bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of Potential Diagnostic and Vaccine Candidates of
Helicobacter pylori
by Two-Dimensional Gel Electrophoresis, Sequence Analysis, and Serum Profiling
Authors: McAtee C. Patrick , Lim Moon Young , Fung Kevin , Velligan Mark , Fry Kirk , Chow Theresa , Berg Douglas E. 
Published: December 2019

ABSTRACT

There is great interest in characterizing the proteins of the gastric pathogen
Helicobacter pylori
, especially those to which humans respond immunologically, because of the potential importance of such proteins in diagnosis and vaccine development. Two-dimensional gel electrophoresis was used to separate and identify potential antigens of
H. pylori
ATCC 43504. Over 30 proteins were reactive in Western blots with pooled sera from 14 infected patients. These proteins were analyzed by N-terminal sequence analysis. Fourteen proteins were determined to be distinct from any proteins previously described from
H. pylori
; the others were previously isolated and characterized proteins. Analysis of eight distinct
H. pylori
strains showed that most of these antigens were produced by all of the strains. We propose that collection of new antigens such as those recognized here will be useful in serologic tests for detecting and monitoring
H. pylori
infection and may also serve as potential targets for antimicrobial agent or vaccine development.
Assignment of Immunoglobulin G1 and G2 Concentrations to Pneumococcal Capsular Polysaccharides 3, 6B, 14, 19F, and 23F in Pneumococcal Reference Serum 89-SF
Authors: Soininen Anu , Seppälä Ilkka , Wuorimaa Tomi , Käyhty Helena 
Published: December 2019

ABSTRACTWe describe standardization of an enzyme-linked immunosorbent assay (ELISA) for measuring immunoglobulin G1 (IgG1) and IgG2 concentrations of antibodies to pneumococcal capsular polysaccharide (Pnc PS). The ELISA uses a human pneumococcal reference serum pool, lot 89-SF, as a reference. IgG1 and IgG2 concentrations were assigned to antibodies to Pnc PS serotypes 3, 6B, 14, 19F, and 23F in 89-SF by ELISA using affinity-purified monoisotypic IgG1 and IgG2 preparations. The sum of IgG1 and IgG2 concentrations in 89-SF agrees well with the previously assigned IgG concentrations. The IgG1 and IgG2 values in 89-SF were used to measure antibodies to Pnc PS 6B, 14, and 23F in adult pre- and postimmunization sera and the sum of IgG1 and IgG2 concentrations correlated well with the IgG values. Furthermore, the IgG2/IgG1 ratio did not affect the detection of IgG1, the isotype usually represented by a lower concentration.
Development of a Colony Lift Immunoassay To Facilitate Rapid Detection and Quantification of
Escherichia coli
O157:H7 from Agar Plates and Filter Monitor Membranes
Authors: Ingram David T. , Lamichhane Chinta M. , Rollins David M. , Carr Lewis E. , Mallinson Edward T. , Joseph Sam W. 
Published: December 2019

ABSTRACT

E. coli
O157:H7 is a food-borne adulterant that can cause hemorrhagic ulcerative colitis and hemolytic uremic syndrome. Faced with an increasing risk of foods contaminated with
E. coli
O157:H7, food safety officials are seeking improved methods to detect and isolate
E. coli
O157:H7 in hazard analysis and critical control point systems in meat- and poultry-processing plants. A colony lift immunoassay was developed to facilitate the positive identification and quantification of
E. coli
O157:H7 by incorporating a simple colony lift enzyme-linked immunosorbent assay with filter monitors and traditional culture methods. Polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, Mass.) were prewet with methanol and were used to make replicates of every bacterial colony on agar plates or filter monitor membranes that were then reincubated for 15 to 18 h at 36 ± 1°C, during which the colonies not only remained viable but were reestablished. The membranes were dried, blocked with blocking buffer (Kirkegaard and Perry Laboratories [KPL], Gaithersburg, Md.), and exposed for 7 min to an affinity-purified horseradish peroxidase-labeled goat anti-
E. coli
O157 antibody (KPL). The membranes were washed, exposed to a 3,3′,5,5′-tetramethylbenzidine membrane substrate (TMB; KPL) or aminoethyl carbazole (AEC; Sigma Chemical Co., St. Louis, Mo.), rinsed in deionized water, and air dried. Colonies of
E. coli
O157:H7 were identified by either a blue (via TMB) or a red (via AEC) color reaction. The colored spots on the PVDF lift membrane were then matched to their respective parent colonies on the agar plates or filter monitor membranes. The colony lift immunoassay was tested with a wide range of genera in the family
Enterobacteriaceae
as well as different serotypes within the
E. coli
genus. The colony lift immunoassay provided a simple, rapid, and accurate method for confirming the presence of
E. coli
O157:H7 colonies isolated on filter monitors or spread plates by traditional culture methods. An advantage of using the colony lift immunoassay is the ability to test every colony serologically on an agar plate or filter monitor membrane simultaneously for the presence of the
E. coli
O157 antigen. This colony lift immunoassay has recently been successfully incorporated into a rapid-detection, isolation, and quantification system for
E. coli
O157:H7, developed in our laboratories for retail meat sampling.
Analysis of Immunoglobulin A Antibodies to
Helicobacter pylori
in Serum and Gastric Juice in Relation to Mucosal Inflammation
Authors: Hayashi Shunji , Sugiyama Toshiro , Yokota Kenji , Isogai Hiroshi , Isogai Emiko , Oguma Keiji , Asaka Masahiro , Fujii Nobuhiro , Hirai Yoshikazu 
Published: December 2019

ABSTRACT

Helicobacter pylori
is a major etiologic agent in gastroduodenal disorders. In this study, immunoglobulin A (IgA) antibodies to
H. pylori
antigens were evaluated in serum and gastric juice specimens obtained from patients with gastritis or peptic ulcers by utilizing antibody capture enzyme-linked immunosorbent assays (ACELISAs). Urease α subunit (UA), urease β subunit (UB), the 66-kDa heat shock protein (HSP), and the 25-kDa protein (25K) were used as antigens for the ACELISAs. The antibody titers of the ACELISAs reflect the ratio of
H. pylori
-specific IgA to total IgA. The ratio is stable, although the antibody concentration fluctuates in gastric juice. By using ACELISAs it was possible to evaluate quantitatively not only serum IgA antibodies but also gastric juice secretory IgA (S-IgA) antibodies. In both serum IgA and gastric juice S-IgA ACELISAs, the titers of antibody to HSP and 25K were remarkably correlated with the histologic grade of gastritis, whereas those to UA and UB were not strongly correlated with histologic grade. Thus, it is useful for estimating the histologic grade of gastritis to quantify serum IgA and gastric juice S-IgA antibodies to HSP and 25K.
Age-Related Changes in Blood Lymphocyte Subsets of Saudi Arabian Healthy Children
Authors: Shahabuddin S. , Al-Ayed I. , El-Rab M. O. Gad , Qureshi M. I. 
Published: December 2019

ABSTRACTThe age-related changes in absolute and percentage values of lymphocyte subsets in the peripheral blood of healthy children of different ages (1 month to 13 years) were studied by flow cytometry. The absolute and percentage values for most lymphocyte subpopulations differed substantially with age. Comparisons among age groups from infants through adults revealed progressive declines in the absolute numbers of leukocytes, total lymphocytes, and T, B, and natural killer (NK) cells. The percentages of T cells increased with age. Within the T-lymphocyte population, the CD8+subset increased but the CD4+subset decreased, resulting in a declining CD4+/CD8+ratio. The percentage of B cells declined, but that of NK cells remained unchanged. The percentage of HLA-DR+T cells increased over time, but their number changed inconsistently. Our findings confirm and extend earlier reports on age-related changes in lymphocyte subpopulations. These data should be useful in the interpretation of disease-related changes, as well as therapy-dependent alterations, in lymphocyte subsets in children of different age groups.
Serodiagnosis of Neosporosis in Individual Cows and Dairy Herds: A Comparative Study of Three Enzyme-Linked Immunosorbent Assays
Authors: Wouda W. , Brinkhof J. , van Maanen C. , de Gee A. L. W. , Moen A. R. 
Published: December 2019

ABSTRACTThe performance of three enzyme-linked immunosorbent assays (ELISA) for detection of antibodies toNeospora caninumin bovine sera was evaluated by using various categories of sera. Two commercial ELISA methods, one based on chemically fixed intact tachyzoites and one based on a sonicate lysate of whole tachyzoites, were compared with an in-house ELISA based on a detergent lysate of whole tachyzoites. A brief description of the development of the latter ELISA is also given. There was good agreement among all three tests with regard to postabortion sera. By using acute-phase abortion sera from cows with confirmedN. caninum-induced and non-N. caninum-induced abortions, satisfactory levels of sensitivity and specificity were calculated for all tests. In addition, similar test results were obtained with postpartum samples from dams and calves. However, considerable differences were found between test results of sequential samples and cross-sectional and total-herd samples. Apparently, these discrepancies were due to different sensitivities of the tests for detection of low antibody levels in chronically infected animals. It is suggested that these differences were primarily due to the use of different antigens and different test sample dilutions. It is concluded that all tests are applicable as an additional diagnostic tool in cases of abortion in cattle and for monitoring of congenitally infected calves. For herd screening, the lysate-based ELISAs appear to be more adequate because of their higher sensitivities.
rK39 Enzyme-Linked Immunosorbent Assay for Diagnosis of
Leishmania donovani
Infection
Authors: Zijlstra E. E. , Daifalla N. S. , Kager P. A. , Khalil E. A. G. , El-Hassan A. M. , Reed S. G. , Ghalib H. W. 
Published: December 2019

ABSTRACT

The rK39 enzyme-linked immunosorbent assay (ELISA) was compared with the direct agglutination test (DAT) for
Leishmania donovani
infection in the Sudan. rK39 ELISA proved more sensitive than DAT in diagnosis of kala-azar (93 and 80%, respectively); both tests may remain positive up to 24 months after treatment. For patients with post-kala-azar dermal leishmaniasis and individuals with subclinical infection, rK39 ELISA performed as well as DAT but could detect infection 6 months earlier in ∼40% of patients. Conversion in DAT and rK39 ELISA also occurred in leishmanin skin test (LST)-positive individuals, suggesting active parasite replication (rK39 is an amastigote antigen) in these presumably immune individuals. In contrast to DAT, rK39 ELISA also detected infection in randomly selected LST-positive individuals (in four of six) and endemicity (LST-negative) controls (in one of five). rK39 ELISA appears more sensitive than DAT and may prove an important tool in epidemiological studies.
Antigenic Variation in
Bacteroides forsythus
Detected by a Checkerboard Enzyme-Linked Immunosorbent Assay
Authors: Sims Tom J. , Mancl Lloyd A. , Braham Pamela H. , Page Roy C. 
Published: December 2019

ABSTRACT

Evidence indicating that multiple serotypes of
Bacteroides forsythus
participate in rapidly progressing periodontal infections has not been reported previously. Our aim was to develop an assay for detecting subsets of
B. forsythus
clinical isolates which differ in serogroup membership and subsets of patients with immunoglobulin G (IgG) responses which differ in serogroup recognition. A checkerboard enzyme-linked immunosorbent assay (ELISA) was used to assess variation in the IgG binding profiles of 22 clinical isolates in sera from 28 patients with early-onset rapidly progressive periodontitis. To accommodate the maximum number of isolates and sera in a given assay run, a multiplate assay grid with standard 96-well microtest plates was established. Single dilutions of individual sera were placed in rows crossing columns of isolate-coated wells, and antigen-specific IgG immobilized in the wells was measured as ELISA absorbance. Pooled sera and isolates were assayed in parallel to serve as negative controls for variation in IgG binding profiles. Correlation and hierarchical cluster analysis of the absorbance data matrix showed that the isolates could be sorted into at least four clusters based on variations in their IgG binding profiles across different sera. Furthermore, at least two patient clusters were defined by variations in their serum IgG antigen recognition profiles across different isolates. We conclude that multiple serogroups of
B. forsythus
exist and that different serogroups are dominant in the antibody response of different patients. The method applied here could be used to serologically classify clinical isolates of other species which evoke a serum antibody response in patients.
Levels of Cytokines and Immune Activation Markers in Plasma in Human Immunodeficiency Virus Infection: Quality Control Procedures
Authors: Aziz Najib , Nishanian Parunag , Fahey John L. 
Published: December 2019

ABSTRACTProcedures for quality control (QC) in a laboratory that concentrates on cytokine and soluble marker measurements in biological fluids are outlined. Intra-assay, interassay, and interlaboratory experiences are presented. Plasma and serum β2-microglobulin (β2M) and neopterin test data are presented in greatest detail, along with substantial tumor necrosis factor alpha (TNF-α), gamma interferon, soluble interleukin-2 receptor-α (sIL-2Rα), sTNF-RII, IL-4, and IL-6 data. Recommended QC procedures for cytokine and soluble-marker testing include replicate testing of two or more reference samples provided by the kit manufacturer, replicate testing of in-house frozen reference QC samples that represent normal and abnormal analyte contents, retesting 15 to 20% of randomly selected samples, and comparing normal reference ranges each year. Also, eight cytokines and soluble markers were evaluated in human immunodeficiency virus (HIV)-seronegative and HIV-seropositive individuals stratified on the basis of CD4 T-cell numbers. Levels of some but not all cytokines in serum increased in HIV infection. There was a tendency for cytokines to increase with more advanced disease, defined by reduced CD4 T-cell numbers. Cytokine changes did not relate closely to CD4 level, indicating that separate information was provided by the measurements of TNF-α, sTNF-RII, sIL-2Rα, β2M, and neopterin. Serum IL-4 and TNF-α levels were not increased. The quality of laboratory data can impact on clinical relevance. Interlaboratory comparisons revealed substantial differences at some sites and documented the need for external proficiency-testing quality assurance programs.
Development of a Fluorescent Focus Identification Assay Using Serotype-Specific Monoclonal Antibodies for Detection and Quantitation of Rotaviruses in a Tetravalent Rotavirus Vaccine
Authors: Yang D. P. , Goldberg K. M. , Ma Xiang-Dong , Magargle W. , Rappaport R. 
Published: December 2019

ABSTRACT
A fluorescent focus identification assay (FFIDA) was developed for use in experimental studies and for quantitation of the components in a tetravalent live oral rotavirus vaccine. The assay utilizes four serotype-specific neutralizing monoclonal antibodies (MAb) to detect and quantify individual rotaviruses by immunofluorescence staining of fixed virus-infected monkey kidney cells. In mixed virus infections, all four MAb, W1 (serotype 1), 1C10 (serotype 2), R1 (serotype 3), and S4 (serotype 4), specifically stain the relevant homologous serotype without exhibiting any cross-reactivity against the other serotypes. Furthermore, the test is sensitive enough to differentiate at least twofold (0.3 log) differences in virus titer. The results of testing four individual experimental vaccine lots three or more consecutive times showed that all four lots contained similar proportions of the four vaccine strains as detected by the classical plaque neutralization identification test. The rapidity and efficiency of the FFIDA are desirable attributes that make it suitable for use in studies requiring identification and quantitation of one or more of the four major rotavirus serotypes.
Accumulation of Acid-Fast Lipochrome Bodies in Glial Cells of the Midbrain Nigral Lesion in Parkinson’s Disease
Authors: Kohbata Shunro , Tamura Tomokazu , Hayashi Ryoichi 
Published: December 2019

ABSTRACT
To confirm or refute the proposed link between nocardiae and Parkinson’s disease (PD), we investigated the presence of acid-fast spherical structures similar to filterable nocardiae at the midbrain nigral lesions of three patients with PD. Many clusters of acid-fast lipochrome bodies were dense around blood vessels in the two patients with Hoehn and Yahr stage II and III PD. These clusters were present in the vicinity of melanin-pigmented neurons in the three PD patients studied. Examination of adjacent hematoxylin-and-eosin-stained sections indicated that they consisted of yellow-green granules, bodies, and aggregates in ballooned glial cells. On the other hand, no clusters of acid-fast lipochrome bodies were observed at the compacta region of three control patients. Our results suggest that the immunological and genetic relationship between the acid-fast lipochrome bodies and filterable nocardiae should be investigated.
Angiocentric CD3+T-Cell Infiltrates in Human Immunodeficiency Virus Type 1-Associated Central Nervous System Disease in Children
Authors: Katsetos Christos D. , Fincke John E. , Legido Agustin , Lischner Harold W. , de Chadarevian Jean-Pierre , Kaye Edward M. , Platsoucas Chris D. , Oleszak Emilia L. 
Published: December 2019

ABSTRACTA significant proportion of brain tissue specimens from children with AIDS show evidence of vascular inflammation in the form of transmural and/or perivascular mononuclear-cell infiltrates at autopsy. Previous studies have shown that in contrast to inflammatory lesions observed in human immunodeficiency virus type 1 (HIV-1) encephalitis, in which monocytes/macrophages are the prevailing mononuclear cells, these infiltrates consist mostly of lymphocytes. Perivascular mononuclear-cell infiltrates were found in brain tissue specimens collected at autopsy from five of six children with AIDS and consisted of CD3+T cells and equal or greater proportions of CD68+monocytes/macrophages. Transmural (including endothelial) mononuclear-cell infiltrates were evident in one patient and comprised predominantly CD3+T cells and small or, in certain vessels, approximately equal proportions of CD68+monocytes/macrophages. There was a clear preponderance of CD3+CD8+T cells on the endothelial side of transmural infiltrates. In active lesions of transmural vasculitis, CD3+T-cell infiltrates exhibited a distinctive zonal distribution. The majority of CD3+cells were also CD8+and CD45RO+. Scattered perivascular monocytes/macrophages in foci of florid vasculitis were immunoreactive for the p24 core protein. In contrast to the perivascular space, the intervening brain neuropil was dominated by monocytes/macrophages, microglia, and reactive astrocytes, containing only scant CD3+CD8+cells. Five of six patients showed evidence of calcific vasculopathy, but only two exhibited HIV-1 encephalitis. One patient had multiple subacute cerebral and brainstem infarcts associated with a widespread, fulminant mononuclear-cell vasculitis. A second patient had an old brain infarct associated with fibrointimal thickening of large leptomeningeal vessels. These infiltrating CD3+T cells may be responsible for HIV-1-associated CNS vasculitis and vasculopathy and for endothelial-cell injury and the opening of the blood-brain barrier in children with AIDS.
Anticytomegalovirus (Anti-CMV) Immunoglobulin G Avidity in Identification of Pregnant Women at Risk of Transmitting Congenital CMV Infection
Authors: Lazzarotto Tiziana , Spezzacatena Patrizia , Varani Stefania , Gabrielli Liliana , Pradelli Paola , Guerra Brunella , Landini Maria Paola 
Published: December 2019

ABSTRACT
In this work, we show that the determination of the anticytomegalovirus antibody avidity carried out before week 18 of gestation is a helpful tool to identify women for enrollment in prenatal diagnosis. This procedure can identify all pregnant women who will give birth to an infected newborn.
Dysregulated Synthesis of Intracellular Type 1 and Type 2 Cytokines by T Cells of Patients with Cutaneous T-Cell Lymphoma
Authors: Lee Bang-Ning , Duvic Madeleine , Tang Chih-Kwang , Bueso-Ramos Carlos , Estrov Zeev , Reuben James M. 
Published: December 2019

ABSTRACTMycosis fungoides (MF) and Sezary syndrome (SS) are the two main clinical entities of cutaneous T-cell lymphoma (CTCL). As the disease progresses from MF to SS, a switch from a type 1 (interleukin [IL]-2 and gamma interferon [IFN-γ]) to a type 2 (IL-4) cytokine production profile occurs. Although roles for type 1 and type 2 cytokines in the pathogenesis of CTCL have been proposed, the cellular origins of these cytokines are unclear. Using flow cytometry to identify individual T-cell subsets, we studied cytokine synthesis by the T cells of 13 patients with SS and 12 with MF and 9 hematologically healthy donors. Upon activation with phorbol 12-myristate 13-acetate (PMA), the numbers of T cells synthesizing IL-2 were similar for all study groups. Whereas the predominant T-cell producing IL-2 in healthy donors and in those with MF was CD7+, in patients with SS, it was CD7−. Although the number of IL-4+CD4+T cells was low for all study groups, there was a significantly higher number of IL-4+CD8+T cells in patients with MF than in those with SS or healthy donors. There was a decline in the number of IFN-γ-producing T cells in CTCL donors compared to that in healthy donors. More importantly, there was a significant decrease in the number of IFN-γ-producing T cells with disease progression from MF to SS. The inability of these T cells to synthesize IFN-γ may have prognostic value in CTCL, since it may be responsible for the progression of the disease from MF to SS.
Protection against Influenza Virus Infection of Mice Fed
Bifidobacterium breve
YIT4064
Authors: Yasui Hisako , Kiyoshima Junko , Hori Tetuji , Shida Kan 
Published: December 2019

ABSTRACT

Mice fed
Bifidobacterium breve
YIT4064 and immunized orally with influenza virus were more strongly protected against influenza virus infection of the lower respiratory tract than ones immunized with influenza virus only. The number of mice with enhanced anti-influenza virus immunoglobulin G (IgG) in serum upon oral administration of
B. breve
YIT4064 and oral immunization with influenza virus was significantly greater than that upon oral immunization with influenza virus only. These findings demonstrated that the oral administration of
B. breve
YIT4064 increased anti-influenza virus IgG antibodies in serum and protected against influenza virus infection. The oral administration of
B. breve
YIT4064 may enhance antigen-specific IgG against various pathogenic antigens taken orally and induce protection against various virus infections.