bluuebunny/crossref_metadata_2025_split huggingface dataset:
Recombinant Protein-Based Enzyme-Linked Immunosorbent Assay and Immunochromatographic Tests for Detection of Immunoglobulin G Antibodies to Severe Acute Respiratory Syndrome (SARS) Coronavirus in SARS Patients
Authors: Guan Ming , Chen Hsiao Ying , Foo Shen Yun , Tan Yee-Joo , Goh Phuay-Yee , Wee Shock Hwa 
Published: March 2004

ABSTRACT

An enzyme-linked immunosorbent assay (ELISA) and a rapid immunochromatographic test for detection of immunoglobulin G (IgG) antibodies in severe acute respiratory syndrome (SARS) patients were developed by utilizing the well-characterized recombinant proteins Gst-N and Gst-U274. The ELISA detected IgG antibodies to SARS-CoV in all 74 convalescent-phase samples from SARS patients while weakly cross-reacting to only 1 of the 210 control sera from healthy donors. This finding thus led to a kit sensitivity, specificity, and accuracy of 100, 99.5, and 99.6%, respectively. The test thus provided a positive predictive value (PPV) of 98.7% and a negative predictive value (NPV) of 100%. In addition, the ELISA gave a positive delta of 5.4 and a negative delta of 3.6, indicating an excellent differentiation between positives and negatives. The same recombinant proteins were also applied to a newly developed platform for the development of a 15-min rapid test. The resulting rapid test has an excellent agreement of 99.6%, with a kappa value of 1.00, with the ELISA. Again, this rapid test was able to detect 100% of the samples tested (
n
= 42) while maintaining a specificity of 99.0% (
n
= 210). The PPV and NPV for the rapid test thus reached 95.3 and 100%, respectively.
Specific HLA-DRB and -DQB Alleles and Haplotypes Confer Disease Susceptibility or Resistance in Bahraini Type 1 Diabetes Patients
Authors: Al-Harbi Einas M. , Abbassi Abdul-Jabbar , Tamim Hala , al-Jenaidi Fayza , Kooheji Mariam , Kamal Madeeha , al-Mahroos Salwa , al-Nasir Faisal , Motala Ayesha A. , Almawi Wassim Y. 
Published: March 2004

ABSTRACT

Insofar as genetic susceptibility to type 1 diabetes is associated with HLA class II genes, with certain allelic combinations conferring disease susceptibility or resistance, this study assessed the distributions of HLA-DR and -DQ among 107 unrelated patients with type 1 diabetes and 88 healthy controls from Bahrain, all of Arab origin. The HLA
-
DRB and -DQB genotypes were determined by PCR-sequence-specific priming. The following alleles showed the strongest association with type 1 diabetes among patients versus controls according to their frequencies:
DRB1
*
030101
(0.430 versus 0.097;
P
< 0.001),
DRB1
*
040101
(0.243 versus 0.034;
P
< 0.001),
DQB1
*
0201
(0.467 versus 0.193;
P
< 0.001), and
DQB1
*
0302
(0.229 versus 0.091;
P
< 0.001). When the frequencies of alleles in controls were compared to those in patients, negative associations were seen for
DRB1
*
100101
(0.085 versus 0.014;
P
< 0.001),
DRB1
*
110101
(0.210 versus 0.060;
P
< 0.001),
DQB1
*
030101
(0.170 versus 0.075;
P
= 0.006), and
DQB1
*
050101
(0.335 versus 0.121;
P
< 0.001). In addition, the
DRB1
*
030101
-
DQB1
*
0201
(70.1 versus 22.7%;
P
< 0.001) and
DRB1
*
030101
-
DQB1
*
0302
(21.5 versus 0.0%;
P
< 0.001) genotypes were more prevalent among patients, thereby conferring disease susceptibility, whereas the
DRB1
*
100101
-
DQB1
*
050101
(20.5 versus 2.8%;
P
< 0.001),
DRB1
*
110101
-
DQB1
*
030101
(28.4 versus 8.4%;
P
< 0.001), and
DRB1
*
110101
-
DQB1
*
050101
(30.7 versus 0.9%;
P
< 0.001) genotypes were more prevalent among controls, thus assigning a protective role. These results confirm the association of specific HLA-DR and -DQ alleles and haplotypes with type 1 diabetes and may underline several characteristics that distinguish Bahraini patients from other Caucasians patients.
Innate Immune Responses of Calves during Transient Infection with a Noncytopathic Strain of Bovine Viral Diarrhea Virus
Authors: Müller-Doblies Doris , Arquint Adrian , Schaller Patrick , Heegaard Peter M. H. , Hilbe Monika , Albini Sarah , Abril Carlos , Tobler Kurt , Ehrensperger Felix , Peterhans Ernst , Ackermann Mathias , Metzler Alfred 
Published: March 2004

ABSTRACTIn this study, six immunocompetent calves were experimentally infected with a noncytopathic strain of bovine viral diarrhea virus (BVDV), and the effects of the viral infection on parameters of the innate immune response of the host were analyzed. Clinical and virological data were compared with the temporal activation of the alpha/beta interferon-regulated Mx gene in white blood cells (WBC) and skin as well as the upregulation of the acute-phase serum proteins haptoglobin (Hp) and serum amyloid A (SAA). The viral strain used did provoke transient health impairment, namely, fever and leukopenia that were associated with viremia, viral shedding with nasal secretions, and antiviral seroconversion. Complete recovery was observed within 3 weeks. Elevated levels of SAA and Hp were apparent from days 4 to 13 and 8 to 11, respectively. In WBC, the levels of Mx mRNA and Mx protein were elevated from days 2 to 15. In the context of this study with BVDV, the level of Mx protein expression in WBC provided the most telling diagnostic window to monitor the host's ongoing innate immune response.
Combined Use of Enzyme-Linked Immunosorbent Assay and Flow Cytometry To Detect Antibodies toTrypanosoma cruziin Domestic Canines in Texas
Authors: Shadomy Sean V. , Waring Stephen C. , Chappell Cynthia L. 
Published: March 2004

ABSTRACTCanines may be sentinels and/or reservoirs for humanTrypanosoma cruziexposures. This study adapted a method originally designed for human diagnostics to detect serum immunoglobulin G toT. cruziin canines. The method combined an enzyme-linked immunosorbent assay (ELISA) for screening and flow cytometry detection of anti-live trypomastigote antibodies (ALTA) for confirmation. The assays were optimized by using known positive and negative control canine sera, and cutoff values were established. The ELISA and ALTA assay easily distinguished between reactive (positive controls) and nonreactive (negative controls) sera and were used to test sera collected in a cross-sectional seroprevalence survey of 356 domestic canines from Harris County, Tex., and the surrounding area. Fifty-three (14.9%) of 356 asymptomatic canines in the survey were positive by ELISA, and 5 (1.4%) were confirmed positive with the ALTA assay, with an additional 4 (1.1%) canines classified as “suspect positive.” Thus, the overall prevalence ofT. cruziantibodies in this population was 2.6%. This is the first U.S. study to use the combination of ELISA and ALTA to detect serum antibodies toT. cruziand the first report of the prevalence ofT. cruziinfection in domestic canines in the Houston, Tex. (Harris County), region. Our results demonstrate that the combination of ELISA and ALTA has been successfully adapted for use in testing canines for serological evidence ofT. cruziinfection. Seroprevalence survey results suggest thatT. cruziantibody-positive domestic canines in the peridomestic setting are present in the Houston, Tex., region and further suggest thatT. cruziis enzootic in the region.
Molecular Cloning and Characterization of Tropomyosin, a Major Allergen of
Chironomus kiiensis
, a Dominant Species of Nonbiting Midges in Korea
Authors: Jeong Kyoung Yong , Yum Hye-Yung , Lee In-Yong , Ree Han-Il , Hong Chein-Soo , Kim Dong Soo , Yong Tai-Soon 
Published: March 2004

ABSTRACT

Chironomids are widely and abundantly distributed in the vicinity of standing waters. Larvae of
Chironomus
and some other genera are known to contain hemoglobins, which have been described as a major allergen, and the adults that have no hemoglobins also have been reported to contain allergens. In this study, we tried to establish the role of chironomid allergy and characterize the allergen of
Chironomus kiiensis
adults. Skin tests using
C. kiiensis
adult extracts were performed on patients with allergic symptoms. A cDNA library of
C. kiiensis
adults was screened with
C. kiiensis
immune mouse sera to identify allergens, and results were confirmed using skin test-positive human sera. Recombinant allergen was expressed in
Escherichia coli
and purified by affinity chromatography using nickel-nitrilotriacetic acid agarose to investigate its allergenic properties. Out of 275 allergic patients 14.2% showed a positive reaction to
C. kiiensis
adult crude extracts in the skin test. The tropomyosin was cloned by immunoscreening and expressed in
Escherichia coli. C. kiiensis
tropomyosin has a high homology at the amino acid level with tropomyosins which were previously known to be allergens in various arthropods (
Periplaneta americana
, 86.3%;
Panulirus stimpson
, 78.9%;
Dermatophagoides pteronyssinus
, 76.5%). Specific immunoglobulin E antibodies reacting to recombinant tropomyosin were detected in 17 (81%) of 21 patients whose skin test results were positive. Cross-reactivity against house dust mites and other insects was noticed with mouse anti-recombinant tropomyosin immune serum.
C. kiiensis
adults were shown to be an important source of inhalant allergens in Korea. Molecular cloning of
C. kiiensis
tropomyosin was performed and IgE reactivity was demonstrated using skin test-positive human sera. Recombinant tropomyosin will be useful for further studies or clinical applications.
D8/17 and CD19 Expression on Lymphocytes of Patients with Acute Rheumatic Fever and Tourette's Disorder
Authors: Weisz Julie L. , McMahon William M. , Moore Jill C. , Augustine Nancy H. , Bohnsack John F. , Bale James F. , Johnson Michael B. , Morgan Jubel F. , Jensen Jenise , Tani Lloyd Y. , Veasy L. George , Hill Harry R. 
Published: March 2004

ABSTRACTD8/17, an alloantigen found on B lymphocytes, has been reported to be elevated in patients susceptible to rheumatic fever and may be associated with autoimmune types of neuropsychiatric disorders. The pediatric-autoimmune-neuropsychiatric-disorders-associated-with-streptococci model is a putative model of pathogenesis for a group of children whose symptoms of obsessive-compulsive disorder and Tourette's disorder (TD) are abrupt and may be triggered by an infection with group A streptococci. As a test of this model, we have examined D8/17 levels on the B cells of patients with TD and acute rheumatic fever (ARF) along with those on the B cells of normal controls by flow cytometry. We have utilized several different preparations of D8/17 antibody along with a variety of secondary antibodies but have been unable to show an association with an elevated percentage of D8/17-positive, CD19-positive B cells in either ARF or TD. We did find, however, that the percentages of CD19-positive B cells in ARF and TD patients were significantly elevated compared to those in normal controls. Group A streptococcal pharyngitis patients also had an elevated percentage of CD19 B cells, however. These studies failed to confirm the utility of determining the percentage of B cells expressing the D8/17 alloantigen in ARF patients or our sample of TD patients. In contrast, the percentage of CD19-positive B cells was significantly elevated in ARF and TD patients, as well as group A streptococcal pharyngitis patients, suggesting a role for inflammation and/or autoimmunity in the pathogenesis of these disorders.
Effects of Infection with Transmissible Gastroenteritis Virus on Concomitant Immune Responses to Dietary and Injected Antigens
Authors: Bailey Michael , Haverson Karin , Miller Bevis , Jones Philip , Sola Isabel , Enjuanes Luis , Stokes Christopher R. 
Published: March 2004

ABSTRACTNormal piglets weaned onto soy- or egg-based diets generated antibody responses to fed protein. Concurrent infection with transmissible gastroenteritis virus (TGEV) did not affect the responses to dietary antigens at weaning, nor did it affect the subsequent development of tolerance. However, TGEV infection did enhance the primary immunoglobulin M (IgM) and IgG1, but not IgG2, antibody responses to injected soy in comparison to those of uninfected animals. Paradoxically, TGEV-infected animals showed an enhanced primary IgG1 antibody response to injected soy at 4 weeks of age, but they subsequently showed a reduced secondary response after an intraperitoneal challenge at 9 weeks of age in comparison to uninfected animals. The results suggest that an enteric virus, either used as a vaccine vector or present as a subclinical infection, may not have significant effects on the development of dietary allergies but may have effects both on the primary response and on the subsequent recall response to systemic antigens to which the animal is exposed concurrently with virus antigens.
Prevalence of Neutralizing Antibodies to Adenoviral Serotypes 5 and 35 in the Adult Populations of The Gambia, South Africa, and the United States
Authors: Nwanegbo Edward , Vardas Eftyhia , Gao Wentao , Whittle Hilton , Sun Huijie , Rowe David , Robbins Paul D. , Gambotto Andrea 
Published: March 2004

ABSTRACT
One of the major limitations of the use of adenoviruses as gene therapy vectors is the existence of preformed immunity in various populations. Recent studies have linked failure of adenoviral gene therapy trials to the presence of antiadenoviral neutralizing antibodies (NAb). Understanding the distribution and specificity of such antibodies will assist in the design of successful recombinant adenoviral gene therapies and vaccines. To assess the prevalence of NAb to adenovirus serotypes 5 and 35 (Ad5 and Ad35), we analyzed serum samples from adult immunocompetent individuals living in The Gambia, South Africa, and the United States by using a neutralization assay. Serum samples were incubated with A549 lung carcinoma cells and adenoviruses encoding enhanced green or yellow fluorescent proteins; results were analyzed by fluorescence microscopy and flow cytometry. Using this technique, we found a high prevalence of NAb against Ad5 in Gambian, South African, and U.S. subjects at both low and high titers. Conversely, all subjects displayed a low prevalence of NAb to Ad35; when present, anti-Ad35 NAb were seen at low titers. Because of the ability of adenoviruses to elicit systemic and mucosal immune responses, Ad35 with its low NAb prevalence appears to be an attractive candidate vector for gene therapy applications.
Sustained Lipopolysaccharide-Induced Lung Inflammation in Mice Is Attenuated by Functional Deficiency of the Fas/Fas Ligand System
Authors: Matute-Bello Gustavo , Winn Robert K. , Martin Thomas R. , Liles W. Conrad 
Published: March 2004

ABSTRACTTo determine whether the Fas/Fas ligand (FasL) (CD95/CD178) system contributes to the development of an inflammatory response in vivo, 2.5 μg of bacterial lipopolysaccharide (LPS; endotoxin) per g was administered intranasally to healthy mice (C57BL/6) and mutant mice deficient in either Fas (lprmice) or FasL (gldmice). Sustained LPS-induced neutrophilic inflammation in the lungs was attenuated in bothlprandgldmice. These observations provide further evidence of a proinflammatory role for the Fas/FasL system in the lungs.
Profiles of Antibody Responses against Severe AcuteRespiratory Syndrome Coronavirus Recombinant Proteins andTheir Potential Use as DiagnosticMarkers
Authors: Tan Yee-Joo , Goh Phuay-Yee , Fielding Burtram C. , Shen Shuo , Chou Chih-Fong , Fu Jian-Lin , Leong Hoe Nam , Leo Yee Sin , Ooi Eng Eong , Ling Ai Ee , Lim Seng Gee , Hong Wanjin 
Published: March 2004

ABSTRACT
A
new coronavirus (severe acute respiratory syndrome
coronavirus [SARS-CoV]) has been identified to
be the etiological agent of severe acute respiratory syndrome. Given
the highly contagious and acute nature of the disease, there is an
urgent need for the development of diagnostic assays that can detect
SARS-CoV infection. For determination of which of the viral proteins
encoded by the SARS-CoV genome may be exploited as diagnostic antigens
for serological assays, the viral proteins were expressed individually
in mammalian and/or bacterial cells and tested for reactivity with sera
from SARS-CoV-infected patients by Western blot analysis. A total of 81
sera, including 67 from convalescent patients and seven pairs from two
time points of infection, were analyzed, and all showed
immunoreactivity towards the nucleocapsid protein (N). Sera from some
of the patients also showed immunoreactivity to U274 (59 of 81
[73%]), a protein that is unique to SARS-CoV. In
addition, all of the convalescent-phase sera showed immunoreactivity to
the spike (S) protein when analyzed by an immunofluorescence method
utilizing mammalian cells stably expressing S. However, samples from
the acute phase (2 to 9 days after the onset of illness) did not react
with S, suggesting that antibodies to N may appear earlier than
antibodies to S. Alternatively, this could be due to the difference in
the sensitivities of the two methods. The immunoreactivities to these
recombinant viral proteins are highly specific, as sera from 100
healthy donors did not react with any of them. These results suggest
that recombinant N, S, and U274 proteins may be used as antigens for
the development of serological assays for
SARS-CoV.
Preventive Effects ofEscherichia coliStrain Nissle 1917 on Acute and Chronic Intestinal Inflammation in Two Different Murine Models of Colitis
Authors: Schultz Michael , Strauch Ulrike G. , Linde Hans-Jörg , Watzl Sonja , Obermeier Florian , Göttl Claudia , Dunger Nadja , Grunwald Nicole , Schölmerich Jürgen , Rath Heiko C. 
Published: March 2004

ABSTRACTEscherichia colistrain Nissle 1917 (EcN) is as effective in maintaining remission in ulcerative colitis as is treatment with mesalazine. This study aims to evaluate murine models of acute and chronic intestinal inflammation to study the antiinflammatory effect of EcN in vivo. Acute colitis was induced in mice with 2% dextran-sodium sulfate (DSS) in drinking water. EcN was administered from day −2 to day +7. Chronic colitis was induced by transfer of CD4+CD62L+T lymphocytes from BALB/c mice in SCID mice. EcN was administered three times/week from week 1 to week 8 after cell transfer. Mesenteric lymph node (MLN) cytokine secretion (of gamma interferon [IFN-γ], interleukin 5 [IL-5], IL-6, and IL-10) was measured by enzyme-linked immunosorbent assay. Histologic sections of the colon were analyzed by using a score system ranging from 0 to 4. Intestinal contents and homogenized MLN were cultured, and the number ofE. coli-like colonies was determined. EcN was identified by repetitive extragenic palindromic (REP) PCR. EcN administration to DSS-treated mice reduced the secretion of proinflammatory cytokines (IFN-γ, 32,477 ± 6,377 versus 9,734 ± 1,717 [P= 0.004]; IL-6, 231 ± 35 versus 121 ± 17 [P= 0.02]) but had no effect on the mucosal inflammation. In the chronic experimental colitis of the transfer model, EcN ameliorated the intestinal inflammation (histology score, 2.7 ± 0.2 versus 1.9 ± 0.3 [P= 0.02]) and reduced the secretion of proinflammatory cytokines. Translocation of EcN and residentE. coliinto MLN was observed in the chronic colitis model but not in healthy controls. Administration of EcN ameliorated acute and chronic experimental colitis by modifying proinflammatory cytokine secretion but had no influence on the acute DSS-induced colitis. In this model, preexisting colitis was necessary for translocation of EcN and residentE. coliinto MLN.
Immunoglobulin M Antibody Responses toMycobacterium ulceransAllow Discrimination between Cases of Active Buruli Ulcer Disease and Matched Family Controls in Areas Where the Disease Is Endemic
Authors: Okenu Daniel M. N. , Ofielu Lazarus O. , Easley Kirk A. , Guarner Jeannette , Spotts Whitney Ellen A. , Raghunathan Pratima L. , Stienstra Ymkje , Asamoa Kwame , van der Werf Tjip S. , van der Graaf Winette T. A. , Tappero Jordan W. , Ashford David A. , King C. Harold 
Published: March 2004

ABSTRACTBuruli ulcer disease (BUD) is an emerging disease caused byMycobacterium ulcerans. In the present study we have characterized the serological reactivities of sera from volunteer case patients with laboratory-confirmed BUD and controls living in three different regions of Ghana where the disease is endemic to determine if serology may be useful for disease confirmation. Our results showed highly reactive immunoglobulin G (IgG) responses among patients with laboratory-confirmed disease, healthy control family members of the case patients, and sera from patients with tuberculosis from areas where BUD is not endemic. These responses were represented by reactivities to multiple protein bands found in theM. ulceransculture filtrate (CF). In contrast, patient IgM antibody responses to theM. ulceransCF (MUCF) proteins were more distinct than those of healthy family members living in the same village. A total of 84.8% (56 of 66) of the BUD patients exhibited strong IgM antibody responses against MUCF proteins (30, 43 and 70 to 80 kDa), whereas only 4.5% (3 of 66) of the family controls exhibited such responses. The sensitivity of the total IgM response for the patients was 84.8% (95% confidence interval [CI], 74.3 to 91.6%), and the specificity determined with sera from family controls was 95.5% (95% CI, 87.5 to 98.4%). These studies suggest that the IgM responses of patients with BUD will be helpful in the identification and production of theM. ulceransrecombinant antigens required for the development of a sensitive and specific serological assay for the confirmation of active BUD.
High Prevalence of Antibodies to Hepatitis A and E Viruses and Viremia of Hepatitis B, C, and D Viruses among Apparently Healthy Populations in Mongolia
Authors: Takahashi Masaharu , Nishizawa Tsutomu , Gotanda Yuhko , Tsuda Fumio , Komatsu Fumio , Kawabata Terue , Hasegawa Kyoko , Altankhuu Murdorjyn , Chimedregzen Ulziiburen , Narantuya Luvsanbasaryn , Hoshino Hiromi , Hino Kunihiko , Kagawa Yasuo , Okamoto Hiroaki 
Published: March 2004

ABSTRACT
The prevalence of infection with hepatitis A virus (HAV), HBV, HCV, HDV, and HEV was evaluated in 249 apparently healthy individuals, including 122 inhabitants in Ulaanbaatar, the capital city of Mongolia, and 127 age- and sex-matched members of nomadic tribes who lived around the capital city. Overall, hepatitis B surface antigen (HBsAg) was detected in 24 subjects (10%), of whom 22 (92%) had detectable HBV DNA. Surprisingly, HDV RNA was detectable in 20 (83%) of the 24 HBsAg-positive subjects. HCV-associated antibodies were detected in 41 (16%) and HCV RNA was detected in 36 (14%) subjects, none of whom was coinfected with HBV, indicating that HBV/HCV carriers account for one-fourth of this population. Antibodies to HAV and HEV were detected in 249 (100%) and 28 (11%) subjects, respectively. Of 22 HBV DNA-positive subjects, genotype D was detected in 21 subjects and genotype F was detected in 1 subject. All 20 HDV isolates recovered from HDV RNA-positive subjects segregated into genotype I, but these differed by 2.1 to 11.4% from each other in the 522- to 526-nucleotide sequence. Of 36 HCV RNA-positive samples, 35 (97%) were genotype 1b and 1 was genotype 2a. Reflecting an extremely high prevalence of hepatitis virus infections, there were no appreciable differences in the prevalence of hepatitis virus markers between the two studied populations with distinct living place and lifestyle. A nationwide epidemiological survey of hepatitis viruses should be conducted in an effort to prevent de novo infection with hepatitis viruses in Mongolia.