bluuebunny/crossref_metadata_2025_split huggingface dataset:
Method of separation and concentration of fetal nucleated red blood cells in maternal blood and its application to fetal diagnosis
Authors: Wada Seiji , Kitagawa Michihiro 
Published: May 2004

ABSTRACT  A fetal diagnostic method that is without risk to the embryo has been long awaited in the field of gene diagnosis. Establishment of non‐invasive fetal diagnosis using maternal peripheral blood will greatly contribute to perinatal medical care. The lectin method that we have studied and developed selectively recovers nucleated red blood cells (NRBC) among fetal cells mixed in maternal peripheral blood.Maternal blood, 7 mL, was collected with ethylene diamine tetraacetic acid (EDTA) treatment in each week of gestation and subjected to preliminary concentration by density centrifugation (Histopaque [Histopaque‐1077; Sigma Diagnostics, MO, USA], specific gravity: 1.095), and NRBC were separated and collected on slide glasses by the lectin method (soybean agglutinin [SBA]: 50 µg/mL).To investigate selective adhesion of the erythrocyte fraction, the SBA concentration was set to 50 µg/mL, and the cells were labeled with CD11a and CD33 (anti‐white blood cell antibodies) and investigated by flowcytometry. Erythrocytes adhered at a high rate (87.0 ± 9.7%) while the adhesion rates of granulocytes, monocytes, and lymphocytes were low, confirming the usefulness of the method for separation and recovery of the erythrocyte fraction.When recovery of NRBC was investigated using this method, a mean of 6.57 ± 7.12 cells were recovered from 1 mL of maternal blood (May–Gluüwald–Giemsa stain). The number of recovered NRBC increased slowly with pregnancy, but differences were not significant. To confirm that recovered NRBC were derived from the fetus, NRBC were recovered by the lectin method in four patients suspected of 18 trisomy by echography and analyzed by fluorescence in situ hybridization. Three hybridization signals were detected in NRBC at a high frequency, showing that most cells were derived from the fetus and thus, fetal diagnosis may be possible.Since the procedure of the lectin method we have developed is simple, and high concentration efficiency can be obtained at a low cost, it may be clinically applicable.
S Protein of Severe Acute Respiratory Syndrome-Associated Coronavirus Mediates Entry into Hepatoma Cell Lines and Is Targeted by Neutralizing Antibodies in Infected Patients
Authors: Hofmann Heike , Hattermann Kim , Marzi Andrea , Gramberg Thomas , Geier Martina , Krumbiegel Mandy , Kuate Seraphin , Überla Klaus , Niedrig Matthias , Pöhlmann Stefan 
Published: May 2004

ABSTRACT
The severe acute respiratory syndrome-associated coronavirus (SARS-CoV) causes severe pneumonia with a fatal outcome in approximately 10% of patients. SARS-CoV is not closely related to other coronaviruses but shares a similar genome organization. Entry of coronaviruses into target cells is mediated by the viral S protein. We functionally analyzed SARS-CoV S using pseudotyped lentiviral particles (pseudotypes). The SARS-CoV S protein was found to be expressed at the cell surface upon transient transfection. Coexpression of SARS-CoV S with human immunodeficiency virus-based reporter constructs yielded viruses that were infectious for a range of cell lines. Most notably, viral pseudotypes harboring SARS-CoV S infected hepatoma cell lines but not T- and B-cell lines. Infection of the hepatoma cell line Huh-7 was also observed with replication-competent SARS-CoV, indicating that hepatocytes might be targeted by SARS-CoV in vivo. Inhibition of vacuolar acidification impaired infection by SARS-CoV S-bearing pseudotypes, indicating that S-mediated entry requires low pH. Finally, infection by SARS-CoV S pseudotypes but not by vesicular stomatitis virus G pseudotypes was efficiently inhibited by a rabbit serum raised against SARS-CoV particles and by sera from SARS patients, demonstrating that SARS-CoV S is a target for neutralizing antibodies and that such antibodies are generated in SARS-CoV-infected patients. Our results show that viral pseudotyping can be employed for the analysis of SARS-CoV S function. Moreover, we provide evidence that SARS-CoV infection might not be limited to lung tissue and can be inhibited by the humoral immune response in infected patients.
Early Detection of a Two-Long-Terminal-Repeat Junction Molecule in the Cytoplasm of Recombinant Murine Leukemia Virus-Infected Cells
Authors: Serhan Fatima , Penaud Magalie , Petit Caroline , Leste-Lasserre Thierry , Trajcevski Stéphane , Klatzmann David , Duisit Ghislaine , Sonigo Pierre , Moullier Philippe 
Published: May 2004

ABSTRACT
We showed that a U5-U3 junction was reproducibly detected by a PCR assay as early as 1 to 2 h postinfection with a DNase-treated murine leukemia virus (MLV)-containing supernatant in aphidicolin-arrested NIH 3T3 cells, as well as in nonarrested cells. Such detection is azidothymidine sensitive and corresponded to neosynthesized products of the reverse transcriptase. This observation was confirmed in two additional human cell lines, TE671 and ARPE-19. Using cell fractionation combined with careful controls, we found that a two-long-terminal-repeat (two-LTR) junction molecule was detectable in the cytoplasm as early as 2 h post virus entry. Altogether, our data indicated that the neosynthesized retroviral DNA led to the early formation of structures including true two-LTR junctions in the cytoplasm of MLV-infected cells. Thus, the classical assumption that two-LTR circles are a mitosis-dependent dead-end product accumulating in the nucleus must be reconsidered. MLV-derived products containing a two-LTR junction can no longer be used as an exclusive surrogate for the preintegration complex nuclear translocation event.
PB1-F2, an Influenza A Virus-Encoded Proapoptotic Mitochondrial Protein, Creates Variably Sized Pores in Planar Lipid Membranes
Authors: Chanturiya A. N. , Basañez G. , Schubert U. , Henklein P. , Yewdell J. W. , Zimmerberg J. 
Published: May 2004

ABSTRACT
A frameshifted region of the influenza A virus PB1 gene encodes a novel protein, termed PB1-F2, a mitochondrial protein that can induce cell death. Many proapoptotic proteins are believed to act at the mitochondrial outer membrane to form an apoptotic pore with lipids. We studied the interaction of isolated, synthetic PB1-F2 (sPB1-F2) peptide with planar phospholipid bilayer membranes. The presence of nanomolar concentrations of peptide in the bathing solution induced a transmembrane conductance that increased in a potential-dependent manner. Positive potential on the side of protein addition resulted in a severalfold increase in the rate of change of membrane conductance. sPB1-F2-treated membranes became permeable to monovalent cations, chloride, and to a lesser extent, divalent ions. Despite various experimental conditions, we did not detect the distinctive conductance levels typical of large, stable pores, protein channels, or even pores that are partially proteinaceous. Rather, membrane conductance induced by sPB1-F2 fluctuated and visited almost all conductance values. sPB1-F2 also dramatically decreased bilayer stability in an electric field, consistent with a decrease in the line tension of a lipidic pore. Since similar membrane-destabilizing profiles are seen with proapoptotic proteins (e.g., Bax) and the cytoplasmic helix of human immunodeficiency virus gp41, we suggest that the basis for sPB1-F2-induced cell death may be the permeabilization and destabilization of mitochondrial membranes, leading to macromolecular leakage and apoptosis.
Cross-Packaging of Human Immunodeficiency Virus Type 1 Vector RNA by Spleen Necrosis Virus Proteins: Construction of a New Generation of Spleen Necrosis Virus-Derived Retroviral Vectors
Authors: Parveen Zahida , Mukhtar Muhammad , Goodrich Adrienne , Acheampong Edward , Dornburg Ralph , Pomerantz Roger J. 
Published: May 2004

ABSTRACT

The ability of the nonlentiviral retrovirus spleen necrosis virus (SNV) to cross-package the genomic RNA of the distantly related human immunodeficiency virus type 1 (HIV-1) and vice versa was analyzed. Such a model may allow us to further study HIV-1 replication and pathogenesis, as well as to develop safe gene therapy vectors. Our results suggest that SNV can cross-package HIV-1 genomic RNA but with lower efficiency than HIV-1 proteins. However, HIV-1-specific proteins were unable to cross-package SNV RNA. We also constructed SNV-based
gag-pol
chimeric variants by replacing the SNV integrase with the HIV-1 integrase, based on multiple sequence alignments and domain analyses. These analyses revealed that there are conserved domains in all retroviral integrase open reading frames (
orf
), despite the divergence in the primary sequences. The transcomplementation assays suggested that SNV proteins recognized one of the chimeric variants. This demonstrated that HIV-1 integrase is functional in the SNV
gag-pol orf
with a lower transduction efficiency, utilizing homologous (SNV) RNA, as well as the heterologous vector RNA of HIV-1. These findings suggest that homology in the conserved sequences of the integrase protein may not be fully competent in the replacement of protein(s) from one retrovirus to another, and there are likely several other factors involved in each of the steps related to replication, integration, and infection. However, further studies to dissect the
gag-pol
region will be critical for understanding the mechanisms involved in the cleavage of reverse transcriptase, RNase H, and integrase. These studies should provide further insight into the design and development of novel molecular approaches to block HIV-1 replication and to construct a new generation of SNV-based vectors.
Role of an Arbovirus Nonstructural Protein in Cellular Pathogenesis and Virus Release
Authors: Owens Randall J. , Limn Chang , Roy Polly 
Published: May 2004

ABSTRACT

The insect-borne
Bluetongue virus
(BTV) is considered the prototypic
Orbivirus
, a member of the
Reovirus
family. One of the hallmarks of
Orbivirus
infection is the production of large numbers of intracellular tubular structures of unknown function. For BTV these structures are formed as the polymerization product of a single 64-kDa nonstructural protein, NS1, encoded by the viral double-stranded RNA genome segment 6. Although the NS1 protein is the most abundant viral protein synthesized in infected cells, its function has yet to be determined. One possibility is that NS1 tubules may be involved in the translocation of newly formed viral particles to the plasma membrane, and NS1-specific monoclonal antibodies have been shown to react with viral particles leaving infected cells. In the present study we generated a mammalian cell line that expresses a recombinant single-chain antibody fragment (scFv) derived from an NS1-specific monoclonal antibody (10B1) and analyzed the effect that this intracellular antibody has on BTV replication. Normally, BTV infection of mammalian cells in culture results in a severe cytopathic effect within 24 to 48 h postinfection manifested by cell rounding, apoptosis, and lytic release of virions into the culture medium. However, infection of scFv-expressing cells results in a marked reduction in the stability of NS1 and formation of NS1 tubules, a decrease in cytopathic effect, an increased release of infectious virus into the culture medium, and budding of virions from the plasma membrane. These results suggest that NS1 tubules play a direct role in the cellular pathogenesis and morphogenesis of BTV.
Postattachment Events Associated with Viral Entry Are Necessary for Induction of Interferon-Stimulated Genes by Human Cytomegalovirus
Authors: Netterwald James R. , Jones Thomas R. , Britt William J. , Yang Shao-Jun , McCrone Ian P. , Zhu Hua 
Published: May 2004

ABSTRACT
Utilizing a human cytomegalovirus-specific fusion inhibitor and an antiglycoprotein H antibody, we studied the role of virion fusion in interferon-stimulated gene (ISG) induction. Our results indicate that ISG induction does not occur when virion-mediated, post-high-affinity attachment events are inhibited by either reagent. Thus, virion-mediated postattachment events, such as fusion, are required for ISG induction.
Sur la continuité automatique des épimorphismes dans les ☆‐algèbres de Banach
Authors: Oukhtite L. , Tajmouati A. , Tidli Y. 
Published: May 2004

Nous étudions les problèmes de continuité automatique dans des algèbres de Banach avec involutions.
Nous obtenons aussi des nouveoux résultats concernant ☆‐idéals des ☆‐algèbres.We study the automatic continuity problems for Banach algebras with involutions. We also obtain some new results concerning ☆‐ideals of ☆‐algebras.
Uncertainties of Derived Dewpoint Temperature and Relative Humidity
Authors: Lin X. , Hubbard K. G. 
Published: May 2004

Abstract
This paper presents an evaluation of derived dewpoint temperature and derived relative humidity, in which the dewpoint temperature is calculated using measured ambient air temperature and measured relative humidity variables and the derived relative humidity is calculated from measured dewpoint temperature. The derived dewpoint temperature and relative humidity are calculated using algorithms provided by the World Meteorological Organization. The method of uncertainty analysis, provided by the National Institute of Standards and Technology, is applied to calculate the uncertainties of an indirect measurement of derived dewpoint temperature and derived relative humidity. The results from the uncertainty analyses of derived and observed variables suggest that the use of derived dewpoint temperature and derived relative humidity involves risk because the uncertainties of modern dewpoint temperature and relative humidity sensors can create several degrees Celsius of error in the derived dewpoint temperature and several percent in the derived relative humidity.
Improvement in quality of life of patients with AL amyloidosis treated with high-dose melphalan and autologous stem cell transplantation
Authors: Seldin David C. , Anderson Jennifer J. , Sanchorawala Vaishali , Malek Karim , Wright Daniel G. , Quillen Karen , Finn Kathleen T. , Berk John L. , Dember Laura M. , Falk Rodney H. , Skinner Martha 
Published: May 2004

Abstract
Treatment of AL amyloidosis patients with high-dose melphalan chemotherapy followed by autologous peripheral blood stem cell transplantation (HDM/SCT) can produce hematologic complete responses (CRs) and improvement in organ function. To determine whether these responses are accompanied by improvement in quality of life (QOL), we employed the Medical Outcomes Study (MOS) 36-item Short Form General Health Survey (SF-36) questionnaire for 544 patients evaluated between 1994 and 2002. At baseline, the scores were significantly lower on all 8 SF-36 scales compared with age-matched population norms: the composite physical component summary (PCS) for the AL patients was 34.5 versus the population norm of 46.8, and the mental component summary (MCS) was 45.0 versus the norm of 51.5. All SF-36 scores improved at 1 year, with the MCS reaching the population norm. The PCS, though improved, was still lower than normal but was greater in the subgroup of patients who achieved a hematologic CR; the PCS normalized at 2 years in these patients. Thus, treatment of AL amyloidosis patients with HDM/SCT produces measurable and sustained improvements in quality of life, particularly in those patients who achieve hematologic CR.
Experimental rhesus lymphocryptovirus infection in immunosuppressed macaques: an animal model for Epstein-Barr virus pathogenesis in the immunosuppressed host
Authors: Rivailler Pierre , Carville Angela , Kaur Amitinder , Rao Pasupuleti , Quink Carol , Kutok Jeffery L. , Westmoreland Susan , Klumpp Sherry , Simon Meredith , Aster Jon C. , Wang Fred 
Published: May 2004

Abstract
To develop a model for Epstein-Barr virus (EBV) pathogenesis in immunosuppressed hosts, we studied experimental infections of immunocompetent versus SHIV 89.6P–infected, immunosuppressed rhesus macaques with the EBV-related rhesus lymphocryptovirus (LCV). Primary LCV infection after oral inoculation of 4 immunocompetent animals was characterized by an acute viremia and seroconversion followed by asymptomatic LCV persistence. Four immunosuppressed macaques infected orally with LCV failed to develop an LCV-specific humoral response and viremia was more pronounced, but there was no evidence of LCV-induced lymphoproliferative disease. A more aggressive primary challenge was administered by intravenous inoculation of 108 autologous, LCV-immortalized B cells in 4 additional immunosuppressed animals. Two animals with modest immunosuppression remained asymptomatic, and 1 of 2 severely immunosuppressed animals developed an aggressive, monoclonal LCV-positive lymphoma. These studies demonstrate the potential for lymphomagenesis in an experimental model system for EBV infection and underscore the strength and depth of immune control in limiting LCV-induced lymphoproliferative disease.
Transduction of NO-bioactivity by the red blood cell in sepsis: novel mechanisms of vasodilation during acute inflammatory disease
Authors: Crawford Jack H. , Chacko Balu K. , Pruitt Heather M. , Piknova Barbora , Hogg Neil , Patel Rakesh P. 
Published: May 2004

Abstract
Sepsis is an acute inflammatory disease characterized by dysfunctional blood flow and hypotension. Nitric oxide (NO) is elevated during sepsis and plays an integral role in the associated vascular pathology. However, precise mechanisms and functions of NO in sepsis remain unclear. In this study, we show that red blood cells (RBCs) are foci for nitrosative reactions during acute inflammation, resulting in the formation of cells that can promote systemic vascular relaxation in an uncontrolled manner. Specifically, using experimental models of endotoxemia and surgical sepsis, NO adducts were found in the RBCs, including S-nitrosohemoglobin (SNOHb). These RBCs, referred to as septic RBCs, spontaneously stimulated vasodilation in a manner consistent with elevated SNOHb concentrations. Moreover, relaxation was cyclic guanosine monophosphate (cGMP) dependent and was inhibited by RBC lysis and glutathione but not by the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5 tetramethylimidazoline 1-oxyl 3-oxide (C-PTIO). The potential mechanism of septic RBC–mediated vasorelaxation is discussed and may involve the intermediate, nitroxyl (HNO). Coupled with data showing that NO adducts in septic RBCs were dependent on the inducible nitric oxide synthase and correlated with plasma nitrite, these findings provide a novel framework to understand mechanisms underlying dysfunctional blood flow responses during sepsis. Specifically, the concept that RBCs directly mediate systemic hypotension through NO-dependent mechanisms is discussed.
Layered and laterally constrained 2D inversion of resistivity data
Authors: Auken Esben , Christiansen Anders Vest 
Published: May 2004

In a sedimentary environment, quasi‐layered models often can represent the actual geology more accurately than smooth minimum‐structure models. We present a 2D inversion scheme with lateral constraints and sharp boundaries (LCI) for continuous resistivity data. All data and models are inverted as one system, producing layered solutions with laterally smooth transitions. The models are regularized through lateral constraints that tie interface depths or thicknesses and resistivities of adjacent layers. A priori information, used to resolve ambiguities and to add, for example, geological information, can be added at any point of the profile and migrates through the lateral constraints to parameters at adjacent sites. Similarly, information from areas with well‐resolved parameters migrates through the constraints to help resolve areas with poorly constrained parameters. The estimated model is complemented by a full sensitivity analysis of the model parameters supporting quantitative evaluation of the inversion result.A simple synthetic model proves the need for a quasi‐layered, 2D inversion when compared with a traditional 2D minimum‐structure inversion. A 2D minimum‐structure inversion produces models with spatially smooth resistivity transitions, making identification of layer boundaries difficult.A continuous vertical electrical sounding field example from Sweden with a depression in the depth to bedrock supports the conclusions drawn from the synthetic example. A till layer on top of the bedrock, hidden in the traditional inversion result, is identified using the 2D LCI scheme. Furthermore, the depth to the bedrock surface is easily identified for most of the profile with the 2D LCI model, which is not the case with the model from the traditional minimum‐structure inversion.
Multiple-stage screening and mortality in the Multiple Risk Factor Intervention Trial
Authors: Eberly Lynn E , Neaton James D , Thomas Avis J , Dai Yu ,   Multiple Risk Factor Intervention Trial Research Group
Published: May 2004

Background During the design phase of a clinical trial, sample size estimates should take into account medical screening criteria, the ‘healthy volunteer’ effect, consequences of run-in phases, and secular trends in the event rate of interest. All of these have been shown to relate to subsequent event rates, and hence trial power to detect intervention effects. The Multiple Risk Factor Intervention Trial (MRFIT) used three successive screenings of 361 662 men to enroll 12 866; observed coronary heart disease (CHD) mortality after a mean of 6.9 years was substantially lower than projected during design. We explore factors which may have contributed to these mortality differences and whether they persisted throughout follow-up. Methods Proportional hazards models were used to compare 25-year mortality according to trial eligibility, self-exclusions, medical exclusions, and participation. Results After adjustment for baseline risk factors and age, there was higher mortality among men excluded for presence of disease [hazard ratio (HR) 1.52, 95% confidence interval (CI) 1.43–1.61, for total; HR 1.92, CI 1.75–2.11, for CHD] compared to those not excluded which persisted throughout follow-up. Volunteers had lower total (HR 0.82, CI 0.76–0.87) and CHD (HR 0.79, CI 0.70–0.88) mortality than those discontinuing participation. Men excluded with characteristics deemed likely to interfere with adherence had higher total (HR 1.19, CI 1.07–1.33) and noncardiovascular disease (CVD) (HR 1.32, CI 1.14–1.53) mortality but no higher CVD (HR 1.04, CI 0.88–1.23) or CHD (HR 0.98, CI 0.80–1.20) mortality compared to those not excluded. Differences in mortality were stronger during the first five years, but declined only slightly over 25 years. Conclusions 25-year mortality was significantly higher for non-volunteers and exclusions. Differences between observed and predicted six-year total mortality for trial participants were largely attributable to volunteers and exclusions, but there were additional differences for CHD mortality, which were likely due to downward secular trends. These results emphasize the importance of anticipating these factors during clinical trial design, even for trials of short duration.