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Objective: To determine whether synovial expression of triggering receptor expressed on myeloid cells 1 (TREM-1) is upregulated in patients with distinct types of inflammatory or non-inflammatory arthritis.
Methods: Synovial fluid (SF) samples were analysed for levels of soluble TREM-1 (sTREM; n = 132), tumour necrosis factor alpha (TNFα, n = 78) and leucocyte TREM-1 messenger RNA (n = 48). Synovial tissue from four rheumatoid arthritis (RA) patients, two patients with Crohn's-associated arthritis, one patient with ankylosing spondylitis and one patient with osteoarthritis were examined for TREM-1 expression by immunohistology, and three of the RA samples were also analysed by Western blotting.
Results: Synovial fluid sTREM-1 levels in septic arthritis and RA were similar to each other and were each greater than those in gouty arthritis, non-septic/non-RA inflammatory arthritis and non-inflammatory arthritis. Synovial fluid TNFα and sTREM-1 levels correlated with each other, and sTREM-1 and leucocyte TREM-1 mRNA levels each correlated with SF leucocyte counts. TREM-1 in RA was expressed in situ in synovial tissue by cells of myelomonocytic lineage but was not detectably expressed in control osteoarthritis synovial tissue.
Conclusions: Synovial TREM-1 expression is increased in septic arthritis and RA. In patients with acute inflammatory arthritis, elevated SF sTREM-1 levels may point the clinician to a diagnosis of septic arthritis or RA. In RA patients, targeting TREM-1 may have therapeutic benefits by reducing local proinflammatory cytokine and chemokine release.
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Many conditions of widely differing aetiologies give rise to effusions within joint cavities. Inflammatory synovial effusions, regardless of aetiology, are characterised by the recruitment of numerous inflammatory cells into the synovial space mediated by, and resulting in, elevated levels of multiple cytokines and/or chemokines. Differences in synovial fluid (SF) cytokine concentrations among patients with discrete inflammatory and non-inflammatory arthritides have been described but have not been universally appreciated.1 2 Identification of some cytokines that could reliably distinguish among the various arthritides could potentially have great diagnostic utility and might afford new insights into disease pathogenesis.
Triggering receptor expressed on myeloid cells 1 (TREM-1) is a recently described member of the immunoglobulin superfamily expressed on neutrophils and a subset of monocytes and tissue macrophages.3 4 5 When bound to its as yet unidentified ligand(s), the transmembrane domain of TREM-1 associates with DAP12, and in turn triggers the secretion of inflammatory mediators such as tumour necrosis factor alpha (TNFα), IL-8, granulocyte macrophage colony-stimulating factor, monocyte chemotactic protein 1 and myeloperoxidase.3 4 5 6 Whereas crosslinking of TREM-1 alone induces modest cellular activation and proinflammatory cytokine secretion, TREM-1 acts synergistically with receptors for pathogen-associated molecular patterns.3 4 5 7 This results in considerably greater cytokine secretion than the sum of the responses induced by either TREM-1 or pathogen-associated molecular pattern-mediated activation alone. Although early studies did not reveal increased TREM-1 expression in tissue samples from subjects with non-infectious inflammatory conditions,4 5 more recent publications have implicated TREM-1-mediated inflammation in both acute and chronic non-infectious inflammatory disorders.8 9 10
In addition to the membrane-bound form, a soluble form of TREM-1 (sTREM-1) has been detected in mouse and human sera,11 12 and is suggested to arise from the proteolytic cleavage of membrane-bound TREM-1 by matrix metalloproteinases.13 In humans, sTREM-1 has been detected in plasma from volunteers injected with lipopolysaccharide and from patients with sepsis.14 15 Moreover, sTREM-1 has been advocated as a diagnostic marker of infection for patients with pneumonia and as a prognostic marker for patients with septic shock.16 17
In this study, we examined SF sTREM-1 levels in patients with infectious or distinct non-infectious inflammatory arthritis and in patients with non-inflammatory synovial effusions. As expected, we detected elevated SF sTREM-1 levels in septic arthritis. Unexpectedly, we also detected similarly elevated SF sTREM-1 levels in rheumatoid arthritis (RA) patients along with substantial in-situ TREM-1 expression on myelomonocytic cells in synovial tissue.
Materials and methods
The study protocol was approved by the Institutional Review Board of the University of Southern California Keck School of Medicine, and all subjects provided informed consent.
Patients requiring diagnostic and/or therapeutic arthrocentesis at the Los Angeles County + University of Southern California Medical Center were recruited for this study. A joint effusion was considered to be "inflammatory" if the SF leucocyte count was greater than 2000 cells/mm3. Underlying causes of joint effusions were determined by the medical history, physical examination and SF analyses (cell count, Gram stain, culture and crystal examination). Each patient's demographics at the time of the arthrocentesis and the final diagnoses are listed in table 1. Four patients with osteoarthritis and one patient with a traumatic effusion had SF leucocyte counts modestly greater than 2000 cells/mm3 ("minimally inflammatory") but were, nonetheless, included in the non-inflammatory cohort due to the underlying clinical nature of the joint effusion. Of the 18 SF samples from patients diagnosed as having septic arthritis, 14 were caused by Staphylococcus aureus, two by group A Streptococcus, one by Escherichia coli and one by Propionibacterium acnes. A definitive diagnosis could not be made in five patients with inflammatory arthritis and were labelled as unknown.
SF sTREM-1 level determination
SF was centrifuged within 1–2 h following arthrocentesis to pellet the cells. Cell-free SF was analysed for sTREM-1 levels by a recently developed sandwich ELISA, the methods for which have previously been described in detail.17 All measurements were performed in duplicate and in a blinded fashion.
Western blotting for SF sTREM-1
Protein samples were resolved by electrophoresis on a denaturating 4–12% acrylamide gel, followed by transfer to a PVDF membrane and probing with an anti-TREM-1 primary antibody (monoclonal mouse IgG1, 0.5 μg/ml, clone 193015; R&D, Minneapolis, Minnesota, USA). Polyclonal horseradish peroxidase (HRP)-labelled goat anti-mouse Ig (1/2000; Dako, Carpinteria, California, USA) was used as a secondary antibody. Binding was detected by chemoluminescence using a SuperSignal West Pico Kit (Pierce, Rockford, Illinois, USA).
SF leucocyte TREM-1 mRNA level determination
Total RNA/messenger RNA was extracted from the SF pellet using RNA STAT-60 isolation reagent (Tel-Test, Friendswood, Texas, USA) and complementary DNA was generated using the First Strand cDNA Synthesis Kit (Fermentas, Hanover, Maryland, USA). Real-time PCR analysis was performed with the Cyber Green-based real-time quantitative PCR technique using SuperArray (Frederick, Maryland, USA) TREM-1 and β-actin (control) primer sets and assay conditions and was analysed on the ABI PRISM 7900HT system (Applied Biosystems, Foster City, California, USA). Results are presented as the ratio of TREM-1 mRNA to β-actin mRNA, which was calculated by the formula: (2 exp (Ctβ-actin − CtTREM-1)).
SF TNFα level determination
Cell-free SF was analysed for TNFα levels by the solid-phase ELISA method (BD Biosciences, Le Pont de Claix, France). The sensitivity of the technique allows the detection of levels as low as 2 pg/ml. All measurements were performed in duplicate and in a blinded fashion.
Synovial tissue from four RA patients, two patients with Crohn's-associated arthritis, one patient with ankylosing spondylitis and one patient with knee osteoarthritis were collected during joint surgery, snap-frozen in liquid nitrogen, and stored at −80°C. Five micrometre-thick frozen serial sections were cut at −30°C, collected on clean glass slides and fixed for 30 s in ethanol. Immunohistochemistry was performed with the Dako Immunocytomaton (Envision HRP kit; Dako, Glostrup, Denmark) protocol and reagents. After rehydration in phosphate buffered saline (PBS), endogenous peroxidase was blocked for 15 minutes at room temperature. The sections were then incubated with unconjugated monoclonal antibodies to CD33, CD14 and HLA-DR (Beckman Coulter, Miami, Florida, USA) or TREM-1 (R&D) for 45 minutes at room temperature in a moist chamber. After three washes in PBS, a second incubation was performed with a polyclonal HRP anti-mouse antibody for 45 minutes at room temperature. After another three washes in PBS, the sections were developed with a diaminobenzidine solution in substrate buffer for 5 minutes at room temperature, washed in distilled water and ammonia and counterstained with haematoxylin.
Western blotting of synovial tissue extracts
Synovial tissue from three RA patients was extracted with the RIPA lysis buffer (Santa Cruz Biotechnology, Santa Cruz, California, USA) supplemented with protease inhibitors. The clear protein extract was subjected to sodium dodecylsulphate–polyacrylamide gel electrophoresis and Western blotting by blocking the membrane with 5% bovine serum albumin and incubating for 1 h with anti-TREM-1 (R&D) in PBS-Tween followed by HRP anti-mouse (Dako). Chemoluminescence substrate was used to reveal the bands (Chemoluminescence reagent Plus; NEN Life Science, Boston, Massachusetts, USA) for 5 minutes, which were then impressed in the dark on X-Omat blue film (Sigma-Aldrich, St Louis, Missouri, USA). After developing, the film was read with a Biorad Gel Doc XR (Biorad, Hercules, California, USA).
Statistical analysis
All analyses were performed using SigmaStat software (SPSS). Results that did not follow a normal distribution were log-transformed to achieve normality. Parametric testing between two matched or unmatched groups was performed by the paired or unpaired t test, respectively. When log transformation failed to generate normally distributed data or the equal variance test was not satisfied, non-parametric testing was performed between two groups by the Mann–Whitney rank sum test. Correlations were determined by Pearson product moment correlation for interval data and by Spearman rank order correlation for ordinal data or for interval data that did not follow a normal distribution. Analyses are presented for each cohort of patients described in table 1. Additional analyses were also performed with exclusion of the five "non-inflammatory" patients who had "minimally inflammatory" effusions and with re-assignment of these patients to the "other inflammatory" cohort. In neither case did changes in statistical significance emerge.
Elevated SF sTREM-1 levels in septic arthritis and RA
As illustrated in fig 1A, SF sTREM-1 levels in septic arthritis were greater than those in gout, non-septic/non-RA inflammatory arthritis and non-inflammatory arthritis (p = 0.003, p<0.001 and p<0.001, respectively). These results are consistent with previously published observations of increased sTREM-1 in infectious conditions compared with those in similar non-infectious states.14 17 Unexpectedly, SF sTREM-1 levels in RA were also greater than those in gout, non-septic/non-RA inflammatory arthritis and non-inflammatory arthritis (p = 0.002, p = 0.001 and p<0.001, respectively) and were very similar to SF sTREM-1 levels in septic arthritis.
As rheumatoid factor in biological fluids (including SF) can confound determinations by ELISA by promoting multimeric complexes of unpredictable sizes in the presence of the assay's detecting antibody, we serially diluted RA SF and measured sTREM-1 levels in the diluted samples. Linear responses with serial dilutions of RA SF were obtained (data not shown), suggesting little interference from rheumatoid factor in these samples. To confirm further that bona fide sTREM-1 was being detected, Western blotting of SF was performed and verified the presence of sTREM-1 (fig 1B). Its molecular weight of approximately 28 kDa strongly suggests that the measured sTREM-1 in our ELISA is a product of cleaved membrane-bound TREM-1 rather than being an alternative splice variant lacking exon 3, which would have a lower molecular mass of approximately 15 kDa.13
SF TNFα levels
One of the inflammatory cytokines secreted in response to TREM-1 ligation is TNFα.3 4 In our cohort of patients, SF TNFα levels were markedly greater in septic arthritis than in gout, non-septic/non-RA inflammatory arthritis, or non-inflammatory arthritis (p = 0.005, p = 0.040 and p<0.001, respectively) and trended towards greater levels than in RA (p = 0.079; fig 1C). SF TNFα levels in RA were similar to those in gout and non-septic/non-RA inflammatory arthritis (p = 0.243 and p = 0.547, respectively) and trended towards greater levels than in non-inflammatory arthritis (p = 0.052). These observations agree well with other studies of SF TNFα levels in patients with various types of arthritis.2 18 Of the 39 RA SF samples, 14 were from patients on anti-TNFα therapy at the time of arthrocentesis. SF TNFα levels were actually higher in these patients than in RA patients who were not being treated with an anti-TNFα agent (p = 0.031). Although no statistically significant difference in SF sTREM-1 levels between these two sub-cohorts of RA could be detected (p = 0.783, data not shown), there was a positive correlation among all patients tested between SF sTREM-1 and SF TNFα levels (r = 0.366, p = 0.001, fig 1D), suggesting a relationship between the two.
As SF from septic arthritis patients harboured the highest levels of both TNFα and sTREM-1, we excluded these patients from a secondary analysis. Among the non-septic arthritis patients, the correlation between SF sTREM-1 and TNFα levels persisted (r = 0.343, p = 0.005). When only samples with detectable levels of TNFα within this cohort were assessed, the correlation remained strong (r = 0.542, p = 0.024), demonstrating that even in the absence of infection, a relationship exists between sTREM-1 and TNFα in inflammatory effusions.
Relationship between SF leucocyte counts and SF sTREM-1 levels
As neutrophils and monocytes each express TREM-1,3 4 19 20 these inflammatory cells are candidate sources of SF sTREM-1. Given that SF leucocytes positively correlated with SF TNFα levels in our patients (r = 0.269, p = 0.018) and that TREM-1 ligation is associated with inflammatory cytokine release, we hypothesised that a relationship would exist between SF leucocytes and SF sTREM-1 levels. Indeed, among all patients, SF sTREM-1 levels did positively correlate with synovial leucocyte counts (r = 0.357, p<0.001, fig 2A). In addition, SF neutrophils, lymphocytes and monocytes individually also correlated significantly with SF sTREM-1 levels (r = 0.285, p = 0.001; r = 0.299, p<0.001; r = 0.206, p = 0.021, respectively; data not shown).
As the SF leucocyte counts in septic arthritis patients were markedly greater than those in any of the other patient cohorts (fig 2B) and may have skewed the overall results, we separately analysed the non-septic arthritis patients. The correlation between total SF leucocyte count and the sTREM-1 level persisted (r = 0.314, p<0.001; fig 2C) as well as the SF leucocyte differentials (neutrophils r = 0.227, p = 0.018; lymphocytes r = 0.345, p<0.001; and monocytes r = 0.218, p = 0.024; data not shown), suggesting that even in non-infectious effusions, recruited leucocytes contribute substantially to the elevation in SF sTREM-1 levels.
SF leucocyte TREM-1 mRNA levels
Despite the significant differences in both SF sTREM-1 levels and SF leucocyte counts among the individual patient cohorts, there were no significant differences in SF leucocyte TREM-1 mRNA levels among them (fig 3A). The median SF leucocyte TREM-1 mRNA levels, nevertheless, were greater in the septic arthritis and RA groups, so significant differences may have emerged with a greater number of samples. Indeed, SF sTREM-1 levels positively correlated with SF leucocyte TREM-1 mRNA levels (r = 0.344, p = 0.017; fig 3B) suggesting that the measured SF sTREM-1 arises at least partly from leucocytes that have migrated into the SF.
In-situ studies of TREM-1 expression in RA synovial tissue
The likely contribution of SF leucocytes to the increased SF sTREM-1 levels in RA did not preclude other potential sources of sTREM-1. In addition to TREM-1 expression by SF leucocytes, TREM-1 was strongly expressed in RA synovial tissue in a fine punctate pattern along the surface membrane of large cells located among lymphoid infiltrates in the subintimal layer (fig 4A). That TREM-1-expressing cells were of myelomonocytic lineage was established by CD14, CD33 and HLA-DR staining of serial sections (data not shown). In contrast, non-inflammatory osteoarthritis synovial tissue demonstrated essentially no surface expression of TREM-1 (fig 4B), indicating that TREM-1 is not constitutively expressed in synovial tissue. Western blotting of synovial tissue protein extracts demonstrated in each tested RA sample a strong TREM-1 band with the expected molecular weight of 30 kDa for the full non-cleaved protein (fig 4C), confirming the immunostaining results. We also observed binding of anti-TREM-1 to synovial macrophages in each sample from the two patients with Crohn's-associated arthritis and from the patient with ankylosing spondylitis (data not shown), suggesting that macrophages may contribute to synovial inflammation in other inflammatory arthritides.
In this study, we show that SF sTREM-1 levels are significantly elevated in septic arthritis patients compared with those in patients with non-septic/non-RA inflammatory arthritis or non-inflammatory arthritis (fig 1A). This preferential elevation of SF sTREM-1 levels in septic arthritis is reminiscent of the preferential elevations of sTREM-1 levels in bronchoalveolar lavage fluids from patients with infectious pneumonia and in sera from patients with sepsis.14 17
Unexpectedly, sTREM-1 levels were also elevated in SF from RA patients (fig 1A). In fact, the levels were similar to those seen in septic arthritis, and the degree to which SF sTREM-1 levels are elevated appears to be unique to RA among the various causes of non-infectious inflammatory arthritis. The sTREM-1 levels measured by ELISA in RA SF are not spuriously elevated, as evidenced by the detection of sTREM-1 bands by Western blotting (fig 1B) and the linear diminution of measured sTREM-1 levels in serially diluted SF samples (data not shown). Our finding adds to a recently growing list of non-infectious inflammatory conditions associated with increased sTREM-1 levels or surface TREM-1 expression, including inflammatory bowel disease (IBD),9 21 paraneoplastic pleural effusions,22 acute pancreatitis23 and Helicobacter pylori-negative peptic ulcer disease.24
The mechanistic underpinnings of increased SF sTREM-1 levels in RA remain uncertain. TNFα can modestly upregulate TREM-1 expression,3 raising the possibility that elevated SF sTREM-1 levels in RA were consequent to elevated SF TNFα levels. However, inasmuch as TNFα levels in RA were not significantly elevated compared with non-septic/non-RA inflammatory arthritis despite increased sTREM-1 concentrations in the former (fig 1A, C), the elevated SF sTREM-1 levels in RA cannot solely be passive reflections of SF TNFα concentrations. In addition, even though RA patients treated with an anti-TNFα agent had higher levels of SF TNFα compared with the other RA patients, the use of anti-TNFα therapy did not appear to influence the expression of SF sTREM-1.
Indeed, it is possible that heightened synovial expression of TREM-1 actually promotes increased SF TNFα levels in addition to being upregulated itself by TNFα. Ligation of TREM-1 leads to the release of several pro-inflammatory cytokines, including TNFα.3 4 5 6 The correlation between SF sTREM-1 and TNFα levels in our patients (fig 1D) suggests that local TNFα production is linked to levels of SF sTREM-1 and, by inference, to the upregulation of surface TREM-1 expression. Schenk et al21 demonstrated marked reductions in TNFα and TNFα mRNA in mice with colitis following treatment with a TREM-1 antagonist peptide, suggesting a direct link between TREM-1 expression and TNFα production. Additional studies are needed to test this possibility in RA.
Given the role that TREM-1 ligation plays in the upregulation of inflammatory cytokines and chemokines, the individual cells that express TREM-1 and release sTREM-1 in each of the discrete inflammatory arthritis conditions could have important consequences for disease pathogenesis and/or maintenance. Whereas the exact cellular source of sTREM-1 in the SF of patients with inflammatory arthritis is unknown, the positive correlation between SF leucocytes and both SF sTREM-1 levels and TREM-1 mRNA expression (figs 2A and 3B) suggests that inflammatory leucocytes recruited into the SF are, at least partly, responsible for sTREM-1 production. Whereas the significantly elevated SF sTREM-1 levels were expected in septic arthritis, the similarly increased levels observed in RA SF may reflect additional sources of sTREM-1 independent of recruited leucocytes. In our study, the mean SF leucocyte differentials for each cohort were as follows: RA 67% neutrophils (N), 21% lymphocytes (L), 12% monocytes (M); septic 92% N, 3% L, 5% M; gout 87% N, 4% L, 9% M; other inflammatory 73% N, 14% L, 13% M; and non-inflammatory 34% N, 43% L, 23% M. The differences seen in leucocyte differentials within each cohort do not appear to explain the differences seen in SF sTREM-1 levels. For example, whereas gout had a total SF leucocyte count similar to that of RA, the SF leucocyte differential in gout (relatively rich in neutrophils with the potential to express high levels of TREM-1) would seemingly have favoured higher sTREM-1 levels compared with RA (relatively rich in lymphocytes, which are relatively poor expressers of TREM-1). This, however, was empirically not observed, suggesting that alternative cellular sources of synovial sTREM-1 substantially contributed to the marked elevation seen in RA.
One possibility is that a significant contributor of sTREM-1 in RA SF arises from myelomonocyte cells within synovial tissue (fig 4A). In RA, resident tissue macrophages and fibroblasts produce the majority of cytokines and chemokines found in the affected joints, and the observed spectrum is similar to that which follows TREM-1 ligation.3 4 Supportive of this argument is the dramatic increase in the frequency and number of TREM-1-expressing intestinal macrophages in humans with ulcerative colitis or Crohn's disease and in experimental mouse models of IBD.21 Increased TREM-1 expression correlated with disease activity in both clinical and experimental IBD, and TREM-1 crosslinking resulted in marked increases in proinflammatory cytokines, including TNFα, IL-8 and monocyte chemotactic protein 1.21 Indeed, in samples from two patients with inflammatory arthritis associated with Crohn's disease, immunoflorescence staining revealed the presence TREM-1-expressing synovial macrophages (data not shown). Furthermore, the lack of TREM-1 expression in non-inflammatory synovial tissue from osteoarthritis (fig 4B) suggests that TREM-1 is not constitutively expressed in synovial tissue. TREM-1 expression is thus probably an important mediator of inflammation in several inflammatory arthritis conditions, including IBD-associated arthritis and gout. The abundance of sTREM-1 in RA may merely be a reflection of the hypercellular synovium of RA patients relative to that in patients with other types of inflammatory arthritis. Accordingly, increased TREM-1 expression in RA synovial tissue may play an important contributory role in the ongoing inflammatory response. Additional experiments utilising increased numbers of surgical samples are needed to determine if relationships exist between synovial TREM-1 expression, quantitative synovial monomyelocytic assessments and levels of SF sTREM-1.
In conclusion, sTREM-1 levels are increased in the SF from patients with septic arthritis compared with those in SF from patients with other types of arthritis, with the noteworthy exception of RA. The realisation of increased SF TREM-1 expression in RA may not only provide additional insight into the pathogenesis of RA, but may potentially suggest an additional therapeutic target. Although the endogenous ligand of TREM-1 has yet to be determined, blocking TREM-1 with a monoclonal antibody in a murine model of lipopolysaccharide-induced shock and peritonitis reduced hyperresponsiveness and lethality.6 In addition, interfering with TREM-1 engagement in murine colitis resulted in reductions of several proinflammatory cytokines and amelioration of disease.21 Accordingly, blockade of TREM-1 in an active RA patient could potentially result in the downstream reduction of excessive chemokine and cytokine release. Further studies assessing sTREM-1 and TREM-1 expression from various sites are needed in RA as well as the determination of TREM-1 expression in other autoimmune inflammatory diseases.
The authors thank all the subjects for their participation in this study, Professors D Mainard and F Sirveaux for providing RA synovial tissue samples, and Anne Sophie Didier for her technical help with the studies on the synovial tissue samples.
Competing interests None.
Ethics approval The study protocol was approved by the Institutional Review Board of the University of Southern California Keck School of Medicine.
Patient consent Obtained.
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I am writing to show my support for Robert Imperato as Branford's state representative for the 102nd Assembly District.
I met Bob six years ago when I joined the RTM. My first impression of Bob Imperato was that he has a lot of integrity. He has the unique ability to see all sides of an issue and can determine what the next course of action should be to get the job done. His integrity speaks volumes about his character and demeanor.
Bob has many years of public service to the people of Branford and has more than 40 years of expertise in finance and banking for the corporate sector. Bob Imperato is what we need in Hartford. Connecticut is not in a good place right now and Hartford doesn't know how to add, subtract or how to say "No" to bad ideas. Bob Imperato knows how important it is to stay within budget and be fiscally responsible to the taxpayer. Bob Imperato has the integrity and tenacity we need in Hartford now.
I encourage your readers to vote for Bob Imperato as our state representative for the 102nd District. He is the right person at the right time!
Tricia Anderson
Republican Tricia Anderson is a 7th District representative on the RTM.
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10122 Windy Pointe
|Why wait to build? Beautiful Lennox model situated on one of the largest lots in Marina Bay. 5 bedrooms and 4 full baths of luxury living. Stunning finishing touches like crown mold and trim packages, many lighting upgrades, gorgeous kitchen with top-of-the-line appliances and massive pantry. Master suite and 3 of the bedrooms on the second floor, 5th bedroom on the first floor is perfect for guest or live-in help. Large loft/den on the second floor is perfect for a playroom or office. Located on a quiet cul-de-sac with a 20,669 square foot lot for endless outdoor play. Marina Bay offers the amenity-rich lifestyle dreams are made of, close to dining, shopping, and the International Airport as well as Florida Gulf Coast University. MARINA BAY PLAT TWO AS DESC IN INST# 2016000103335 LOT 556
Status: A
Area: FM22 - Fort Myers City Limits
Subdivision: MARINA BAY
Acreage: .47
Year Built: 2017
Taxes: $5,013.00
SqFt: 3,437
Lot Size: 33x286x213x141
Grade School: SCHOOL OF CHOICE
Middle School: SCHOOL OF CHOICE
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Listing Office: BHHS Florida Realty - MLS ID: 219036529
The data relating to real estate for sale or rent on this website comes in part from the Broker Reciprocity Program (BR Program) of Multiple Listing Service of Naples, Inc.
Those properties marked with the BR Program Icon and/or BR House Icon are provided courtesy of the M.L.S. of Naples, Inc.'s, BR Database.
The data relating to real estate for sale/lease on this web site come in part from a cooperative data exchange program of the multiple listing service (MLS) in which this real estate firm (Broker) participates. Detailed information about such properties includes the name of the listing Brokers. No warranties, expressed or implied, are provided for the data herein, or for their use or interpretation by the user. The Florida Association of REALTORS® and its cooperating MLSs do not create, control or review the property data displayed herein and take no responsibility for the content of such records. Federal law prohibits discrimination on the basis of race, color, religion, sex, handicap, familial status or national origin in the sale, rental or financing of housing. Listings last updated:. Updating occurs nightly. Some properties that appear for sale on this website may subsequently have sold and may no longer be available.
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Ilusion Salon de Bellza
Listed in:(770) 739-5719
1588 Veterans Memorial Hwy
Austell, GA 30168
- Beauty and Spas in Austell
- Beauty Salons and Services in Austell
1588 Veterans Memorial Hwy
Austell, GA 30168
general information
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The Socceroos quest for AFC Asian Cup glory on home soil in January begins with a tricky group phase against Kuwait, Oman and Korea Republic.
We take a look at the danger-men who could hurt the Aussies in our Asian Cup group.
Name: Heung-Min Son
Age: 22
Club: Bayer Leverkusen (Germany)
Position: Winger
A club-mate of Socceroo winger Robbie Kruse, Son possesses loads of pace and is equally devastating with both feet. It means he can play anywhere along the front line and has the ability to get in behind defences with his speed and clever runs. Son, who has also played for Hamburg in the Bundesliga, has scored 34 goals in 113 appearances in the German competition. His strike-rate at international level is also pretty good, scoring seven times for the senior national team in 32 caps.
Other players to watch: Sung-Yong Ki (Swansea City), Gyo-Won Han (Jeonbuk Motors), Chung-Yong Lee (Bolton Wanderers)
Name: Abdulaziz Al-Muqbali
Age: 25
Club: Fanja (Oman)
Position: StrikerA real x-factor in Paul Le Guen's side, Al Muqbali (commonly known as Abdul Aziz Humaid Mubarak) should have the Socceroos defence on high-alert as Oman's leading attacker. Al Muqbali scored against Australia in the 2-2 draw in Sydney in a World Cup qualifier in 2013 and has amassed 10 goals in 33 internationals. Not only good with his feet, Al Muqbali is also a threat in the air.
Other players to watch: Ali Al Habsi (Brighton), Hussain Al Hadhri (Al Raed), Ali Al Jabri (Al Nadha)
Name: Bader Al-Mutawa
Age: 29
Club: Al Qadsia (Kuwait)
Position: Attacking midfielder
The most decorated of the Kuwaiti squad, Al-Mutawa is one of the survivors from Kuwait's shock 1-0 win over the Socceroos in Canberra back in 2009. He is considered a legend at his club side Al Qadsia due to his phenomenal strike-rate in front of goal and was voted as Asia's second-best player in 2006. Al-Mutawa, who wears jersey number 17 for both club and country, has scored 46 times from 141 internationals and will have to play a leading role if Jorvan Vieira's side is to go far in January's tournament.
Other players to watch: Abdulaziz Mashaan (Al Qadsia), Fahad Al Enezi (Al Kuwait), Yousef Al Sulaiman (Mazma)
The Socceroos will be looking to win their first AFC Asian Cup when Asia's biggest football tournament heads to Australia in January 2015. 32 matches over 23 days will be played in Sydney, Melbourne, Brisbane, Canberra and Newcastle with the Socceroos kicking off the tournament against Kuwait in Melbourne on January 9.
Featured News
Aussies Abroad: Leckie deployed in new role in Hertha's thrilling Bundesliga clash with Bayern Munich
Matthew Leckie was handed a new role by Hertha Berlin head coach Ante Covic in a high-octane 2-2 draw with Bundesliga champions Bayern Munich. The Caltex Socceroo was regularly deployed in an attacking position by former chief Pal Dardai but the Hungarian was replaced in the o
Van Egmond pleased with Young Socceroos' progress ahead of Myanmar Semi-Final
Gary van Egmond says his Young Socceroos deserve plenty of credit for the way they are performing and developing at the 2019 AFF U-18 Championship. The young Aussies booked their spot in the Semi-Finals of the tournament on Thursday night after a dominant 5-0 win over Singapor
Turkish Super Lig preview: Behich and Basaksehir mean business
Aziz Behich is back in Turkey with Istanbul Basaksehir, and the club at the nation?s heart well and truly mean business. ...elbourne-born Behich joined Istanbul in May, returning to the nation he was once eligible to represent through family lineage, and where he played for Burs
Bundesliga preview: Leckie and Borrello walk with giants
Australians love a challenge, and there is hardly a bigger one in the world of football than taking down Bundesliga giants Bayern Munich. But you can be sure that Caltex Socceroo attackers Mathew Leckie and Brandon Borrello will give it a red-hot crack for both Hertha Berlin a


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KPL Transfers: New signing Ochan will be a hit for AFC Leopards - Juma
AFC Leopards team manager Tom Juma believes goalkeeper Benjamin Ochan will be an instant hit in the Kenyan Premier League (KPL).
The Ugandan has massive experience having played for KCCA FC, Sports Villa SC and Kabwe Warriors, among others.
Juma admits there is real quality in the 30-year-old which will help Ingwe do well in the 2019/20 season.
Editors' Picks
"Ochan has shown us what he can do in training, he is agile and flexible and has shown real potential. I believe he will have a vital role to play for the team next season and help us reach our objectives," Juma told Goal on Wednesday.
"He commands his area well and knows what to do when on and off the ball. I think he will be an instant hit in the league if he reciprocates what he is doing here."
Ochan will take the place of Jairus Adira who is rumoured to be on his way out of the club.
The Ugandan is among seven players who have been signed by the 13-time league champions ahead of the new season which kicks off on August 31.


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Last Saturday, Ryan Hall took to the Octagon in Sacramento and beat a local favorite, Darren Elkins, by decision. While Hall is known for his world class Brazilian jiu jitsu, …read more
Source:: BJPenn.com
Ryan Hall continued his perfect run inside the UFC with a one-sided unanimous decision victory over the veteran Darren Elkins at UFC Fight Night 155 on Saturday, though his antics …read more
Source:: Sherdog
Hall wants his due after landing a bunch of spinning kicks on Darren Elkins and dropping him multiple times. Jiu jitsu ace Ryan Hall fought on the prelim portion of …read more
Source:: MMAMania
Ryan Hall defeated Darren Elkins on the UFC Sacramento prelims Saturday night, but he found himself once again on the receiving end of some criticism along the way. Even though …read more
Source:: MMANews
SACRAMENTO, Calif. – Ryan Hall beat Darren Elkins with a unanimous decision Saturday night on the preliminary card at UFC on ESPN+ 13 in Sacramento, Calif.
Take a look inside the …read more
Source:: MMAJunkie
Ryan Hall didn't get the finish this time, but what he did against Darren Elkins over three rounds at UFC on ESPN+ 13 was perhaps even more impressive than if …read more
Source:: MMAJunkie
"The Damage" Darren Elkins is known to wear it when he sets out to do battle, but on Saturday night, he faces a unique challenge.
He will take on jiu-jitsu wizard …read more
Source:: MMAJunkie
Ryan Hall is moving back down to featherweight to take on Darren Elkins at UFC Sacramento. The jiu-jitsu ace is fighting for the first time since he submitted B.J. Penn …read more
Source:: Sherdog


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- African News (52)
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Air Canada airlines Airports Alberta Aviation Museum Avcorp BC Boeing Bombardier Bombardier Inc CAE canada Corporate CSeries Edmonton Financial FLYHT Global 6000 Helicopters Manufacturing Military Montreal NAV CANADA NORAD Porter Airlines Pratt & Whitney Canada RCAF SAR SkyTrac Systems Transport Canada WestJet
African News
Ethiopian Airlines is pleased to announce the launching of a new corporate loyalty program, Corporate Bonus, enabling companies to accrue miles for passengers, who travel on their corporate account.
Tagged under
Reflecting their commitment to more comprehensively address runway safety issues worldwide over the next two years, the International Civil Aviation Organization (ICAO) and the Civil Air Navigation Services Organisation (CANSO) were joined by over 100 expert participants in Cape Town, South Africa recently at the joint ICAO/CANSO Eastern and Southern African Regional Runway Safety Seminar.
Published in African NewsRead more...
Written by International Civil Aviation Organization
Written by International Civil Aviation Organization
Ethiopian Airlines is pleased to announce that it has been recognized for its commitment to service excellence after winning two awards from African Travel Quarterly (ATQ) magazine on Monday, October 22, 2012 at an event held in Lagos, Nigeria.
Tagged under
Ethiopian Airlines, the fastest growing airline in Africa, has taken another leap forward by receiving its second Boeing 787 Dreamliner. Ethiopian second Dreamliner was delivered in Seattle on Wednesday, October 3, 2012 and arrived at Addis Ababa Bole International Airport on Friday, October 5, 2012.
Tagged under
In another milestone, Ethiopian Airlines again led the African aviation industry by being the first in the continent to receive and operate the B777 Freighter on 19 September 2012.
Tagged under
Following the lifting of the suspension on Dana Air's operating license by the Federal Government on Wednesday, September 5, 2012, the airline has begun a process of recertification by the Nigerian Civil Aviation Authority (NCAA). Also, all the staff of the airline will shortly be undergoing refresher programmes in preparation for commencement of operations.
Published in African News
Written by Dana Air
Written by Dana Air
Tagged under
Ethiopian Aviation Academy graduated 12 pilots, 125 aviation maintenance technicians and 60 cabin crew at a ceremony held at the Ethiopian Headquarters on Thursday, September 6, 2012.
Tagged under
The management and the over 558 staff of Dana Air wish to express our deepest appreciation to the Federal Government of Nigeria over the lifting of the suspension of the airline's operations with effect from Wednesday, September 5, 2012, following government's satisfaction with the air-worthiness of the airline after a rigorous technical, operational and financial audit.
Published in African News
Written by Dana Air
Written by Dana Air
Tagged under
Ethiopian continues its leadership in Africa being the 1st in the continent to take Delivery of 777F
True to its motto "leading the way past, present and the future", and in another milestone for the African aviation industry, Ethiopian Airlines will receive its first B777 Freighter aircraft on September 19, 2012 from Boeing at Everett, Washington. This will make Ethiopian the first to operate this ultra-modern freighter in Africa.
Tagged under
The International Air Transport Association (IATA) reinforced its commitment to African aviation and called on industry and government leaders in Central West Africa to work together to make aviation an even more integral part of African economic development and integration. Safety, regional cooperation and global standards for infrastructure funding were highlighted as key issues which must be addressed.
Published in African NewsRead more...
Written by IATA
Written by IATA
- Swissport awarded "International Cargo Handler of the Year – Africa" for the second time in a row
- Nigeria Validates Bell 429 Certification
- fastjet responds to claims made by CEO of Fly540 Kenya
- Embraer Defense and Security Delivers the First Three A-29 Super Tucano to the National Air Force of Angola
- Ethiopian Temporarily Pulls the 787 out of Service for Precautionary Inspection per US FAA Directive
United Arab Emirates
- Abu Dhabi welcomes one of the first flights from the new King Abdulaziz International Airport
- Salam Air launches three weekly flights from Abu Dhabi to Salalah, Oman
- Abu Dhabi Airports Wins CIPS Procurement Excellence Gold Award
- Abu Dhabi new airport empowers globetrotters with Etisalat's 5G network
- Access Ramp - E10 to MTC to be Closed Permanently
United States of America
- Press Release - Stephen M. Dickson Sworn in as Administrator of the Federal Aviation Administration
- Speech - FAA Administrator Stephen Dickson Swearing-In Remarks
- News and Updates - Stephen M. Dickson Sworn in as Administrator of the Federal Aviation Administration
- News and Updates - FAA Seeks Stakeholder Input on Drone Tests
- Vietnam Airlines Flies Its First Boeing 787-10 Dreamliner
Other International News
- City all agog over new airline
- Air New Zealand has appointed Calum Laming to the newly created role of General Manager Customer Experience
- CEO of Qatar Airways to Participate in World Routes Strategy Summit
- Rockwell Collins named one of the World's Most Ethical Companies for the fourth consecutive year
- MD Helicopters Announces Next Generation of Technology Advancements for Explorer


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Latest News
VDP sets green Pfandbrief standards
The Association of German Pfandbrief Banks (VDP) has introduced minimum standards for issuing green Pfandbriefe, a development that should help seal Germany's position as a leading jurisdiction for green covered bond issuance.23 Aug 2019
New era for bond pricing as corporates, banks push deeply negative
Europe's bond market reopened for the autumn issuance season this week — but it is a new bond market. The summer's queasy bout of bearishness has pushed government bond yields to unprecedented lows and these are now for the first time being tested as a platform for private sector bond issuance.22 Aug 2019
BHH sets new boundary for negative yielding covered bonds
Berlin Hyp, the German mortgage bank, this week set a new record with the most negative yielding covered bond, a transaction that should bolster the confidence of other borrowers who can no longer put off issuing such deals.22 Aug 2019
Equitable leads next wave of Canadian covereds
Canada's Equitable Bank has announced plans to issue its first covered bond, possibly as early as next year. Its decision follows an increase in Canada's regulatory issuance limit and paves the way for several other smaller Canadian banks to follow suit.22 Aug 2019
HSBC Canada to reopen dollar covered bonds
HSBC Bank Canada is set to meet investors with a view to issuing the first dollar covered bond since the summer break and its second covered bond ever. The plans comes amid a growing euro pipeline and follows the first deeply negative yielding euro trade.21 Aug 2019
Virgin plans inaugural euro covered bond
Virgin Money plc has mandated leads for a European roadshow and hopes to issue its first euro denominated covered bond. The transaction is set to emerge at a challenging time, amid rising concerns that the UK will chaotically leave the European Union without reaching an exit agreement at the end of October.20 Aug 2019
BHH raises €1bn 20bp below ECB deposit rate
BerlinHyp (BHH) reopened the covered bond market after the summer break with a €1bn three year that yielded almost 20bp less than the European Central Bank's deposit rate. The transaction could cause some issuers to reconsider their funding approach as the deal can profitably refinance liquidity borrowed under the last Targeted Longer-Term Refinancing Operation (TLTRO II).20 Aug 2019
ASB tees up September roadshow as issuers stick to 'wait and see' mode
ASB Finance Limited has announced plans to sell either a covered bond or a senior bond next month, as FIG bankers suggested that issuers would be careful about restarting supply following a volatile summer period.19 Aug 2019
Swiss Pfandbrief go negative out to 30 years
Yields continued to tumble in the Swiss franc bond market this week with Pfandbriefbank issuing the first 30 year covered bond to be sold with a negative yield, following Münchener Hypothekenbank (MünHyp) striking a new record low for covered bond yields with a nine year trade.15 Aug 2019
Polish mortgage law has 'significant adverse effects'
A draft Polish law, requiring the written consent of property owners to transfer mortgages, would have "significant adverse effects on Polish banks' funding" said the European Central Bank, with respect to the covered bond and securitization markets. However, a legal expert said there would be no direct impact on covered bonds.15 Aug 2019