bluuebunny/crossref_metadata_2025_split huggingface dataset:
New Generation of Cell Culture Assay for Smallpox Vaccine Potency
Authors: Leparc-Goffart Isabelle , Poirier Bertrand , El Zaouk Annie , Tissier Marie-Hélène , Fuchs Florence 
Published: August 2003

ABSTRACT

The potency of smallpox vaccines produced in the 1970s was tested by titration onto chorioallantoic membranes of fertilized hen eggs (CAM assay). The potency specification commonly approved for these vaccines was a titer above 10
8
pock-forming units per milliliter. We developed and validated a cell culture titration assay to have a more reliable potency test. The cell titration assay and the CAM assay were tested in parallel on 34 first-generation smallpox vaccine lots. These allowed us to demonstrate that a correlation does exist between the two titration techniques and to determine a new in-house specification for the cell titration method. This in vitro potency assay will allow us to test first-generation smallpox vaccines produced on the skin of living animals but will also give a hint of the potency specification that should be assigned for new generations of cell-derived smallpox vaccines.
Monitoring of Cytomegalovirus Infection in Solid-Organ Transplant Recipients by an Ultrasensitive Plasma PCR Assay
Authors: Hadaya Karine , Wunderli Werner , Deffernez Christelle , Martin Pierre-Yves , Mentha Gilles , Binet Isabelle , Perrin Luc , Kaiser Laurent 
Published: August 2003

ABSTRACT

Early and accurate monitoring of cytomegalovirus (CMV) infection in solid-organ transplant recipients is of major importance. We have assessed the potential benefit of an ultrasensitive plasma-based PCR assay for renal transplant recipients. The pp65 CMV antigen (pp65 Ag) assay using leukocytes was employed as a routine test for the monitoring of CMV in 23 transplant recipients. We compared the pp65 antigenemia with the CMV load quantified by an ultrasensitive PCR (US-PCR) with a limit of detection of 20 CMV DNA copies/ml of plasma. CMV infection was detected in 215 (67%) of 321 plasma samples by the US-PCR compared with 124 (39%) of 321 samples by the pp65 Ag assay. The US-PCR assay permitted the detection of CMV infection episodes following transplantation a median of 12 days earlier than the pp65 Ag assay. Moreover, during CMV infection episodes, DNA detection by the US-PCR was consistently positive, whereas false negative results were frequently observed with the pp65 Ag assay. We found a good correlation between the two assays, and the peak viral loads were significantly higher in patients with CMV-related complications (median, 5,000 DNA copies/ml) than in those without symptoms (1,160 DNA copies/ml) (
P
= 0.048). In addition, patients that did not require preemptive therapy based on the results of the pp65 assay had CMV loads significantly lower (median, 36 DNA copies/ml) than those that needed treatment (median, 4,703 DNA copies/ml) (
P
< 0.001). These observations provided cutoff levels that could be applied in clinical practice. The ultrasensitive plasma-based PCR detected CMV infection episodes earlier and provided more consistent results than the pp65 Ag assay. This test could improve the monitoring of CMV infection or reactivation in renal transplant recipients.
Automated Extraction and Quantification of Human Cytomegalovirus DNA in Whole Blood by Real-Time PCR Assay
Authors: Mengelle C. , Sandres-Sauné K. , Pasquier C. , Rostaing L. , Mansuy J.-M. , Marty M. , Da Silva I. , Attal M. , Massip P. , Izopet J. 
Published: August 2003

ABSTRACT

The measurement of human cytomegalovirus (HCMV) DNA in blood is becoming the standard method for monitoring HCMV infection in immune-suppressed and unsuppressed patients. As various blood compartments can be used, we have compared the HCMV DNA measured in whole blood (WB), peripheral blood leukocytes (PBL), and plasma by real-time PCR. We tested 286 samples: HCMV DNA was extracted automatically from WB and PBL with the MagNA Pure instrument (Roche Molecular Biochemicals) and manually from plasma samples. The HCMV DNA from WB, PBL, and plasma was measured by real-time Light Cycler PCR. Primers and probe were located in the UL 83 region. HCMV DNA was detected more frequently in WB (88.5%) than in the PBL (65.7%) (
P
< 0.0001) or the plasma (55.2%) (
P
< 0.0001). There was a good correlation between the positive results in WB and in PBL (
r
= 0.68;
P
< 0.0001), and 3.15 log
10
genome copies in 200,000 PBL, equivalent to the threshold value of 50 pp65-positive polymorphonuclear cells per 200,000 leukocytes, was equivalent to 3.4 log
10
genome copies in 200 μl of WB. WB was shown to be suitable for automated extraction and the quantitation of HCMV DNA by real-time Light Cycler PCR by analysis of serial samples from representative patients of various populations. This system may be very useful for monitoring of immune-suppressed and unsuppressed patients.
Genetic Diversity among
Escherichia coli
O157:H7 Isolates from Bovines Living on Farms in England and Wales
Authors: Liebana Ernesto , Smith Richard P. , Lindsay Elisabeth , McLaren Ian , Cassar Claire , Clifton-Hadley Felicity A. , Paiba Giles A. 
Published: August 2003

ABSTRACT

Pulsed-field gel electrophoresis of
Escherichia coli
O157:H7 isolates (
n
= 228) from 122 healthy animals on 11 farms discriminated 57 types. Most clones were found only on individual farms. Numerous clones were found within each farm, with a prevalent clone normally found in several animals. A variety of clones were found within the different phage types.
Improved Molecular Detection of Dengue Virus Serotype 1 Variants
Authors: Reynes Jean-Marc , Ong Sivuth , Mey Channa , Ngan Chantha , Hoyer Stefan , Sall Amadou A. 
Published: August 2003

ABSTRACT
The dengue virus molecular typing method described by Lanciotti and coworkers (R. S. Lanciotti, C. H. Calisher, D. J. Gubler, G. J. Chang, and A. Vance-Vorndam, J. Clin. Microbiol. 30:545-551, 1992) is used worldwide for diagnosis and surveillance. However, it failed to detect DENV-1 variants in Cambodia due to a point mutation. We describe an improvement of the method that allows the detection of additional DENV-1 strains, including potential variants.
Molecular Cloning, Expression, Self-Assembly, Antigenicity, and Seroepidemiology of a Genogroup II Norovirus Isolated in France
Authors: Nicollier-Jamot Béatrice , Pico Valérie , Pothier Pierre , Kohli Evelyne 
Published: August 2003

ABSTRACT
Virus-like particles of Dijon171/96 virus, a genogroup II norovirus, were expressed in a baculovirus system and were used for a seroepidemiological study of 1,078 age-stratified human sera collected in Dijon, France. The results showed a seroprevalence of 74.1%. Furthermore, we showed that murine antibodies generated against recombinant Dijon171/96 virus, and human antibodies recognized discontinuous epitopes on the particles.
babA2-
and
cagA
-Positive
Helicobacter pylori
Strains Are Associated with Duodenal Ulcer and Gastric Carcinoma in Brazil
Authors: Oliveira Adriana Gonçalves , Santos Adriana , Guerra Juliana Becattini , Rocha Gifone Aguiar , Rocha Andreia Maria Camargos , Oliveira Celso Affonso , Cabral Mônica Maria Demas Álvares , Nogueira Ana Margarida Miguel Ferreira , Queiroz Dulciene Maria Magalhães 
Published: August 2003

ABSTRACT

The
babA2
and
cagA
genes were investigated in 208 Brazilian
Helicobacter pylori
strains. A strong association between
babA2
and duodenal ulcer or gastric carcinoma was observed, even after adjusting for confounding factors, such as age, gender, and
cagA
status.
cagA
-positive strains were also independently associated with
H
.
pylori
-related diseases.
Stereotactic Interstitial Radiosurgery with a Miniature X-Ray Device in the Minimally Invasive Treatment of Selected Tumors in the Thalamus and the Basal Ganglia
Authors: Gallina Pasquale , Francescon Paolo , Cavedon Carlo , Casamassima Franco , Mungai Raffaello , Perrini Paolo , Russo Serenella , Di Lorenzo Nicola 
Published: August 2003

The aim of this study was to evaluate the role of interstitial radiosurgery (IR) using the photon radiosurgery system (PRS) in the treatment of selected tumors within the thalamus and the basal ganglia. The PRS is a miniature X-ray generator that was developed for interstitial irradiation. This series included 14 patients (5 with glioblastomas, 4 with low-grade astrocytomas and 5 with metastases) harboring spheroidal lesions with dimensions ranging from 13 to 42 mm (mean 30 mm). After stereotactic biopsy, a radiation dose ranging from 6 to 15.4 Gy (mean 11.3 Gy) was delivered at the target volume margins. Follow-up varied from 3 to 26 months (mean 10.2 months). In the group of glioblastomas, 3 patients died (3–12 months after the procedure) because of tumor progression, while the remaining had tumor control. Two patients with metastases died from systemic disease (4–9 months after the treatment), and 3 were alive and well at the end of the study. Local control was achieved in all metastases. Patients with low-grade astrocytomas were well and imaging studies showed tumor control PRS IR is a minimally invasive procedure for the treatment of selected glial or secondary brain tumors. Compared to conventional radiosurgery (brachytherapy and external radiosurgery), PRS IR presents dose delivery characteristics useful for the treatment of tumors in the thalamus and basal ganglia, without inconveniences such as handling radioisotopes, the need of expensive facilities and radiation protection measures. Although the clinical value needs further investigations, PRS IR seems to be effective in metastases while it provides less benefit in malignant gliomas. PRS IR could have a major role in the treatment of low-grade astrocytomas.
Bilateral Pallidal Stimulation for Cervical Dystonia
Authors: Goto Satoshi , Mita Shuji , Ushio Yukitaka 
Published: August 2003

We sought to identify optimal paradigms of bilateral globus pallidus internus (GPi) stimulation in 3 subsequent patients with severe cervical dystonia. At low frequency stimulation (50–60 Hz) with wide pulse width (500 µs) and high amplitude (4.5–8.0 V), we observed immediate and consistent improvement of dystonia and dystonia-associated pain. Stimulation of the posteroventral portion of the GPi led to pronounced alleviation of dystonia; stimulation of the anterodorsal portion or at the dorsal border of the GPi resulted in significant worsening of symptoms. The therapeutic benefit obtained by using the optimal stimulation parameters continues and has lasted for at least 1 year in each patient.
In vitro Study of Starling’s Hypothesis in a Cultured Monolayer of Bovine Aortic Endothelial Cells
Authors: Pang Zhengyu , Tarbell John M. 
Published: August 2003

Starling’s hypothesis that fluid movement across the microvascular wall is determined by the transmural differences in hydrostatic and osmotic pressures was tested using an in vitro model comprised of bovine aortic endothelial cells grown on a porous support. In all experiments, a 1% bovine serum albumin (BSA) solution was maintained in the abluminal reservoir and the luminal reservoir contained either a 1 or a 5.5% BSA solution. The global osmotic pressure difference across the endothelial layers was thus either 0 or 20.3 cm H<sub>2</sub>O. When the luminal concentration of BSA was changed from 1 to 5.5% at a hydrostatic pressure differential of 5, 10 or 20 cm H<sub>2</sub>O, no reverse flow (in the reabsorption direction) was observed even though the hydrostatic pressure differential was far below the global osmotic pressure differential. In another case, the hydrostatic pressure differential was dropped quickly from 20 to 5 cm H<sub>2</sub>O, while a constant osmotic pressure differential was maintained by 5.5% BSA in the luminal reservoir. A strong transient reabsorption flow was observed over a 30-second period which diminished to undetectable levels within 2.5 min; then a sustained steady-state filtration flow was observed after 20 min. These in vitro experiments support other studies in capillaries showing transient reabsorption that decays to steady-state filtration at longer times.
Quantification of central metabolic fluxes in the facultative methylotroph methylobacterium extorquens AM1 using 13C‐label tracing and mass spectrometry
Authors: Van Dien Stephen J. , Strovas Tim , Lidstrom Mary E. 
Published: August 2003

AbstractThe metabolic fluxes of central carbon metabolism were measured in chemostat‐grown cultures of Methylobacterium extorquens AM1 with methanol as the sole organic carbon and energy source and growth‐limiting substrate. Label tracing experiments were carried out using 70% 13C‐methanol in the feed, and the steady‐state mass isotopomer distributions of amino acids derived from total cell protein were measured by gas chromatography coupled to mass spectrometry. Fluxes were calculated from the isotopomer distribution data using an isotopomer balance model and evolutionary error minimization algorithm. The combination of labeled methanol with unlabeled CO2, which enters central metabolism in two different reactions, provided the discriminatory power necessary to allow quantification of the unknown fluxes within a reasonably small confidence interval. In wild‐type M. extorquens AM1, no measurable flux was detected through pyruvate dehydrogenase or malic enzyme, and very little flux through α‐ketoglutarate dehydrogenase (1.4% of total carbon). In contrast, the α‐ketoglutarate dehydrogenase flux was 25.5% of total carbon in the regulatory mutant strain phaR, while the pyruvate dehydrogenase and malic enzyme fluxes remained insignificant. The success of this technique with growth on C1 compounds suggests that it can be applied to help characterize the effects of other regulatory mutations, and serve as a diagnostic tool in the metabolic engineering of methylotrophic bacteria. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 84: 45–55, 2003.
Environmental compensation in planning: a review of five different countries with major emphasis on the German system
Authors: Rundcrantz Kristina , Skärbäck Erik 
Published: August 2003

AbstractThe use of different measures to compensate the environment when human development projects take place has been frequently discussed in both Europe and the USA. This paper reviews the use of environmental compensation in Germany, the USA, the Netherlands, the UK and Sweden. In Germany compensation methods have been used since the 1970s. In the USA there are requirements for mitigation and compensation measures for wetland losses. In the Netherlands, the compensation measures are focused on certain protected areas. In the UK there are few legal instruments that formally require environmental compensation. In Sweden the new environmental legislation has made it possible to legally demand environmental compensation measures to gain permits for exploitations. However, these new Swedish requirements have only been available since 1999 and the use of compensation measures needs to be developed. In this article we argue that the German environmental compensation practice is most developed and is worth further study in order to develop the use of environmental compensation methods in the Swedish planning system. Copyright © 2003 John Wiley & Sons, Ltd and ERP Environment.
Top‐down and data‐based mechanistic modelling of rainfall–flow dynamics at the catchment scale
Authors: Young Peter 
Published: August 2003

AbstractThe data‐based mechanistic (DBM) approach to modelling has developed as a stochastic, ‘top‐down’ response to the problems associated with the deterministic, ‘bottom‐up’ approach. As such, it can be compared with the deterministic, top‐down modelling methods that have been attracting attention recently in the hydrological literature. Using catchment‐scale rainfall–flow modelling as an example, this paper compares the inductive DBM approach with its hypothetico‐deductive, deterministic alternative and shows how they can be used to identify and estimate low‐order, nonlinear models of the rainfall–flow dynamics in the River Hodder catchment of northwest England based on a limited set of rainfall–flow data. Copyright © 2003 John Wiley & Sons, Ltd.
Data‐supported robust parameterisations in land surface–atmosphere flux predictions: towards a top‐down approach
Authors: Schulz Karsten , Beven Keith 
Published: August 2003

AbstractThe current tendency in ‘physically based’ soil–vegetation–atmosphere transfer (SVAT) schemes is to use increasingly complex process descriptions to predict evaporative fluxes at both patch and landscape scales. This approach does not take proper account of the heterogeneities that are evident in any landscape. A top‐down approach to sub‐grid‐scale land surface parameterization would suggest that the current complexity may not be supported by the calibration data available. By comparing three SVAT schemes of differing complexity within the generalized likelihood uncertainty estimation framework, we demonstrate the utility of simpler rather than more complex models when calibrated against flux data from various intensive field campaigns. A more robust calibration is achieved for a simple evaporative fraction approach allowing the feasible parameter ranges to be more strongly conditioned by the available data. It is argued that a top‐down (dominant mode) predictive model based on a database of such robustly estimated parameter values would result in no greater uncertainty at the scales of application than trying to form parameter sets for complex models from a variety of sources. Copyright © 2003 John Wiley & Sons, Ltd.
NOESY, HOESY, T1 and solid‐state NMR studies on [RuH(η6‐toluene)(Binap)](CF3SO3): a molecule with a strongly distorted piano‐stool structure
Authors: Geldbach Tilmann J. , Rüegger Heinz , Pregosin Paul S. 
Published: August 2003

AbstractDetailed solution‐NMR studies on the distorted ruthenium hydride complex [RuH(η6‐toluene)(Binap)](CF3SO3) (2) are reported. NOE‐spectroscopy, together with low‐temperature 1H and 31P NMR data, reveals restricted rotation around a P—C bond for a specific axial P—phenyl ring with the activation energy determined via simulation. From 19F, 1H HOESY data, the approach of the triflate anion relative to the hydride ligand is established. Comparison of the quadrupole coupling constant CQF from both solution‐ and solid‐state MAS‐NMR on the deuteride [RuD(η6‐benzene)(Binap)](CF3SO3) (1‐D) provide information on the nature of the Ru—H bond. Copyright © 2003 John Wiley & Sons, Ltd.
Non‐degradative dissolution and acetylation of ball‐milled plant cell walls: high‐resolution solution‐state NMR
Authors: Lu Fachuang , Ralph John 
Published: August 2003

SummaryTwo solvent systems for fully dissolving, and optionally derivatizing, finely ground plant cell wall material at room temperature are described: dimethylsulfoxide (DMSO) and tetrabutylammonium fluoride (TBAF) or N‐methylimidazole (NMI). In situ acetylation produces acetylated cell walls (Ac‐CWs) that are fully soluble in chloroform. Lignin structures tested remain fully intact. The dispersion of 13C–1H correlations afforded by two‐dimensional (2D) nuclear magnetic resonance (NMR) experiments reveals the major lignin units, allowing the whole lignin fraction to be analyzed by high‐resolution solution‐state NMR methods for the first time. Non‐degradative cell wall dissolution offers the potential to analyze polysaccharide components, and improve current cell wall analytical methods by using standard homogeneous solution‐state chemistry.
The plant cyclin‐dependent kinase inhibitor ICK1 has distinct functional domains for in vivo kinase inhibition, protein instability and nuclear localization
Authors: Zhou Yongming , Li Genyi , Brandizzi Federica , Fowke Larry C. , Wang Hong 
Published: August 2003

SummaryInteractor/inhibitor 1 of Cdc2 kinase (ICK1) from Arabidopsis thaliana is the first plant cyclin‐dependent kinase (CDK) inhibitor, and overexpression of ICK1 inhibits CDK activity, cell division and plant growth in transgenic plants. In this study, ICK1 and deletion mutants were expressed either alone or as green fluorescent protein (GFP) fusion proteins in transgenic Arabidopsis plants. Deletion of the C‐terminal 15 or 29 amino acids greatly reduced or completely abolished the effects of ICK1 on the transgenic plants, and recombinant proteins lacking the C‐terminal residues lost the ability to bind to CDK complex and the kinase inhibition activity, demonstrating the role of the conserved C‐terminal domain in in vivo kinase inhibition. In contrast, the mutant ICK1ΔN108 with the N‐terminal 108 residues deleted had much stronger effects on plants than the full‐length ICK1. Analyses demonstrated that this effect was not because of an enhanced ability of ICK1ΔN108 protein to inhibit CDK activity, but a result of a much higher level of ICK1ΔN108 protein in the plants, indicating that the N‐terminal domain contains a sequence or element increasing protein instability in vivo. Furthermore, GFP‐ICK1 protein was restricted to the nuclei in roots of transgenic plants, even with the C‐terminal or the N‐terminal domain deleted, suggesting that a sequence in the central domain of ICK1 is responsible for nuclear localization. These results provide mechanistic understanding about the function and regulation of this cell cycle regulator in plants.
Epidemiological Survey of
Babesia
Species in Japan Performed with Specimens from Ticks Collected from Dogs and Detection of New
Babesia
DNA Closely Related to
Babesia odocoilei
and
Babesia divergens
DNA
Authors: Inokuma Hisashi , Yoshizaki Yukako , Shimada Yojiro , Sakata Yoshimi , Okuda Masaru , Onishi Takafumi 
Published: August 2003

ABSTRACT

Detection and analysis of
Babesia
species from ticks recovered from dogs in Japan were attempted by PCR and nucleotide sequence analysis based on the 18S rRNA gene, respectively. A total of 1,136 ticks were examined for
Babesia
DNA by 18S rRNA-based PCR and nucleotide sequencing. Partial sequences of
Babesia canis vogeli
DNA were detected from six ticks in Aomori, Nara, Hiroshima, Oita, and Okinawa Prefectures; and
Babesia gibsoni
Asia-1 DNA was also detected in four ticks in Osaka, Hiroshima, Miyazaki, and Okinawa Prefectures. Unique sequences of 1,678 bp were also obtained from
Ixodes ovatus
ticks in Akita and Fukui Prefectures. The sequences were similar to those of
Babesia odocoilei
(97.7%) and
Babesia divergens
(97.6%). This is the first report of the detection of DNA belonging to this group in Japan.
Single-Nucleotide-Polymorphism Mapping of the
Pseudomonas aeruginosa
Type III Secretion Toxins for Development of a Diagnostic Multiplex PCR System
Authors: Ajayi Temitayo , Allmond Leonard R. , Sawa Teiji , Wiener-Kronish Jeanine P. 
Published: August 2003

ABSTRACT

We mapped the coding single nucleotide polymorphisms in four toxin genes—
exoS
,
exoT
,
exoU
, and
exoY
—of the
Pseudomonas aeruginosa
type III secretion system among several clinical isolates. We then used this information to design a multiplex PCR assay based on the simultaneous amplification of fragments of these genes. Eight strains of known genotype were used to test our multiplex PCR method, which showed 100% sensitivity and specificity in this small sample size. This assay appears to be promising for the rapid and accurate genotyping of the presence of these genes in clinical strains of
P. aeruginosa
.