bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Powerful DNA Extraction Method and PCR for Detection of Microsporidia in Clinical Stool Specimens
Authors: Müller Andreas , Stellermann Kerstin , Hartmann Pia , Schrappe Matthias , Fätkenheuer Gerd , Salzberger Bernd , Diehl Volker , Franzen Caspar 
Published: December 2019

ABSTRACT

The diagnosis of intestinal microsporidiosis has traditionally depended on direct visualization of the parasite in stool specimens or intestinal biopsy samples by light and/or electron microscopy. Limited information about the specificity and sensitivity of PCR for the detection microsporidia in clinical stool specimens is available. To establish a sensitive and specific method for the detection of microsporidia in clinical samples, we studied clinical stool specimens of 104 randomly selected human immunodeficiency virus-infected patients with diarrhea to compare light microscopy and PCR. Fluorochrome Uvitex 2B staining was used for light microscopy. To raise the sensitivity of PCR, we used a powerful and fast DNA extraction method including stool sedimentation, glass bead disruption, and proteinase K and chitinase digestion. PCR was performed with primer pairs V1-PMP2, V1-EB450, and V1-SI500, and the nature of the PCR products was confirmed by Southern blot hybridization. Microsporidiosis was diagnosed by light microscopy in eight patients. Ten patients tested positive for microsporidiosis by PCR.
Enterocytozoon bieneusi
was found in seven cases, and
Encephalitozoon intestinalis
was found in four cases. In one case a double infection with
E. bieneusi
and
E. intestinalis
was diagnosed by PCR, whereas light microscopy showed only
E. bieneusi
infection. PCR testing of stool specimens is useful for diagnosis and species differentiation of intestinal microsporidiosis in HIV patients.
Alloreactivity and Association of Human Natural Killer Cells with the Major Histocompatibility Complex
Authors: Mavoungou Elie , Sall Aicha , Poaty-Mavoungou Virginie , Toure Fousseyni S. , Yaba Philippe , Delicat Andre , Lansoud-Soukate Joseph 
Published: December 2019

ABSTRACTAll NK cells potentially lytic for autologous cells but not expressing self-major histocompatibility complex (MHC)-reactive receptors could be eliminated by a negative selection mechanism during ontogeny. This idea is based on the existence of a NK cell subset expressing a specific inhibitory receptor for allogeneic MHC alleles. As ancestral haplotypes of the MHC appear to define identical MHC haplotypes in unrelated individuals, unrelated individuals having the same ancestral haplotype should also have the same NK-defined allospecificities that have been shown to map to the human MHC. To test this prediction, multiple cell lines from unrelated individuals having the same ancestral haplotypes were tested for the NK-defined allospecificities. It was found that cells having the same ancestral haplotypes do have the same NK-defined specificities. Furthermore, the NK-defined phenotype of cells that possess two different ancestral haplotypes can be predicted from the NK-defined phenotypes of unrelated cells that are homozygous for the ancestral haplotypes concerned. Although the group 1 and 2 NK-defined allospecificities can be explained to some extent by HLA-C alleles, evidence is presented that additional genes may modify the phenotype conferred by HLA-C.
Variants of aCryptococcus neoformansStrain Elicit Different Inflammatory Responses in Mice
Authors: Chen Lin-Chi , Casadevall Arturo 
Published: December 2019

ABSTRACTThe virulence ofCryptococcus neoformansisolates with high and low extracellular proteolytic activity was investigated in mice. No consistent relationship between proteolytic activity and virulence was observed, but isolates derived from one strain were shown to elicit different inflammatory responses.
A Murine Model of Renal Abscess Formation
Authors: Mussalli George M. , Brunnert Steven R. , Hirsch Emmet 
Published: December 2019

ABSTRACT

We developed a murine model of kidney abscess by direct renal injection of either
Escherichia coli
(1 × 10
6
to 7 × 10
6
organisms) or sterile medium. Bacterial infection produced renal abscesses, bacteremia, and late-onset leukocytosis in all animals. Controls were unaffected. This model may be useful for the study of various sequelae of kidney infection.
Recombinant Human Gamma Interferon in Human Immunodeficiency Virus-Infected Children: Safety, CD4+-Lymphocyte Count, Viral Load, and Neutrophil Function (AIDS Clinical Trials Group Protocol 211)
Authors: Shearer William T. , Kline Mark W. , Abramson Stuart L. , Fenton Terence , Starr Stuart E. , Douglas Steven D. 
Published: December 2019

ABSTRACTNineteen children with human immunodeficiency virus (HIV) infection were treated with recombinant human gamma interferon (rIFN-γ) (50 μg/m2subcutaneously three times each week during weeks 1 through 12 and 100 μg/m2subcutaneously three times each week during weeks 13 through 24) in a phase I/II clinical trial. All children continued to receive previously prescribed therapy with oral zidovudine or didanosine. Children were assessed clinically and with laboratory studies during 24 weeks of study treatment and for 12 weeks after completion of rIFN-γ therapy. In general, rIFN-γ therapy was well tolerated. There were two clinical or laboratory adverse events thought to be possibly or probably study drug associated. One child developed acute pancreatitis; another child developed granulocytopenia. Median CD4+-lymphocyte counts and plasma HIV RNA concentrations did not change significantly during therapy. In vitro neutrophil bactericidal activity againstStaphylococcus aureusand superoxide production were not significantly affected by rIFN-γ therapy. We conclude that rIFN-γ therapy in HIV-infected children receiving single-agent antiretroviral therapy is safe and does not produce consistent changes in CD4+-lymphocyte count, plasma HIV RNA concentration, or in vitro neutrophil function.
RNAs in the Sera of Persian Gulf War Veterans Have Segments Homologous to Chromosome 22q11.2
Authors: Urnovitz Howard B. , Tuite James J. , Higashida Jean M. , Murphy William H. 
Published: December 2019

ABSTRACT
Reverse transcriptase PCR (RT-PCR) was used for polyribonucleotide assays with sera from deployed Persian Gulf War veterans with the Gulf War Syndrome and a cohort of nonmilitary controls. Sera from veterans contained polyribonucleotides (amplicons) that were obtained by RT-PCR and that ranged in size from 200 to ca. 2,000 bp. Sera from controls did not contain amplicons larger than 450 bp. DNA sequences were derived from two amplicons unique to veterans. These amplicons, which were 414 and 759 nucleotides, were unrelated to each other or to any sequence in gene bank databases. The amplicons contained short segments that were homologous to regions of chromosome 22q11.2, an antigen-responsive hot spot for genetic rearrangements. Many of these short amplicon segments occurred near, between, or in chromosome 22q11.2 Alu sequences. These results suggest that genetic alterations in the 22q11.2 region, possibly induced by exposures to environmental genotoxins during the Persian Gulf War, may have played a role in the pathogenesis of the Gulf War Syndrome. However, the data did not exclude the possibility that other chromosomes also may have been involved. Nonetheless, the detection of polyribonucleotides such as those reported here may have application to the laboratory diagnosis of chronic diseases that have a multifactorial etiology.
Immunization with a Peptide Corresponding to Chlamydial Heat Shock Protein 60 Increases the Humoral Immune Response in C3H Mice to a Peptide Representing Variable Domain 4 of the Major Outer Membrane Protein ofChlamydia trachomatis
Authors: Motin Vladimir L. , de la Maza Luis M. , Peterson Ellena M. 
Published: December 2019

ABSTRACTC3H (H-2k) mice are susceptible to a vaginal challenge with human strains ofChlamydia trachomatisand thus are a useful strain for testing potentialChlamydiavaccine candidates. However, C3H mice are fairly poor responders in terms of the level of antibody resulting from immunization with potential protective peptides representing variable domains (VDs) of the major outer membrane protein (MOMP). C57BL/6 (H-2b) mice, on the other hand, are moderately resistant to a vaginal challenge but are good responders to the chlamydial MOMP VDs. Peptides representing universal T-cell helper epitopes were employed to determine whether the antibody response to a peptide representing VD4 of the MOMP, which has been shown to contain neutralizing epitopes, could be enhanced in C3H and C57 mice. Universal T-cell helper peptides from tetanus toxin, the pre-S2 region of hepatitis B virus, and the mouse heat shock protein 60, as well as the corresponding segment of theChlamydiaheat shock protein 60 (hspct), were coadministered with the VD4 peptide. Peptides were coencapsulated in liposomes containing the adjuvant monophosphoryl lipid A and administered by using a combination of mucosal and intramuscular injection. The only T-cell helper peptide that improved the immune response as judged by antibody level, in vitro neutralization assays, and T-cell proliferation was hspct. The response in the C57BL/6 strain was not significantly enhanced with hspctover levels achieved with VD4 alone; however, in C3H mice the levels of serum antibody toC. trachomatisincreased to that seen in C57 mice. However, the molecular specificity and immunoglobulin subclass distribution differed from those of the C57 response, and the neutralizing titers and T-cell proliferation responses were lower. In both strains of mice, titers of vaginal antibody toC. trachomatiswere low. In summary, of the T-helper peptides used, only hspctsignificantly enhanced the immune response of C3H mice to the VD4 peptide, but it had only a modest effect on the immune response of C57 mice.
Use of Protein AG in an Enzyme-Linked Immunosorbent Assay for Screening for Antibodies against Parapoxvirus in Wild Animals in Japan
Authors: Inoshima Yasuo , Shimizu Shinya , Minamoto Nobuyuki , Hirai Katsuya , Sentsui Hiroshi 
Published: December 2019

ABSTRACTUsing protein AG in an enzyme-linked immunosorbent assay (ELISA), we tried to detect antibodies against parapoxvirus in 9 species of wild animals in Japan: the Japanese badger (Meles meles anakuma), Japanese black bear (Ursus thibetanus japonicus), Japanese deer (Cervus nippon centralis), Japanese monkey (Macaca fuscata), Japanese raccoon dog (Nyctereutes procyonoides viverrinus), Japanese serow (Capricornis crispus), Japanese wild boar (Sus scrofa leucomystax), masked palm civet (Paguma larvata), and nutria (Myocastor coypus). A total of 272 serum samples were collected over the period from 1984 to 1995 and were tested by the protein AG-ELISA, the agar gel immunodiffusion test, and an indirect immunofluorescence assay. The protein AG-ELISA was effective in a serological survey for parapoxvirus in wild animals, and antibodies were detected only in Japanese serows. A total of 24 of 66 (36.4%) Japanese serows reacted positively, and they were found in almost all prefectures in all years tested. These results suggest that epizootic cycles of parapoxvirus exist widely in Japanese serows and that they could be reservoirs for the virus in the field in Japan. Moreover, it is probable that they might carry the virus to domestic animals such as cattle, sheep, and goats.
Production, Characterization, and Epitope Mapping of a Monoclonal Antibody against Aspartic Proteinase of
Candida albicans
Authors: Na Byoung-Kuk , Chung Gyung-Tae , Song Chul-Yong 
Published: December 2019

ABSTRACT

A monoclonal antibody (MAb; MAb CAP1) that was reactive with extracellular aspartic proteinase of
Candida albicans
(CAP) was produced. The MAb showed strong sensitivity and reactivity to CAP but not to the aspartic proteinases of
Candida parapsilosis
,
Candida tropicalis
, and
Aspergillus fumigatus
or to human cathepsin D or porcine pepsin. The epitope of the CAP recognized by the MAb was the proteinaseous part of CAP and the putative epitope of the MAb was located in the Asp
77
to Gly
103
sequence. This antibody could be useful for the characterization of CAP and would be a valuable probe for the detection of CAP antigen in the sera of patients with invasive candidiasis.
Inhibition of the Activities of Matrix Metalloproteinases 2, 8, and 9 by Chlorhexidine
Authors: Gendron Renée , Grenier Daniel , Sorsa Timo , Mayrand Denis 
Published: December 2019

ABSTRACT

Matrix metalloproteinases (MMPs) are a host cell-derived proteolytic enzyme family which plays a major role in tissue-destructive inflammatory diseases such as periodontitis. The aim of the present study was to evaluate the inhibitory effect of chlorhexidine (CHX) on MMP-2 (gelatinase A), MMP-9 (gelatinase B), and MMP-8 (collagenase 2) activity. Heat-denatured type I collagen (gelatin) was incubated with pure human MMP-2 or -9 activated with
p
-aminophenylmercuric acetate (APMA), and the proteolytic degradation of gelatin was monitored by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Coomassie blue staining. The effect of CHX on MMP-8 activity was also studied with a cellular model addressing the ability of phorbol myristate acetate (PMA)-triggered human peripheral blood neutrophils (polymorphonuclear leukocytes [PMNs]) to degrade native type I collagen. CHX inhibited the activities of both gelatinases (A and B), but MMP-2 appeared to be more sensitive than MMP-9. Adding calcium chloride to the assay mixtures almost completely prevented the inhibition of MMP-9 activity by CHX, while the inhibition of MMP-2 activity could be reversed only when CHX was used at a low concentration. This observation suggests that CHX may act via a cation-chelating mechanism. CHX dose-dependently inhibited collagenolytic activity of MMP-8 released by PMA-triggered PMNs. MMP-8 without APMA activation was inhibited clearly more efficiently than APMA-activated MMP-8. Our study suggests that the direct inhibition of the MMPs’ activities by CHX may represent a new valuable effect of this antimicrobial agent and explains, at least in part, the beneficial effects of CHX in the treatment of periodontitis.
Cell Surface Display of Human Immunodeficiency Virus Type 1 gp120 on
Escherichia coli
by Using Ice Nucleation Protein
Authors: Kwak Young-Don , Yoo Seung-Ku , Kim Eui-Joong 
Published: December 2019

ABSTRACT

A new system designed for cell surface display of recombinant proteins on
Escherichia coli
has been evaluated for expression of eukaryotic viral proteins. Human immunodeficiency virus type 1 (HIV-1) gp120 was fused to the C terminus of ice nucleation protein (INP), an outer membrane protein of
Pseudomonas syringae
. Western blotting, immunofluorescence microscopy, fluorescence-activated cell-sorting analysis, whole-cell enzyme-linked immunosorbent assay, and ice nucleation activity assay confirmed the successful expression of HIV-1 gp120 on the surface of
Escherichia coli
. This study shows that the INP system can be used for the expression of eukaryotic viral proteins. There is also a possibility that the INP system can be used as an AIDS diagnostic system, an oral vaccine delivery system, and an expression system for various heterologous higher-molecular-weight proteins.
Correlation of Cytokine Elaboration with Mononuclear Cell Adhesion to Platelet Storage Bag Plastic Polymers: a Pilot Study
Authors: ElKattan Ikbal , Anderson James , Yun J. K. , Colton E. , Yomtovian Roslyn 
Published: December 2019

ABSTRACT
The basis for many febrile nonhemolytic transfusion reactions associated with platelet transfusion therapy is cytokine elaboration and accumulation in the storage bag, which correlate with the leukocyte content and the length of platelet storage. We propose that a possible additional variable in the elaboration and accumulation of cytokines is the differential adhesion of mononuclear cells to the plastic substrate of the platelet storage bag. We hypothesize that mononuclear cell adhesion-induced cytokine release is greater in random-donor platelet bags composed of the polyolefin polymer compared to the single-donor apheresis platelet bags composed of the polyvinyl chloride polymer with the tri-(2-ethylhexyl) trimellitate (TEHTM) plasticizer. For four blood donors, we demonstrate preferential mononuclear cell adhesion, in vitro, to discs of polyolefin polymer versus discs of polyvinyl chloride polymer with the TEHTM plasticizer. Scanning electron microscopy corroborates this. In addition, proinflammatory cytokine (interleukin 1β [IL-1β] and tumor necrosis factor alpha [TNF-α]) levels are greater in culture wells containing discs of polyolefin polymer than in those containing discs of polyvinyl chloride polymer with the TEHTM plasticizer, and even more so in storage bags containing polyolefin polymer versus polyvinyl chloride polymer with the TEHTM plasticizer (IL-1β, TNF-α, IL-6, and IL-8). This study suggests, for the first time, that differential plastic substrate mononuclear cell adhesion may contribute to cytokine release during platelet storage. This may represent an additional variable in the pathophysiology of febrile nonhemolytic transfusion reactions in patients receiving stored platelet units.
Immunoglobulin G Avidity in Diagnosis of Toxoplasmic Lymphadenopathy and Ocular Toxoplasmosis
Authors: Paul Malgorzata 
Published: December 2019

ABSTRACT

Traditional serological techniques have some limitations in evaluating the duration of
Toxoplasma gondii
infection in pregnant women, patients with lymphadenopathy, and older children suspected of having congenital toxoplasmosis. In these three groups of patients, two variants of
T. gondii
immunoglobulin G (IgG) avidity tests were used: an EIA Kit (Labsystems) and a noncommercial enzyme-linked immunosorbent assay specially elaborated in the laboratory. The avidity of specific IgG in sera from 23 patients with a known recently acquired infection (mainly pregnant women) was low (less than 30%), whereas that in sera from 19 patients with toxoplasmic lymphadenopathy of 3 weeks to 6 months in duration (mean, 8.3 weeks) covered a large range (between 0.2 and 57.8%; mean, 25.7%); high avidity results were observed for 10 of 19 patients (52.6%). The large range of IgG avidity in patients with toxoplasmic lymphadenopathy suggests various durations of infection in these patients, with a tendency for a chronic phase of toxoplasmosis. According to the avidity marker, five patients with lymphadenopathy for less than 3 months did not have a recent
Toxoplasma
infection. In 6 of 19 patients with lymphadenopathy (31.6%), low IgG avidity values persisted until 5 months after the first serological examination. In all four patients with a documented chronic course of
Toxoplasma
infection (6 months to 8 years after the first positive serology), high IgG avidity values were observed. Among sera from 10 children and young immunocompetent adults suspected of having ocular reactivation of congenital toxoplasmosis, all had high IgG avidity values (over 40%), suggesting congenitally acquired ocular infection rather than noncongenital infection. In conclusion, the avidity of IgG is a valuable marker of recent toxoplasmosis in pregnant women, suggests the duration of invasion in patients with lymphadenopathy, and may be helpful for differentiation between reactivation of congenital infection and recently acquired ocular toxoplasmosis in immunocompetent patients. A low IgG avidity does not always identify a recent case of toxoplasmosis, but a high IgG avidity can exclude primary infections of less than 5 months’ duration.
A Reverse-Sandwich Enzyme-Linked Immunosorbent Assay for Verocytotoxin 1 and 2 Antibodies in Human and Bovine Sera
Authors: Miyazawa H. , Bannai H. , Yanase T. , Morita C. , Satoh S. , Sugiyama J. , Taniguchi S. , Inouye S. 
Published: December 2019

ABSTRACT

A reverse-sandwich enzyme-linked immunosorbent assay (ELISA), in which an antibody is sandwiched by antigens, was established for the titration of antibodies to verocytotoxins (VT) in human and animal sera. This assay has two advantages over a conventional indirect ELISA: (i) higher specificity and sensitivity and (ii) the ability to comparably titrate antibodies from different species. The VT1 (Shiga-like toxin 1) antibody-positive rates were 5% in 202 normal adult humans and 99% in 93 normal cattle at a dairy farm. This ELISA is most suitable for seroepidemiologic studies of infections with VT-producing
Escherichia coli
in humans and various animal species.
Identification of an IS711Element Interrupting thewboAGene ofBrucella abortusVaccine Strain RB51 and a PCR Assay To Distinguish Strain RB51 from OtherBrucellaSpecies and Strains
Authors: Vemulapalli Ramesh , McQuiston John R. , Schurig Gerhardt G. , Sriranganathan Nammalwar , Halling Shirley M. , Boyle Stephen M. 
Published: December 2019

ABSTRACTBrucella abortusvaccine strain RB51 is a natural stable attenuated rough mutant derived from the virulent strain 2308. The genetic mutations that are responsible for the roughness and the attenuation of strain RB51 have not been identified until now. Also, except for an assay based on pulsed-field gel electrophoresis, no other simple method to differentiate strain RB51 from its parent strain 2308 is available. In the present study, we demonstrate that thewboAgene encoding a glycosyltransferase, an enzyme essential for the synthesis of O antigen, is disrupted by an IS711element inB. abortusvaccine strain RB51. Exploiting this feature, we developed a PCR assay that distinguishes strain RB51 from all otherBrucellaspecies and strains tested.
Complement Fixation Test To Assess Humoral Immunity in Cattle and Sheep Vaccinated withBrucella abortusRB51
Authors: Adone Rosanna , Ciuchini Franco 
Published: December 2019

ABSTRACTThe live attenuatedBrucella abortusstrain RB51 is a rifampin-resistant, lipopolysaccharide (LPS) O-chain-deficient mutant of virulentB. abortus2308. The reduced O-chain content in RB51 prevents this bacterium from inducing antibodies detectable by the conventional serologic tests for bovine brucellosis diagnosis that mainly identify antibodies to LPS. The absence of available serologic tests for RB51 also complicates the diagnosis of possible RB51 infections in humans exposed to this strain. The purpose of this study was to evaluate the suitability of a complement fixation (CF) test performed with the rough strainB. abortusRB51, previously deprived of anticomplementary activity, in detecting anti-B. abortusRB51 antibodies in cattle and sheep experimentally vaccinated with this strain. The results of this study showed that a CF test with RB51 as the antigen is able to specifically detect antibodies following RB51 vaccination in cattle and sheep. In addition, this method could be a useful tool for detectingB. abortusRB51 infection in humans.
Exogenous Cyclic AMP, Cholera Toxin, and Endotoxin Induce Expression of the Lipopolysaccharide Receptor CD14 in Murine Bone Marrow Cells: Role of Purinoreceptors
Authors: Pedron Thierry , Girard Robert , Chaby Richard 
Published: December 2019

ABSTRACTLittle is known about the mechanisms of lipopolysaccharide (LPS) signaling in immature cells that do not express the LPS receptor CD14 yet. Bone marrow granulocytes do not constitutively express CD14 but can be stimulated by low doses of LPS in the absence of serum and then express an inducible form of LPS receptor (iLpsR). We show that in addition to LPS, cholera toxin (CT) and various cyclic AMP (cAMP) analogs can also induce the expression of iLpsR, which was identified as CD14. Induction was independent of intracellular cAMP. The hypothesis that cAMP analogs act via a cell surface receptor was suggested by the unresponsiveness of trypsin-treated cells to these inducers and by the specific binding of [3H]cAMP to the cells. This binding was not inhibited by LPS or CT but was inhibited by various purine derivatives. However, the receptor involved is not a conventional purinoreceptor since both an agonist and an antagonist of such receptors were able to induce iLpsR expression. The results suggest that cAMP analogs and other purine derivatives induce iLpsR after interaction with an unconventional, trypsin-sensitive, purinoreceptor distinct from LPS and CT receptors.