Differential expression of the FAK family kinases in rheumatoid arthritis and osteoarthritis synovial tissues

The focal adhesion kinase (FAK) family kinases, including FAK and proline-rich kinase 2 (Pyk)2, are the predominant mediators of integrin αvβ3 signaling events that play an important role in cell adhesion, osteoclast pathology, and angiogenesis, all processes important in rheumatoid arthritis (RA). Using immunohistochemical and western blot analysis, we studied the distribution of phospho (p)FAK, pPyk2, pSrc, pPaxillin and pPLCγ in the synovial tissue (ST) from patients with RA, osteoarthritis (OA) and normal donors (NDs) as well as in RA ST fibroblasts and peripheral blood differentiated macrophages (PB MΦs) treated with tumor necrosis factor-α (TNFα) or interleukin-1β (IL1β). RA and OA STs showed a greater percentage of pFAK on lining cells and MΦs compared with ND ST. RA ST fibroblasts expressed pFAK at baseline, which increased with TNFα or IL1β stimulation. Pyk2 and Src were phosphorylated more on RA versus OA and ND lining cells and MΦs. pPyk2 was expressed on RA ST fibrobasts but not in MΦs at baseline, however it was upregulated upon TNFα or IL1β activation in both cell types. pSrc was expressed in RA ST fibroblasts and MΦs at baseline and was further increased by TNFα or IL1β stimulation. pPaxillin and pPLCγ were upregulated in RA versus OA and ND lining cells and sublining MΦs. Activation of the FAK family signaling cascade on RA and OA lining cells may be responsible for cell adhesion and migration into the diseased STs. Therapies targeting this novel signaling pathway may be beneficial in RA

Expression of mucin 3 and mucin 5AC in arthritic synovial tissue

Objective Rheumatoid arthritis (RA) is a chronic inflammatory disease that is characterized by hypertrophy of the synovial tissue, leukocyte infiltration, angiogenesis, and ultimately joint destruction. Mucins (MUCs) are a family of heavily glycosylated proteins that protect epithelial membranes and are used as ligands for cell adhesion. MUC gene expression has been found to be altered in many cancers and inflammatory states. This study was undertaken to examine its expression in synovial tissue (ST) and role in arthritis. Methods We performed immunohistochemistry, Western blotting, and reverse transcriptase–polymerase chain reaction to determine expression patterns of MUC1, MUC2, MUC3, and MUC5AC in RA, osteoarthritic (OA), and normal human ST. Results MUC3 was expressed in synovial lining cells, macrophages, and fibroblasts. Significantly more RA (n = 12) and OA (n = 13) synovial lining cells expressed MUC3 than did normal synovial lining cells (n = 7) (22% and 24% versus 0.4%, respectively; P < 0.05). Additionally, macrophages in RA and OA ST expressed significantly more MUC3 than did macrophages in normal ST (50% and 51% versus 10%, respectively; P < 0.05). MUC5AC was expressed at low levels in synovial lining cells, macrophages, and endothelial cells in RA and OA ST, and was barely expressed in normal ST. MUC1 and MUC2 proteins were not detected in ST. Messenger RNA (mRNA) for MUC3 and MUC5AC was detected in ST, and mRNA for MUC3 was detected in cultured ST fibroblasts. Conclusion These data demonstrate up-regulated MUC expression by ST cells and suggest a novel role of MUC3 and MUC5AC in the pathogenesis of arthritis.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/57898/1/23174_ftp.pd

Doping V2O5 on ZnO via wet incipient method and investigation crystallization and physical properties of undoped and doped ZnO nanparticles

In this contribution, ZnO nanoparticles and vanadium doped ZnO were synthesized by wet chemical and wet incipient method, respectively. ZnO nanoparticles were heated at 650 and 750°C for 3h in air and then characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), energy dispersive (EDX) analysis and photoluminescence (PL). The XRD patterns showed that the ZnO samples have a wurtzite structure (hexagonal phase) and vanadium doped is in V2O5 crystalline structure. Their structural characteristics and physical properties were investigated and compared. XRD and SEM data show that the size of nanoparticles increased from 24.3 to 32.6 nm when the annealing temperature was increased. Also, the results indicate that increasing the degree of crystalline improved the physical properties of the nanoparticles

Risk analysis of BOT contracts using soft computing

Build-Operate-Transfer (BOT) contracts have been widely implemented in developing countries facing budget constraints. Analysing the expected variability in project viability requires extensive risk analysis. An objective analysis of various risk variables and their influence on a BOT project evaluation requires study and integration of many sce­narios into the concession terms, which is complicated and time-consuming. If the process of negotiating the financial parameters and uncertainties of a BOT project could be automated, this would be a milestone in objective decision-mak­ing from various stakeholders’ points of view. A soft computing model would let the user incorporate as many scenarios as could be provided. Extensive risk analysis could then be easily performed, leading to more accurate and dependable results. In this research, an artificial neural network model with correlation coefficient of 0.9064 has been used to model the relationship between important project parameters and risk variables. This information was extracted from sensitiv­ity analysis and Monte Carlo simulation results obtained from conventional spreadsheet data. The resulting consensus would yield to fair contractual agreements for both the government and the concession company. First published online: 01 Jul 201

Gene expression programming approach to cost estimation formulation for utility projects

This article utilizes gene expression programming (GEP) technique to develop a prediction model in order to automate estimating the construction cost of water and sewer replacement/rehabilitation projects. A database gathered for developing the model was established on the basis of data related to 210 actual water and sewer projects obtained from the City of San Diego, California, USA. To verify the predictability of the GEP model, it was examined to estimate the cost of the projects that were not included in the modelling process. Sensitivity analysis technique and professional experiences were employed to determine the contributions of the qualitative factors and quantifiable parameters affecting the cost estimate. The proposed model with correlation coefficient of 0.8467 is adequately capable of estimating the cost of water and sewer replacement/rehabilitation projects. The GEP-based design equation can easily be used for predesign purposes to help allocate budgets and available limited resources effectively

Ten Important Tips in Treating a Patient with Lumbar Disc Herniation

Lumbar disc herniation is a common spinal disorder that usually responds favorably to conservative treatment. In a small percentage of the patients, surgical decompression is necessary. Even though lumbar discectomy constitutes the most common and easiest spine surgery globally, adverse or even catastrophic events can occur. Appropriate patient selection and effective neural decompression constitute the most important points for better surgical outcomes and avoidance of unpleasant complications. Other important tips include timely performance of magnetic resonance imaging, correct interpretation of scan data, preoperative detection of underlying instability, exclusion of non-discogenic sciatica, determination of the main cause of clinical pathology, avoidance of the wrong side or level, and being sure that the more detailed procedure does not necessarily mean the more effective procedure

Interleukin-18 as an in vivo mediator of monocyte recruitment in rodent models of rheumatoid arthritis

Abstract Introduction The function of interleukin-18 (IL-18) was investigated in pertinent animal models of rodent rheumatoid arthritis (RA) to determine its proinflammatory and monocyte recruitment properties. Methods We used a modified Boyden chemotaxis system to examine monocyte recruitment to recombinant human (rhu) IL-18 in vitro. Monocyte recruitment to rhuIL-18 was then tested in vivo by using an RA synovial tissue (ST) severe combined immunodeficient (SCID) mouse chimera. We defined monocyte-specific signal-transduction pathways induced by rhuIL-18 with Western blotting analysis and linked this to in vitro monocyte chemotactic activity. Finally, the ability of IL-18 to induce a cytokine cascade during acute joint inflammatory responses was examined by inducing wild-type (Wt) and IL-18 gene-knockout mice with zymosan-induced arthritis (ZIA). Results We found that intragraft injected rhuIL-18 was a robust monocyte recruitment factor to both human ST and regional (inguinal) murine lymph node (LN) tissue. IL-18 gene-knockout mice also showed pronounced reductions in joint inflammation during ZIA compared with Wt mice. Many proinflammatory cytokines were reduced in IL-18 gene-knockout mouse joint homogenates during ZIA, including macrophage inflammatory protein-3α (MIP-3α/CCL20), vascular endothelial cell growth factor (VEGF), and IL-17. Signal-transduction experiments revealed that IL-18 signals through p38 and ERK½ in monocytes, and that IL-18-mediated in vitro monocyte chemotaxis can be significantly inhibited by disruption of this pathway. Conclusions Our data suggest that IL-18 may be produced in acute inflammatory responses and support the notion that IL-18 may serve a hierarchic position for initiating joint inflammatory responses.http://deepblue.lib.umich.edu/bitstream/2027.42/112330/1/13075_2010_Article_2890.pd

The parasitic helminth product ES-62 suppresses pathogenesis in collagen-induced arthritis by targeting the interleukin-17–producing cellular network at multiple sites

Among many survival strategies, parasitic worms secrete molecules to modulate host immune responses. One such product, ES-62, is protective in the collagen-induced arthritis (CIA) model of rheumatoid arthritis. As IL-17 has been reported to play a pathological role in the development of rheumatoid arthritis, we investigated whether targeting of IL-17 may explain the protection afforded by ES-62 in the CIA model. DBA/1 mice progressively display arthritis following immunization with type-II collagen. The protective effects of ES-62 were assessed by determination of cytokine levels, flow cytometric analysis of relevant cellular populations and in situ analysis of joint inflammation. ES-62 was found to downregulate IL-17 responses in the CIA model. Firstly, it acts to inhibit priming and polarisation of IL-17 responses by targeting a complex IL-17-producing network, involving signalling between dendritic cells and γδ or CD4+ T cells. In addition, ES-62 directly targets Th17 cells by downregulating MyD88 expression to suppress responses mediated by IL-1 and TLR ligands. Moreover, ES-62 modulates migration of γδ T cells and this is reflected by direct suppression of CD44 upregulation and, as evidenced by in situ analysis, dramatically reduced levels of IL-17-producing cells, including lymphocytes, infiltrating the joint. Finally, there is strong suppression of IL-17 production by cells resident in the joint, such as osteoclasts within the bone areas. Such unique multi-site manipulation of the initiation and effector phases of the IL-17 inflammatory network could be exploited in the development of novel therapeutics for rheumatoid arthritis