soy isoflavones supplementation alleviates disease severity in autoimmune-prone MRL-lpr/lpr mice
Authors: Hong YH , Wang TC , Huang CJ , Cheng WY , Lin BF 
Published: August 2008

Soy isoflavones supplements, which are phyto-oestrogens widely used as alternatives to alleviate menopausal syndromes or prevent chronic diseases, may exert oestrogenic and anti-oestrogenic activities. This study aimed to investigate the effects of soy isoflavones supplement on oestrogen-related autoimmune disease, such as systemic lupus erythematosus, using autoimmune-prone female MRL- lpr/lpr mice. Eighty mice of 8 weeks were divided into five groups: 0 (Control), 2 (Isf 2), 10 (Isf 10) and 20 (Isf 20) mg/kg BW/day Phyto SoyaTM isoflavones or 0.375 mg/kg BW/day tamoxifen (TAM) as the positive control, by tube-feeding. Some mice were killed at age 15 weeks for cellular cytokine secretion. The data suggested that the Isf 20 and TAM groups had higher weight gain and survival compared with the control group. At age 22 weeks, the Isf 20 group still had 75% survival comparable to mice treated with TAM. At age 14 weeks, the TAM group showed significantly lower serum anti-double-stranded (ds) DNA IgG and anti-cardiolipin IgG. The mice in the Isf 10 and Isf 20 groups also had lower anti-dsDNA IgG and anti-cardiolipin IgG. The interferon (IFN)-γ secretion from mitogen-stimulated T cells in the Isf 20 and TAM groups were significantly lower than those of control mice. Furthermore, the oestrogenic activity of the methanol extracts of soy isoflavones for oestrogen receptor (ER)β, but not ERα, significantly increased, suggesting that soy isoflavones have a selective modulation of ER activation. Thus, soy isoflavone supplementation did not aggravate murine lupus, but apparently ameliorated the disease.
The ethics of Community Empowerment: tensions in health promotion theory and practice
Authors: Braunack-Mayer Annette , Louise Jennie 
Published: August 2008

The concepts of community participation, empowerment and capacity building are central tenets of contemporary health promotion theory. They reflect the view that health and well-being are shaped by a wide range of social, economic, political and organisational forces that are outside the control of individuals. Despite its theoretical appeal, the practice of Community Empowerment is ethically contentious and can produce ethical dilemmas for health promotion practitioners. In this paper we relate these dilemmas to theoretical considerations, and argue that the empowerment of communities should be understood as a means rather than an end . This leads us to argue for the adoption of what we call a Reflective Equilibrium Community Empowerment approach, which draws on both “top—down” and “bottom—up” methods to help resolve the ethical tensions in health promotion programmes. (Promot Educ 2008;15(3): 5-8)