text:
Chemokine receptors in the rheumatoid synovium: upregulation of CXCR5
DOI: 10.1186/ar1475
© Schmutz et al., licensee BioMed Central Ltd. 2004
Received: 9 August 2004
Accepted: 12 November 2004
Published: 16 December 2004
In patients with rheumatoid arthritis (RA), chemokine and chemokine receptor interactions play a central role in the recruitment of leukocytes into inflamed joints. This study was undertaken to characterize the expression of chemokine receptors in the synovial tissue of RA and non-RA patients. RA synovia (n = 8) were obtained from knee joint replacement operations and control non-RA synovia (n = 9) were obtained from arthroscopic knee biopsies sampled from patients with recent meniscal or articular cartilage damage or degeneration. The mRNA expression of chemokine receptors and their ligands was determined using gene microarrays and PCR. The protein expression of these genes was demonstrated by single-label and double-label immunohistochemistry. Microarray analysis showed the mRNA for CXCR5 to be more abundant in RA than non-RA synovial tissue, and of the chemokine receptors studied CXCR5 showed the greatest upregulation. PCR experiments confirmed the differential expression of CXCR5. By immunohistochemistry we were able to detect CXCR5 in all RA and non-RA samples. In the RA samples the presence of CXCR5 was observed on B cells and T cells in the infiltrates but also on macrophages and endothelial cells. In the non-RA samples the presence of CXCR5 was limited to macrophages and endothelial cells. CXCR5 expression in synovial fluid macrophages and peripheral blood monocytes from RA patients was confirmed by PCR. The present study shows that CXCR5 is upregulated in RA synovial tissue and is expressed in a variety of cell types. This receptor may be involved in the recruitment and positioning of B cells, T cells and monocytes/macrophages in the RA synovium. More importantly, the increased level of CXCR5, a homeostatic chemokine receptor, in the RA synovium suggests that non-inflammatory receptor–ligand pairs might play an important role in the pathogenesis of RA.
Keywordschemokine receptors CXCR5 microarrays rheumatoid synovium
Rheumatoid arthritis (RA) is a chronic inflammatory condition that affects multiple joints, and it results in the accumulation of leukocytes within the synovial tissue (ST) and synovial fluid (SF). The inflammatory infiltrate consists predominantly of B lymphocytes, T lymphocytes and macrophages in the ST, whereas neutrophils are mainly found in the SF. The lymphocyte infiltration is organized in lymphoid-like microstructures in just under 50% of the RA patients; however, the patients present germinal centre reactions in only 20% of cases [1]. The pathogenesis of the RA is still largely unknown but leukocytes and their products play an important role in the development of inflammation, joint destruction and pain [2, 3]. The attraction of leukocytes into the joints is controlled by chemokines, a family of small chemotactic cytokine-like molecules that act as potent mediators of inflammation [4].
Chemokine activity is dependent on the presence of and interaction with chemokine receptors on the leukocyte surface. Indeed, chemokines and their receptors are involved together in the development and perpetuation of inflammation [5]. In vitro and in vivo experiments have indicated that blocking chemokines or their receptors could potentially provide an effective treatment of inflammatory diseases [5, 6]. The 19 receptors so far identified belong to a super-family of G-protein-coupled receptors with seven transmembrane domains [7]. Chemokine receptors have a regulatory effect on the maturation and traffic of leukocytes, and they are implicated in several disease states [8]. There have been several reports on chemokine receptor expression on T cells from RA ST, RA SF and RA peripheral blood (PB) [9–13]. The expression of some chemokine receptors on monocytes/macrophages, dendritic cells and neutrophils has also been reported [14–17], and the importance of the role of chemokine receptors in RA is emerging [18, 19].
CXCR5 is a chemokine receptor highly expressed in recirculating B cells, in subsets of CD4+ and CD8+ T cells and monocytes [20, 21]. It also has been identified on B-cell infiltrates in Sjogren's syndrome [22, 23]. CXCR5 is involved in the immune-system homeostasis and in lymphoid organogenesis [24]. Several morphological and functional studies suggest that lymphoid neogenesis takes place in RA [1, 25, 26]. Furthermore, an important disturbance of follicle and germinal centre formation in the spleen and Peyer's patches is observed in CXCR5-deficient mice [27]. CXCL13, the unique ligand of CXCR5, is also involved in follicular homing, as observed in CXCL13-deficient mice [28].
In view of the role of chemokine receptors in leukocyte traffic, the aim of the present study was to compare their expression in inflamed and non-inflamed tissue to shed light on which chemokine receptors may be involved in the recruitment and retention of leukocytes in ST. We examined chemokine receptor expression in ST taken from RA and non-RA patients using microarray technology, RT-PCR and immunohistochemistry. The microarray and RT-PCR experiments demonstrated the differential expression of CXCR5, and immunohistochemistry showed that this receptor is expressed in B-cell and T-cell infiltrates, on macrophages and blood vessels. Our study identifies CXCR5 as a potentially interesting therapeutic target in RA and points to the use of antagonists to this receptor as a treatment strategy in the disease.
Materials and methods
Tissue and cell source
Details of rheumatoid arthritis (RA) and non-RA patients
Patient (sex, age [years])
Disease duration (years)
1 (male, 69)
Auranofin, NSAID
2 (female, 41)
3 (female, 51)
NSAID, analgesic
4 (female, 79)
Methotrexate, NSAID, steroid
5 (female, 70)
Penicillamine, steroid, NSAID
6 (female, 63)
RA + secondary osteoarthritis
Methotrexate, steroid, analgesic
7 (female, 33)
juvenile chronic arthritis
8 (female, 66)
NSAID, analgesic
1 (female, 50)
Articular cartilage damage
NSAID, analgesic
2 (male, 44)
Meniscal tear
3 (male, 42)
Meniscal tear and articular cartilage damage
4 (male, 60)
Meniscal degeneration
5 (male, 52)
Articular cartilage degeneration
6 (male, 46)
Meniscal tear
7 (male, 53)
Meniscal degeneration
8 (male, 38)
Meniscal tear
9 (male, 43)
Articular cartilage degeneration
1 (female, 91)
2 (female, 56)
NSAID, steroid, methotrexate
3 (male, 67)
4 (male, 67)
Analgesic, NSAID, methotrexate
Monocytes/macrophages were isolated from the PB and SF of another four RA patients (Table 1). In brief, the blood and hyaluronidase-treated SF were centrifuged over a ficoll cushion (Amersham Biosciences, Chalfont St Giles, UK). The macrophages were isolated from the buffy coat layer (lymphocytes, macrophages) by adherence onto a glass dish.
RNA extraction
Total RNA was extracted from frozen blocks of synovia or from isolated monocytes/macrophages using TRIreagent solution (Sigma, Poole, UK) according to the manufacturer's recommendation. The quantity recovered was determined by spectrophotometry and the integrity was assessed by gel electrophoresis. For microarray experiments, equal quantities (7 μg) of RNA from each RA or non-RA patient were pooled and the messenger RNA was extracted using the mRNA GeneElute Kit (Sigma). The quantity recovered was determined by fluorometry using SYBR Green II (Molecular Probes, Leiden, The Netherlands). RNA had to be pooled since only small biopsies could be obtained from non-RA patients.
Microarray technology
The panorama human cytokine gene array (Sigma-Genosys, Pampisford, UK) was used. This array contains 375 different cDNAs including 16 chemokine receptors and 33 chemokines, each printed in duplicate on nylon membranes.
The probe labelling and hybridization were carried out according to the manufacturer's instructions. Briefly, 33P-radiolabelled cDNA probes were prepared from 0.5 μg mRNA (see earlier) using human cytokine cDNA labelling primers (Sigma-Genosys) and AMV reverse transcriptase at 42°C, and were purified on a Sephadex® G-25 spin column (Sigma-Genosys). The arrays were hybridized for 17–18 hours at 65°C, washed and subjected to autoradiography for various lengths of time using Kodak BioMax MR X-ray film.
The intensity of hybridization signals was quantified using the ArrayVision, version 6.0, software (Imaging Research Inc., Haverhill, UK). The intensity of each spot was corrected for background levels using the 'corners between spots' (set to 3 pixels) with or without 'segmentation' protocols, and were normalized for differences in labelling using the average values of seven housekeeping genes: β 2 -microglobulin, β-actin, cyclophilin A, glyceraldehyde-3-phosphate dehydrogenase, HLA-A 0201 heavy chain, human hypoxanthine phosphoribosyl transferase, and α-tubulin. The remaining two housekeeping genes, L19 and transferrin R, were excluded because of signal saturation and differential expression, respectively. The software performs the normalization automatically.
Reverse transcription-polymerase chain reaction
Total RNA aliquots from individual patients were reverse transcribed using oligo(dT18) (MWG Biotech, Ebersberg, Germany) and MMLV reverse transcriptase (Promega, Southampton, UK) at 37°C for 1 hour. The reactions were terminated at 70°C for 10 min and were diluted to 80 μl with H2O. For two of the non-RA patients no more RNA was available for RT-PCR following microarray analysis.
Sequences of the primers used for RT-PCR
Size (base pairs)
Accession number
Immunohistochemistry for CXCR5
The ST from the patients that had been examined at the transcription level was also available for protein expression analysis. Paraffin embedded sections were cut 4 μm thick on 3-aminopropyltriethoxysilane-coated slides. Sections were deparaffinized and rehydrated before blocking endogenous peroxidase activity with H2O2 (0.3%) in methanol. The slides were rinsed with PBS and incubated with normal serum (1:67 in PBS) for 10 min before applying anti-human CXCR5 monoclonal antibody (1:666; R&D, Abingdon, UK) and the respective IgG control (Dako, Ely, UK). The sections were rinsed with PBS and incubated with biotinylated secondary antibody. The antibody binding was detected using reagents in the Vectastain ABC Elite kit (Vector, Peterborough, UK) and the chromogen 3,3'-diaminobenzidine (DAB) (Vector). Sections were rinsed and counter stained in Mayer's haematoxylin.
B cells and macrophages were localized using anti-human CD20 antibodies (1:100; Dako) and CD68 antibodies (clone PG-M1, 1:75; Dako), respectively. CD20 required antigen demasking by 15 min microwaving in citrate buffer (pH 6.0), but H2O2 treatment was not necessary. CD68 antigen was demasked using 0.05% pronase in Tris-buffered saline (pH 7.2) for 10 min.
Double immunohistochemistry was performed with anti-human CD3 rabbit monoclonal antibodies (Labvision) and CXCR5 antibodies. The slides were deparaffinized, rehydrated and microwaved for 15 min in citrate buffer pH 6.0 before being treated with H2O2 in methanol. The slides were incubated with 2.5% normal swine serum for 20 min before applying CD3 diluted 1:60 in 2.5% serum for 30 min. The sections were rinsed with PBS and were incubated with swine anti-rabbit antibody linked to alkaline phosphatase (1:40; Dako). CD3 binding was detected using Vector Red substrate (Vector). Sections were rinsed and were either counter stained in Methyl Green (Vector) or subjected to a second round of immunohistochemistry. CXCR5 was used as for single immunohistochemistry except that blocking and antibody dilutions were made in 2.5% normal horse serum and CXCR5 was revealed with DAB-Nickel (Vector). No counter stain was performed for double immunohistochemistry sections.
Patients and tissue selection
Synovia were obtained from knee joints as this allowed the use of arthroscopic samples of non-RA (normal) as controls instead of osteoarthritic tissue, which can show more enhanced inflammatory changes. The histology of H&E-stained RA synovial sections demonstrated classic signs of inflammation. Mononuclear cell infiltrates were visible in seven out of eight patients and consisted of aggregate structures; one of these seven patients also contained more germinal-like centre structures. In addition one patient revealed a diffuse infiltration. The synovium of the non-RA patients showed minimal signs of inflammation. In eight out of nine patients no mononuclear infiltrates were observed, and in one case only a small infiltrate was seen. No thickening of the intima was observed in the non-RA compared with the RA samples.
Microarray analysis of chemokine receptor expression
To allow rapid preliminary screening of a large number of chemokines and their receptors in RA ST and non-RA ST, chemokine expression was investigated using microarrays.
Chemokine and chemokine receptor expression data analysis
Regulation (ratio RA/non-RA)
Array column 3 (Fig. 1, C3)
Up (2.2 ± 0.2)
0.012 ○●
Up (2.7 ± 0.2)
0.000 ○
0.000 ○
Not visible (NA)
0.000 ○
0.000 ○
Not visible (NA)
0.003 ○
0.000 ○
Not visible (NA)
0.000 ○●
Up (NA)
0.001 ○
0.000 ○
Not visible (NA)
0.022 ○●
0.011 ○
Not visible (2.5 ± 0.9)
0.001 ○●
0.000 ○
Not visible (NA)
0.028 ○●
0.019 ○●
Not visible (1.5 ± 0.1)
0.036 ○●
0.008 ○●
Not visible (9.6 ± 5.6)
0.000 ○●
Up (NA)
Up (3.5 ± 0.1)
Up (22.6 ± 0.7)
Not visible (0.7 ± 0.0)
0.000 ○
Up (NA)
Array column 1 (Fig. 1, C1)
CCL21 (6Ckine)
0.019 ○
Up (1.6 ± 0.1)
0.020 ○
Up (2.8 ± 0.4)
0.019 ○●
Up (1.6 ± 0.1)
0.015 ○●
Up (3.5 ± 0.3)
CCL11 (eotaxin)
Not visible (1.0 ± 0.0)
CCL24 (eotaxin-2)
0.013 ○
Up (2.5 ± 0.3)
CX3CL1 (fractalkine)
0.030 ○●
0.012 ○
Not visible (2.5 ± 0.2)
CXCL1 (GRO-α)
Up (1.6 ± 0.0)
CXCL2 (GRO-β)
Up (1.5 ± 0.0)
CXCL3 (GRO-γ)
Up (1.8 ± 0.0)
Down (0.7 ± 0.0)
0.006 ○●
0.000 ○
Not visible (NA)
0.007 ○●
0.000 ○
Not visible (NA)
Up (3.3 ± 0.2)
0.005 ○
0.000 ○
Not visible (NA)
CCL15 (MIP-1δ)
0.009 ○●
0.008 ○
Not visible (11 ± 13)
XCL1 (lymphotactin)
Up (1.6 ± 0.0)
Not visible (0.9 ± 0.0)
Not visible(0.9 ± 0.0)
Up (1.4 ± 0.1)
0.000 ○
0.002 ○
Not visible (0.0)
0.000 ○●
0.008 ○
Not visible (0.0)
Array column 2 (Fig. 1, C2)
Up (1.9 ± 0.0)
Up (3.6 ± 0.1)
CCL3 (MIP-1α)
Not visible (1.0 ± 0.0)
CCL4 (MIP-1β)
0.018 ○●
Up (1.3 ± 0.2)
CCL20 (MIP-3α)
0.000 ○
0.013 ○
Not visible(0.0)
CCL19 (MIP-3β)
Not visible (0.6 ± 0.0)
Up (0.9 ± 0.0)
Up (4.5 ± 0.4)
Up (1.2 ± 0.1)
Down (0.7 ± 0.0)
0.000 ○
0.000 ○
Not visible (NA)
0.000 ○
0.000 ○
Not visible (NA)
The most obvious differences between RA samples and non-RA samples were for the chemokine receptors CXCR5 and CXCR2, and to a lesser extent CXCR4, which gave stronger signals in RA samples (Fig. 1). In order to quantify the differential expression of these receptors the densities of autoradiographic spots were measured using ArrayVision software (Table 3). The criteria we set for a gene to be considered as upregulated or downregulated were a RA/non-RA ratio higher than 3 or lower than 0.3, respectively, and a 95% confidence interval below 10% (criteria as [30]). In the present study the expression of CXCR5 and CXCR4 was 22.6 ± 0.7-fold higher and 3.5 ± 0.1-fold higher in RA tissue than in non-RA tissue, respectively. These results indicated that, of the chemokine receptors studied, CXCR5 was the most upregulated in RA (Table 3). The upregulation of CXCR2 could not be calculated for mathematical reasons because the signal intensity of CXCR2 in non-RA tissue after correction for the background was zero. CXCR2 was only visible on the non-RA autoradiogram upon prolonged exposure, at which point the housekeeping genes were saturated and were therefore unsuitable for quantification purposes.
Out of the 33 chemokines present on the arrays, 29 of these ligands were visible on the RA membranes and 21 on the non-RA membranes (Fig. 1 and Table 3). These included CXCL13, CXCL12, CXCL8, CXCL1-3 and CXCL5, which are ligands for the chemokine receptors CXCR5, CXCR4 and CXCR2. Several chemokines were visible on RA microarrays but not on non-RA microarrays (namely CXCL13, CCL21 and CCL24), suggesting that these genes might be induced in the inflamed synovium. In contrast, there were no chemokine signals that were present on non-RA membranes and were absent on RA membranes. Where chemokine signals were detectable on RA and non-RA microarrays, it was possible to quantify the degree of upregulation or downregulation using the criteria described earlier for chemokine receptors. Of these chemokines, the following showed upregulation: CCL18 (4.5 ± 0.4-fold increase), CXCL9 (3.6 ± 0.1-fold increase), CXCL5 (3.5 ± 0.3-fold increase), and CXCL8 (3.3 ± 0.2-fold increase). No chemokines displayed a downregulation with a RA/non-RA ratio less than 0.3. The upregulation of CXCL9 in RA synovia is in agreement with the only microarray study of RA synovia, in which this chemokine was also shown to be increased [31]. In our study only five chemokines (CXCL7, CCL13, CCL20, CCL17 and CCL25) could not be detected at all, whether in RA or non-RA samples.
The rapid screening of several genes at once made array technology a very attractive method. Its use has revealed disadvantages, however, including the requirement for large amounts of RNA (which are not always available from human tissue biopsies), a susceptibility to experimental variability and a lack of standard optimum methods for statistical analysis [32]. Arrays also present the risk of cross-hybridization leading to false positive or negative results [31]. However, the array approach remains a valuable tool if the samples can be pooled and if it is used in conjunction with alternative methods such as RT-PCR.
RT-PCR analysis of CXCR5
To identify the cell types expressing CXCR5, and since RNA expression and protein expression do not always correlate, the protein expression of this receptor and three specific cell markers (CD20, CD3 and CD68) was investigated by immunohistochemistry of paraffin-embedded sections.
For all immunohistochemistry experiments in this study, the use of isotype-matched immunoglobulin controls or sera instead of primary antibodies resulted in negative staining of RA sections and non-RA sections (Figs 3b,3d,3g,3h, 4c,4d,4f and 5c,5d,5f).
RT-PCR on isolated RA monocytes/macrophages
The major finding of the present study is that CXCR5 is upregulated in the RA synovium. The cells expressing this chemokine receptor are B lymphocytes, T lymphocytes, macrophages and endothelial cells. The increased numbers of B lymphocytes, T lymphocytes and macrophages producing CXCR5 in the RA synovium are probably responsible for the increased expression of the receptor in this chronically inflamed tissue. The majority (seven out of eight) of the RA synovia included in this study contained substantial lymphoid aggregates but only one out of nine non-RA patients presented a very small infiltrate. These cell aggregates contained CD20+ B cells that expressed CXCR5. The expression of CXCR5 has been reported in mature B cells and secondary lymphoid organs but as far as the authors are aware this is the first report of a chemokine receptor expressed by B cells in the RA synovium and its ectopic lymphoid structures.
Our findings are particularly interesting in view of the functional role of B cells in RA. This includes autoantibody production, antigen presentation, a role in lymphoid follicle and germinal centre formation, and the promising results of the anti-CD20 treatment in RA [33, 34]. The microarrays showed that the mRNA for CXCL13, the only known ligand of CXCR5, was present in the RA synovium and not in the non-RA synovium. Furthermore, other reports have shown a CXCL13 message in RA synovia, together with its protein that localizes to follicular dendritic cells, endothelial cells and synovial fibroblasts, suggesting that these cells produce the chemokine [1, 25]. Taken together with our data, this indicates that CXCR5 on B cells may be important in the recruitment of these cells into the RA synovium, in addition to their positioning and retention within the synovial infiltrates. In this regard, the role of CXCR5 on B cells in secondary lymphoid organs has been well documented [35, 36]. CXCR5 guides B cells into the B-cell follicles and also directly promotes the recruitment of these cells into Peyer's patches via high endothelial venules [27, 28, 37, 38]. In addition CXCR5-deficient mice exhibit impaired development of lymph nodes and Peyer's patches, and the tissue architecture of these organs is severely disturbed showing a lack of B-cell follicles [27, 28].
Our double immunohistochemistry data indicate that there is a population of CXCR5+CD3+ T cells present in the RA synovium. CXCR5+ T cells have been shown in secondary lymphoid tissue where some of these cells localize within germinal centres [20, 39], and it is proposed that CXCR5 enables them to enter B-cell follicles guided by CXCL13 [36]. Within these follicles they may provide B-cell help and have therefore been named follicular B helper T cells, since purified human tonsillar CD4+CXCR5+ T cells efficiently stimulate the production of immunoglobulins by B cells [39, 40]. These follicular B helper T cells are CD57+, whereas the majority of the CXCR5+ T cells that are present in interfollicular and T-cell areas of the lymphoid tissue are CD57- and are poor B-cell helpers [41]. Since lymphoid neogenesis occurs in the RA synovium it is possible that the CXCR5 expression on T cells as shown in the present study is involved in the positioning of these cells within the synovium and in providing B-cell help, although further studies are required to characterize this synovial T-cell population. Whether the two populations of CXCR5+CD57+ and CXCR5+CD57- T cells are present in the RA synovium and what their role could be is still unknown. However, CD57+ T cells are reported to be present in the RA synovium and SF, where levels of this marker are elevated compared with controls [42, 43]. Furthermore, an involvement of CD57+ T cells has been shown in disease activity of RA [44].
Immunohistochemical experiments indicated that CD68+ cells in the synovial intima express CXCR5. Intimal cells comprise two cell types: macrophage-like cells and fibroblast-like cells. In RA the former macrophage-like cells are numerous, comprising up to 80% of this cell layer [45]. It has been reported that in the RA synovium anti-CD68 reacts strongly with intimal macrophages, but fibroblasts also show some reactivity with this antibody [45]. Therefore, since macrophages are abundant in the RA intima and because of their strong reactivity with anti-CD68, it is likely that intimal macrophages are positive for CXCR5. In the normal non-RA intima, macrophages are positive for CD68 and fibroblasts are negative, making it more certain that macrophages express CXCR5 in this cell layer [45]. Consequently, RT-PCR was performed to verify that RA macrophages/monocytes can express CXCR5. The RT-PCR did indeed demonstrate CXCR5 mRNA in macrophages from RA SF, as well as PB monocytes from the same RA patients. Since the CXCR5 mRNA is expressed at similar levels in RA PB and RA SF it is suggested that the contribution of monocytes/macrophages to the upregulation of CXCR5 in the RA synovium is due to their increased number, rather than due to an increased abundance of CXCR5 transcripts per cell. CXCR5 mRNA has also been found in normal human PB monocytes by RT-PCR [21]. Studies by ourselves and other workers have shown that monocytes/macrophages express several other CXC chemokine receptors in RA, including CXCR1, CXCR2 and CXCR4 [15, 16, 46]. Furthermore, RA monocytes/macrophages express CC chemokine receptors such as CCR1, CCR2, CCR3 and CCR5 [14], illustrating their broad profile of chemokine receptor expression.
Endothelial cells are another cell type expressing CXCR5 in the synovium. There have been several reports of endothelial cells in the RA synovium expressing chemokine receptors, including CXCR3 and CXCR4, in addition to the Duffy antigen that is a non-signalling chemokine receptor [18, 47–49]. In the RA synovium there is increased turnover of blood vessels with enhanced formation of new blood vessels together with enhanced vascular regression [50, 51]. These mechanisms are regulated by the balance of angiogenic and angiostatic factors, and these factors include chemokines. Some chemokines are angiogenic (e.g. CXCL8, CXCL12, CCL1 and CCL2) and other chemokines are angiostatic (e.g. CXCL9 and CXCL10), and activation of their respective chemokine receptors results in the stimulation of or inhibition of endothelial cell proliferation [47, 52–57]. CXCL13 has been shown to have an angiostatic function, inhibiting the angiogenic effects of FGF-2 on human umbilical vein endothelial cells [58]. In addition, the presence of CXCR5 in a variety of cultured human endothelial cells – from umbilical and saphenous veins, for example – may mediate the angiostatic effects of CXCL13 [58, 59]. Our data showing the presence of CXCR5 on endothelial cells in the synovium and the presence of its ligand in this tissue [1, 25]suggest that CXCR5 may play an angiostatic role in RA pathophysiology, although the angiostatic effects of CXCL13 could potentially be acting through CXCR3, which is also expressed by the RA synovial endothelium [48, 60].
In the present study mRNA for other chemokine receptors were detected in the RA synovium, such as CXCR1, CXCR2, CXCR4, Bob, CCR1, CCR2a, CCR7, CCR9 and CX3CR1 (CXCR3 was not on the microarray). All of these showed variable degrees of increased mRNA expression in RA, although the upregulation was less compared with that of CXCR5. Several previous reports have shown the expression of chemokine receptors by leukocytes from RA joints. These have included CCR4–CCR6, CXCR3, CXCR4 and CX3CR1 by T lymphocytes [9, 12, 13, 19] and CCR1–CCR5 and CXCR1–CXCR4 by monocytes/macrophages [14–16, 18]. Such reports mainly focused on selected cell types and certain chemokine receptors. The present study took a different approach. Ours was primarily a whole-tissue study examining the mRNA expression of a wide range of chemokine receptors in RA and control synovia. While our study is in general accord with previous reports, differences may in part be due to the RA ST used. This tissue was highly infiltrated and, in all but one sample, had extensive lymphoid follicles bearing resemblance to those of secondary lymphoid organs. This feature may be responsible for the particular upregulation of the constitutive chemokine receptor CXCR5. In addition, our RA patients had long-standing disease (Table 1) and the patient sample may also have influenced the types of chemokine receptors expressed.
Our study demonstrates the expression of CXCR5 on B cells, on T cells, on monocytes/macrophages and on endothelial cells in the RA synovium. The expression of a marker shared by cells that are known to play a central role in the process of chronic inflammation is of particular interest and suggests that targeting CXCR5 could provide a powerful new treatment for RA.
haematoxylin and eosin
peripheral blood
phosphate-buffered saline
polymerase chain reaction
rheumatoid arthritis
reverse transcription
synovial fluid
synovial tissue.
The authors are indebted to the patients who kindly agreed to take part in this study. They thank Mr C McGeoch, Mr D Rees, Mr L van Niekerk and Mr S White and the theatre teams for their help in obtaining synovial tissue. They are also very grateful to P Evans, M Pritchard and N Harness for their histological expertise and to J Menage for helpful immunohistochemistry discussion. Finally, the authors thankfully acknowledge the Henry Smith Charity, Droitwich Medical Trust Ltd and the Orthopaedic Institute Ltd for their financial support.
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This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is cited.


text:
It was set to be standard campaign fare – a final rally in a hotel ballroom in a downtown Newark hotel with confetti, balloons and a Hollywood star to make sure enough folks showed up. The candidate is Cory Booker, and the celebrity Eva Longoria, former cast member of 'Desperate Housewives'.
Nothing else about New Jersey's special election for a US Senate seat is standard however. Certainly there is nothing desperate about it for Mr Booker, mayor of Newark since 2006. Few doubt he will emerge the winner in primary elections on Tuesday or that he will also prevail on 16 October when voters will pick between him and a Republican opponent. (The state hasn't sent a Republican to the Senate since 1972.)
Mayor Booker can "start looking for a Washington apartment," right now concedes Maurice Carroll, who directed a Quinnipiac University survey that last week showed him steamrolling his rivals for the Democratic nomination. The polls showed 54 per cent supporting Mr Booker. His nearest rival was US Congressman Frank Pallone with the backing of only 17 per cent of voters. In the race as well is Rush Holt, also a member of Congress, and Sheila Oliver, the speaker of the state legislature who had hopes of becoming the first woman and African American elected to the US Senate from New Jersey.
For all three the primary campaign of the last several weeks has been an exercise in frustration (and possibly futility). Even new revelations of Mr Booker's involvement in a video-sharing internet start-up called Waywire that has earned him around $1 million (£646,000) have done nothing to slow his momentum. When they gathered for a first TV debate last month, the mayor didn't show up; he preferred to attend a fancy fund-raising event on his behalf on the Hudson River across from Manhattan hosted by Oprah Winfrey.
That Mr Booker had a bigger future has been clear to national Democrats for a long while. Some in the party had hoped he would challenge Chris Christie, the Republican Governor of New Jersey and Republican heavyweight, who is seeking re-election in November. Mr Booker had already telegraphed his interest in seeking a US Senate seat instead when one of the state's sitting senators, Frank Lautenberg, died. That meant Mr Booker could pursue his ambition sooner than he had thought.
It is not that his stewardship of Newark has been beyond criticism. While Mr Booker has done much to shine attention on its multiple problems and attract some small amounts of private investment – including a $100 million persona gift from Facebook founder Mark Zuckerberg to help city schools – the scars of its past problems, many dating back to race riots that ravaged its urban-scape in 1967, very much remain. Today 26 per cent of Newark's residents are living in poverty, nearly three times the level nationally.
Where he has succeeded is in making himself into a political brand. A tall and imposing 44-year-old African-American bachelor with a shaved head and ready smile, Mr Booker commanded a prime-time spot at the Democratic convention last year. He attained almost hero status last year after dashing into flames to save a neighbour from a house fire. He is famously prolific on Twitter with words of compassion. "Be kind to one another because we are all a bit more fragile than we let on," he posted today.
When David Cameron, the Prime Minister, visited New York last year, he chose to spend several hours across the river with Mr Booker in New Jersey. Mr Cameron said he wanted to "exchange some information on schooling, on crime, on police accountability, on regeneration". The mayor, we also were reminded, studied at Oxford University as a Rhodes Scholar. Bill Clinton did the same.
When Mr Booker did turn up for a final debate last week, his rivals did what they could to puncture him. "The mayor talks about how he's hands-on, but he's not even in Newark at least 20 per cent of the time and now he's got this new Internet start up," Mr Pallone attempted.
- More about:
- Democrats (US)
- Investment
- New Jersey
- New York City
- Schools
- US Senate

subset: commoncrawl


text:
In a message dated 6/2/1999 6:24:19 PM Central Daylight Time,
email@example.com writes:
<< Any good ways to control them? >>
I find that using Sevin (either 5% or 10%) serves double duty. It kills the
cutworms and deters the slugs. Be very careful using the vinegar solution--I
nearly killed 4 hostas using a 6:1 solution. Some people use an ammonia
solution. Both of these require direct hits on the slugs, however.
The best slug bait available in my neck of the woods is Spectracide
Sluggit.Also, the paste is effective.
There is a British product that is even better. It is Bio something are
other. Perhaps it is now available in the states, but not in Indy.
Clyde Crockett z5
To sign-off this list, send email to firstname.lastname@example.com with the
message text UNSUBSCRIBE HOSTA-OPEN


text:
Susan Hekman
|Susan Hekman
February 28, 1949
Grand Rapids, Michigan
|Academic background
|Alma mater
|University of Washington
|Academic work
|Main interests
|Postmodern feminism
|Notable ideas
|Criticism of standpoint feminism
Susan Jean Hekman (born February 28, 1949),[1] is a postmodern feminist and the professor of political science and director of the graduate humanities program at the University of Texas at Arlington (UTA).[2] Hekman's research has been critical of standpoint feminist theory.[3]
Hekman was born in Grand Rapids, Michigan, USA, in 1949. Hekman earned her Ph.D from the University of Washington in Seattle.[4] While Hekman stayed at NIAS, she completed the first chapter of her proposed book, Subject Matters: The Evolution of the Subject in Feminist Theory. Hekman stated that, "the book will be an analysis of the major contributions to the development of the subject in feminist theory from the mid-twentieth century to the present. The first chapter, an analysis of the path-breaking work of Simone de Beauvoir, defines the themes that will be pursued in subsequent chapter. After completing this chapter, I sketched out the outline of the entire book, mapping the organization of the chapters and the theses that I will develop."[4]
Selected works[edit]
- Hekman, Susan; Alaimo, Stacy (2008). Material feminisms. Bloomington: Indiana University Press. ISBN 9780253219466.
Journal articles[edit]
- Hekman, Susan (Winter 1997). "Truth and method: feminist standpoint theory revisited". Signs: Journal of Women in Culture and Society. The University of Chicago Press via JSTOR. 22 (2): 341–365. JSTOR 3175275.
- See also: Hartsock, Nancy (Winter 1997). "Comment on Hekman's "Truth and Method: Feminist Standpoint Theory Revisited": truth or justice?". Signs: Journal of Women in Culture and Society. The University of Chicago Press via JSTOR. 22 (2): 367–374. JSTOR 3175277.
- ^ "Hekman, Susan J.". Library of Congress. Retrieved 16 January 2015.
(Susan J. Hekman) data sht. (b. 02-28-49)
- ^ "Dr. Susan Hekman: Professor of Political Science; Director, Graduate Humanities Program". Department of Political Science, University of Texas at Arlington. Retrieved 16 January 2015.
- ^ Flynn, Elizabeth A. (2002). "Introduction". In Flynn, Elizabeth A. Feminism beyond modernism. Carbondale: Southern Illinois University Press. p. 12. ISBN 9780809324354.
- ^ a b "Hekman, S.J.". Netherlands Institute for Advanced Study. Retrieved 15 January 2015.
External links[edit]
- Profile page: Susan Hekman, University of Texas at Arlington
|This feminism-related article is a stub. You can help Wikipedia by expanding it.
|This biography of an American political scientist is a stub. You can help Wikipedia by expanding it.


text:
Home > Ways to Give > Volunteer > Recognition
Volunteers share their heart, mind and soul each and every day at Genesis. We celebrate their service and dedication during Genesis Volunteer Week, April 5 to 11.
Our volunteers are always ready to lend a helping hand ... always ready to share the compassion in their hearts. Volunteers are a part of our team and are an important part of an exceptional patient experience at Genesis.
During the week, a banquet is held to thank and honor all of our volunteers. We also award several volunteers for their years or hours of service. Following are this year's award recipients.
Linda Beader
Janet Bone
Kristy Corbin
Isabelle Dawson
Cheryl Dreier
Barney Emmert
Becky Hancher
Celeste Jennings
Sister Bernadette Selinsky
Lisa Smith
Carolyn Taylor
Molly Tharp
Lynda Wagner
Ruth Ware
Jerry Wolfe
Beth Anders
Penny Boyer
Jack Campbell
Pauline Davis
Ron Davis
Beverly Elekes
Lois Gruenebaum
Martha Hardesty
Jim Hinze
Scotti Hinze
Alice Jones
Michael Kieffer
William Klein
Gen Maniaci
Tony Maniaci
Robin Martin
Lynn McGlade
Pat Payne
Sue Wolf
Jerry Zwelling
Marie Campbell
Nancy Clever
Anne Kuhlman
Rose Ann Lee
Sue Matson
Shirley Paxton
Charlotte Sayles
Karen Spriegel
Ron Swope
Judy Trout
Ed Yetter
Wanda Carmichael
Linda Dicks
Sister Mary Ann Nugent
Bill Porter
Martha Ross
Margene Ryan
Jim Snider
Chelsea Connar
Malcolm Frye
Mary Evelyn Parkinson
Ron Patterson
Tom Porter
Shirley Sands
Walter Sands
Charlotte Bregnard
Wayne Cunningham
Tom Dusenbery
Bill Timmons
Pat Dusenbery
Melvina Larrick
Rosalie Snider
Ken Wells
Karen Curtin
Jay Wiseman
Mellvina Gibbs
Wanda Hagley
George Kackley
Ada Spillman
Bev Little
Sally Sowers
Patti Frame
Linda Kuhn
Lorraine Sealover
Whitey Mauk
Jane Willey
Evelyn Bates
Charlotte McGuire
Terri Carpenter
Pat Frederick
George "Bud" Hardesty
John Schaumleffel
Donna Fischer
Carol Hoffman
Joseph Walter
Carolyn Fox
Harry Freeman
Kay Steen
Dayle Zechiel
Charles White
Janet Coffey
Melody White
Marlene Work
Help Genesis provide top-quality health care to our community by volunteering at the hospital or for our hospice services.
Apply online today
Download our application [PDF] to become a patient and family advisor, and learn more about what the role requires.


text:
$ 19.99
The shirt says it all. Any self respecting Cardinals fan can't like those Seahawks! Make sure everyone knows which side you're on. All designs are the protected intellectual property of Smack Apparel; © 2016 Smack Apparel Inc.
It's your first amendment right to stay in the know.
© 2017 Smack Apparel.
Make sure your Smack Apparel t-shirt fits appropriately:


text:
Each month, Rated People connects over 50,000 homeowners with quality tradesmen - simply enter your job details in the form above to get started.
We'll match your job to the relevant tradesmen in your area - those interested will pay to see your contact details and call you to quote.
As each tradesman expresses interest in quoting on your job, we'll send you their profile and ratings to help you choose the right one for you.
After the work is complete, return to the site and rate your tradesman based on their quality of work, reliability and value for money.
Added 22/05/2016
sunroom extension that has to be built up and taking wall out of kitchen to fit a central...
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Reconfiguration / Renovation Services required: electrical, painting / decorating, tiling...
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Unsold lead, updated details: Reconfiguration / Renovation Services required: bathroom installation,...
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fitting out a barn conversion, to include new staircase ,windows x7, garage doors x3,internal...
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We are in the process of purchasing a house in Castle Cary and wish to get some quotes for...
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The results speak for themselves...really good job done
Very pleased with the job. Substantial internal wall removed, steal beam installed, boarded and plastered all in one day. Would recommend.
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Highly recommend using this tradesman. Very reliable and did a superb job!
I was very pleased work, they were always quick to respond to any emails and they showed up on time as well!
Managing your job is easier with the Rated People Homes App, download it today.


text:
Half-life of EPA
Half-life of EPA
For sheer mass gains it sounds like low fish oil intake and high inflammatory oil intake seems best.
However I'm not about to abstain from fish oil, so my question is how long until the fish oil is "out of your system" to maximize muscle gain?
I've read that the half-life can be 1.5 to almost 3 days, but I know HL depends on dose.
Any ideas how long it takes for high doses for it to get out of the system enough for stuff like X-Factor to work optimally?
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text:
Results 1 to 7 of 7
May 6th, 2008, 01:24 PM #1Delay in updates
Panic set in when I checked my Adsense this afternoon but it seems I was not the only who had problem seeing updates. Quickly checked WW forums and took a deep sigh....
Anyone here having probs?
May 6th, 2008, 01:30 PM #2
Same here - low figures have not changed since logged in this morning.Since June 10, 2012 a vegan aarf but still writing the Hound Dawg Sports Blog
"If you don't have time to do it right, when will you have time to do it over?" -John Wooden;
"Raj, theres no place for truth on the internet." -Howard Wolowitz[/SIZE]
May 6th, 2008, 01:37 PM #3
- Join Date
- January 18th, 2005
- Location
- West Coast USA
- Posts
- 3,043
Same here - low figures have not changed since logged in this morning.
ADWORDS I have sent a trouble ticket, for what that is worth
May 6th, 2008, 01:39 PM #4Off Topic - no Google impressions today
Has anyone else noticed that Google Adsense reporting is broken?
No impressions or clicks are being recorded today.
<Merged thread>
Last edited by Kevin; May 6th, 2008 at 06:34 PM. Reason: Merged thread
May 6th, 2008, 01:40 PM #5
Mines been blank all day, - No data available -
May 6th, 2008, 02:05 PM #6
Yeah - 2:00 p.m. and I'm only at 15 cents. Ouch, that's less than 1/200th of where I normally am at this time of day. My sites are all working, so evidently the Big G is not having a good day.Bill / Marketing Blog @ 12PM - Current project: Resurrecting my "baby" at South Baltimore..
Cute Personal Checks and Business Checks
If you are too busy to laugh you are too busy.
May 6th, 2008, 06:53 PM #7
Sorry guys. I goofed up a merge/move. Should be all set now.Kevin Webster
twitter: levelanalytics
Kayak Fishing
Web Analytics and Affiliate Marketing
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off topic but interesting: GoogleBy Roland in forum Midnight Cafe'Replies: 4Last Post: May 19th, 2004, 07:25 AM
Google Page Rank is Off Topic?By jaskemr in forum Commission Junction - CJReplies: 3Last Post: November 4th, 2001, 12:17 AM


text:
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This review is from: Nikon 2137 SLR Lenses AF NIKKOR 50mm f/1.8D Lens Black
Pros: Very fast lens
Bokeh is great
Very inexpensive
Works on FX and DX sensors
Fast focus
Cons: No internal AF motor, so you need a camera that has a AF motor or be okay with manual focus
Does not come with lens hood
Focusing is loud
Other Thoughts: Make sure your camera has an AF motor built in (higher end Nikons, such as D7000, D7100, any FX camera) otherwise the lens will be completely manual.
Also remember if you have a DX sensor, your lens won't be f/1.8 @ 50mm, but something like f/2.7 @ 75mm.
I would say this is probably the best starter lens if you want to get into photography. It's very inexpensive, works on FX and DX cameras just fine (and older Nikon film cameras--probably as old as 1960's Nikons), crystal clear and absolutely sharp in all areas of the image, and very small and lightweight. Plus, since it is prime, you cannot zoom and are forced to move around to compose your photograph.
This review is from: Remote Shutter Release Switch MC-Dc2 For Nikon D90 D3100
Pros: • It has a button on one end and a 10-pin Nikon connector on the other
• You press the button, the camera takes a photo
Cons: There's a "hold" feature on this device. There's a plastic slide that goes over the button to hold it down. However, this item is very cheaply built and the slide gets in the way.
Other Thoughts: If you're looking for a cheap shutter release, then this is the item for you! It works, it's cheap, and it feels cheap. I don't know if I would like to use this if my camera is in "bulb" mode due to the slide, but for any timed or mirror up release, this is fine.
The description says this works on a Nikon D90 and D3000 (if I recall correctly), but also works on a Nikon D7000 and D7100 (and should work with any camera with a Nikon 10-pin connector).
This review is from: Nikon D7000 16.2MP DX-Format CMOS Digital SLR Camera - Body Only
Pros: • High build quality (magnesium alloy, as opposed to plastic), feels "right" (hard to describe good build quality, but you know it when you see it)
• Good value for "low level professional" camera (cheaper than D300S at the time of this post, has more features)
• Dual SD card slots can expand the number of shots you can take or use as an instant backup
• Excellent image quality if paired with good glass
• Light weight
• Offers the same features as professional level cameras (i.e., mirror up, takes a cable release, 6 frames per second, takes excellent video)
• Commonly-used settings (ISO, white balance, exposure compensation, flash compensation) are buttons, not options in the menu
• Body-only version, so you don't need to have the kit lenses (which aren't bad, but it's better to save your money and buy good glass)
Cons: • Slow buffer (it goes 6 frames per second, but only for a second or two in raw--it has to write to the card before it will let you take more photos) As I don't do sports photography (at least for a living), this does not matter to me
• Crop sensor--for a few hundred dollars more, you can get a full-frame sensor with similar features. But you'll need full-frame lenses.
Other Thoughts: If you haven't upgraded to full-frame lenses but need a new camera body, this (or the D7100) is the choice.READ FULL REVIEW


text:
Raspberry Crumble Muffins
|Plain flour
|6 Ounce (170 Gram)
|Caster sugar
|2 Ounce (55 Gram)
|Light brown sugar
|1 3⁄4 Ounce (50 Gram)
|Baking powder
|2 Teaspoon
|1⁄8 Teaspoon
|Ground cinnamon
|1 Teaspoon
|4 Ounce, melted (115 Gram)
|4 Fluid Ounce (125 Milliliter)
|Fresh raspberries
|5 Ounce (140 Gram)
|Grated lemon rind
|1 (Rind Of 1 Lemon)
|Finely chopped pecans/Walnut
|1 Ounce (30 Gram)
|Dark brown sugar
|2 Ounce (55 Gram)
|Plain flour
|3 Tablespoon
|3 Tablespoon, melted
1. Preheat a 350°F/180°C/ Gas 4 oven Grease 12 deep muffin tins or use paper cases.
2. Sift the flour into a bowl. Add the sugars, baking powder, salt and cinnamon and stir to blend.
3. Make a well in the centre. Place the butter, egg and milk in the well and mix until just combined. Stir in the raspberries and lemon rind. Spoon the mixture into the prepared tins, filling the cups almost to the top.
4. For the crumble topping, mix the nuts, dark brown sugar, flour and cinnamon in a bowl. Add the melted butter and stir to blend.
5. Spoon some of the crumble over each bun. Bake until browned, about 25 minutes. Transfer to a rack to cool slightly. Serve warm.
Recipe Summary
Rate It
Nutrition Rank
Nutrition Facts
Serving size
Calories 497 Calories from Fat 236
% Daily Value*
Total Fat 27 g41.5%
Saturated Fat 14.6 g73.1%
Trans Fat 0 g
Cholesterol 94 mg31.3%
Sodium 194.7 mg8.1%
Total Carbohydrates 62 g20.6%
Dietary Fiber 3.4 g13.5%
Sugars 29.6 g
Protein 5 g9.6%
Vitamin A 14.7% Vitamin C 12.2%
Calcium 17.8% Iron 4.6%
*Based on a 2000 Calorie diet


text: I started my music career at age 10 in the church, traveling throughout the island of Jamaica up until the age of 15 when I migrated to the United States. I took a break for a while and seriously started writing, arranging music and doing gigs with the band, Musical Disciples. I have what it takes to work any stage. I bring real music to diverse audiences with a unique flavor. God is my inspiration. I put Him first, then music, then people. I respect life and nature and hope for mankind to live together as one. Love and respect to all living creatures. One love.


text:
Shongum Playground Partnership Inc
Our Mission+ Edit Mission Statement
Add a mission statement for "Shongum Playground Partnership Inc." What do they do and what makes them unique?
Start a Conversation
Shongum Playground Partnership Inc is a Central Corporation. For Tax purposes Contributions are deductible. The organization is a Organization which receives a substantial part of its support from a governmental unit or the general public 170(b)(1)(A)(vi) They currently have assets between over $50,000,000 and income between over $50,000,000 .
Financial information in-depth
Form 990 documents
We do not have Form 990 documents for this nonprofit yet. Check back periodically.
Assets and Revenues reported None
- Asset Amount:
- Income Amount:
- Form 990 Revenue Amount:
Organization Information
- NTEE Code:
- Recreation & Sports
- Physical Fitness & Community Recreational Facilities
- Parks & Playgrounds
- Ruling Date:
- Dec. 1, 1998
- Group Exemption Number:
- Subsection Code:
- Affiliation Code:
- Central
- Classification Code:
- Deductibility Code:
- Contributions are deductible.
- Foundation Code:
- Organization which receives a substantial part of its support from a governmental unit or the general public 170(b)(1)(A)(vi)
- Activity Code:
- Organization Code:
- Corporation
- Exempt Organization Status Code:
- Organization that did not respond to an IRS CP 140 notice requesting information on its continued exempt status
- Advanced Ruling Expiration Date:
- Filing Regulation Code:
- 990 - Required to file Form 990-N - Income less than $25,000 per year
- Filing Regulation Code:
- No 990-PF return
- Asset Code:
- over $50,000,000
- Income Code:
- over $50,000,000
- Account Period:
- September
This is my nonprofit
Become accredited today and earn the trust of your donors.Get Started


text:
Bone Drawing Fail14
By jason in Design Fail on June 15, 2011
i didnt know Snookie was endorsed by treatco
I love subliminal messages!
the fuck wrong with you its obvious the dogs got a boner.
thus there's no fail (being sarcastic)
Doggy Viagra?
Feed this to your dog for a bigger bone. Women love this treat.
omg keith your comments are filling up the "recent comments" lol
Knick knack paddy whack, son!
and the cat treats are a pussy holding a vagina
You've got great insights about drawing, keep up the good work!
lawl it's a double fail as far as i'm concerned. Looks like it says Neggars 'n Jewsers… hahahah and then the boner… soooooo FAIL!


text: Amazing new 913 fairways & hybrids!!!! Tony H 14 January 2013 By Tony H Well I was one of the lucky ones to have a fitting for the new 913 fairways/hybrids this morning....after my excellent fitting last year for the 913 driver I was so impressed I had to go back!!! Once again, an unbelievable experience at the Celtic manor fitting centre and I highly recommend anyone to go to a fitting centre for their next clubs! Turned out to be an expensive visit as I'm looking at my 4 iron & 3 iron hybrid to be replaced with the new 913H and they were SOOOO good!!! I am now debating on the 3 wood as on the day the numbers and consistency from my RBZ 3 wood were beating the 913 fd setup that we looked at, decision time!! A great experience and the service I've received from titleist is nothing but first class....Rory what have you done...lol slowly making the switch though, just the irons and putter left that aren't titleist!!!! :-) Quote Reply


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dc showcase presents green arrow free download PWc8 superfile
Publisher's Description
Title: dc showcase presents green arrow
Languages: Multiple languages
Last week:
Last updated:
03/22/2015 01:53:23
OS support:
Windows (all)
User rating:
[4.2 out of 5] (193 votes)
TOP Downloads dc showcase presents green arrow:
03/22/2015 01:53:23
DC Showcase: Green Arrow is a 2010 short animated film, directed by Joaquim Dos Santos and written by Greg Weisman, featuring Neal McDonough as Green Arrow, …7/18/2013 · Green Arrow protects a young princess from assassins while waiting for Black Canary at the airport. For more DC Comics, subscribe NOW: http://bit.ly ...Directed by Joaquim Dos Santos. With Neal McDonough, Malcolm McDowell, Steve Blum, Grey Griffin. At an airport, the Emerald Archer must protect a young princess …You are going to Watch DC Showcase Green Arrow Movie online free episodes with HQ / high quality. Stream Anime DC Showcase Green Arrow Count Vertigo hires Merlyn …Showcase Presents is a collection of trade paperbacks featuring various characters from the Silver Age and Bronze Age era of DC Comics publishing.DCComics.com: Welcome to the Official Site for DC Comics. DC Comics is home to the "World's Greatest Super Heroes," including SUPERMAN, BATMAN, WONDER WOMAN, GREEN ...6/30/2014 · Sipnosis: Catwoman intenta rastrear un envío de carga misteriosa que está vinculado a un jefe criminal de Gotham City llamado Rough Cut. DC Showcase : es ...Green Arrow is a fictional superhero who appears in comic books published by DC Comics. Created by Morton Weisinger and designed by George Papp, he first …Directed by Joaquim Dos Santos. With Gary Cole, Alyssa Milano, Jeff Bennett, Rob Paulsen. As Detective Jim Corrigan investigates a murder, The Spectre delivers ...DC Trade Paperbacks (TP) InStockTrades ... In this volume collecting issues #84-88 of the award-winning series, more members of The Trust fall, and one family head ...
dc showcase presents green arrow
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text:
Danny Boy
By Joseph Flummerfelt
SATB choir (a cappella) - Difficulty: Medium (Adult)
Traditional. Arranged by Joseph Flummerfelt. Joseph Flummerfelt A Choral Series. Main Key: E Flat Major. Folk, Secular. Octavo. 4 pages. Duration 1 minute 40 seconds. Published by Hinshaw Music Inc. (HI.HPC7077).
Item Number: HI.HPC7077
Key: E flat major. Text: Frederick E. Wetherby.


text:
Is Creative Advertising incorrect or missing key business information? Update your Hotfrog listing today.
Contact details
Are these details correct?
Is Creative Advertising in 4, Videsh Viram Building, Near Shivmurti, Kankhal Road, Mayapur Niranjani Akhara Haridwar H O, Haridwar Uttar Pradesh 249401 your business? Claim your listing and attract more leads by adding more content, photos and other business details.
We have more Advertising services in Haridwar Uttar Pradesh available on Hotfrog INDIA. You can update your search for Haridwar Advertising by location, keyword or service options.
Peeplan Peeth Bazar HARIDWAR Uttar Pradesh 249407
7th KM Haridwar-Delhi Road Jwalapur HARIDWAR Uttar Pradesh 249407
2 bilkeshwar colony HARIDWAR Uttar Pradesh 249401
Plot No.51,52,53 sector 8A,IIE sidcul HARIDWAR Uttar Pradesh 249404
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Write a review!
0 reviews of Goodfellas Pizza & Wings, first by you?
Goodfellas Pizza & Wings Articles — Things To Know Before You Go
Photo by redpeyton Pizza is a food so beloved in the U.S. it seems like it's been around forever, but there are some immediate clues that pizza as we know it today is a relatively new invention. ... read more
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Is Gul Technologies Singapore incorrect or missing key business information? Update your Hotfrog listing today.
Contact details
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Hotfrog Singapore provides information regarding Gul Technologies Singapore in Singapore. Gul Technologies Singapore is located at 9 Oxley Rise #03-02, Singapore 238697 and provides Automotive services. Contact them on 6223 7211.
Is Gul Technologies Singapore in 9 Oxley Rise #03-02, Singapore 238697 your business? Claim your listing and attract more leads by adding more content, photos and other business details.
We have more Automotive services in Singapore available on Hotfrog Singapore. You can update your search for Singapore Automotive by location, keyword or service options.
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text:
Domain Cos. has broken ground on the $48.4 million Paramount at South Market, the first structure in its $200 million mixed-use project in New Orleans. Plans for the project include 600 apartment units and thousands of square feet of retail space, as well as a parking garage. Developers are still seeking financing for the project.
Full Story:The Times-Picayune (New Orleans)


text:
- Type
- Commercial
- Pets
- No
- Baths
- 1.00
- Ft2
- 900
- Lease
- Flexible
General Amenities
Interior Amenities
High Traffic Exposure
900 Sq. Ft.
Includes all Utilities
ADA Compliant Bathroom
Newly Remodeled
Flexible Lease Terms
$12.00 Sq. Ft. ($900.00 Per Month)
For more information contact:
Bob Leach
1901 Broadwater Ave, Ste.1
Billings MT 59102
406-254-2468 or 406-670-6358
Billings, Montana 59105
United States
Western Managment
If you would like to send this property to a friend that you think may be interested, please complete the form below. To send this property to multiple friends, enter each email separated by a comma in the 'Friends Email' field.
Billings, Montana 59105
United States
Western Managment


text:
Content as my skin goes numb...
from the chilling uncertainty standing on railroad tracks.
The noise is deafening. The blinding light ahead,
be it a beautiful omen suggesting bloody dilapidation.
Jumping before the heart panics me into misperception.
© Copyright 2017 Carly Hill. All rights reserved.
Status: Finished
Genre: Thrillers
hey welcome to Booksie
while reading this i got goose bumps
well expressed your feeling on words
keep writing
would love to see more of your works
dont forget to update me your work
Poem / Mystery and Crime
Miscellaneous / Thrillers
Paste the link to picture in the entry below:
Paste the link to Youtube video in the following entry:
Cannot annotate a non-flat selection. Make sure your selection starts and ends within the same node.
An annotation cannot contain another annotation.
There was an error uploading your file.


text:
'I can hope and try, not predict'
Hyderabad: Indian athletes are projected to win medals in a handful of disciplines at the London Olympics, but none is closer to the goal than iconic badminton star Saina Nehwal.
Saina spoke to IANS on her Olympic chances with measured equanimity.
"It is the highest level of the game and a dream for any sportsperson; I can only hope and try, cannot predict.
"Let's wait and see what London holds for me. My coach Gopi sir (Pullela Gopichand) and the team are doing their best to help me with my game and build confidence level," said the badminton star.
Four years ago when she played at the Beijing Olympics she was a teen - 18 - and she became the first Indian to enter the badminton quarterfinals at the Olympics. Now she is heading for London with confidence, experience and with several titles in the run-up to boot. She has emerged a clear threat to the badminton superpowers.
Those who have followed the Hyderabadi's badminton from close quarters insist the 22-year-old is India's best bet for a medal in London. Coach Syed Mohammed Arif, who has seen Saina's rise, is one who is willing to put his money on her to come back with a medal.
The expectations from Saina have risen after her splendid showing on the circuit in recent months. After bagging the Thailand Open and Indonesia Super Series titles last month, the world number four stood up to the domination of the Chinese girls and proved they can be beaten.
Saina has defeated all top players of the world except current No.1 Wang Yihan, one of the three Chinese who will be in action in the women's singles at the London Olympics.
"Let us see what happens. I will give my 100 percent in all of the matches," she said when asked if she is confident of beating Wang this time.
Saina believes all opponents are going to be tough as every player is working very hard for the prestigious competition.
Will she be under pressure of expectations?
"No, it all boils down to what happens on that particular day against an opponent. I believe in hard work and leave the rest to the Almighty. "
The Chinese are not the only ones she has to deal with. She has other difficult customers from other countries, too, on her way to the medal round.
As for the Chinese, she was beaten by four Chinese five times. Only Wang Yihan is a definite threat to her as she could not beat her even once in her five meetings. The other player who has harassed her is third-ranked Li Xuerui.
Should China prefer to field Asian champion and third-placed Li Xuerui ahead of the second ranked Wang Xin or fourth-placed Wang Shixian, it will add to Saina's worries.
Saina has an identical record against Li Xuerui and World No.2 Wang Xin, winning twice in six meetings. She has won thrice against fourth-ranked Wang Shixian in four encounters.
Has Saina and her coach worked out a strategy to tackle the Chinese. "It is all spontaneous and on that particular day how our strategy will work; we are concentrating on skills, stamina and my speed."
The Haryana-born champion is going through intensive practice sessions with four other Olympics-bound shuttlers at the Gopichand Badminton Academy. She said her preparations were "going in the right direction under the watchful eyes of my coach."
She is physically and mentally geared up for the challenge. "Yes, I am and looking forward to London."
On leading the Indian badminton challenge, Saina said: "It is indeed a matter of pride to represent my country in a second Olympics. I hope my other four friends will also be elated like me to be the part of Indian team."
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text:
Catch it! Children's Event
Reigate Deanery Churches
Saturday, 7 January 2017 from 18:00 to 22:15 (GMT)
Who's Going
San Francisco, California
London, United Kingdom
Florence, Italy
Event Details
Catch it! Is a children's event for everyone in school year 3-7 from around the Reigate Deanery area.
With fun and games, sport, art and activities all under one roof!
Encouraging children to worship God with their whole life and everything they do in it. There will be workshops and gathered worship for all to join in with.
*Catch it! may include sporting and art activites, we advise wearing approriate clothing.
What is the date and time of the event?
Saturday 13th May 2017
Where can I contact the organiser with any questions?
Contact Anita and Richard Colpus on 01737 243846
Do I have to bring my printed ticket to the event?
No you do not need to print a ticket.
Are there an age limit to enter the event?
The guideline for age is school years 3 to 7, we realise that there will be exceptions so please contact for more info.
Can I stay with my child?
Yes you can stay with a child if you would like to, please speak to the organiser for advice.
Can my childs friend come along too?
Yes the event is open to all.
Is there a cost?
The cost of the event will be £5.00 per person for the whole afternoon including activities, equipment and refreshments which can be made on the day.
When & Where
St Luke's Church
Church Road
RH2 8HY Reigate
United Kingdom
Saturday, 7 January 2017 from 18:00 to 22:15 (GMT)
Add to my calendar


text:
Welcome to the keynote forum, plenary speakers, delegates, collaboration, exhibitors, sponsors, the press, and other participants to the International summit on Nursing & Health care. It's an exciting time for attending Innovate Nursing 2017 as we continue to grow and adapt, remaining always adaptable, motivated and responsive for "Advanced Research and scientific approach in Nursing and Healthcare". Our organization is confronting a time of many changes and we're meeting these changes during a time of larger nation-wide and global change. The world of Nursing and healthcare is an exciting area in which to work on clinical case studies, and we'll continue to meet and bring inspired people together in forums like this, to ensure our Innovate conferences remains at the cutting edge.
We would like to give you an idea of what you can expect and what we hope to achieve.
- Category: Health
- Tags: health care, nursing


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text: Collegiate Flags > Marshall Thundering Herd Flag Marshall University Banner Quantity $38.95 Out of Stock Add to Cart IN STOCK Standard & Express Shipping Available Order by 2pm CST (M-F) — Ships Same Day Item ID#: UB-Banner8-MTH Tweet Wish Share View All Marshall Thundering Herd Flag ITEMS Our Marshall University Banner measures 2x8 feet, is constructed of thick 100% nylon and has screen printed banner logos for Marshall University as shown. The Marshall University Banner is bordered with ten (10) reinforced metal grommets for hanging at your next tailgate party, in your game room, or as an interior or exterior gameday decoration.All college banners are officially licensed by the NCAA and selected university. Customers Also Bought... Flag & Pennant Accessories USA 3x5 Ft. Flag $28.95 Out of Stock Add To Cart


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Arts, Design, Fashion & Beauty, Society & Culture, TV & FilmFollow
|Fashion & Beauty
MODTV® Fashion Video Podcast takes you backstage at fashion week in New York, Milan and Paris with the world's top models, fashion designers, beauty pros and celebrities. Created by Karen Morrison, Executive Producer & Director MODTV. For more information write us at [email protected] or visit our website MOD-TV.COM
March 18th, 2013
Episode 161 of 346 episodes
Actress Emma Stone takes us backstage at Calvin Klein Collection for Fall 2013. Designer Francisco Costa tells us why he loves to celebrate the female form.
You'll need to sign in to write a comment.
Welcome to the daily audio workout for your head.


text:
HSN focuses on its digital and mobile future
Digital sales generate 55% of HSN's overall sales, and the retailer is looking to new platforms, such as Facebook Live, to acquire customers.
HSN focuses on its digital and mobile future
Digital sales generate 55% of HSN's overall sales, and the retailer is looking to new platforms, such as Facebook Live, to acquire customers.
Wal-Mart gives Jet shoppers a taste of its house brands
Sam's Choice and Great Value are among the Wal-Mart brands now available on Jet.com.
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Sponsored by 2013 Social Media 300
With anticipation that web sales will grow by 133% in 2006, The Fruit Company is rolling out a new shopping cart, order management system, and site design changes.
The Fruit Company's e-commerce business is bearing so much fruit that the online retailer is overhauling and updating its infrastructure to keep pace with demand.
In addition to rolling out a new shopping cart and order management system, The Fruit Company, No. 485 in the Internet Retailer Top 500 Guide to Retail Web Sites , is also redesigning major portions of its web store.
Working with Aivea Corp., a Portland, OR, e-commerce and systems development company, Fruit Company will soon introduce a new shopping cart that reduces the check-out process by at least two steps. The new shopping cart also features new enhancements for customers to track their order status and shopping history and other information.
Fruit Company is using Aviea to help with systems integration and programming, but most of the work is being designed and implemented internally, says vice president of marketing and technology Matt Stuewe. The upgrades are needed to keep pace with online sales, which could top $10 million in 2006, an increase of 133% over web revenue of $4.3 million in 2005.
With rising sales, Fruit Company sees an excellent opportunity to carve out a greater niche in the competitive online gifts market. But the key to winning new customers and retaining more repeat business is better customer service. To obtain better customer service, Fruit Company is designing and implementing a new order management and fulfillment system. The new system will require less employee training and help with expediting order management. "With the new system we can do a better job of picking, packing and shipping our East Coast orders and our West Coast orders," Stuewe says. "The delivery companies have earlier East Coast deadlines and now we can do a better job with adjusting orders and meeting schedules."
Fruit Company also is updating its site design with new drop-down menus at the top of the page. The goal is to get customers to the specific merchandising categories they are interested in, such as fruit baskets or gift towers. "We've been working on the systems upgrades for about one year," Stuewe says. "It's been worth the effort and we will reap the benefits for a long time to come."
Reason one: They're constantly trying to sell their customer, rather than to help and engage ...
Posted 02/23/17
Consumers will engage with products and brands in new ways online in the year ahead.
Posted 02/23/17
The Supreme Court is surely going to find this question right back on their doorstep next year."


text:
This project received 9 bids from talented freelancers with an average bid price of $2469 USD.Get free quotes for a project like this
Project BudgetN/A
Total Bids9
Project Description
My company needs to hire a coder capable of developing successfully a social network website focused in fashion and photography.
We are looking to accomplish a quality website that could bring its users a lot of services that will be specified later. the main features the website need to include are:
-Private accounts.
-blogging tool
-sharing pictures tool (similar to [url removed, login to view])
-virtual shop
-premium tools
The objetive is to create the web's largest fashion community site
specifications will be given later.
Thanks for the attention.
## Deliverables
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Results 16 to 20 of 21
Thread: Is Luca/Luka too feminine?
June 23rd, 2013 04:19 PM #16Simon, Eloise/Louise, Faye, Judah, Thea, Felix, Iris, Cordelia, Roscoe, Lydia, Jasper, Phaedra, Adrian, Lucinda, Jane, Conrad, Wallace, Finnegan, Sylvie, Charlie, Dashiell, Juniper, Atlas, Matilda, Julian, Alice, Marlowe, Octavia, Jack, Marigold, Archer, Gabriel, Persephone, Raphael, Dov
Just a grad student, dreaming ahead...
June 23rd, 2013 05:09 PM #18Member
- Join Date
- May 2013
- Posts
- 44
June 23rd, 2013 05:33 PM #20Senior Member
- Join Date
- Jan 2010
- Posts
- 495
I love Luca (since the old days of the show ER). It is a beautiful name and is absolutely all male to me. It would be a handsome, strong choice!
June 23rd, 2013 06:08 PM #22Senior Member
- Join Date
- May 2012
- Posts
- 1,151
Definitely masculine to me. In a sexy, dark-haired, Euro-accented kind of way.
June 23rd, 2013 06:11 PM #24
all boy to me. It makes me think of a character from ER who's name was Luka.♥Mckinley Mayfair, Christian Wayne & Sir Angus Fionn♥
Favorites June 2016Elizabeth, Adelaide, Juniper, Amelia, Arabella, Beatrice, Clara, Charlotte, Cordelia, Hermionethe rest of the girls
William, Jasper, Edgar, Gilbert, Morgan, Henry, Sebastian, Arthur, Monroe, Basilthe rest of the boys
Lyra Primrose, Amelia Hyacinth, Emmeline Bronte/ Magnus Frey, Julian Mackenzie, Sebastian Lakethe rest of my combos


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An appliqu t-shirt with a stretchy rib neck and long layered sleeves in a striped finish. Get your little hero kitted out in this Camper Van top for fun days out.
Thank you for visiting Frugi Boys Camper Van Print Layered T-Shirt, Navy on shoppingfind.co.uk
We bring the products from many websites to one website for you to search, save and compare.


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Study Abroad Programs
Architectural engineering is an international network, as building design, planning, and construction know no boundaries. We provide our AE students with the education, expertise, and tools they need to excel as a globally engaged engineer.
In addition to the study abroad programs offered through the College of Engineering Global Education and Penn State Global Programs, the Department of Architectural Engineering has partnered with the Pantheon Institute in Rome and the University of Hong Kong and Tsinghua University in China to provide AE students with a summer of global learning and experience.
Why study abroad?
More than 40 percent of our undergraduate architectural engineering students participate in the China and Rome study abroad programs.
Participating in the AE study abroad programs allows students to:
- enrich their studies with instruction in various disciplines within the context of other cultures
- expand their knowledge of the people and culture of a country or region
- receive an experience that will enhance their academic and professional development
- increase their international understanding


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Favorite Movies: Zombie Movies, Horror, Pride and Prejudice, Miyazaki, etc.
Favorite Shows: The Walking Dead, Archer, Sherlock, Community, and Pretty Little Liars
Favorite Music: Anything except country and rap. I mostly listen to Indie-pop, Alternative, and Acoustic Indie. Some artists: Breathe Carolina, Three Days Grace, Panic at the Disco, Sleeping with Sirens, Chidos, Hot Chelle Rae, Gentlemen Hall, The Ready Set, Atreyu, Aiden, Anberlin, Passion Pit, MGMT, Jens Lekmen, Ellie Goulding, Ra Ra Riot, Fall Out Boy, Owl City, Explosions in the Sky, Go Periscope, Pierce The Veil, Escape the Fate, Sky Sailing, Maroon 5, Awolnation, Flux Pavilion, Flight Facilities, The Cure, Usher, Mayday Parade, Dizzy Gilespie, Miles Davis, and many others. Just ask or give suggestions. :)
Favorite Food: Pasta, Sushi, and Tacos <----(I love taco bell)
Favorite Video Games: Resident Evil 4 and Prop Hunt
2. Mountains
3. Sight
4. Love
5. God
6. Hearing


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Now we'll find out how new GM John Idzik handles his first potentially tough negotiation.
The Jets, trying to create additional salary-cap space, want to renegotiate Santonio Holmes' contract, ESPN's Adam Schefter reported Sunday night. You could see this coming a mile away.
Holmes has a team-high $11 million base salary (his cap figure is $12.5 million), and he's coming off a serious foot injury that could imperil his chances of being 100 percent for the season. In fact, he was due to have a second surgery this month to have plates removed from his surgically repaired foot.
Plus, Holmes can be a pain in the rear end, and you can bet Idzik would like to start fresh, removing some of the old stink from the locker room.
As of Sunday night, there was no word on whether the Jets want him to take a pay cut or simply restructure the contract. Holmes has a lot of leverage here because he has a $7.5 million guarantee and there would be an $11 million cap hit if they cut him.
Ah, but take a closer look; it's not as much leverage as you might think. There's off-set language in the contract, meaning Holmes -- if released -- can't earn more than $7.5 million in 2013. In other words, he can't double dip. He's maxed out at $7.5 million, so it might behoove him to take a haircut on that $11 million, knowing he'd make the most money by remaining with the Jets.
Holmes is a proud, stubborn and prickly guy, so there's no telling how he'll react to this. He could tell the Jets to jump in the lake. If he does, as much as it would hurt cap-wise, I think there's a chance they could cut him.
Of course, a healthy Holmes is the only playmaker on the Jets' offense. As weird as this may sound, Rex Ryan believes Holmes' season-ending injury hurt the team more than Darrelle Revis' season-ending injury.