bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ten‐Year Follow‐Up of a Patient with Metastatic Ewing′s Sarcomaof the Pelvis
Authors: Ashford Robert U. , McCloskey Eugene V. , Purohit O. Prakash , Ingram Christine E. , Grimer Robert J. , Coleman Robert E. 
Published: January 2003

Patient: We report a 32‐year‐old women with a pelvic Ewing′s sarcoma, who developed skeletal metastases within 20 months
of diagnosis but following treatment remains disease‐free at 10 years.Discussion: Ewing′s sarcoma is a highly malignant tumour of bone. Presentation of a pelvic tumour over the age of 30 years
is extremely rare and associated with a poor prognosis. Early recurrence is also associated with an extremely grave prognosis.
In our patient there was little or no response to salvage chemotherapy, but, against the odds, remission was induced
with pamidronate and palliative radiotherapy to some but not all sites of disease.This remission has been maintained without
additional therapy for a further 5 years.
Differential Expression of IRS‐1 and IRS‐2 in UterineLeiomyosarcomas with Distinct Oncogenic Phenotypes: Lack ofCorrelation with Downstream Signaling Events
Authors: Colombatti Alfonso , Russo Pietro , Cervi Marta , Bogetto Laura , Wassermann Bruna , Mainiero Fabrizio , Spessotto Paola 
Published: January 2003

Purpose: Insulin receptor substrates (IRSs) are essential for insulin‐induced mitogenic effects on several cell types but they
also are involved in cell transformation.We investigated whether the differential constitutive expression and potential distinct
downstream signaling events of IRS‐1 and IRS‐2 might be related to discrete tumourigenic phenotypes of three human
uterine leiomyosarcoma cell lines, one of which was specifically isolated for the present study.Methods and results: SK‐UT‐1B egressed effectively from a gellyfied Matrigel matrix and grew as did DMR cells in an
anchorage‐independent manner in agar and induced rapidly growing tumours in nude mice. On the contrary, SK‐LMS‐1
cells did not emigrate from Matrigel, neither grew in agar nor were they tumourigenic. IRS‐2 was highly expressed in the
more malignant cell lines, whereas IRS‐1 was present only in SK‐LMS‐1 cells. However, upon insulin stimulation both IRS‐
1 and IRS‐2 were tyrosine phosphorylated with a similar kinetic in the respective cell lines; furthermore, after 1 min of
insulin stimulation PI3‐kinase associated with IRSs and after 2 min Shc was phosphorylated and associated with Grb2 with
minor differences detectable among the various cell lines in the duration of phosphorylation and/or in their association irrespective
of whether IRS‐1 or IRS‐2 were expressed.Discussion: Our findings tend to exclude that the malignancy displayed by uterine leiomyosarcomas might be directly linked
to the activation of distinct IRS‐1‐ or IRS‐2‐dependent pathways.
Statistical Classification Strategy for Proton Magnetic ResonanceSpectra of Soft Tissue Sarcoma: An Exploratory Study withPotential Clinical Utility
Authors: Bezabeh Tedros , El-Sayed Samy , Patel Rakesh , Somorjai Ray L. , Bramwell Vivien , Kandel Rita , Smith Ian C. P. 
Published: January 2003

Purpose: Histological grading is currently one of the best predictors of tumor behavior and outcome in soft tissue sarcoma.
However, occasionally there is significant disagreement even among expert pathologists. An alternative method that gives
more reliable and non‐subjective diagnostic information is needed. The potential use of proton magnetic resonance spectroscopy
in combination with an appropriate statistical classification strategy was tested here in differentiating normal
mesenchymal tissue from soft tissue sarcoma.Methods: Fifty‐four normal and soft tissue sarcoma specimens of various histological types were obtained from 15 patients.
One‐dimensional proton magnetic resonance spectra were acquired at 360 MHz. Spectral data were analyzed by using both
the conventional peak area ratios and a specific statistical classification strategy.Results: The statistical classification strategy gave much better results than the conventional analysis. The overall classification
accuracy (based on the histopathology of the MRS specimens) in differentiating normal mesenchymal from soft tissue
sarcoma was 93%, with a sensitivity of 100% and specificity of 88%.The results in the test set were 83, 92 and 76%, respectively.
Our optimal region selection algorithm identified six spectral regions with discriminating potential, including those
assigned to choline, creatine, glutamine, glutamic acid and lipid.Conclusion: Proton magnetic resonance spectroscopy combined with a statistical classification strategy gave good results in
differentiating normal mesenchymal tissue from soft tissue sarcoma specimens ex vivo. Such an approach may also differentiate
benign tumors from malignant ones and this will be explored in future studies.
Identification of B- and T-Cell Epitopes of BB, a Carrier Protein Derived from the G Protein ofStreptococcusStrain G148
Authors: Goetsch Liliane , Haeuw Jean Francois , Champion Thierry , Lacheny Christine , N’Guyen Thien , Beck Alain , Corvaia Nathalie 
Published: January 2003

ABSTRACTMost conventional vaccines consist of killed organisms or purified antigenic proteins. Such molecules are generally poorly immunogenic and need to be coupled to carrier proteins. We have identified a new carrier molecule, BB, derived from the G protein ofStreptococcusstrain G148. We show that BB is able to induce strong antibody responses when conjugated to peptides or polysaccharides. In order to localize T and B cell epitopes in BB and match them with the albumin-binding region of the molecule, we immunized mice with BB, performed B and T pepscan analyses, and compared the results with pepscan done with sera and cells from humans. Our results indicate that BB has two distinct T helper epitopes, seven linear B-cell epitopes, and one conformational B-cell epitope in BALB/c mice. Four linear B-cell epitopes were identified from human sera, three of which overlapped mouse B-cell epitopes. Finally, three human T-cell epitopes were detected on the BB protein. One of these T-cell epitopes is common to BALB/c mice and humans and was localized in the region that contains the albumin-binding site. These data are of interest for the optimization of new carrier molecules derived from BB.
Use of Vesicular Stomatitis Virus Pseudotypes Bearing Hantaan or Seoul Virus Envelope Proteins in a Rapid and Safe Neutralization Test
Authors: Ogino Michiko , Ebihara Hideki , Lee Byoung-Hee , Araki Koichi , Lundkvist Åke , Kawaoka Yoshihiro , Yoshimatsu Kumiko , Arikawa Jiro 
Published: January 2003

ABSTRACT

A vesicular stomatitis virus (VSV) pseudotype bearing hantavirus envelope glycoproteins was produced and used in a neutralization test as a substitute for native hantavirus. The recombinant VSV, in which the enveloped protein gene (G) was replaced by the green fluorescent protein gene and complemented with G protein expressed in
trans
(VSVΔG*G), was kindly provided by M. A. Whitt. 293T cells were transfected with plasmids for the expression of envelope glycoproteins (G1 and G2) of HTNV or SEOV and were then infected with VSVΔG*G. Pseudotype VSV with the Hantaan (VSVΔG*-HTN) or Seoul (VSVΔG*-SEO) envelope glycoproteins were harvested from the culture supernatant. The number of infectious units (IU) of the pseudotype VSVs ranged from 10
5
to 10
6
/ml. The infectivity of VSVΔG*-HTN and VSVΔG*-SEO was neutralized with monoclonal antibodies, immune rabbit sera, and sera from patients with hemorrhagic fever with renal syndrome, and the neutralizing titers were similar to those obtained with native hantaviruses. These results show that VSVΔG*-HTN and -SEO can be used as a rapid, specific, and safe neutralization test for detecting hantavirus-neutralizing antibodies as an effective substitute for the use of native hantaviruses. Furthermore, the IU of VSVΔG*-HTN and -SEO did not decrease by more than 10-fold when stored at 4°C for up to 30 days. The stability of the pseudotype viruses allows distribution of the material to remote areas by using conventional cooling boxes for use as a diagnostic reagent.
Infection Exclusion of the Rickettsial PathogenAnaplasma marginalein the Tick VectorDermacentor variabilis
Authors: de la Fuente José , Blouin Edmour F. , Kocan Katherine M. 
Published: January 2003

ABSTRACTAnaplasma marginaleis a tick-borne, rickettsial cattle pathogen that is endemic in several areas of the United States. Recent studies (J. de la Fuente, J. C. Garcia-Garcia, E. F. Blouin, J. T. Saliki, and K. M. Kocan, Clin. Diagn. Lab. Immunol. 9:658-668, 2002) demonstrated that infection of cultured tick cells and bovine erythrocytes with one genotype ofA. marginaleexcluded infection with other genotypes, a phenomenon referred to as infection exclusion. The present study was undertaken to confirm the phenomenon of infection exclusion ofA. marginalegenotypes in a tick vector,Dermacentor variabilis. Only one genotype ofA. marginale(Virginia isolate) was detected by PCR in ticks that fed first on a calf infected with a Virginia isolate and second on a calf infected with an Oklahoma isolate. These studies demonstrate that infection exclusion ofA. marginalegenotypes also occurs in naturally infected ticks, as well as in cattle and cultured tick cells, and results in establishment of only one genotype per tick.
Detection of Low-Avidity Immunoglobulin G in Oral Fluid Samples: New Approach for Rubella Diagnosis and Surveillance
Authors: Akingbade D. , Cohen B. J. , Brown D. W. G. 
Published: January 2003

ABSTRACT
Low-avidity rubella immunoglobulin G (IgG) was detected in oral fluid samples from 30 of 32 rubella IgM-positive patients (sensitivity, 94%) and from 4 of 34 IgM-negative patients (specificity, 88%). Measuring IgG avidity in oral fluid samples could improve the reliability of rubella surveillance when the incidence of the disease and the positive predictive value of IgM tests are low.
Competitive Enzyme-Linked Immunosorbent Assay Based on a Rhoptry-Associated Protein 1 Epitope Specifically Identifies
Babesia bovis
-Infected Cattle
Authors: Goff Will L. , McElwain Terry F. , Suarez Carlos E. , Johnson Wendell C. , Brown Wendy C. , Norimine Junzo , Knowles Donald P. 
Published: January 2003

ABSTRACT

The competitive enzyme-linked immunosorbent assay (cELISA) format has proven to be an accurate, reliable, easily standardized, and high-throughput method for detecting hemoparasite infections. In the present study, a species-specific, broadly conserved, and tandemly repeated B-cell epitope within the C terminus of the rhoptry-associated protein 1 of the hemoparasite
Babesia bovis
was cloned and expressed as a histidine-tagged thioredoxin fusion peptide and used as antigen in a cELISA. The assay was optimized with defined negative and positive bovine sera, where positive sera inhibited the binding of the epitope-specific monoclonal antibody BABB75A4. The cELISA accurately differentiated animals with
B. bovis
-specific antibodies from uninfected animals and from animals with antibodies against other tick-borne hemoparasites (98.7% specificity). In addition,
B. bovis
-specific sera from Australia, Argentina, Bolivia, Puerto Rico, and Morocco inhibited the binding of BABB75A4, confirming conservation of the epitope. The assay first detected experimentally infected animals between 13 and 17 days postinfection, and with sera from naturally infected carrier cattle, was comparable to indirect immunofluorescence (98.3% concordance). The assay appears to have the characteristics necessary for an epidemiologic and disease surveillance tool.
Alterations in T-Cell Receptor Vβ Repertoire of CD4 and CD8 T Lymphocytes in Human Immunodeficiency Virus-Infected Children
Authors: Kharbanda Monica , McCloskey Thomas W. , Pahwa Rajendra , Sun Mei , Pahwa Savita 
Published: January 2003

ABSTRACTPerturbations in the T-cell receptor (TCR) Vβ repertoire were assessed in the CD4 and CD8 T lymphocytes of human immunodeficiency virus (HIV)-infected children who were receiving therapy during the chronic phase of infection by flow cytometry (FC) and PCR analysis. By FC, representation of 21 TCR Vβ subfamilies was assessed for an increased or decreased percentage in CD4 and CD8 T cells, and by PCR, 22 TCR Vβ subfamilies of CD4 and CD8 T cells were analyzed by CDR3 spectratyping for perturbations and reduction in the number of peaks, loss of Gaussian distribution, or clonal dominance. The majority of the TCR Vβ subfamilies were examined by both methods and assessed for deviation from the norm by comparison with cord blood samples. The CD8-T-lymphocyte population exhibited more perturbations than the CD4 subset, and clonal dominance was present exclusively in CD8 T cells. Of the 55 total CD8-TCR Vβ families classified with clonal dominance by CDR3 spectratyping, only 18 of these exhibited increased expression by FC. Patients with high numbers of CD8-TCR Vβ families with decreased percentages had reduced percentages of total CD4 T cells. Increases in the number of CD4-TCR Vβ families with increased percentages showed a positive correlation with skewing. Overall, changes from normal were often discordant between the two methods. This study suggests that the assessment of HIV-induced alterations in TCR Vβ families at cellular and molecular levels yields different information and that our understanding of the immune response to HIV is still evolving.
Serum Interferon (IFN)-Neutralizing Antibodies and Bioactivities of IFNs in Patients with Severe Type II Essential Mixed Cryoglobulinemia
Authors: Scagnolari Carolina , Casato Milvia , Bellomi Francesca , De Pisa Francesca , Turriziani Ombretta , Coviello Rossella , Pirro Maria Rosaria , Dianzani Ferdinando , Antonelli Guido 
Published: January 2003

ABSTRACT
The efficacy of alpha interferon (IFN-α) in the treatment of severe type II essential mixed cryoglobulinemia (EMC) has been reported previously. In some patients, the development of neutralizing antibodies to recombinant IFN-α (rIFN-α) can affect the clinical response achieved with rIFN-α; a second treatment with natural IFN-α preparations may reinduce the clinical response. In the present study the ability of leukocyte IFN (LeIFN) to restore the response was investigated from a pharmacodynamic viewpoint. Specifically, the pharmacodynamic profiles of different IFN-α preparations were studied by measuring the serum neopterin levels and the levels of expression of protein MxA mRNA in in vivo peripheral blood mononuclear cells in two patients with EMC whose resistance to rIFN-α2a treatment increased concomitantly with the development of neutralizing antibodies. These markers were measured before injection and at 24 and 48 h after a single injection of rIFN-α2a, consensus IFN [(C)IFN], or LeIFN. No increase or only a slight increase in MxA mRNA levels was detectable after administration of rIFN-α2a or (C)IFN, whereas a significant increase (≥10-fold) in MxA mRNA expression was recorded following administration of LeIFN. The neutralizing antibodies to rIFN-α2a cross-react with (C)IFN. Sera from these patients neutralized most but not all of the subtypes present in the natural IFN-α (LeIFN) mixture, and no significant increase in neopterin levels was observed after these patients were switched to LeIFN treatment. In summary, the data demonstrate that the problem of neutralizing antibodies still exists and that LeIFN may induce an increase in the level of MxA mRNA expression but not an increase in neopterin levels in patients who are resistant to treatment with rIFN-α2a or (C)IFN.
Analysis of Linear B-Cell Epitopes of the Nucleoprotein of Ebola Virus That Distinguish Ebola Virus Subtypes
Authors: Niikura Masahiro , Ikegami Tetsuro , Saijo Masayuki , Kurata Takeshi , Kurane Ichiro , Morikawa Shigeru 
Published: January 2003

ABSTRACT
Ebola virus consists of four genetically distinguishable subtypes. We developed monoclonal antibodies (MAbs) to the nucleoprotein (NP) of Ebola virus Zaire subtype and analyzed their cross-reactivities to the Reston and Sudan subtypes. We further determined the epitopes recognized by these MAbs. Three MAbs reacted with the three major subtypes and recognized a fragment containing 110 amino acids (aa) at the C-terminal extremity. They did not show specific reactivities to any 10-aa short peptides in Pepscan analyses, suggesting that these MAbs recognize conformational epitope(s) located within this region. Six MAbs recognized a fragment corresponding to aa 361 to 461 of the NP. Five of these six MAbs showed specific reactivities in Pepscan analyses, and the epitopes were identified in two regions, aa 424 to 430 and aa 451 to 455. Three MAbs that recognized the former epitope region cross-reacted with all three subtypes, and one that recognized the same epitope region was Zaire specific. One MAb, which recognized the latter epitope region, was reactive with Zaire and Sudan subtypes but not with the Reston subtype. These results suggest that Ebola virus NP has at least two linear epitope regions and that the recognition of the epitope by MAbs can vary even within the same epitope region. These MAbs showing different subtype specificities might be useful reagents for developing an immunological system to identify Ebola virus subtypes.
Measurement of Human Cytomegalovirus Loads by Quantitative Real-Time PCR for Monitoring Clinical Intervention in Transplant Recipients
Authors: Li Haijing , Dummer J. Stephen , Estes Wray R. , Meng Shufang , Wright Peter F. , Tang Yi-Wei 
Published: January 2003

ABSTRACT

Quantitative monitoring of human cytomegalovirus (HCMV) infection is helpful in determining appropriate antiviral management of transplant recipients. Quantitative PCR technologies have demonstrated accuracy in measuring systemic HCMV loads. A total of 298 consecutive whole-blood specimens submitted to the Clinical Virology Laboratory at Vanderbilt University Medical Center from 15 February to 31 October 1999 were included in the study. In addition to a qualitative colorimetric microtiter plate PCR assay (MTP-PCR) and a semiquantitative pp65 antigenemia assay, each specimen was measured for HCMV loads by a quantitative PCR assay performed on an ABI PRISM 7700 Sequence Detection System (TaqMan). Compared to results of the MTP-PCR, the sensitivity, specificity, positive predictive value, and negative predictive value were 70.5, 97.5, 87.8, and 92.8% for the antigenemia assay and were 96.7, 92.0, 75.6, and 99.1% for the TaqMan assay, respectively. There was a high correlation between antigenemia values and HCMV loads as determined by the TaqMan (
r
= 0.989;
P
< 0.001). Antigenemia values of 0, 1 to 10, 11 to 100, 101 to 1,000, and over 1,000 positive cells per 2 × 10
5
leukocytes corresponded to median HCMV loads measured by TaqMan of 125, 1,593, 5,713, 16,825, and 5,425,000 copies/ml, respectively. Corresponding to antigenemia values of 1 to 2, 10, and 50 positive cells per 2 × 10
5
leukocytes, HCMV viral loads of 1,000, 4,000, and 10,000 copies/ml are proposed as cutoff points for initiating antiviral therapy in patient groups with high, intermediate, and low risk of CMV diseases.
Commercial Assay for Detection of
Giardia lamblia
and
Cryptosporidium parvum
Antigens in Human Fecal Specimens by Rapid Solid-Phase Qualitative Immunochromatography
Authors: Garcia Lynne S. , Shimizu Robyn Y. , Novak Susan , Carroll Marilyn , Chan Frank 
Published: January 2003

ABSTRACT

The ImmunoCard STAT!
Cryptosporidium
/
Giardia
rapid assay (Meridian Bioscience, Inc.) is a solid-phase qualitative immunochromatographic assay that detects and distinguishes between
Giardia lamblia
and
Cryptosporidium parvum
in aqueous extracts of human fecal specimens (fresh, frozen, unfixed, or fixed in 5 or 10% formalin or sodium acetate-acetic acid-formalin). By using specific antibodies, antigens specific for these organisms are isolated and immobilized on a substrate. After the addition of appropriate reagents, a positive test is detected visually by the presence of a gray-black color bar (regardless of the intensity) next to the organism name printed on the test device. A control is included in the device. Steps include tube preparation (buffer, patient specimen, conjugates A and B), testing (addition of sample onto the test device), and visual reading (total time, 12 min). Test performance was evaluated with known positive and negative stool specimens (170 specimens positive for
Giardia
and 231 specimens negative for
Giardia
) (85 specimens positive for
Cryptosporidium
and 316 specimens negative for
Cryptosporidium
); they were tested with trichrome, iron-hematoxylin, or modified acid-fast stains or the Meridian Bioscience, Inc.,
Giardia
/
Cryptosporidium
Merifluor combination reagent; specimens with discrepant results were retested by using the Merifluor combination reagent. On the basis of the results of the reference methods, the sensitivities, specificities, and positive and negative predictive values were as follows: for
G. lamblia
, 93.5, 100, 100, and 95.5%, respectively; for
C. parvum
, 98.8, 100, 100, and 99.7%, respectively. False-negative results for
G. lamblia
were obtained with specimens with low parasite numbers (
n
= 7) or specimens containing trophozoites only (
n
= 3); one specimen with a false-negative result contained numerous cysts. The one specimen false negative for
C. parvum
was confirmed to be positive by immunofluorescence. No cross-reactivity was seen with 10 different protozoa (152 challenges), nine different helminths (35 challenges), or human cells (4 challenges) found in fecal specimens. This rapid test system may be very beneficial in the absence of trained microscopists; however, for patients who remain symptomatic after a negative result, the ova and parasite examination and special stains for other coccidia and the microsporidia should always remain options.
Detection of
Mycoplasma genitalium
by PCR Amplification of the 16S rRNA Gene
Authors: Jensen Jørgen Skov , Borre Martin B. , Dohn Birthe 
Published: January 2003

ABSTRACT

In order to develop a species-specific PCR for the detection of
Mycoplasma genitalium
, the sequence of 1,490 bases of the 16S rRNA gene was determined for
M. genitalium
G37 (type strain) and four Danish isolates of
M. genitalium
. The sequences of the four Danish strains, mutually different with respect to their MgPa gene, were 100% homologous, although they carried a single common base substitution compared to the type strain. Among members of the
Mycoplasma pneumoniae
phylogenetic cluster,
M. genitalium
showed the most-prominent homology to the 16S rRNA sequence of
M. pneumoniae
(98% homology). From regions showing the least homology to the
M. pneumoniae
16S rRNA gene sequence, primers were chosen to amplify DNA from
M. genitalium
only. Two sets of primers were selected for their ability to detect <10 to 50
M. genitalium
genome copies without cross-reactions with
M. pneumoniae
. The performance of these primers was compared to the performance of two pairs of primers amplifying parts of the MgPa adhesin gene; 1,030 randomly selected specimens submitted for
Chlamydia trachomatis
culture were screened with one of the 16S rRNA gene primer sets. A total of 41 specimens were found to be positive for this gene; 40 of these could be confirmed by one of the MgPa primer sets, whereas the other MgPa primer set detected only 21 positive specimens out of 40. These results indicate that estimates of the prevalence of
M. genitalium
in various populations using MgPa PCR primers could be incorrectly low if the PCR primers are located in variable regions of the MgPa gene.