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Article ; Online: Cost-effectiveness of a 3-antigen versus single-antigen vaccine for the prevention of hepatitis B in adults in the United States.

Talbird, Sandra E / Anderson, Seri A / Nossov, Misha / Beattie, Nell / Rak, Aaron T / Diaz-Mitoma, Francisco

2023 Volume 41, Issue 23, Page(s) 3506–3517

Abstract: Objectives: The first 3-antigen hepatitis B vaccine was approved ... Engerix-B: Methods: A cost-effectiveness model was developed using a combined decision-tree and Markov ... effective intervention for preventing HBV infection and addressing the long-standing burden of hepatitis B ...

Abstract	Objectives: The first 3-antigen hepatitis B vaccine was approved by the United States (US) Food and Drug Administration in November 2021 and was recommended by the Centers for Disease Control and Prevention in 2022. We estimated the cost-effectiveness of this 3-antigen vaccine (PreHevbrio™) relative to the single-antigen vaccine, Engerix-B Methods: A cost-effectiveness model was developed using a combined decision-tree and Markov structure to follow 100,000 adults over their remaining lifetimes after vaccination with either the 3-antigen or single-antigen vaccine. Outcomes from societal and healthcare sector perspectives were calculated for adults aged 18-44, 45-64, and ≥65 years; adults with diabetes; and adults with obesity. Seroprotection rates were obtained from the phase3, head-to-head PROTECT trial (NCT03393754). Incidence, vaccine costs, vaccine adherence rates, direct and indirect costs, utilities, transition probabilities, and mortality were obtained from published sources. Health outcomes and costs (2020USD) were discounted 3% annually and reported by vaccine and population. One-way sensitivity and scenario analyses were conducted. Results: In the model, the 3-antigen vaccine led to fewer HBV infections, complications, and deaths compared with the single-antigen vaccine in all modeled populations due to higher rates and faster onset of seroprotection. Compared with the single-antigen vaccine, the 3-antigen vaccine had better health outcomes, more quality-adjusted life-years (QALYs), and lower costs in adults aged 18-64 years, adults with diabetes, and adults with obesity (dominant strategy). For adults aged ≥65 years, the 3-antigen vaccine was cost-effective compared with the single-antigen vaccine ($26,237/QALY gained) below common willingness-to-pay thresholds ($50,000-$100,000/QALY gained). In sensitivity analyses, results were sensitive to vaccine cost per dose, incidence, and age at vaccination. Conclusion: The recently approved 3-antigen vaccine is a cost-saving or cost-effective intervention for preventing HBV infection and addressing the long-standing burden of hepatitis B among US adults.
MeSH term(s)	Adult ; Humans ; United States/epidemiology ; Cost-Benefit Analysis ; Hepatitis B/epidemiology ; Hepatitis B/prevention & control ; Vaccination ; Hepatitis B virus ; Hepatitis B Vaccines ; Diabetes Mellitus ; Quality-Adjusted Life Years
Chemical Substances	Hepatitis B Vaccines
Language	English
Publishing date	2023-05-03
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	605674-x
ISSN	1873-2518 ; 0264-410X
ISSN (online)	1873-2518
ISSN	0264-410X
DOI	10.1016/j.vaccine.2023.04.022
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Article ; Online: A novel digital PCR-based method to quantify (switched) B cells reveals the extent of allelic involvement in different recombination processes in the IGH locus.

Zoutman, Willem H / Nell, Rogier J / Versluis, Mieke / Pico, Ingrid / Khanh Vu, T H / Verdijk, Robert M / van der Burg, Mirjam / Langerak, Anton W / van der Velden, Pieter A

Molecular immunology

2022 Volume 145, Page(s) 109–123

Abstract: B cells fulfill an important role in the adaptive immunity. Upon activation and immunoglobulin (IG ... applications, it can be important to quantify (switched) B cells accurately in a variety of body fluids and ... both depend on the accessibility of B cell epitopes and therefore require intact (fixed) cells ...

Abstract	B cells fulfill an important role in the adaptive immunity. Upon activation and immunoglobulin (IG) class switching, these cells function in the humoral immunity compartment as plasma cells. For clinical applications, it can be important to quantify (switched) B cells accurately in a variety of body fluids and tissues of benign, inflammatory and malignant origin. For decades, flow cytometry and immunohistochemistry (IHC) have been the preferred methods for quantification. Although these methods are widely used, both depend on the accessibility of B cell epitopes and therefore require intact (fixed) cells. Whenever samples are low in quantity and/or quality, accurate quantification can be difficult. By shifting the focus from epitopes to DNA markers, quantification of B cells remains achievable. During differentiation and maturation, B cells are subjected to programmed genetic recombination processes like VDJ rearrangements and class switch recombination (CSR), which result in deletion of specific sequences of the IGH locus. These cell type-specific DNA "scars" (loss of sequences) in IG genes can be exploited as B cell markers in digital PCR (dPCR) based quantification methods. Here, we describe a novel, specific and sensitive digital PCR-based method to quantify mature and switched B cells in DNA specimens of benign and (copy number unstable) malignant origin. We compared this novel way of B cell quantitation with flow cytometric and immunohistochemical methods. Through cross-validation with flow cytometric sorted B cell subpopulations, we gained quantitative insights into allelic involvement in different recombination processes in the IGH locus. Our newly developed method is accurate and independent of the cellular context, offering new possibilities for quantification, even for (limited) small samples like liquid biopsies.
MeSH term(s)	B-Lymphocytes ; DNA ; Genes, Immunoglobulin Heavy Chain/genetics ; Immunoglobulin Class Switching/genetics ; Polymerase Chain Reaction
Chemical Substances	DNA (9007-49-2)
Language	English
Publishing date	2022-03-23
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	424427-8
ISSN	1872-9142 ; 0161-5890
ISSN (online)	1872-9142
ISSN	0161-5890
DOI	10.1016/j.molimm.2022.03.003
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Article ; Online: B.1.1.7 and B.1.351 SARS-CoV-2 variants display enhanced Spike-mediated fusion

Rajah, Maaran Michael / Hubert, Mathieu / Bishop, Elodie / Saunders, Nell / Robinot, Rémy / Grzelak, Ludivine / Planas, Delphine / Zivaljic, Marija / Planchais, Cyril / Guivel-Benhassine, Florence / Porrot, Françoise / Mouquet, Hugo / Chakrabarti, Lisa / Buchrieser, Julian / Schwartz, Olivier

bioRxiv

Abstract: SARS-CoV-2 B.1.1.7 (variant Alpha) and B.1.351 (variant Beta) have supplanted pre-existing strains ... type-I interferon (IFN) sensitivity of the variants remain poorly characterized. Here, we assessed B.1 ... 1.7 and B.1.351 spread and fusion in cell cultures. B.1.1.7 and B.1.351 replicated similarly ...

Abstract	SARS-CoV-2 B.1.1.7 (variant Alpha) and B.1.351 (variant Beta) have supplanted pre-existing strains in many countries. Severe COVID-19 is characterized by lung abnormalities, including the presence of syncytial pneumocytes. Syncytia form when infected cells fuse with adjacent cells. The fitness, cytopathic effects and type-I interferon (IFN) sensitivity of the variants remain poorly characterized. Here, we assessed B.1.1.7 and B.1.351 spread and fusion in cell cultures. B.1.1.7 and B.1.351 replicated similarly to D614G reference strain in Vero, Caco-2, Calu-3 and primary airway cells and were similarly sensitive to IFN. The variants formed larger and more numerous syncytia. Variant Spikes, in the absence of any other viral proteins, resulted in faster fusion relative to D614G. B.1.1.7 and B.1.351 fusion was similarly inhibited by interferon induced transmembrane proteins (IFITMs). Individual mutations present in the variant Spikes modified fusogenicity, binding to ACE2 and recognition by monoclonal antibodies. Also, B.1.1.7 and B.1.351 variants remain sensitive to innate immunity components. The mutations present in the two variants globally enhance viral fusogenicity and allow for antibody evasion.
Keywords	covid19
Language	English
Publishing date	2021-06-11
Publisher	Cold Spring Harbor Laboratory
Document type	Article ; Online
DOI	10.1101/2021.06.11.448011
Database	COVID19

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Article ; Online: Role for ZAP-70 Signaling in the Differential Effector Functions of Rituximab and Obinutuzumab (GA101) in Chronic Lymphocytic Leukemia B Cells.

Skopelja-Gardner, Sladjana / Jones, Jonathan D / Hamilton, B JoNell / Danilov, Alexey V / Rigby, William F C

Journal of immunology (Baltimore, Md. : 1950)

2017 Volume 199, Issue 4, Page(s) 1275–1282

Abstract: Rituximab (RTX) has been the hallmark anti-CD20 mAb for the treatment of B cell neoplasms ... including B cell chronic lymphocytic leukemia (B-CLL). Recently, a novel humanized anti-CD20 mAb ... antigenic modulation), as well as a process that alters B cell phenotype and function. This study investigates ...

Abstract	Rituximab (RTX) has been the hallmark anti-CD20 mAb for the treatment of B cell neoplasms, including B cell chronic lymphocytic leukemia (B-CLL). Recently, a novel humanized anti-CD20 mAb obinutuzumab (GA101) has been implemented as first-line CLL therapy. Treatment of CLL patients with RTX is associated with CD20 loss via an FcγR-mediated process, trogocytosis. RTX-induced trogocytosis has been characterized as both the means of resistance to therapy, via loss of cell surface target proteins (antigenic modulation), as well as a process that alters B cell phenotype and function. This study investigates the nature and clinical relevance of GA101-mediated trogocytosis. In this study, we demonstrate that GA101 is a more potent mediator of trogocytosis than RTX in vitro in both normal B cells and B-CLL cells. Qualitative differences in the effector function of these anti-CD20 Abs appear specific to B-CLL cells. GA101-mediated CD19 and CD20 trogocytosis from B-CLL cells is associated with its ability to induce homotypic adhesion (HA). The degree of HA varies between CLL patients and positively correlates with the expression of ZAP-70, a BCR-associated kinase. Deregulation of ZAP-70 using tyrosine kinase inhibitors, gefitinib or ibrutinib, diminishes HA formation and trogocytosis by GA101. Taken together, these findings elucidate the differences in trogocytosis and HA formation mediated by anti-CD20 mAbs RTX and GA101, as well as provide a novel link between ZAP-70 expression and these effector functions.
MeSH term(s)	Antibodies, Monoclonal, Humanized/pharmacology ; Antibodies, Monoclonal, Murine-Derived/pharmacology ; Antigens, CD20/immunology ; Antineoplastic Agents/pharmacology ; B-Lymphocytes/drug effects ; B-Lymphocytes/immunology ; B-Lymphocytes/metabolism ; Cell Adhesion ; Cell Adhesion Molecules/metabolism ; Gefitinib ; Humans ; Leukemia, Lymphocytic, Chronic, B-Cell/drug therapy ; Leukemia, Lymphocytic, Chronic, B-Cell/immunology ; Leukemia, Lymphocytic, Chronic, B-Cell/metabolism ; Monocytes/drug effects ; Monocytes/immunology ; Monocytes/metabolism ; Pyrazoles/pharmacology ; Pyrimidines/pharmacology ; Quinazolines/pharmacology ; Receptors, IgG/immunology ; Rituximab/pharmacology ; Signal Transduction/drug effects ; ZAP-70 Protein-Tyrosine Kinase/genetics ; ZAP-70 Protein-Tyrosine Kinase/metabolism
Chemical Substances	Antibodies, Monoclonal, Humanized ; Antibodies, Monoclonal, Murine-Derived ; Antigens, CD20 ; Antineoplastic Agents ; Cell Adhesion Molecules ; Pyrazoles ; Pyrimidines ; Quinazolines ; Receptors, IgG ; ibrutinib (1X70OSD4VX) ; Rituximab (4F4X42SYQ6) ; ZAP-70 Protein-Tyrosine Kinase (EC 2.7.10.2) ; ZAP70 protein, human (EC 2.7.10.2) ; obinutuzumab (O43472U9X8) ; Gefitinib (S65743JHBS)
Language	English
Publishing date	2017-07-14
Publishing country	United States
Document type	Journal Article
ZDB-ID	3056-9
ISSN	1550-6606 ; 0022-1767 ; 1048-3233 ; 1047-7381
ISSN (online)	1550-6606
ISSN	0022-1767 ; 1048-3233 ; 1047-7381
DOI	10.4049/jimmunol.1602105
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Article ; Online: Induction of interleukin-6 production by rituximab in human B cells.

Jones, Jonathan D / Hamilton, B JoNell / Skopelja, Sladjana / Rigby, William F C

Arthritis & rheumatology (Hoboken, N.J.)

2014 Volume 66, Issue 11, Page(s) 2938–2946

Abstract: ... antineutrophil cytoplasmic antibody-associated vasculitis is not easily related to B cell depletion alone. Prior studies have shown ... that RTX mediates a rapid stripping of CD20 and CD19 from the human B cell through a process known ... as trogocytosis. The aim of the present study was to investigate whether changes in B cell phenotype resulting ...

Abstract	Objective: Rituximab (RTX), an anti-CD20 monoclonal antibody, is highly effective in the treatment of several autoimmune diseases. The mechanism by which RTX treatment improves rheumatoid arthritis and antineutrophil cytoplasmic antibody-associated vasculitis is not easily related to B cell depletion alone. Prior studies have shown that RTX mediates a rapid stripping of CD20 and CD19 from the human B cell through a process known as trogocytosis. The aim of the present study was to investigate whether changes in B cell phenotype resulting from trogocytosis would diminish the ability of B cells to promote autoimmune disease. Methods: Human peripheral blood mononuclear cells were cultured with RTX under conditions that permitted trogocytosis. Changes in B cell phenotype and cytokine production were measured in the basal state and under conditions of activation with interleukin-4 (IL-4)/anti-CD40. The effects of RTX were characterized in terms of a requirement for interaction with the Fcγ receptor (FcγR) and other Fc-dependent interactions. Results: Trogocytosis induced a marked loss of surface CD19, IgD, CD40, and B cell-activating factor receptor, but did not alter induction of CD86 expression on purified B cells following IL-4/anti-CD40 treatment. Unexpectedly, RTX-dependent trogocytosis of normal human B cells in vitro led to a rapid up-regulation of IL-6 production, with no effect on the production of tumor necrosis factor α, IL-1β, interferon-γ, or IL-10. This effect was Fc-dependent and required the presence of an FcγR-bearing cell. Moreover, this effect involved the release of preformed intracellular IL-6 protein, as well as marked increases in IL-6 messenger RNA levels. Conclusion: RTX-mediated trogocytosis of B cells in vitro results in acute production and release of IL-6. The nature of this effect and how it is related to the acute infusion reactions seen with RTX administration remain to be determined.
MeSH term(s)	Antibodies, Monoclonal, Murine-Derived/pharmacology ; Antirheumatic Agents/pharmacology ; B-Lymphocytes/drug effects ; B-Lymphocytes/metabolism ; CD40 Antigens/metabolism ; Cells, Cultured ; Humans ; Immunoglobulin D/metabolism ; In Vitro Techniques ; Interleukin-10/metabolism ; Interleukin-1beta/metabolism ; Interleukin-4/metabolism ; Interleukin-6/metabolism ; Receptors, IgG/metabolism ; Rituximab ; Tumor Necrosis Factor-alpha/metabolism ; Up-Regulation/drug effects
Chemical Substances	Antibodies, Monoclonal, Murine-Derived ; Antirheumatic Agents ; CD40 Antigens ; Immunoglobulin D ; Interleukin-1beta ; Interleukin-6 ; Receptors, IgG ; Tumor Necrosis Factor-alpha ; Interleukin-10 (130068-27-8) ; Interleukin-4 (207137-56-2) ; Rituximab (4F4X42SYQ6)
Language	English
Publishing date	2014-07-31
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2756371-6
ISSN	2326-5205 ; 2326-5191
ISSN (online)	2326-5205
ISSN	2326-5191
DOI	10.1002/art.38798
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Article: A novel digital PCR-based method to quantify (switched) B cells reveals the extent of allelic involvement in different recombination processes in the IGH locus

Zoutman, Willem H. / Nell, Rogier J. / Versluis, Mieke / Pico, Ingrid / Khanh Vu, T.H. / Verdijk, Robert M. / van der Burg, Mirjam / Langerak, Anton W. / van der Velden, Pieter A.

Molecular immunology. 2022 May, v. 145

2022

Abstract	B cells fulfill an important role in the adaptive immunity. Upon activation and immunoglobulin (IG) class switching, these cells function in the humoral immunity compartment as plasma cells. For clinical applications, it can be important to quantify (switched) B cells accurately in a variety of body fluids and tissues of benign, inflammatory and malignant origin. For decades, flow cytometry and immunohistochemistry (IHC) have been the preferred methods for quantification. Although these methods are widely used, both depend on the accessibility of B cell epitopes and therefore require intact (fixed) cells. Whenever samples are low in quantity and/or quality, accurate quantification can be difficult. By shifting the focus from epitopes to DNA markers, quantification of B cells remains achievable. During differentiation and maturation, B cells are subjected to programmed genetic recombination processes like VDJ rearrangements and class switch recombination (CSR), which result in deletion of specific sequences of the IGH locus. These cell type-specific DNA “scars” (loss of sequences) in IG genes can be exploited as B cell markers in digital PCR (dPCR) based quantification methods. Here, we describe a novel, specific and sensitive digital PCR-based method to quantify mature and switched B cells in DNA specimens of benign and (copy number unstable) malignant origin. We compared this novel way of B cell quantitation with flow cytometric and immunohistochemical methods. Through cross-validation with flow cytometric sorted B cell subpopulations, we gained quantitative insights into allelic involvement in different recombination processes in the IGH locus. Our newly developed method is accurate and independent of the cellular context, offering new possibilities for quantification, even for (limited) small samples like liquid biopsies.
Keywords	DNA ; epitopes ; flow cytometry ; genetic recombination ; humoral immunity ; immunoglobulins ; immunohistochemistry ; liquids ; loci ; polymerase chain reaction
Language	English
Dates of publication	2022-05
Size	p. 109-123.
Publishing place	Elsevier Ltd
Document type	Article
ZDB-ID	424427-8
ISSN	1872-9142 ; 0161-5890
ISSN (online)	1872-9142
ISSN	0161-5890
DOI	10.1016/j.molimm.2022.03.003
Database	NAL-Catalogue (AGRICOLA)

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Zs.A 166: Show issues

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Article ; Online: Rituximab mediates loss of CD19 on B cells in the absence of cell death.

Jones, Jonathan D / Hamilton, B JoNell / Rigby, William F C

Arthritis and rheumatism

2012 Volume 64, Issue 10, Page(s) 3111–3118

Abstract: Objective: To evaluate loss of the B cell-specific marker CD19 after the addition of rituximab ... Evidence of B cell death was evaluated by measuring messenger RNA (mRNA) levels as well ... This reduction occurred in the absence of complement. Despite the decrease in CD19 expression, B cell death did ...

Abstract	Objective: To evaluate loss of the B cell-specific marker CD19 after the addition of rituximab (RTX) to healthy donor blood and to determine the role of complement-mediated cytotoxicity in these cells. Methods: Whole blood and peripheral blood mononuclear cells (PBMCs) from healthy donors were evaluated for the loss of CD19 in the presence of RTX using flow cytometry. The effect of complement on CD19 loss was examined using serum-free media, C3- and C5-deficient sera, and a C5-blocking antibody. Evidence of B cell death was evaluated by measuring messenger RNA (mRNA) levels as well as by flow cytometry. Transfer of CD19 antigen to monocytes and neutrophils was evaluated by flow cytometry and confocal microscopy. Results: RTX induced a rapid decrease in CD19 count (mean 51%; n = 37) in PBMCs. This reduction occurred in the absence of complement. Despite the decrease in CD19 expression, B cell death did not occur, as evidenced by a lack of change in CD19 or CD20 mRNA levels and a lack of change in CD19 levels determined by intracellular staining and through the use of viability dyes. The CD19 antigen was shown to be transferred to monocytes and neutrophils in an Fc-dependent manner. Conclusion: Our findings indicate that the addition of RTX to healthy donor PBMCs in vitro results in complement-independent loss of CD19 without causing B cell death. CD19 is transferred from B cells to monocytes and neutrophils during shaving of the RTX-CD20 complex in an Fc-dependent manner. These data suggest that monitoring the effect of RTX by measuring the CD19+ cell count may be compromised by this activity.
MeSH term(s)	Antibodies, Monoclonal, Murine-Derived/pharmacology ; Antigens, CD19/metabolism ; Antirheumatic Agents/pharmacology ; B-Lymphocytes/drug effects ; B-Lymphocytes/immunology ; B-Lymphocytes/metabolism ; Cell Death/drug effects ; Cell Death/immunology ; Humans ; Leukocytes, Mononuclear/drug effects ; Leukocytes, Mononuclear/immunology ; Leukocytes, Mononuclear/metabolism ; Rituximab
Chemical Substances	Antibodies, Monoclonal, Murine-Derived ; Antigens, CD19 ; Antirheumatic Agents ; Rituximab (4F4X42SYQ6)
Language	English
Publishing date	2012-06-07
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	127294-9
ISSN	1529-0131 ; 0004-3591 ; 2326-5191
ISSN (online)	1529-0131
ISSN	0004-3591 ; 2326-5191
DOI	10.1002/art.34560
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Article: Malignant B-cell lymphoma of the Harder's gland in a rabbit.

Volopich, Sabine / Gruber, Andrea / Hassan, Jasmin / Hittmair, Katharina M / Schwendenwein, Ilse / Nell, Barbara

Veterinary ophthalmology

2005 Volume 8, Issue 4, Page(s) 259–263

Abstract: ... lymph nodes. The rabbit was euthanized and histopathology identified the retrobulbar mass as B-cell malignant ...

Abstract	A 22-month-old, female rabbit was presented with a 1-day history of acute unilateral exophthalmos. Ultrasonography and computed tomography (CT) of the orbit revealed an orbital mass. Retrobulbar lymphoma was diagnosed following fine-needle aspiration biopsy (FNAB). Thoracic radiographs were normal, and ultrasonography of the abdomen showed focal hypoechoic thickening of the bowel wall and hypoechoic enlarged lymph nodes. The rabbit was euthanized and histopathology identified the retrobulbar mass as B-cell malignant lymphoma of the Harder's gland. Mesenteric lymph nodes, caecum, and both kidneys were also affected. This is the first documented case of malignant lymphoma of the Harder's gland in a rabbit.
MeSH term(s)	Animals ; Biopsy, Fine-Needle/veterinary ; Diagnosis, Differential ; Female ; Harderian Gland ; Kidney Neoplasms/diagnosis ; Kidney Neoplasms/diagnostic imaging ; Kidney Neoplasms/secondary ; Lymphoma, B-Cell/diagnosis ; Lymphoma, B-Cell/diagnostic imaging ; Lymphoma, B-Cell/pathology ; Neoplasm Metastasis ; Orbital Neoplasms/diagnosis ; Orbital Neoplasms/diagnostic imaging ; Orbital Neoplasms/pathology ; Rabbits ; Tomography, X-Ray Computed/veterinary ; Ultrasonography
Language	English
Publishing date	2005-07
Publishing country	England
Document type	Case Reports ; Journal Article ; Review
ZDB-ID	2011043-1
ISSN	1463-5224 ; 1463-5216
ISSN (online)	1463-5224
ISSN	1463-5216
DOI	10.1111/j.1463-5224.2005.00400.x
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Article ; Online: Dynamics of Lewis b binding and sequence variation of the babA adhesin gene during chronic Helicobacter pylori infection in humans.

Nell, Sandra / Kennemann, Lynn / Schwarz, Sandra / Josenhans, Christine / Suerbaum, Sebastian

mBio

2014 Volume 5, Issue 6

Abstract: ... BabA, which mediates binding to the blood group antigen Lewis b [Le(b)]. To study the dynamics of Le(b ... from chronically infected individuals. A complete loss or significant reduction of Le(b) binding was observed ... in strains from 5 out of 23 individuals, indicating that the Le(b) binding phenotype is quite stable during ...

Abstract	Unlabelled: Helicobacter pylori undergoes rapid microevolution during chronic infection, but very little is known about how this affects host interaction factors. The best-studied adhesin of H. pylori is BabA, which mediates binding to the blood group antigen Lewis b [Le(b)]. To study the dynamics of Le(b) adherence during human infection, we analyzed paired H. pylori isolates obtained sequentially from chronically infected individuals. A complete loss or significant reduction of Le(b) binding was observed in strains from 5 out of 23 individuals, indicating that the Le(b) binding phenotype is quite stable during chronic human infection. Sequence comparisons of babA identified differences due to mutation and/or recombination in 12 out of 16 strain pairs analyzed. Most amino acid changes were found in the putative N-terminal extracellular adhesion domain. One strain pair that had changed from a Le(b) binding to a nonbinding phenotype was used to study the role of distinct sequence changes in Le(b) binding. By transformations of the nonbinding strain with a babA gene amplified from the binding strain, H. pylori strains with mosaic babA genes were generated. Recombinants were enriched for a gain of Le(b) binding by biopanning or for BabA expression on the bacterial surface by pulldown assay. With this approach, we identified several amino acid residues affecting the strength of Le(b) binding. Additionally, the data showed that the C terminus of BabA, which is predicted to encode an outer membrane β-barrel domain, plays an essential role in the biogenesis of this protein. Importance: Helicobacter pylori causes a chronic infection of the human stomach that can lead to ulcers and cancer. The bacterium can bind to gastric epithelial cells with specialized outer membrane proteins. The best-studied protein is the BabA adhesin which binds to the Lewis b blood group antigen. Since H. pylori is a bacterium with very high genetic variability, we asked whether babA evolves during chronic infection and how mutations or recombination in babA affect binding. We found that BabA-mediated adherence was stable in most individuals but observed a complete loss of binding or reduced binding in 22% of individuals. One strain pair in which binding was lost was used to generate babA sequences that were mosaics of a functional allele and a nonfunctional allele, and the mosaic sequences were used to identify amino acids critically involved in binding of BabA to Lewis b.
MeSH term(s)	Adhesins, Bacterial/genetics ; Adhesins, Bacterial/metabolism ; Bacterial Adhesion ; DNA, Bacterial/chemistry ; DNA, Bacterial/genetics ; Genetic Variation ; Helicobacter Infections/microbiology ; Helicobacter pylori/genetics ; Helicobacter pylori/isolation & purification ; Helicobacter pylori/metabolism ; Helicobacter pylori/physiology ; Humans ; Lewis Blood Group Antigens/metabolism ; Molecular Sequence Data ; Mutation, Missense ; Protein Binding ; Sequence Analysis, DNA ; Sequence Deletion
Chemical Substances	Adhesins, Bacterial ; BabA protein, Helicobacter pylori ; DNA, Bacterial ; Lewis Blood Group Antigens
Language	English
Publishing date	2014-12-16
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2557172-2
ISSN	2150-7511 ; 2161-2129
ISSN (online)	2150-7511
ISSN	2161-2129
DOI	10.1128/mBio.02281-14
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Article ; Online: Erratum for Nell et al., Dynamics of Lewis b Binding and Sequence Variation of the babA Adhesin Gene during Chronic Helicobacter pylori Infection in Humans.

Nell, Sandra / Kennemann, Lynn / Schwarz, Sandra / Josenhans, Christine / Suerbaum, Sebastian

mBio

2015 Volume 6, Issue 5, Page(s) e01233–15

Language	English
Publishing date	2015-09-15
Publishing country	United States
Document type	Published Erratum
ZDB-ID	2557172-2
ISSN	2150-7511 ; 2161-2129
ISSN (online)	2150-7511
ISSN	2161-2129
DOI	10.1128/mBio.01233-15
Database	MEDical Literature Analysis and Retrieval System OnLINE

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