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  1. Article ; Online: Proteomics-based strategies to identify proteins relevant to chronic lymphocytic leukemia.

    Alsagaby, Suliman A / Khanna, Sanjay / Hart, Keith W / Pratt, Guy / Fegan, Christopher / Pepper, Christopher / Brewis, Ian A / Brennan, Paul

    Journal of proteome research

    2014  Volume 13, Issue 11, Page(s) 5051–5062

    Abstract: Chronic lymphocytic leukemia (CLL), a malignant B-cell disorder, is characterized by a heterogeneous clinical course. Two-dimensional nano liquid chromatography (2D-nano-LC) coupled with matrix-assisted laser desorption/ionization time-of-flight tandem ... ...

    Abstract Chronic lymphocytic leukemia (CLL), a malignant B-cell disorder, is characterized by a heterogeneous clinical course. Two-dimensional nano liquid chromatography (2D-nano-LC) coupled with matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF/TOF MS) (LC-MALDI) was used to perform qualitative and quantitative analysis on cellular extracts from 12 primary CLL samples. We identified 728 proteins and quantified 655 proteins using isobaric tag-labeled extracts. Four strategies were used to identify disease-related proteins. First, we integrated our CLL proteome with published gene expression data of normal B-cells and CLL cells to highlight proteins with preferential expression in the transcriptome of CLL. Second, as CLL's outcome is heterogeneous, our quantitative proteomic data were used to indicate heterogeneously expressed proteins. Third, we used the quantitative data to identify proteins with differential abundance in poor prognosis CLL samples. Fourth, hierarchical cluster analysis was applied to identify hidden patterns of protein expression. These strategies identified 63 proteins, and 4 were investigated in a CLL cohort (39 patients). Thyroid hormone receptor-associated protein 3, T-cell leukemia/lymphoma protein 1A, and S100A8 were associated with high-risk CLL. Myosin-9 exhibited reduced expression in CLL samples from high-risk patients. This study shows the usefulness of proteomic approaches, combined with transcriptomics, to identify disease-related proteins.
    MeSH term(s) Aged ; Aged, 80 and over ; Biomarkers, Tumor/analysis ; Biomarkers, Tumor/blood ; Calgranulin A/metabolism ; Cluster Analysis ; Follow-Up Studies ; Humans ; Kaplan-Meier Estimate ; Leukemia, Lymphocytic, Chronic, B-Cell/blood ; Leukemia, Lymphocytic, Chronic, B-Cell/mortality ; Leukemia, Lymphocytic, Chronic, B-Cell/pathology ; Leukemia, Lymphocytic, Chronic, B-Cell/therapy ; Leukemia, T-Cell/metabolism ; Microarray Analysis ; Middle Aged ; Molecular Motor Proteins/metabolism ; Myosin Heavy Chains/metabolism ; Prognosis ; Proteins/analysis ; Proteins/metabolism ; Proteomics/methods ; Reproducibility of Results ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods ; Tandem Mass Spectrometry/methods
    Chemical Substances Biomarkers, Tumor ; Calgranulin A ; MYH9 protein, human ; Molecular Motor Proteins ; Proteins ; Myosin Heavy Chains (EC 3.6.4.1)
    Language English
    Publishing date 2014-11-07
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2078618-9
    ISSN 1535-3907 ; 1535-3893
    ISSN (online) 1535-3907
    ISSN 1535-3893
    DOI 10.1021/pr5002803
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article: Proteomics-Based Strategies To Identify Proteins Relevant to Chronic Lymphocytic Leukemia

    Alsagaby, Suliman A / Brennan Paul / Brewis Ian A / Fegan Christopher / Hart Keith W / Khanna Sanjay / Pepper Christopher / Pratt Guy

    Journal of Proteome Research. 2014 Nov. 07, v. 13, no. 11

    2014  

    Abstract: Chronic lymphocytic leukemia (CLL), a malignant B-cell disorder, is characterized by a heterogeneous clinical course. Two-dimensional nano liquid chromatography (2D-nano–LC) coupled with matrix-assisted laser desorption/ionization time-of-flight tandem ...

    Abstract Chronic lymphocytic leukemia (CLL), a malignant B-cell disorder, is characterized by a heterogeneous clinical course. Two-dimensional nano liquid chromatography (2D-nano–LC) coupled with matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF/TOF MS) (LC–MALDI) was used to perform qualitative and quantitative analysis on cellular extracts from 12 primary CLL samples. We identified 728 proteins and quantified 655 proteins using isobaric tag-labeled extracts. Four strategies were used to identify disease-related proteins. First, we integrated our CLL proteome with published gene expression data of normal B-cells and CLL cells to highlight proteins with preferential expression in the transcriptome of CLL. Second, as CLL’s outcome is heterogeneous, our quantitative proteomic data were used to indicate heterogeneously expressed proteins. Third, we used the quantitative data to identify proteins with differential abundance in poor prognosis CLL samples. Fourth, hierarchical cluster analysis was applied to identify hidden patterns of protein expression. These strategies identified 63 proteins, and 4 were investigated in a CLL cohort (39 patients). Thyroid hormone receptor-associated protein 3, T-cell leukemia/lymphoma protein 1A, and S100A8 were associated with high-risk CLL. Myosin-9 exhibited reduced expression in CLL samples from high-risk patients. This study shows the usefulness of proteomic approaches, combined with transcriptomics, to identify disease-related proteins.
    Keywords B-lymphocytes ; disease course ; gene expression ; liquid chromatography ; lymphocytic leukemia ; lymphoma ; patients ; prognosis ; protein synthesis ; proteins ; proteome ; proteomics ; quantitative analysis ; tandem mass spectrometry ; thyroid hormones ; T-lymphocytes ; transcriptome ; transcriptomics
    Language English
    Dates of publication 2014-1107
    Size p. 5051-5062.
    Publishing place American Chemical Society
    Document type Article
    ZDB-ID 2078618-9
    ISSN 1535-3907 ; 1535-3893
    ISSN (online) 1535-3907
    ISSN 1535-3893
    DOI 10.1021%2Fpr5002803
    Database NAL-Catalogue (AGRICOLA)

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  3. Article ; Online: Characterization of platelet aminophospholipid externalization reveals fatty acids as molecular determinants that regulate coagulation.

    Clark, Stephen R / Thomas, Christopher P / Hammond, Victoria J / Aldrovandi, Maceler / Wilkinson, Gavin W / Hart, Keith W / Murphy, Robert C / Collins, Peter W / O'Donnell, Valerie B

    Proceedings of the National Academy of Sciences of the United States of America

    2013  Volume 110, Issue 15, Page(s) 5875–5880

    Abstract: Aminophospholipid (APL) trafficking across the plasma membrane is a key event in cell activation, apoptosis, and aging and is required for clearance of dying cells and coagulation. Currently the phospholipid molecular species externalized are unknown. ... ...

    Abstract Aminophospholipid (APL) trafficking across the plasma membrane is a key event in cell activation, apoptosis, and aging and is required for clearance of dying cells and coagulation. Currently the phospholipid molecular species externalized are unknown. Using a lipidomic method, we show that thrombin, collagen, or ionophore-activated human platelets externalize two phosphatidylserines (PSs) and five phosphatidylethanolamines (PEs). Four percent of the total cellular PE/PS pool (∼300 ng/2 × 10(8) cells, thrombin), is externalized via calcium mobilization and protease-activated receptors-1 and -4, and 48% is contained in microparticles. Apoptosis and energy depletion (aging) externalized the same APLs in a calcium-dependent manner, and all stimuli externalized oxidized phospholipids, termed hydroxyeicosatetraenoic acid-PEs. Transmembrane protein-16F (TMEM-16F), the protein mutated in Scott syndrome, was required for PE/PS externalization during thrombin activation and energy depletion, but not apoptosis. Platelet-specific APLs optimally supported tissue factor-dependent coagulation in human plasma, vs. APL with longer or shorter fatty acyl chains. This finding demonstrates fatty acids as molecular determinants of APL that regulate hemostasis. Thus, the molecular species of externalized APL during platelet activation, apoptosis, and energy depletion were characterized, and their ability to support coagulation revealed. The findings have therapeutic implications for bleeding disorders and transfusion therapy. The assay could be applied to other cell events characterized by APL externalization, including cell division and vesiculation.
    MeSH term(s) Aging ; Annexin A5/chemistry ; Apoptosis ; Biotinylation ; Blood Coagulation ; Blood Platelets/metabolism ; Calcium/metabolism ; Cell Membrane/metabolism ; Dose-Response Relationship, Drug ; Fatty Acids/chemistry ; Gene Expression Regulation ; Humans ; Phospholipids/chemistry ; Thrombin/chemistry ; Thrombin/metabolism ; Time Factors
    Chemical Substances Annexin A5 ; Fatty Acids ; Phospholipids ; Thrombin (EC 3.4.21.5) ; Calcium (SY7Q814VUP)
    Language English
    Publishing date 2013-03-25
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    DOI 10.1073/pnas.1222419110
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    Zs.A 1148: Show issues Location:
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  4. Article: Analysis of CD4(+) T-cell responses to human papillomavirus (HPV) type 11 L1 in healthy adults reveals a high degree of responsiveness and cross-reactivity with other HPV types.

    Williams, O Martin / Hart, Keith W / Wang, Eddie C Y / Gelder, Colin M

    Journal of virology

    2002  Volume 76, Issue 15, Page(s) 7418–7429

    Abstract: Human papillomavirus type 11 (HPV-11) infection causes genital warts and recurrent respiratory papillomatosis. While there is compelling evidence that CD4(+) T cells play an important role in immune surveillance of HPV-associated diseases, little is ... ...

    Abstract Human papillomavirus type 11 (HPV-11) infection causes genital warts and recurrent respiratory papillomatosis. While there is compelling evidence that CD4(+) T cells play an important role in immune surveillance of HPV-associated diseases, little is known about human CD4(+) T-cell recognition of HPV-11. We have investigated the CD4(+) T-cell responses of 25 unrelated healthy donors to HPV-11 L1 virus-like particles (VLP). CD4(+) T-cell lines from 21 of 25 donors were established. Cell sorting experiments carried out on cells from six donors demonstrated that the response was located in the CD45RA(low) CD45RO(high) memory T-cell population. To determine the peptide specificity of these responses, epitope selection was analyzed by using 95 15-mer peptides spanning the entire HPV-11 L1 protein. No single region of L1 was immunodominant; responders recognized between 1 and 10 peptides, located throughout the protein, and peptide responses fell into clear HLA class II restricted patterns. Panels of L1 peptides specific for skin and genital HPV were used to show that the L1 CD4(+) T-cell responses were cross-reactive. The degree of cross-reactivity was inversely related to the degree of L1 sequence diversity between these viruses. Finally, responses to HPV-11 L1 peptides were elicited from ex vivo CD45RO(+) peripheral blood mononuclear cells, demonstrating that recognition of HPV-11 was a specific memory response and not due to in vitro selection during tissue culture. This is the first study of CD4(+) T-cell responses to HPV-11 in healthy subjects and demonstrates marked cross-reactivity with other skin and genital HPV types. This cross-reactivity may be of significance for vaccine strategies against HPV-associated clinical diseases.
    MeSH term(s) Amino Acid Sequence ; Blood Donors ; CD4-Positive T-Lymphocytes/immunology ; Capsid Proteins ; Cells, Cultured ; Cross Reactions ; Epitope Mapping ; Epitopes, T-Lymphocyte/chemistry ; Epitopes, T-Lymphocyte/immunology ; Humans ; Immunologic Memory ; Lymphocyte Activation ; Molecular Sequence Data ; Oncogene Proteins, Viral/immunology ; Papillomaviridae/classification ; Papillomaviridae/immunology ; Peptides/chemical synthesis ; Peptides/chemistry ; Peptides/immunology
    Chemical Substances Capsid Proteins ; Epitopes, T-Lymphocyte ; L1 protein, Human papillomavirus type 11 ; Oncogene Proteins, Viral ; Peptides
    Language English
    Publishing date 2002-08
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 80174-4
    ISSN 1098-5514 ; 0022-538X
    ISSN (online) 1098-5514
    ISSN 0022-538X
    DOI 10.1128/jvi.76.15.7418-7429.2002
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
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  5. Article: Associations between human leukocyte antigens and nonresponsiveness to influenza vaccine.

    Gelder, Colin M / Lambkin, Rob / Hart, Keith W / Fleming, Douglas / Williams, O Martin / Bunce, Mike / Welsh, Kenneth I / Marshall, Sara E / Oxford, John

    The Journal of infectious diseases

    2002  Volume 185, Issue 1, Page(s) 114–117

    Abstract: Influenza remains a major cause of morbidity and mortality, particularly in at-risk groups where vaccination reduces complications of infection but is not universally protective. In order to determine whether human leukocyte antigen (HLA) class II ... ...

    Abstract Influenza remains a major cause of morbidity and mortality, particularly in at-risk groups where vaccination reduces complications of infection but is not universally protective. In order to determine whether human leukocyte antigen (HLA) class II polymorphisms modulate anti-influenza antibody responses to vaccination, a cohort of HLA-typed at-risk donors was investigated. The subjects were recruited from a single urban family practice. Hemagglutination-inhibition (HAI) titers were measured immediately before and 28 days after subunit vaccination. Nonresponsiveness was defined as failure to mount an HAI response to any component of the trivalent influenza vaccine. When the nonresponders and responders with HLA class II were compared, the nonresponder group had more HLA-DRB1*07-positive donors (13/32 vs. 6/41 responders; P=.016, Fisher's exact test) and fewer HLA-DQB1*0603-9/14-positive donors (2/32 vs. 14/41 responders; P=.0045). Thus, polymorphisms in HLA class II molecules appear to modulate antibody responses to influenza vaccination.
    MeSH term(s) Adult ; Aged ; Antibodies, Viral/blood ; Antibody Formation ; Genes, MHC Class II/genetics ; HLA-D Antigens ; Histocompatibility Testing ; Humans ; Influenza Vaccines/immunology ; Influenza, Human/prevention & control ; Middle Aged ; Polymorphism, Genetic ; Vaccination
    Chemical Substances Antibodies, Viral ; HLA-D Antigens ; Influenza Vaccines
    Language English
    Publishing date 2002-01-01
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 3019-3
    ISSN 1537-6613 ; 0022-1899
    ISSN (online) 1537-6613
    ISSN 0022-1899
    DOI 10.1086/338014
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    Uh II Zs.50: Show issues Location:
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    Zs.MO 445: Show issues

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  6. Article: HLA class II polymorphisms and susceptibility to recurrent respiratory papillomatosis.

    Gelder, Colin M / Williams, O Martin / Hart, Keith W / Wall, Siôn / Williams, Gareth / Ingrams, Duncan / Bull, Peter / Bunce, Mike / Welsh, Ken / Marshall, Sara E F / Borysiewicz, Leszek

    Journal of virology

    2001  Volume 77, Issue 3, Page(s) 1927–1939

    Abstract: Recurrent respiratory papillomatosis (RRP) is characterised by multiple laryngeal papillomas. Left untreated, the lesions enlarge, spread, and endanger the airway. Medical treatments are unsatisfactory, and repeated surgery remains the mainstay of ... ...

    Abstract Recurrent respiratory papillomatosis (RRP) is characterised by multiple laryngeal papillomas. Left untreated, the lesions enlarge, spread, and endanger the airway. Medical treatments are unsatisfactory, and repeated surgery remains the mainstay of therapy. RRP is caused by human papillomavirus (HPV) infection. However, since oral HPV infection is common and RRP is rare, other host and/or viral factors may contribute to pathogenesis. In an attempt to identify such factors, we have investigated 60 patients. The patients were HLA class I, II, and tumor necrosis factor TNF typed by sequence-specific primer PCR, and the results compared to those for 554 healthy controls by using Fisher's exact test. Peripheral blood mononuclear cell proliferative responses of 25 controls and 10 patients to HPV-11 L1 virus-like particles (VLP) were compared. Short-term VLP-specific T-cell lines were established, and recognition of L1 was analyzed. Finally, the L1 open reading frames of HPV isolates from four patients were sequenced. Susceptibility to RRP was associated with HLA DRB1*0301 (33 of 60 patients versus 136 of 554 controls, P < 0.0001). The three most severely affected patients were homozygous for this allele. A range of T-cell proliferative responses to HPV-11 VLP were observed in DRB1*0301-positive healthy donors which were comparable to those in DRB1*0301-negative controls. Individuals with juvenile-onset RRP also mounted a range of VLP responses, and their magnitude was negatively correlated with the clinical staging score (P = 0.012 by the Spearman rank correlation). DRB1*0301-positive patients who responded to L1 recognized the same epitope as did matched controls and produced similar cytokines. Sequencing of clinical isolates excluded the possibility that nonresponsiveness was the result of mutation(s) in L1.
    MeSH term(s) Adolescent ; Adult ; Capsid Proteins ; Child ; Child, Preschool ; Female ; Genes, MHC Class II ; Genetic Predisposition to Disease ; Genotype ; Humans ; Infant ; Laryngeal Neoplasms/genetics ; Laryngeal Neoplasms/immunology ; Male ; Middle Aged ; Neoplasm Recurrence, Local/genetics ; Neoplasm Recurrence, Local/immunology ; Oncogene Proteins, Viral/immunology ; Papilloma/genetics ; Papilloma/immunology ; Papillomaviridae/immunology ; Papillomavirus Infections/genetics ; Papillomavirus Infections/immunology ; Polymorphism, Genetic ; Tumor Virus Infections/genetics ; Tumor Virus Infections/immunology ; Virion/immunology
    Chemical Substances Capsid Proteins ; L1 protein, Human papillomavirus type 11 ; Oncogene Proteins, Viral
    Language English
    Publishing date 2001-09-18
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 80174-4
    ISSN 1098-5514 ; 0022-538X
    ISSN (online) 1098-5514
    ISSN 0022-538X
    DOI 10.1128/jvi.77.3.1927-1939.2003
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    Zs.A 609: Show issues Location:
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