Junctional adhesion molecule C mediates leukocyte adhesion to rheumatoid arthritis synovium

Objective Leukocyte infiltration into the rheumatoid arthritis (RA) synovium is a multistep process in which leukocytes leave the bloodstream and invade the synovial tissue (ST). Leukocyte transendothelial migration and adhesion to RA ST requires adhesion molecules on the surface of endothelial cells and RA ST fibroblasts. This study was undertaken to investigate the role of junctional adhesion molecule C (JAM-C) in mediating leukocyte recruitment and retention in the RA joint. Methods Immunohistologic analysis was performed on RA, osteoarthritis (OA), and normal ST samples to quantify JAM-C expression. Fibroblast JAM-C expression was also analyzed using Western blotting, cell surface enzyme-linked immunosorbent assay, and immunofluorescence. To determine the role of JAM-C in leukocyte retention in the RA synovium, in vitro and in situ adhesion assays and RA ST fibroblast transmigration assays were performed. Results JAM-C was highly expressed by RA ST lining cells, and its expression was increased in OA ST and RA ST endothelial cells compared with normal ST endothelial cells. JAM-C was also expressed on the surface of OA ST and RA ST fibroblasts. Furthermore, we demonstrated that myeloid U937 cell adhesion to both OA ST and RA ST fibroblasts and to RA ST was dependent on JAM-C. U937 cell migration through an RA ST fibroblast monolayer was enhanced in the presence of neutralizing antibodies against JAM-C. Conclusion Our results highlight the novel role of JAM-C in recruiting and retaining leukocytes in the RA synovium and suggest that targeting JAM-C may be important in combating inflammatory diseases such as RA.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/61229/1/23867_ftp.pd

bcl-2 expression is not associated with survival in metastatic cutaneous melanoma: A historical cohort study

<p>Abstract</p> <p>Background</p> <p>Programmed cell death (apoptosis) has been implicated in tumor development and may affect the metastatic potential of tumor cells. The role of bcl-2, a proto-oncogene that inhibits apoptosis, has been studied in several malignancies, including cutaneous melanoma (CM). The purpose of this study was to evaluate the immunohistochemical expression of bcl-2 in 35 regional lymph node, 28 subcutaneous and 17 visceral CM metastases, correlating the findings with patient survival.</p> <p>Methods</p> <p>In a historical cohort study patient survival was correlated with the expression of bcl-2 in regional lymph node, subcutaneous and visceral metastases of CM. Eighty slides containing surgical specimens from 50 patients diagnosed with stage III and IV CM, 28 male (56%) and 22 female (44%), were analyzed. Mean age at diagnosis was 43 years (16–74 years; median = 42 years). Mean Breslow depth was 5.01 mm (0.4–27.5 mm). The slides were submitted to immunohistochemical reaction using anti-bcl-2 monoclonal antibody and classified according to the degree of staining (< 5%; 5 to 50%; or > 50% of tumor cells stained). The relationship between bcl-2 protein expression and survival for each type of metastasis, gender and age at initial diagnosis was analyzed.</p> <p>Results</p> <p>Mean overall survival was 33.9 months after the diagnosis of the initial metastatic lesion (range: 0 to 131 months). Twenty-four out of 50 patients (48%) had died from CM by the end of the study period. bcl-2 expression was detected in 74.3, 85.7 and 82.4% of lymph node, subcutaneous and visceral metastases, respectively. After univariate and multivariate analyses, no correlation was found between positive bcl-2 expression and overall survival for the types of metastases evaluated.</p> <p>Conclusion</p> <p>The immunohistochemical expression of bcl-2 in metastasis alone is not a prognostic marker for CM.</p

Adaptation and Preadaptation of Salmonella enterica to Bile

Bile possesses antibacterial activity because bile salts disrupt membranes, denature proteins, and damage DNA. This study describes mechanisms employed by the bacterium Salmonella enterica to survive bile. Sublethal concentrations of the bile salt sodium deoxycholate (DOC) adapt Salmonella to survive lethal concentrations of bile. Adaptation seems to be associated to multiple changes in gene expression, which include upregulation of the RpoS-dependent general stress response and other stress responses. The crucial role of the general stress response in adaptation to bile is supported by the observation that RpoS− mutants are bile-sensitive. While adaptation to bile involves a response by the bacterial population, individual cells can become bile-resistant without adaptation: plating of a non-adapted S. enterica culture on medium containing a lethal concentration of bile yields bile-resistant colonies at frequencies between 10−6 and 10−7 per cell and generation. Fluctuation analysis indicates that such colonies derive from bile-resistant cells present in the previous culture. A fraction of such isolates are stable, indicating that bile resistance can be acquired by mutation. Full genome sequencing of bile-resistant mutants shows that alteration of the lipopolysaccharide transport machinery is a frequent cause of mutational bile resistance. However, selection on lethal concentrations of bile also provides bile-resistant isolates that are not mutants. We propose that such isolates derive from rare cells whose physiological state permitted survival upon encountering bile. This view is supported by single cell analysis of gene expression using a microscope fluidic system: batch cultures of Salmonella contain cells that activate stress response genes in the absence of DOC. This phenomenon underscores the existence of phenotypic heterogeneity in clonal populations of bacteria and may illustrate the adaptive value of gene expression fluctuations

Responsiveness of human T lymphocytes to bacterial superantigens presented by cultured rheumatoid arthritis synoviocytes

Objective . Type B fibroblastic synoviocytes are abundant in inflamed joints of patients with rheumatoid arthritis (RA), and can secrete cytokines and other mediators of inflammation. The aim of this study was to determine whether cell lines derived from RA type B synoviocytes could also serve as accessory cells for T lymphocyte activation. Methods . Cells from RA synoviocyte lines, with or without preculture in interferon-Γ (IFNΓ), were cultured with purified peripheral blood T cells, in the presence or absence of superantigens or other accessory cell–dependent T cell mitogens. T cell proliferation was measured by thymidine incorporation, and synoviocyte surface markers were analyzed by flow cytometry. Results . RA type B synoviocyte lines were potent accessory cells for T cell responses to bacterial superantigens or lectins, and direct cell-cell contact was required. Preculture in IFNΓ augmented synoviocyte expression of major histocompatibility complex (MHC) class II molecules and of ligands for some T cell costimulatory receptors, but synoviocyte accessory cell function was evident even in the absence of IFNΓ. Blocking studies using monoclonal antibodies supported the notion of a role for CD2, CD11a/CD18 and MHC class II molecules in synoviocyte-dependent T cell activation. Monoclonal antibodies against IFNΓ, interleukin-1Β (IL-1Β), IL-6, IL-8, and tumor necrosis factor Α failed to block the T cell proliferative responses, but anti–IL-2 was strongly inhibitory. Conclusion . Cultured RA type B synoviocytes can perform some of the functions of professional antigen-presenting cells. If such cells have similar properties in vivo, they may be important participants in activation of immune responses, in addition to their previously described synthetic and proinflammatory roles. If RA synovial tissue T cells, like normal peripheral blood T cells, can respond to superantigens presented by synoviocytes, this interaction could be important in the pathogenesis of RA.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/37807/1/1780390117_ftp.pd

The VCAM-1 gene that encodes the vascular cell adhesion molecule is a target of the Sry-related high mobility group box gene, Sox18

VCAM-1 (vascular cell adhesion molecule-1) and Sox18 are involved in vascular development. VCAM-1 is an important adhesion molecule that is expressed on endothelial cells and has a critical role in endothelial activation, inflammation, lymphatic pathophysiology, and atherogenesis. The Sry-related high mobility group box factor Sox18 has previously been implicated in endothelial pathologies. Mutations in human and mouse Sox18 leads to hypotrichosis and lymphedema. Furthermore, both Sox18 and VCAM-1 have very similar spatio-temporal patterns of expression, which is suggestive of crosstalk. We use biochemical techniques, cell culture systems, and the ragged opossum (RaOP) mouse model with a naturally occurring mutation in Sox18 to demonstrate that VCAM-1 is an important target of Sox18. Transfection, site-specific mutagenesis, and gel shift analyses demonstrated that Sox18 directly targeted and trans-activated VCAM-1 expression. Importantly, the naturally occurring Sox18 mutant attenuates the expression and activation of VCAM-1 in vitro. Furthermore, in vivo quantitation of VCAM-1 mRNA levels in wild type and RaOP mice demonstrates that RaOP animals show a dramatic and significant reduction in VCAM-1 mRNA expression in lung, skin, and skeletal muscle. Our observation that the VCAM-1 gene is an important target of SOX18 provides the first molecular insights into the vascular abnormalities in the mouse mutant ragged and the human hypotrichosis-lymphedematelangiectasia disorder

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No abstract.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/34298/1/210_ftp.pd

Seismic and aseismic slip during the 2006 Copiapó swarm in North-Central Chile

International audienceEarthquake swarms commonly occur along the Chilean subduction zone, witnessing fast seismic and slow aseismic slip behavior at the plate interface. However, the largest seismic swarms observed in Chile, particularly in the Copiapó-Atacama region, remain poorly documented, and the underlying processes have yet to be understood. Here, we perform seismological and geodetic analyses to investigate the 2006 Copiapó swarm, which developed in April and May 2006. The swarm began on April 19, with a magnitude Ml 5.3 earthquake. During the nine following days, we observe a migration of seismicity along the plate interface, the occurrence of doublets events, and a potential slow slip event in the GPS time series at site Copiapó. Then, on April 30, a first earthquake with Mw 6.6 occurred at 15 km depth at the plate contact. It likely triggered a second earthquake of magnitude Mw 6.5, which occurred 144 min later, 10 km northwest of the first earthquake. Using InSAR, we determined the slip distribution associated with these two earthquakes and detailed the postseismic slip they triggered in the next days and weeks. This “postseismic” phase appears to be predominantly aseismic, while the moment released during the “coseismic” phase is comparable to other seismic crises that occurred in Atacama. Although we did not find a larger seismic and aseismic ratio than in other swarms in South America, we suggest a similar mechanism of slow deformation as a driver of seismicity during seismic swarms. Finally, we propose that the slow and fast behavior of the 2006 Copiapó swarm is a consequence of the subduction of the Copiapó Ridge seamounts, which affects both the plate interface and the overriding plate by inducing complex interactions between seismic and aseismic processes

Seismic and aseismic slip during the 2006 Copiapó swarm in North-Central Chile

International audienceEarthquake swarms commonly occur along the Chilean subduction zone, witnessing fast seismic and slow aseismic slip behavior at the plate interface. However, the largest seismic swarms observed in Chile, particularly in the Copiapó-Atacama region, remain poorly documented, and the underlying processes have yet to be understood. Here, we perform seismological and geodetic analyses to investigate the 2006 Copiapó swarm, which developed in April and May 2006. The swarm began on April 19, with a magnitude Ml 5.3 earthquake. During the nine following days, we observe a migration of seismicity along the plate interface, the occurrence of doublets events, and a potential slow slip event in the GPS time series at site Copiapó. Then, on April 30, a first earthquake with Mw 6.6 occurred at 15 km depth at the plate contact. It likely triggered a second earthquake of magnitude Mw 6.5, which occurred 144 min later, 10 km northwest of the first earthquake. Using InSAR, we determined the slip distribution associated with these two earthquakes and detailed the postseismic slip they triggered in the next days and weeks. This “postseismic” phase appears to be predominantly aseismic, while the moment released during the “coseismic” phase is comparable to other seismic crises that occurred in Atacama. Although we did not find a larger seismic and aseismic ratio than in other swarms in South America, we suggest a similar mechanism of slow deformation as a driver of seismicity during seismic swarms. Finally, we propose that the slow and fast behavior of the 2006 Copiapó swarm is a consequence of the subduction of the Copiapó Ridge seamounts, which affects both the plate interface and the overriding plate by inducing complex interactions between seismic and aseismic processes