bluuebunny/crossref_metadata_2025_split huggingface dataset:
Performance of Competitive and Indirect Enzyme-Linked Immunosorbent Assays, Gel Immunoprecipitation with Native Hapten Polysaccharide, and Standard Serological Tests in Diagnosis of Sheep Brucellosis
Authors: Marín C. M. , Moreno E. , Moriyón I. , Díaz R. , Blasco J. M. 
Published: December 2019

ABSTRACTCompetitive and standard enzyme-linked immunosorbent assays (ELISAs), rose bengal (RB), complement fixation, and agar gel immunoprecipitation with native hapten (AGID-NH) were compared by using sera fromBrucella-free,Brucella melitensis-infected, andB. melitensisRev1-vaccinated sheep. The most sensitive tests were indirect ELISA and RB, and the most specific tests were AGID-NH and competitive ELISA. We show that RB followed by AGID-NH is a simple and effective system for diagnosing sheep brucellosis.
Impaired Interleukin-8-Induced Degranulation of Polymorphonuclear Neutrophils from Human Immunodeficiency Virus Type 1-Infected Individuals
Authors: Meddows-Taylor Stephen , Martin Desmond J. , Tiemessen Caroline T. 
Published: December 2019

ABSTRACTDegranulation of peripheral blood polymorphonuclear leukocytes (PMNLs) was monitored in human immunodeficiency virus (HIV) type 1 (HIV-1)-infected individuals with or without pulmonary tuberculosis (HIV/TB and HIV groups, respectively) by measuring the release of β-glucuronidase induced by interleukin-8 (IL-8). This was increased in a dose-dependent manner in the control groups consisting of healthy blood donors and patients with pulmonary tuberculosis. In contrast, PMNLs from the HIV and HIV/TB groups responded reciprocally in the same assay; that is, higher IL-8 input concentrations resulted in the release of less enzyme than lower IL-8 input concentrations. The degranulation response of PMNLs from HIV-1-infected individuals was similarly altered for another agonist,N-formyl-methionyl-leucyl-phenylalanine, suggesting that impairment of the nonoxidative armature of PMNL was a more generalized phenomenon. However, impaired IL-8-induced degranulation was found to be associated with the reduced expression of both IL-8 receptors, A and B, on whole-blood PMNLs from HIV-1-infected patients compared with that on whole-blood PMNLs from healthy persons. The density of IL-8RA, in particular, was most reduced on the surfaces of PMNLs from those patients with the poorest degranulation in response to IL-8. Inefficient agonist-induced degranulation may contribute to the increased susceptibility of HIV-1-infected persons to secondary microbial infections, this being further exacerbated in HIV/TB patients who, in addition, display defects in phagocytosis and oxidative burst.
Preparation of Recombinant Human Monoclonal Antibody Fab Fragments Specific forEntamoeba histolytica
Authors: Tachibana Hiroshi , Cheng Xun-Jia , Watanabe Katsuomi , Takekoshi Masataka , Maeda Fumiko , Aotsuka Satoshi , Kaneda Yoshimasa , Takeuchi Tsutomu , Ihara Seiji 
Published: December 2019

ABSTRACTGenes coding for human antibody Fab fragments specific forEntamoeba histolyticawere cloned and expressed inEscherichia coli. Lymphocytes were separated from the peripheral blood of a patient with an amebic liver abscess. Poly(A)+RNA was isolated from the lymphocytes, and then genes coding for the light chain and Fd region of the heavy chain were amplified by a reverse transcriptase PCR. The amplified DNA fragments were ligated with a plasmid vector and were introduced intoEscherichia coli. Three thousand colonies were screened for the production of antibodies toE. histolyticaHM-1:IMSS by an indirect fluorescence-antibody (IFA) test. Lysates from fiveEscherichia coliclones were positive. Analysis of the DNA sequences of the five clones showed that three of the five heavy-chain sequences and four of the five light-chain sequences differed from each other. When the reactivities of theEscherichia colilysates to nine reference strains ofE. histolyticawere examined by the IFA test, three Fab fragments with different DNA sequences were found to react with all nine strains and another Fab fragment was found to react with seven strains. None of the four human monoclonal antibody Fab fragments reacted withEntamoeba disparreference strains or with other enteric protozoan parasites. These results indicate that the bacterial expression system reported here is effective for the production of human monoclonal antibodies specific forE. histolytica. The recombinant human monoclonal antibody Fab fragments may be applicable for distinguishingE. histolyticafromE. disparand for use in the serodiagnosis of amebiasis.
Molecular Cloning of the Gene for a Conserved Major Immunoreactive 28-Kilodalton Protein ofEhrlichia canis: a Potential Serodiagnostic Antigen
Authors: McBride Jere W. , Yu Xue-jie , Walker David H. 
Published: December 2019

ABSTRACTA gene encoding a 28-kDa protein ofEhrlichia caniswas cloned, sequenced, and expressed, and a comparative molecular analysis with homologous genes ofE. canis,Cowdria ruminantium, andEhrlichia chaffeensiswas performed. The complete gene has an 834-bp open reading frame encoding a protein of 278 amino acids with a predicted molecular mass of 30.5 kDa. An N-terminal signal sequence was identified, suggesting that the protein undergoes posttranslational modification to a mature 27.7-kDa protein (P28). TheE. canis p28gene has significant nucleic acid and amino acid sequence homologies with theE. chaffeensisouter membrane protein-1 (omp-1) gene family, with theCowdria ruminantium map-1gene, and with otherE. canis28-kDa-protein genes. Southern blotting revealed the presence of at least two additional homologousp28gene copies in theE. canisgenome, confirming thatp28is a member of a polymorphic multiple-gene family. Amino acid sequence analysis revealed thatE. canisP28 has four variable regions, and it shares similar surface-exposed regions, antigenicity, and T-cell motifs withE. chaffeensisP28. Thep28genes from seven differentE. canisisolates were identical, indicating that the gene for this major immunoreactive protein is highly conserved. In addition, reactivity of sera from clinical cases of canine ehrlichiosis with the recombinant P28 demonstrated that the recombinant protein may be a reliable serodiagnostic antigen.
A New Cell Enzyme-Linked Immunosorbent Assay Demonstrates Gamma Interferon Suppression by Beta Interferon in Multiple Sclerosis
Authors: Bakhiet Moiz , Özenci Volkan , Withagen Carin , Mustafa Maha , Fredrikson Sten , Link Hans 
Published: December 2019

ABSTRACTMultiple sclerosis (MS) is a demyelinating disorder of the central nervous system of unknown etiology. Immune mechanisms involving the proinflammatory cytokine gamma interferon (IFN-γ) are believed to play an important role in the pathogenesis of MS. IFN-β-1b has been introduced as a treatment for MS and was found to reduce the number and severity of clinical exacerbations. To examine the influence of IFN-β-1b on myelin basic protein (MBP)-specific and phytohemagglutinin-induced IFN-γ production, we developed a cell-released capturing enzyme-linked immunosorbent assay (CRC-ELISA), which rapidly measures spontaneous and antigen- or mitogen-induced cellular IFN-γ production. CRC-ELISA documented a significant MBP-specific T-cell response in the blood of untreated MS patients, as assessed by IFN-γ production. This response was suppressed in MS patients treated with IFN-β-1b. The present work confirms in vivo the in vitro suppressive effects of IFN-β-1b on IFN-γ production in MS. Moreover, it provides a powerful new technique for detection of cytokines.
Estimation of Growth Rates of
Escherichia coli
BJ4 in Streptomycin-Treated and Previously Germfree Mice by In Situ rRNA Hybridization
Authors: Rang Camilla U. , Licht Tine Rask , Midtvedt Tore , Conway Patricia L. , Chao Lin , Krogfelt Karen A. , Cohen Paul S. , Molin Søren 
Published: December 2019

ABSTRACT

The growth physiology of
Escherichia coli
during colonization of the intestinal tract was studied with four animal models: the streptomycin-treated mouse carrying a reduced microflora, the monoassociated mouse with no other microflora than the introduced strain, the conventionalized streptomycin-treated mouse, and the conventionalized monoassociated mouse harboring a full microflora. A 23S rRNA fluorescent oligonucleotide probe was used for hybridization to whole
E. coli
cells fixed directly after being taken from the animals, and the respective growth rates of
E. coli
BJ4 in the four animal models were estimated by correlating the cellular concentrations of ribosomes with the growth rate of the strain. The growth rates thus estimated from the ribosomal content of
E. coli
BJ4 in vivo did not differ in the streptomycin-treated and the monoassociated mice. After conventionalization there was a slight decrease of the bacterial growth rates in both animal models.
Effect of Early Antibiotic Treatment on the Antibody Response to Cytoplasmic Proteins of
Brucella melitensis
in Mice
Authors: Bowden Raul A. , Racaro Graciela C. , Baldi Pablo C. 
Published: December 2019

ABSTRACT

To test whether antibiotic therapy hampers the antibody response to
Brucella
antigens, 30 BALB/c mice were infected with
Brucella melitensis
H38 and randomized for treatment with doxycycline administered intraperitoneally for 42 days starting at 7 or 28 days postinfection (p.i.) (groups DOX7 and DOX28, respectively) or for no treatment (control group). Antibodies to smooth lipopolysaccharide (LPS) reached peak levels (mean optical density [OD] = 2.618) between days 56 and 70 p.i. in the control group, and similar peak levels (mean OD = 2.486) were observed in the DOX28 group, but significantly lower peak levels (mean OD = 0.821) were observed at 28 days p.i. in the DOX7 group. The antibody response against cytoplasmic proteins depleted of LPS (CPs) reached maximal levels (mean OD = 2.402) between days 56 and 70 p.i. in the control group, but no response was detected in the DOX7 group. Anti-CP antibodies were detected in only three animals from the DOX28 group, at levels significantly lower than those in the control group (mean maximal OD = 0.791). The pattern of antibody response to an 18-kDa cytoplasmic protein of
Brucella
spp. was similar to that against the CP antigen. This study shows that early antibiotic treatment affects the antibody response of mice to cytoplasmic proteins of
Brucella
and, to a lesser extent, to LPS.
CD4-Positive and CD8-Positive Cytotoxic T Lymphocytes Contribute to Human Papillomavirus Type 16 E6 and E7 Responses
Authors: Nakagawa Mayumi , Stites Daniel P. , Palefsky Joel M. , Kneass Zachary , Moscicki Anna-Barbara 
Published: December 2019

ABSTRACTCytotoxic T-lymphocyte (CTL) responses to E6 and E7 were previously shown to be more commonly detectable in human papillomavirus type 16 (HPV-16)-positive women without squamous intraepithelial neoplasia (SIL) than in HPV-16-positive women with SIL (M. Nakagawa, D. P. Stites, S. Farhat, J. R. Sisler, B. Moss, F. Kong, A. B. Moscicki, and J. M. Palefsky, J. Infect. Dis. 175:927–931, 1997). The objective of this study was to characterize the phenotype(s) of the effector cell population responsible for HPV-16 E6- and E7-specific cytotoxic responses. Peripheral blood mononuclear cells were stimulated with HPV-16 E6 or E7 fusion protein. Cells from an autologous B-lymphoblastoid cell line, infected with vaccinia virus expressing E6 or E7, served as target cells. The effector cells were characterized by using natural-killer-cell removal, antibody blocking, and T-cell subset separation. Our results suggest that both CD4 and CD8 T lymphocytes contribute to HPV-16 E6- and E7-specific CTL responses although their relative contributions vary from individual to individual. On the other hand, natural killer cells in the effector cell population contribute to background activities but not to HPV-specific responses in this assay system.
Similar Humoral and Cellular Immunological Reactivities to Human Herpesvirus 6 in Patients with Multiple Sclerosis and Controls
Authors: Enbom Malin , Wang Fu-Zhang , Fredrikson Sten , Martin Claes , Dahl Helena , Linde Annika 
Published: December 2019

ABSTRACT
Several studies have suggested an association between human herpesvirus 6 (HHV-6) and multiple sclerosis (MS). We have previously studied intrathecal production of antibody to lymphotropic herpesviruses in MS patients and the presence of human herpesvirus 1 to 7 DNAs in cerebrospinal fluid (CSF). In the present study anti-HHV-6 immunoglobulin M (IgM) in serum and anti-HHV-6 IgG subclasses in serum and CSF were examined and the lymphoproliferative response to HHV-6 was analyzed. The PCR examination was refined by purifying DNA from CSF and retesting the samples for HHV-6 DNA. There were no statistically significant differences between the groups concerning IgM positivity, distribution of IgG subclasses, or lymphoproliferative response to HHV-6. The purification of DNA increased the number of PCR-positive samples from 0 of 71 to 4 of 68. The study does not give additional support to the possibility that HHV-6 is a common cause of MS, but a role for the virus in a subset of patients cannot be excluded.
Use of a Monoclonal Antibody against anEscherichia coliO26 Surface Protein for Detection of Enteropathogenic and Enterohemorrhagic Strains
Authors: Kerr Paul , Ball Hywel , China Bernard , Mainil Jacques , Finlay David , Pollock David , Wilson Ian , Mackie Dermot 
Published: December 2019

ABSTRACTA monoclonal antibody (MAb) was obtained from a mouse immunized with solubilized outer membrane proteins extracted from a bovine enterohemorrhagic strain ofEscherichia coli(EHEC), O26. The MAb produced a strong immunoblot reaction at approximately 21 kDa for an O26 strain containing the intimin gene (eae) and verocytotoxin (VT), but not with an O26eae- and VT-negative strain, or O157eae- and VT-positive strains. The MAb was used in a sandwich enzyme-linked immunosorbent assay (ELISA) format to screen strains from animal and human sources, and all reactive strains were characterized for the presence ofeaeand the gene encoding VT factors by PCR. The antigen was detected in a group of strains containing a high proportion of O26, the majority of which wereeaepositive with or without VT; these were isolated mostly from animal enteritis cases but included a small number of human enteric isolates. Nonreactors includedeae-positive (with or without VT) O157 strains and one O26 strain. In a survey of mixed cultures from both animal and human enteric disease, ELISA-positive reactions were obtained from 7.1 to 11.2% of samples from bovine, porcine, ovine, and human sources. The two human O8 and ten animal O26 ELISA-reactive pure strains obtained from these samples contained sixeae- and/or VT-positive strains; the other six strains lost their ELISA positivity following storage at −70°C, after which none were found to contain eithereaeor VT factors. The association of the antigen detected by the MAb with significant enteropathogenicE. coliand EHEC virulence factors in isolates from both animal and human enteric infections indicates a diagnostic potential for the assay developed.
A Novel Enzyme-Linked Immunosorbent Assay Using the RecombinantActinobacillus pleuropneumoniaeApxII Antigen for Diagnosis of Pleuropneumonia in Pig Herds
Authors: Leiner Gero , Franz Burkart , Strutzberg Katrin , Gerlach Gerald-F. 
Published: December 2019

ABSTRACTFor the surveillance of pig herds infected with porcine pleuropneumonia, an enzyme-linked immunosorbent assay (ELISA) using the recombinantActinobacillus pleuropneumoniaeApxII protein as species- but not serotype-specific antigen was developed. Using this ELISA, 243 of 400 animals from 22A. pleuropneumoniae-infected herds were classified as seropositive.
A New Sensitive Serological Assay for Detection of Lentivirus Infections in Small Ruminants
Authors: Saman Eric , Van Eynde Geertrui , Lujan Luis , Extramiana Belén , Harkiss Gordon , Tolari Francesco , Gonzàlez Lorenzo , Amorena Beatriz , Watt Neil , Badiola Juan 
Published: December 2019

ABSTRACT

Lentivirus infections in small ruminants represent an economic problem affecting several European countries with important sheep-breeding industries. Programs for control and eradication of these infections are being initiated and require reliable screening assays. This communication describes the construction and evaluation of a new serological screening enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to maedi-visna virus (MVV) in sheep and to caprine arthritis encephalitis virus (CAEV) in goats. The solid phase is sensitized with a combination of the major core protein p25 of MVV produced in
Escherichia coli
and a peptide derived from the immunodominant region of the viral transmembrane protein gp46. The peptide carries an N-terminal biotin residue and is complexed with streptavidin prior to being coated. The new assay was evaluated with 2,336 sheep serum samples from different European countries with large differences in the levels of prevalence of MVV infections, and the results have been compared to those of the standard agar gel immunodiffusion test. Discrepant samples were analyzed by Western blotting with viral lysate, and most sera could be classified unambiguously. The estimated overall sensitivity of the new ELISA was 99.4% (95% confidence interval [CI], 98.4 to 99.8%) and the specificity was 99.3% (95% CI, 98.7 to 99.6%). A limited set of goat sera (
n
= 212) was also analyzed, with similar results. These data indicate that the new assay is a reliable tool that can be used in control and eradication programs for small ruminant lentivirus infections.
Inducible and Constitutive In Vitro Neutrophil Chemokine Expression by Mammary Epithelial and Myoepithelial Cells
Authors: Barber Michele R. , Pantschenko Alexander G. , Hinckley Lynn S. , Yang T. J. 
Published: December 2019

ABSTRACTPreviously, our laboratory showed that bovine and caprine mammary secretions are chemotactic and that chemoattractants found in these secretions are qualitatively different according to infection status and/or lactation stage. However, the cellular source of the chemoattractants has not been defined. In this study we used a modified Boyden chamber assay to examine the ability of previously established caprine mammary epithelial cell (CMEC) and myoepithelial cell (CMMyoEC) lines to produce chemoattractants for neutrophils. We found that CMEC culture supernatants, but not those of CMMyoEC cultures, induced in vitro neutrophil chemotaxis. Further characterization showed that chemotactic activity was produced when the cells underwent contact-induced differentiation. Neutrophil migration was chemotactic, not chemokinetic, and was augmented when the epithelial and myoepithelial cells were cocultured. Additionally, chemotactic activity was inducible byStaphylococcus aureusplus alpha-toxin,Escherichia coli, and interleukin-1β (IL-1β) in CMEC cultures. However, CMMyoEC cultures could not be induced to produce chemotactic activity. Anti-IL-8 antibody was able to block some constitutively produced chemotactic activity and chemotactic activity induced by IL-1β andS. aureusplus alpha-toxin. These results indicate that epithelial cells may play a major role in producing chemoattractants, specifically IL-8, in the mammary gland.
Regeneration and Tolerance Factor: a Correlate of Human Immunodeficiency Virus-Associated T-Cell Activation
Authors: Givens Tara S. , DuChateau Brian K. , Boomer Jonathan S. , Westerman Maxwell P. , Gilman-Sachs Alice , Beaman Kenneth D. 
Published: December 2019

ABSTRACTHuman immunodeficiency virus (HIV) infection causes extensive phenotypic alterations in lymphocytes. Cellular markers that are normally absent or expressed at low levels on quiescent cells are upregulated throughout the disease course. The transmembrane form of regeneration and tolerance factor (RTF) is expressed at negligible levels on resting T cells but is quickly upregulated following in vitro stimulation and activation. Recently, we reported that expression of RTF was significantly higher in cells from HIV-seropositive (HIV+) individuals than in cells from HIV-seronegative (HIV−) individuals. Because T cells from HIV+individuals express markers reflecting chronic activation, we hypothesized that these in vivo-activated cells would coexpress RTF. Flow cytometry was used to assess RTF expression on activated (CD38+and HLA-DR+) CD4+and CD8+T cells. HIV+individuals had higher percentages of RTF+CD38+(P< 0.0001) or RTF+HLA-DR+(P= 0.0001) CD4+T cells than HIV−individuals. In HIV+individuals, increased percentages of CD4+T cells that were RTF+, RTF+CD38+, and RTF+HLA-DR+correlated inversely with the absolute number and percentage of CD4+T cells and correlated positively with plasma β2-microglobulin concentrations. HIV+individuals had higher percentages of CD8+T cells that were RTF+CD38+(P= 0.0001) or RTF+HLA-DR+(P= 0.0010). In HIV+individuals, increased percentages of CD8+T cells that were RTF+HLA-DR+correlated inversely with the percentage of CD4+T cells, and high percentages of CD8+T cells that were RTF+CD38+correlated positively with plasma β2-microglobulin levels. These findings strongly suggest that increased RTF expression is a correlate of HIV-associated immune system activation.
Detection of Borreliacidal Antibodies in Lyme Borreliosis Patient Sera Containing Antimicrobial Agents
Authors: Jobe Dean A. , Rawal Nenoo , Schell Ronald F. , Callister Steven M. 
Published: December 2019

ABSTRACT

The borreliacidal-antibody test has been used for the serological detection and confirmation of Lyme borreliosis. However, the presence of antimicrobial agents in serum can confound the accurate detection of borreliacidal antibodies. In this study, we developed a
Bacillus subtilis
agar diffusion bioassay to detect small concentrations of antimicrobial agents in serum. We also used XAD-16, a nonionic polymeric resin, to adsorb and remove high concentrations of amoxicillin, cefotaxime, ceftriaxone, cefuroxime, doxycycline, and erythromycin without significantly affecting even small concentrations of immunoglobulin M (IgM) or IgG borreliacidal antibodies. High concentrations of penicillin could also be removed by adding 1 U of penicillinase without significantly influencing the levels of borreliacidal antibodies. These simple procedures greatly enhance the clinical utility of the borreliacidal-antibody test.
Declining Immunoglobulin A Production in Prostates of Men with AIDS
Authors: Carson Henry J. , Lueck Nathan E. 
Published: December 2019

ABSTRACT
We studied immunoglobulin A production (IgA) in prostates of men with AIDS. Prostate sections from AIDS patients and human immunodeficiency virus-negative men were stained for IgA with immunoperoxidase. Prostate sections from nondiseased men were positive for IgA, while prostate sections from AIDS patients were essentially negative for IgA. Diminishing secretory IgA production may represent a characteristic of AIDS.
bca
, Beta Gene, and Gene Product Divergency in Reference and Prototype Strains of
Streptococcus agalactiae
Authors: Maeland Johan A. , Bevanger Lars , Iversen Grethe , Lyng Randi Valsoe 
Published: December 2019

ABSTRACT

Reference and prototype strains of
Streptococcus agalactiae
(GBS) were originally selected on the basis of phenotypic traits which, however, do not always mirror genotypic traits. A total of 14 reference and prototype GBS strains were examined by PCR designed to detect the
bca
and beta genes, encoding the c proteins c
α
and c
β
, respectively. The cognate proteins were detected by whole-cell-based fluorescent antibody testing and Western blotting. The PCR for beta gene detection and the antibody-based c
β
protein detection showed concordant results with all of the isolates, whereas 7 of 14 strains which did not express c
α
protein at detectable levels contained
bca
gene elements, consistent with
bca
gene and gene product divergency in these strains. The results emphasize the importance of genetic characterization of reference and prototype strains of GBS which, in the past, have been selected on the basis of phenotypic traits.