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  • 1
    Language: English
    In: Blood, 27 January 2011, Vol.117(4), pp.1102-3
    Description: In this issue of Blood, Roger and colleagues present data on the magnitude of influence that broad-spectrum HDAC inhibitors exert on TLR-driven immune responses, thus demonstrating that HDAC inhibitors are immunosuppressive drugs.
    Keywords: Medicine ; Biology ; Chemistry ; Anatomy & Physiology;
    ISSN: 00064971
    E-ISSN: 1528-0020
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  • 2
    Language: English
    In: FASEB journal : official publication of the Federation of American Societies for Experimental Biology, March 2011, Vol.25(3), pp.863-74
    Description: Suppressor of cytokine signaling (SOCS) proteins are inhibitors of cytoplasmic Janus kinases (Jak) and signal transducer and activator of transcription (STAT) signaling pathways. Previously the authors surprisingly observed that SOCS1 translocated into the nucleus, which was because of the presence of a nuclear localization sequence. This report now hypothesizes that SOCS1 mediates specific functions within the nuclear compartment because it is instantly transported into the nucleus, as shown by photoactivation and live cell imaging in human HEK293 cells. The NFκB component p65 is identified as an interaction partner for SOCS1 but not for other members of the SOCS family. SOCS1 bound to p65 only within the nucleus. By means of its SOCS box domain, SOCS1 operated as a ubiquitin ligase, leading to polyubiquitination and proteasomal degradation of nuclear p65. Thus, SOCS1 limited prolonged p65 signaling and terminated expression of NFκB inducible genes. Using mutants that lack either nuclear translocation or a functional SOCS box, this report identifies genes that are regulated in a manner dependent on the nuclear availability of SOCS1. Data show that beyond its receptor-proximal function in Jak/STAT signaling, SOCS1 also regulates the duration of NFκB signaling within the cell nucleus, thus exerting a heretofore unrecognized function.
    Keywords: Suppressor of Cytokine Signaling Proteins ; Gene Expression Regulation -- Physiology ; Signal Transduction -- Physiology ; Transcription Factor Rela -- Metabolism
    ISSN: 08926638
    E-ISSN: 1530-6860
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  • 3
    Language: English
    In: Journal of clinical microbiology, June 2013, Vol.51(6), pp.1906-8
    Description: We evaluated the fully automated molecular BD MAX Cdiff assay (BD Diagnostics) and the Xpert C. difficile test (Cepheid) for rapid detection of Clostridium difficile infection. Culture was done on chromogenic agar followed by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry identification and toxin detection. Repeat testing was required for 1.8% and 6.0% of the BD MAX and Xpert tests, respectively. Sensitivities, specificities, positive predictive values (PPV), and negative predictive values (NPV) were 90.5%, 97.9%, 89.3%, and 98.1%, respectively, for BD MAX and 97.3%, 97.9%, 90.0%, and 99.5%, respectively, for Xpert.
    Keywords: Bacteriological Techniques -- Methods ; Clostridium Infections -- Diagnosis ; Clostridium Difficile -- Isolation & Purification ; Feces -- Microbiology ; Molecular Diagnostic Techniques -- Methods
    E-ISSN: 1098-660X
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  • 4
    Language: English
    In: Journal of clinical microbiology, July 2013, Vol.51(7), pp.2337-43
    Description: Pneumocystis jirovecii is an opportunistic pathogen in immunocompromised and AIDS patients. Detection by quantitative PCR is faster and more sensitive than microscopic diagnosis yet requires specific infrastructure. We adapted a real-time PCR amplifying the major surface glycoprotein (MSG) target from Pneumocystis jirovecii for use on the new BD MAX platform. The assay allowed fully automated DNA extraction and multiplex real-time PCR. The BD MAX assay was evaluated against manual DNA extraction and conventional real-time PCR. The BD MAX was used in the research mode running a multiplex PCR (MSG, internal control, and sample process control). The assay had a detection limit of 10 copies of an MSG-encoding plasmid per PCR that equated to 500 copies/ml in respiratory specimens. We observed accurate quantification of MSG targets over a 7- to 8-log range. Prealiquoting and sealing of the complete PCR reagents in conical tubes allowed easy and convenient handling of the BD MAX PCR. In a retrospective analysis of 54 positive samples, the BD MAX assay showed good quantitative correlation with the reference PCR method (R(2) = 0.82). Cross-contamination was not observed. Prospectively, 278 respiratory samples were analyzed by both molecular assays. The positivity rate overall was 18.3%. The BD MAX assay identified 46 positive samples, compared to 40 by the reference PCR. The BD MAX assay required liquefaction of highly viscous samples with dithiothreitol as the only manual step, thus offering advantages for timely availability of molecular-based detection assays.
    Keywords: Automation, Laboratory -- Methods ; Microbiological Techniques -- Methods ; Molecular Diagnostic Techniques -- Methods ; Pneumocystis Infections -- Diagnosis ; Pneumocystis Carinii -- Isolation & Purification ; Real-Time Polymerase Chain Reaction -- Methods
    E-ISSN: 1098-660X
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  • 5
    Language: English
    In: Journal of clinical microbiology, October 2012, Vol.50(10), pp.3365-7
    Description: We evaluated the new, fully automated molecular BD Max methicillin-resistant Staphylococcus aureus (MRSA) assay for detection of methicillin-resistant S. aureus in a low-prevalence (4.1%) setting. Sensitivity, specificity, and positive and negative predictive values were 93.9%, 99.2%, 83.8%, and 99.7%, respectively. The assay reported fewer unresolved results than the BD GeneOhm MRSA ACP assay.
    Keywords: Methicillin-Resistant Staphylococcus Aureus -- Isolation & Purification ; Molecular Diagnostic Techniques -- Methods ; Staphylococcal Infections -- Microbiology
    E-ISSN: 1098-660X
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  • 6
    Language: English
    In: Journal of clinical microbiology, December 2014, Vol.52(12), pp.4343-6
    Description: We evaluated the performance of the BD Max MRSA XT assay for use with different swab types. The 90% detection rates (95% confidence intervals) were 387 (97 to 1,551), 877 (238 to 3,230), 986 (183 to 5,287), 1,292 (328 to 5,078), 2,400 (426 to 13,518), and 5,848 (622 to 55,021) CFU/swab for Liquid Stuart, Liquid Amies, dry, Amies Gel without charcoal, ESwab collection, and Amies gel with charcoal swabs (Becton Dickinson), respectively. Amies Gel without charcoal, ESwab collection, and Amies gel with charcoal swabs had a tendency to be less sensitive, but none of the differences was statistically significant.
    Keywords: Carrier State -- Diagnosis ; Methicillin-Resistant Staphylococcus Aureus -- Isolation & Purification ; Molecular Diagnostic Techniques -- Methods ; Specimen Handling -- Methods ; Staphylococcal Infections -- Diagnosis
    ISSN: 00951137
    E-ISSN: 1098-660X
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  • 7
    Language: English
    In: Journal of Clinical Microbiology, 05/01/2014, Vol.52(5), pp.1809-1809
    Description: Volume 51, no. 7, p. 2337–2343, 2013. Page 2338, column 1, lines 2–3: The sample process probe that was used and published for the study, SPC probe Cy5-TTG ATG CCT CTT ACA TTG CTC CAC CTT TCC T-BHQ2, contains an error. The correct sequence, which fully fits the sample process control plasmid, contains one additional nucleotide and is as follows: Cy5-TTG ATG CCT CTT CAC ATT GCT CCA CCT TTC CT-BHQ2.
    Keywords: Medicine ; Biology;
    ISSN: 0095-1137
    E-ISSN: 1098660X
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  • 8
    Language: English
    In: The Journal of biological chemistry, 23 March 2012, Vol.287(13), pp.9923-30
    Description: Suppressor of cytokine signaling (SOCS)3 belongs to a family of proteins that are known to exert important functions as inducible feedback inhibitors and are crucial for the balance of immune responses. There is evidence for a deregulated immune response in chronic inflammatory skin diseases. Thus, it was the aim of this study to investigate the regulation of SOCS proteins involved in intracellular signaling pathways occurring during inflammatory skin diseases and analyze their impact on the course of inflammatory responses. Because we and others have previously described that the cytokine IL-27 has an important impact on the chronic manifestation of inflammatory skin diseases, we focused here on the signaling induced by IL-27 in human primary keratinocytes compared with autologous blood-derived macrophages. Here, we demonstrate that SOCS3 is critically involved in regulating the cell-specific response to IL-27. SOCS3 was found to be significantly up-regulated by IL-27 in macrophages but not in keratinocytes. Other STAT3-activating cytokines investigated, including IL-6, IL-22, and oncostatin M, also failed to up-regulate SOCS3 in keratinocytes. Lack of SOCS3 up-regulation in skin epithelial cells was accompanied by prolonged STAT1 and STAT3 phosphorylation and enhanced CXCL10 production upon IL-27 stimulation compared with macrophages. Overexpression of SOCS3 in keratinocytes significantly diminished this enhanced CXCL10 production in response to IL-27. We conclude from our data that keratinocytes have a cell type-specific impaired capacity to up-regulate SOCS3 which may crucially determine the course of chronic inflammatory skin diseases.
    Keywords: Keratinocytes -- Metabolism ; Macrophages -- Metabolism ; Suppressor of Cytokine Signaling Proteins -- Biosynthesis
    E-ISSN: 1083-351X
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  • 9
    Language: English
    In: The Journal of biological chemistry, 2011, Vol.286(31), pp.27278-27287
    Description: RIG-I is a major innate immune sensor for viral infection, triggering an interferon (IFN)-mediated antiviral response upon cytosolic detection of viral RNA. Double-strandedness and 5'-terminal triphosphates were identified as motifs required to elicit optimal immunological signaling. However, very little is known about the response dynamics of the RIG-I pathway, which is crucial for the ability of the cell to react to diverse classes of viral RNA while maintaining self-tolerance. In the present study, we addressed the molecular mechanism of RIG-I signal detection and its translation into pathway activation. By employing highly quantitative methods, we could establish the length of the double-stranded RNA (dsRNA) to be the most critical determinant of response strength. Size exclusion chromatography and direct visualization in scanning force microscopy suggested that this was due to cooperative oligomerization of RIG-I along dsRNA. The initiation efficiency of this oligomerization process critically depended on the presence of high affinity motifs, like a 5'-triphosphate. It is noteworthy that for dsRNA longer than 200 bp, internal initiation could effectively compensate for a lack of terminal triphosphates. In summary, our data demonstrate a very flexible response behavior of the RIG-I pathway, in which sensing and integration of at least two distinct signals, initiation efficiency and double strand length, allow the host cell to mount an antiviral response that is tightly adjusted to the type of the detected signal, such as viral genomes, replication intermediates, or small by-products.
    Keywords: Immunity, Innate ; Dead-Box RNA Helicases -- Physiology;
    ISSN: 1083-351X
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  • 10
    Language: English
    In: Neuroscience Letters, 03 March 2011, Vol.490(3), pp.170-174
    Description: ▶ Intracerebral hemorrhage induces a profound reduction of systemic immune cells. ▶ Despite this cellular immunosupression, T trend to be constant in number. ▶ After large hematomas animals are susceptible to infections. Inflammatory cascades are increasingly recognized as an important pathophysiological mechanism in intracerebral hemorrhage (ICH). In contrast, the effect of ICH on the systemic immune system has barely been investigated. We examined the effects of different hematoma volumes on immune cell subpopulations in experimental murine ICH. In C57BL/6 mice, ICH was induced by striatal injection of autologous blood (10, 30 or 50 μL). Control animals received the respective sham operation. Three days after ICH induction, differential blood leukocyte counting was performed. Lymphocyte subpopulations were further characterized by flow cytometry in blood, spleen, lymph node and thymus. Infectious complications were studied using microbiological cultures of blood and lungs. Only after large ICH a marked decrease of leukocyte counts and most lymphocyte subsets was observed in all organs. Despite this general leukocytopenia, a significant, up to 10-fold increase, was detected in the monocyte population after extensive hemorrhage. After moderate ICH induction, only specific lymphocyte subpopulations were differentially affected. Mature thymic cells were unaffected while immature CD4+CD8+ cells were depleted by over 90% after large ICH. A significant proportion of mice with extensive ICH (36.4%) developed spontaneous pneumonia and/or bacteremia while none of the sham operated mice had infectious complications. The ICH size determines the extent of systemic immunomodulation. Large ICH predisposes animals to infections.
    Keywords: Intracerebral Hemorrhage ; Post Stroke Immunosupression ; Experimental Stroke ; Medicine ; Anatomy & Physiology
    ISSN: 0304-3940
    E-ISSN: 1872-7972
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