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Conceived and designed the experiments: GAP CDP. Performed the experiments: MS MB EC. Analyzed the data: MS GAP. Contributed reagents/materials/analysis tools: SSJ EDZ. Wrote the paper: GAP CP MS.
Mesenchymal stem cells (MSC) preferentially migrate to damaged tissues and, due to their immunomodulatory and trophic properties, contribute to tissue repair. Although MSC express molecules, such as membrane cofactor protein (CD46), complement decay-accelerating factor (CD55), and protectin (CD59), which confer protection from complement-mediated lysis, MSC are recruited and activated by anaphylatoxins after transplantation, potentially causing MSC death and limiting therapeutic benefit. We have previously demonstrated that transduction of MSC with a retrovirus encoding HCMV-US proteins resulted in higher levels of MSC engraftment due to decreased HLA-I expression. Here, we investigate whether engineering MSC to express US2 (MSC-US2), US3 (MSC-US3), US6 (MSC-US6), or US11 (MSC-US11) HCMV proteins can alter complement recognition, thereby better protecting MSC from complement attack and lysis. HCMV-US proteins increased MSC CD59 expression at different levels as determined by flow cytometric evaluation of the median fluorescence intensity ratio (MFI). A significant increase in CD59 expression was seen in MSC-US2, MSC-US3, and MSC-US6, but not in MSC-US11. Only MSC-US2 displayed increased expression of CD46, while US2 and US3 proteins were both able to augment the percentage of MSC expressing this molecule. Regardless of the HCMV protein expressed, none changed CD55 MFI; however, expression of US6, US11, and US2 each increased the percentage of MSC that were positive for this molecule. Because US2 protein was the most efficient in up-regulating all three complement regulatory proteins, we used a functional complement-mediated cytotoxicity assay to investigate whether MSC-US2 were protected from complement-mediated lysis. We demonstrated that over-expression of the US2 protein reduced complement lysis by 59.10±12.89% when compared to untransduced MSC. This is the first report, to our knowledge, describing a role of HCMV-US proteins in complement evasion, and our data shows that over-expression of US2 protein on MSC could serve as a strategy to protect these cells from complement lysis.
The complement system, an integral component of the innate defense mechanism, [1] plays an essential role in the inflammatory process, and serves as a critical bridge between the innate and adaptive arms of the immune response [2], [3], [4]. Upon activation, complement proteins function as chemotactic factors and amplifiers of the inflammatory response, promote the destruction of infectious agents, cause cytolysis of damaged cells and tissues, and play an active role in allograft rejection [5], [6].
Mesenchymal stem cells (MSC) express functional receptors for anaphylatoxins C3a and C5a, and these receptors contribute to the recruitment of MSC to sites of injury [7]. In addition to being recruited to sites of complement activation, MSC can also activate the complement system, which results in production of soluble C3a and C5a and deposition of complement-activated molecules on their surfaces [8]. MSC express soluble factor H, the complement regulatory proteins CD46, CD55, and CD59, allowing MSC to be able to inhibit activation of the complement system to a certain extent [8], [9], [10]. Still, in the presence of an activated complement system, these innate mechanisms of protection are insufficient to prevent cellular damage and death [11]. It is possible that despite displaying molecules which confer protection from complement-mediated lysis, MSC are still able to be damaged by the activated complement proteins after being recruited to wound sites, leading to premature cell death. Indeed, recent studies have demonstrated that complement activation is responsible for MSC injury after infusion, and that inhibition of complement activation could be a novel strategy to improve existing MSC-based therapies [11].
Human cytomegalovirus (HCMV) is a ubiquitous pathogen, usually causing an asymptomatic primary infection, remaining latent throughout life [12]. This virus has evolved several strategies to avoid immune system recognition, allowing life-long co-existence with its host [13]. In order to evade the complement system, upon egression from the infected cell, HCMV incorporates the host-encoded complement inhibitor proteins, CD55 and CD59, into its envelope [14]. Furthermore, HCMV has been shown to up-regulate the host-encoded CD55 and CD46 after infection of human fibroblasts [15]. We and others have previously shown that genetically engineering mesenchymal, and other cell populations, with retroviral vectors encoding HCMV proteins US2, US3, US6, and US11 down-modulated HLA-I expression [16], [17], [18], leading to a decrease in allogeneic cytotoxic T lymphocyte (CTL) activation and natural killer cell (NK) killing [19]. In this study, we investigated the role of the HCMV US proteins US2, 3, 6 and 11 in protecting MSC from complement lysis, and demonstrated that different US proteins up-regulate the expression of complement regulatory proteins at different levels, but that overexpression of US2 protein on MSC enhanced the production of all of the complement regulatory molecules expressed on these cells. Furthermore, using a complement lysis assay, we demonstrated that expression of US2 on MSC protected these cells from complement lysis. Therefore, expression of HCMV US proteins on MSC, particularly US2, might constitute a potential strategy by which, in mismatched recipients, one could increase MSC engraftment and/or persistence in the injured tissue in order to release trophic factors.
Mesenchymal stem cells were purified from fetal liver tissue as previously described [20]. Liver tissue was purchased from Advanced Bioscience Resources (Alameda, CA, USA), homogenized to obtain a single-cell suspension, and Stro-1+ cells were isolated by using anti-Stro-1 antibody (R&D Systems, Minneapolis, MN) and magnetic cell sorting (Miltenyi Biotec, Inc., Auburn, CA) according to manufacturer's guidelines. Stro-1+ cells were then cultured at low density in 0.02% gelatin (Sigma, St Louis MO) on coated flasks using MSCGM™ (Mesenchymal Stem Cell Growth Medium BulletKit® Lonza, Walkersville, Maryland, USA) medium. The RetroPackTM PT67 Packaging Cell Line, purchased from Clontech (Clontech, Mountain View, CA), was used to generate the recombinant retroviral vectors. These studies were performed and approved according to the guidelines from the Office of Human Research Protection at the University of Nevada at Reno.
Construction of the retroviral vectors used in these studies have been previously reported in detail [19]. Briefly, US2, US3, US6, and US11 coding regions were PCR amplified from a clinical HCMV isolate (kindly provided by Dr. Stephen St. Jeor, University of Nevada Reno, Nevada, USA), introducing EcoRI and XhoI restriction sites for US3, US6, and US11, and EcoRI and SalI for US2. The purified PCR products were then ligated into pMSCV-Neo retroviral vector (Clontech, Mountain View, CA) that had been previously digested with EcoRI and XhoI or SalI. These recombinant plasmids were transformed into One Shot Top10 chemically competent cells (Invitrogen, Grand Island, NY), and transformed Top10 cells were selected with ampicillin 50 µg/ml (Sigma, St Louis MO). Positive clones were confirmed by PCR using primers for the US insert, miniprep digestions, and sequencing. For US3, US6, and US11 cloning, digestion was performed with EcoRI and XhoI restriction enzymes. For US2 cloning, EcoRI and BglII restriction sites were digested, as US2 cloning generated a SalI/XhoI hybrid restriction site.
Each US recombinant and an empty plasmid were used to transfect the RetroPackTM PT67 Packaging Cell Line (Clontech, Mountain View, CA) using Lipofectamine 2000 (Gibco-Invitrogen, Carlsbad, CA) according to the manufacturer's instructions. Stable transformants were selected with 500 µg/ml G418 (Invitrogen, Grand Island, NY) at 72 hrs after transfection and for 5 days thereafter. Supernatants were collected and filtered with 0.45 µm low binding protein-syringe filter (Pall Corporation, Port Washington,NY) to eliminate live cells and cell debris.
Subconfluent cultures of MSC were transduced for 6 hours with filtered collected supernatant containing either US recombinant or empty pMSCVneo retrovirus diluted 11 in serum-free QBSF60 medium (Quality Biological Inc., Derwood, MD) and 8 µg/mL protamine sulfate (Calbiochem, La Jolla, CA). MSCGM™ medium (Mesenchymal Stem Cell Growth Medium BulletKit® Lonza, Walkersville, Maryland, USA) was added after the transduction. At 48 hrs, 500 µg/ml of G418 (Invitrogen, Grand Island, NY) was added and MSC were incubated for 5 days, replacing the selection media every two/three days. Stable transduced cells were analyzed for transgene expression by PCR in order to amplify the US gene and neoR, and were designated as MSC-US2, MSC-US3, MSC-US6 and MSC-US11 according to the HCMV US protein that they expressed, or MSC-E for carrying the empty vector. Primers to amplify neomycin were: F-5′ GTGGAGAGGCTATTCGGCTA3′ and R-5′ CCTTGAGCCTGGCGAACAGT3′. Total RNA was purified from transduced and not transduced MSC by using TRIzol® Reagent with the PureLink™ RNA Micro Kit (Gibco – Invitrogen, Carlsbad, CA). 5 µl of RNA were DNase-treated with RQ1 RNase-Free DNase (Promega, Madison, WI). RT was carried out using the SuperScript III first-strand synthesis system and random primers (Gibco – Invitrogen, Carlsbad, CA) according to the manufacturer's protocol. For PCR, the reaction mixture consisted on 1.5 mM MgCl2, 20 mM Tris- HCl (pH 8.3), 50 mM KCl, 0.2 mM dNTP mixture, 0.02 U of Taq DNA recombinant polymerase (Gibco – Invitrogen, Carlsbad, CA), 0.004 µg/µl of each primer and 5 µl of RNA DNase treated. Primers used for cloning and analyzing presence of US genes were the following:
US2 F Eco, 5′ATAGAATTCA-ATGAACAATCTCTGGAAAGC3′;
US2R-SalI, 5′ATTGTCGAC-TCAGCACACGAAAAACCG3′;
US3F-EcoRI 5′ATAGAATTCAA-ATGAAGCCGGTGTTGG3′;
US3R-Xho I 5′GCGCTCGAG-TTAAATAAATCGCAGACGG3′
US6F-EcoRI 5′AAGGAATTC-ATGGATCTCTTGATACGTCTCGG3′
US6R-Xho I 5′ ATTCTCGAG-TCAGGAGCCACAACGTCG3′
US11F-EcoRI 5′ TAAGAATTCA-ATGAACCTTGTAATGCTT3′
US11 R-Xho I T5′ATCTCGAG-TCACCACTGGTCCG3′.
The amplification conditions were initial denaturation at 94°C (3 minutes), followed by 30 cycles of denaturation at 94°C (45 seconds), annealing (30 seconds) at 55°C, elongation at 72°C (1 minute 30 seconds), and a final elongation of 10 minutes at 72°C. The PCR products were electrophoresed on 1% agarose gel in 1× Tris-acetate-EDTA buffer and visualized by UV transillumination (Bio-spectrum Imaging System; UVP, Inc., Upland, CA) and visionWorksLS software.
MSC, MSC-E MSC-US2, MSC-US3, MSC-US6 or MSC-US11, were analyzed for CD46, CD55 and CD59 protein surface expressions. A total of 1×105 to 1×106 cells/sample detached in 1× PBS with a BD Falcon™ scraper (BD Bioscience Pharmingen, San Diego, CA) were incubated for 15 min at RT with 10 µl mouse anti-human CD59:RPE (AbD Serotec USA, Raleigh, NC), 10 µl FITC mouse anti-human CD46 (BD Bioscience Pharmingen, San Diego, CA), or 10 µl FITC mouse anti-human CD55 (BD Bioscience Pharmingen, San Diego, CA). Stained cells were washed with 0.1% azide in 1× PBS (Sigma, St Louis MO), centrifuged at 1500 rpm for 5 min, and fixed with 1% formaldehyde (Fisher Scientific, Pittsburgh, PA) in BD FACSFlow Sheath Fluid (BD Bioscience Pharmingen, San Diego, CA), and analyzed using a FACSort system (Becton Dickinson, Mountain View, CA, USA) with CELLQUEST software (Becton Dickinson, Mountain View, CA, USA). For each sample, 2×104 cells were acquired. Forward and side-scatter plots were used to exclude dead cells and debris from the histogram analysis plots. Median Fluorescence Intensity ratio (MFI) was obtained by dividing the Median Fluorescence Intensity obtained for a specific marker by the Median Fluorescence Intensity of the isotype control staining for the same cell population. The percentage of positive cells for the corresponding marker was obtained by performing side scatter versus FL1 or FL2.
1.5×105 MSC or MSC-US2 cells were washed, scraped, and incubated with 5 µg/ml human IgM (myeloma) (Rockland, Gilbertsville, PA, USA) whole molecule in 1 ml of MesenCult®-ACF Medium (StemCell, Palo Alto, CA, USA) for 15 min at room temperature (experimental condition). The cells were centrifuged and washed once with MesenCult®-ACF Medium. Rabbit serum complement (Innovative Research Inc, Plymouth, MN, USA) was added at a 14 dilution in MesenCult®-ACF Medium and the cells were incubated for 30 min at 37°C. The cells were centrifuged, washed twice with PBS and incubated at RT in the dark for 15 minutes with 0.1 µM calcein AM and 8 µM Ethidium Homodimer-1 (Molecular Probes- Invitrogen, Carlsbad, CA) in 1 ml of PBS. Following staining, 20,000 cells were counted on a FACSort system with CELLQUEST software (Becton Dickinson) and Flowjo (Tree Star, Inc. Ashland, OR) was used to analyze the results. In addition, a control containing rabbit serum complement in the absence of human IgM was included (control condition). Before running the experimental and control samples, parameters on the FACSort System were set using single color staining of live and ethanol-fixed MSC (dead cells). Specifically, MSC were scraped from culture flasks and stained with 0.1 µM calcein AM (Molecular Probes- Invitrogen) in 1 ml of PBS for 15 minutes at RT in the dark, and analyzed with the FACSort system and CELLQUEST software (Becton Dickinson). The resultant gate was set as the "live cells". To set the "dead cell" gate, another aliquot of MSC were fixed with 100% ethanol for 10 minutes, washed with PBS, stained with 8 µM Ethidium Homodimer-1 (Molecular Probes- Invitrogen) in 1 ml of PBS for 15 minutes at RT in the dark, and analyzed with the FACSort system and CELLQUEST software (Becton Dickinson). MSC stained with calcein were counted as viable due to the fact that when calcein freely enters in the cell as a non-fluorescent molecule, it is modified by cell estearases into a fluorescent compound which after excitation at 483 nm emits at 535 nm. Ethidium homodimer-1 is only able to enter cells with membrane damage, i.e., dead or dying cells. Following incorporation into the cell and excitation at 485 nm, this compound emits at 650 nm. Due to the differences in emission spectra between calcein and Ethidium homodimer-1, these compounds can be used simultaneously and allow one to clearly distinguish between live and dead cells. Double-positive cells were counted as having membrane damage, and double-negative cells were excluded from the analysis. The percentage of cytotoxicity was calculated as follows: [(percent of dead cells under experimental conditions – percent of spontaneously dead cells)/(100 – spontaneously dead)]*100 [21]. The percentage of spontaneously dead cells was determined by incubating the cells with 5 µg/ml human IgM (myeloma) whole molecule in MesenCult®-ACF Medium for 15 min at room temperature without adding rabbit serum complement.
Experiments were independently repeated at least three times. Results were presented as mean ± SEM. Unpaired two-tailed student T test was used to analyze the statistical significance of the results obtained with the Flow cytometric complement-mediated cytotoxicity assay.
For the analysis of CD59 surface levels, to compare MFI ratio and percentage of CD59 positive cells, one-way analysis of variance (ANOVA) was performed. After confirming a statistical difference among the groups, Fisher's least significant difference (LSD) test analysis was performed as a post-doc test due to the unequal number of samples between groups. In the case of analysis of CD46 and CD55 MFI ratio on MSC as well as percentage of positive cells, one-way analysis of variance (ANOVA) was performed followed by Tukey's honestly significant difference (HSD) as the post-hoc test due to equal number of samples among the groups.
For all analyses, a p value <0.05 was considered to be statistically significant.
It has previously been shown that an increase in cell surface expression of the complement regulatory protein CD59 will confer protection against complement lysis [22]. In order to investigate the effect of US proteins on the expression of CD59, we transduced MSC with the different MSCVneo retroviruses expressing US2, US3, US6 or US11, and used, as a control, both MSC non-transduced (MSC), or transduced with the empty MSCVneo retrovirus expressing only the NeoR gene (MSC-E). Flow cytometric analysis was then used to analyze the surface expression of CD59 on US transduced- and non-transduced MSC as described in the Material and Methods section. Figure 1A depicts representative results obtained from at least 3 independent experiments. Transduction of MSC with pMSCVneo (MSC-E) did not significantly change CD59 median fluorescence intensity (MFI ratio levels: 138.93±33.20) or the percentage of CD59 positive cells (92.38±2.47) when compared to non-transduced MSC (MFI ratio: 128.80±23.94; percentage of CD59 positive cells: 92.64±3.53) (p>0.05) (Figure 1B). Moreover, MSC-US11 did not produce a significant increase in CD59 expression, when compared with untransduced MSC or MSC-E, as determined by MFI ratio: MSC-US11 MFI ratio: 155.49±54.94 or percentage of CD59 positive cells: 93.92±2.30% (p>0.05). However, expression of US2, US3, or US6 proteins significantly increased the levels of CD59 expression, as measured by a change in MFI ratio and percentage of CD59 positive cells, when compared to untransduced MSC and MSC-E (p<0.05). The MFI ratio of CD59 on MSC-US2 was 273.19±35.77, that of MSC-US3 was 319.82±64.87, and on MSC-US6 265.77±16.22. Moreover, 97.27±1.26% of MSC-US2, 96.80±1.47% of MSC-US3 and 96.87±0.69% of MSC-US6 cells were positive for CD59 surface marker.
Next we studied the expression of membrane cofactor protein (MCP), or CD46, another membrane-bound complement regulatory molecule. This protein protects the host cell from complement damage by inactivating the complement components C3b and C4b [23]. Flow cytometric analysis demonstrated that, before transduction, 11.56±0.57% of MSC expressed CD46, and had a MFI ratio of 3.22±0.03, and that transduction with the empty vector, MSCVneo, resulted in a slight increase in the percentage of cells expressing CD46, 17.90±1.01%, (p<0.05) but a decrease in overall expression of CD46: MFI ratio of 2.72±0.09 (p<0.05), (Figure 2A).
MSC-US6 displayed higher percentage of CD46 positive cells when compared to MSC, but this increase was not statistically significant when compared with MSC-E (p>0.05): 19.07±0.1% of MSC-US6 were positive for CD46 (MFI ratio: 2.73±0.03). In contrast, a significant increase in the percentage of CD46 positive cells was found when US11 (21.28±1.38) or even more markedly US3 (27.76±0.68%), was expressed on MSC (p<0.05), but the MFI ratio of these cells (MSC-US3: 2.67±0.03 and for MSC-US11: 2.42±0.42) remained similar to those of MSC-E (p>0.05). MSC-US2 displayed the highest increase in both the percentage of CD46 positive cells (27.67±1.61%)(p<0.05) and the MFI ratio (MFI ratio: 3.46±0.04)(p<0.05). These results showed that expression of US3 and to an even greatest extend US2 on MSC, resulted in efficient up-regulation of CD46 expression on MSC. Figure 2B depicts a representative result from three independent experiments indicating the CD46 MFI ratio levels, obtained in this particular experiment (black filled histogram) and isotype control staining (unfilled histogram).
Complement decay-accelerating factor (DAF), also known as CD55 prevents the assembly of the C3 as well as C5 convertases and blocks the formation of the membrane attack complex [24]. This molecule is expressed constitutively in 37.79±3.30% of MSC (MFI ratio: 7.71±0.46). Transduction of MSC with a retroviral vector encoding only the neoR gene, (MSC-E) did not change the percentage of CD55 positive cells but reduced the MFI ratio; MSC-E: 42.84±1.03% (p>0.05), MFI ratio: 5.27±0.17 (p<0.05). The percentage of MSC-US3 positive for CD55 was 45.21±1.31% and displayed a MFI ratio of 5.22±0.12. Therefore, expression of US3 on MSC resulted in a pattern of CD55 expression similar to that of MSC-E. By contrast, MSC-US2 exhibited a higher increase in the percentage of CD55 positive cells (49.70±1.08%; p<0.05) when compared to MSC or MSC-E (p<0.05), and an increased MFI ratio when compared to MSC-E levels (MFI ratio: 6.79±0.13; p<0.05). Amongst the US HCMV proteins tested, however US6 and US11 were the most effective at enhancing the percentage of positive CD55 cells when compared to non-transduced MSC (p<0.05) or MSC-E (p<0.05). The percentage of MSC-US6 and US11 positive cells for CD55 were 56.72±0.66% and 60.86±1.03% respectively (Figure 3A). Importantly, the MFI ratio for MSC-US6 and MSC-US11 remained similar to that of non-transduced MSC, and was, respectively, 8.35±0.13 and 8.22±0.30. Figure 3B depicts one representative result from three independent experiments, indicating the MFI ratio levels for CD55 staining (black filled histogram) and isotype control staining (unfilled histogram).
When compared to other US HCMV proteins, US2 protein appeared to exert the broader effect on modulating expression of complement regulatory molecules, since it was either the most, or at least equally, efficient at up-regulating the MFI ratio and/or the percentage of cells expressing CD46, CD55, and CD59 molecules. Therefore, MSC-US2 was chosen to be further tested, using a functional complement-lysing assay, to investigate the role of this US HCMV protein in protection from complement lysis. Figure 4A shows the results of a representative experiment (n=3) depicting the flow cytometry-based complement-mediated cytotoxicity assay, performed as described in detail in the Materials and Methods section. MSC were evaluated in the presence of IgM and rabbit serum complement (experimental) or in the presence of IgM and absence of the rabbit serum complement (control). The same experimental and control conditions were used using MSC-US2. The percentage of cytotoxicity was calculated for both cell populations according to the formula described in the Material and Methods section. Complement cytotoxicity on control MSC was 48.27±2.16% but expression of US2 on MSC reduced the percentage of cytotoxicity to 20.27±6.94%, a 59.10±12.89% reduction in complement lysis (p<0.05) (Figure 4B).
The efficacy of allogeneic MSC as therapeutic tools has been reported in numerous studies, (reviewed in [25], [26], [27]); however, controversy still remains whether these cells, upon transplantation, are able to elicit an immune response in the recipient, lessening their therapeutic potential when compared with their autologous counterpart. Studies showed that some of the factors responsible for the failure of donor MSC to engraft include the mismatch of HLA class I molecules between the donor and the recipient [28], [29] and activation and damage by the complement system [11]. Although MSC display on their surface proteins that inhibit complement system activation such as CD59, CD46, CD55 [8], [10] and inhibitor factor H, [9] the complement system still is able to recognize and injure MSC after their infusion [11].
Our results are in agreement with these prior studies showing that MSC constitutively express high levels of CD59, while CD46 and CD55 are so to a lesser degree. Moreover, we show for the first time that engineering MSC to express US2, US3, US6, or US11 HCMV proteins alters the expression of these regulatory complement recognition molecules and protects MSC from complement attack and lysis.
Overexpression of US2, US3, and US6 (but not US11) HCMV proteins, significantly increased CD59 surface molecule levels and the percentage of CD59 positive cells. US6 and US11 proteins did not alter either the expression or the percentage of cells positive for CD46, while US3 induced only an increase in the percentage of MSC that were CD46 positive. However, US2 protein induced the highest increase in the percentage of CD46 positive cells, and in the level of expression on a per cell basis. The US6 and US11 proteins effectively increased the percentage of MSC positive for CD55 but not its intensity of expression, whereas US3 protein left the pattern of CD55 expression on MSC unaltered. By contrast, MSC-US2 exhibited an increase in both the percentage of CD55 positive cells and MFI ratio. Overall, these results demonstrated that expression of US2 HCMV protein on MSC had the ability to increase expression of all three surface molecules MSC produced for protection against complement activation. Although we did not investigate the mechanism by which US2 was able to alter the expression of CD46, CD55 and CD59, it is possible that the disruption and accumulation of HLA-I complexes in the Endoplasmic Reticulum (ER) caused by over expression of US2 leads to ER stress and consequent increase in the levels of NF-κB [30]. Since CD46, CD55 and CD59 are all known to be induced by NF-κB dependent pathways, this could provide a possible explanation for your findings[31]
To establish the functional significance of the altered expression of CD46, CD55 and CD59 by US2 we performed a FACS-based complement-lysing assay, and demonstrated a 59.10±12.89% reduction in complement lysis in MSC expressing US2 protein, when compared to the control.
While recent studies showed that MSC were able to trigger C3 binding to their surface, and higher C3 binding capacity was correlated with higher suppression of PBMC- proliferation [8], we have also recently shown that expression of US proteins, including US2, reduces HLA-I expression and consequently decreases peripheral blood mononuclear cell recognition of MSC [19]. Furthermore, although MSC-US2 expressed reduced levels of HLA-I expression, they were no more susceptible to NK killing than untransduced MSC [19] due to high expression of CD55 molecules [32]. This recent work, combined with the present findings, show that expression of US2 HCMV on MSC provides multiple layers of protection to these cells, decreasing not only peripheral blood mononuclear cell proliferation, but also lysis of MSC by NK cells and by the complement system. In conclusion our data suggests that use of these genetically-engineered MSC, mainly MSC-US2, would have an enhanced advantage in the presence of an exacerbated inflammatory microenvironment and/or in an allogeneic transplantation setting, resulting in decreased rejection by the immune system, extending the time in survival and thereby enhancing their therapeutic potential.
This work was supported by National Institutes of Health (Bethesda, MD, USA) grants HL73737 and HL97623. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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I am on a scarf knitting spree..and, my closet is overflowing with knitted scarves. The recent one I finished is a simple feather and fan lace scarf also called as Old Shale scarf. Remember the yarn I got as a Christmas gift from Cathy.. I couldn't resist knitting with it..so, I cast on for this scarf as soon as I got back from my Christmas vacation and finally finished it two days back.. The yarn is Madeline Tosh merino light in a colorway called Lichen. I love the colors in it.
It is a mindless pattern and a perfect T.V. knitting material although, I must say that I do not get much T.V. time, thanks to the preschooler in my home. The scarf did look like lichen when it was unblocked. For my non-yarny friends, let me explain blocking in a few words. Blocking is a process of soaking the knitted or crocheted item in water for some time and then letting it dry flat in the shape required. Blocking helps the natural fibers keep the shape its blocked in, evens out the stitches and improves the drape of the finished object(FO). So, if you don't believe the change blocking brings about to an FO, here is visible proof of it.. Here is the "Before" shot of the scarf looking as if a dead wood gathered moss and lichen on it..
And, for the gratuitous "After" shot.. Ta da!
A picture is worth a thousand words,eh?! Blocking really opened up the stitches and the lace. The scarf grew by 10 inches in length after blocking. Yarn Harlot has an excellent tutorial on blocking, for the uninitiated. I am more of a process knitter than a product knitter and blocking is a process I enjoy the most in lace knitting. Its like a fairy tale of the ugly duckling turning into a swan.. And since I like the look of lace against the light , here is a shot of the scarf against the light. And, that's the view from my balcony two days back.
Here are the stats:
Pattern: Old Shale Scarf by Tienne
Yarn: Madeline Tosh merino light; 1 skein
Needle: 4mm
Started: January 5 2011
Completed: January 29 2011
Raveled here: Lichen on Shale!
Incidentally, this scarf is my first knitted FO in year 2011. I hope this is a good start for a year filled with crafts and hobbies.
Peace and love,

subset: commoncrawl


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Mountain View Estate
Colorado, La Plata County, Durango – MLS 730082
Exquisite custom 10,215 SF, 5 bedroom, 6 bath log home in the exclusive Two Dogs community with 9+ lakefront acres. The home was completed in 2005 and consists of three levels with lovely views of Engineer Mountain, Spud Mountain, Twilight Peak, the Needles Mountains and the Hermosa Cliffs. The quality architecture and design masterfully accentuate the exterior and interior use of log construction consisting of a combination of 20" plus individually cut Douglas Fir logs, natural stone, custom ironwork and wood shingle roof. The acreage is predominately forested with large Ponderosa Pines, Aspen trees, native ground cover and landscaped yard surrounding the home, which features meandering stone walkways, retaining walls and a gas fire pit in the backyard. The home enjoys direct access and rights to a private lake on its western boundary and the San Juan National Forest on its northern property line. This is the ultimate Colorado retreat site with easy access to Durango Mountain Resort, The Glacier Club and historic downtown Durango.
Location and Area
Two Dogs is located in the San Juan Mountains on US Hwy 550, 22 miles north of downtown Durango. The entrance gate is just off Highway 550. Two Dogs is conveniently located for recreation, 2 minutes from skiing at Durango Mountain Resort, 10 minutes from The Glacier Club, 20 minutes from Dalton Ranch Golf Club and 30 minutes to Durango. The community is secure, private and quiet with many opportunities for hiking, snowshoeing and horseback riding. The National Forest can also be accessed from the property.
Durango is a town of rural mountain charm and character, and has many big city conveniences, including a state-of-the-art new hospital, many fine restaurants, hotels, shopping, a four year college, golf courses, first class fly fishing, numerous outdoor hiking and biking trails, with endless activities for the outdoor sports enthusiast. It is located at 6,512 feet above sea level, surrounded by scenic mountain views, and over 300 days of sunshine each year. Comfortable summer temperatures reach the upper 80's, while the autumn days are clear, cool and full of vibrant foliage colors. The crisp and sunny winter days often feature abundant powdery snow.
The La Plata County area that includes Durango and Bayfield has a population of 52,000. La Plata airport runs daily commercial service to Denver, Salt Lake City, Albuquerque and Phoenix, with seasonal service to Dallas, and includes facilities for full service general aviation. The property is 105 miles from Telluride, 205 miles from Albuquerque and Santa Fe, 170 miles from Grand Junction, about 300 miles from Denver and Colorado Springs, and 450 miles from Phoenix.
Two Dogs Community Amenities
- Gated stone and overhang entrance with beautiful landscaping
- Paved entrance area - all roads in Two Dogs are paved
- Lighted mail and parcel boxes at the entrance by the gate
- Strong design covenants to protect quality of homes in the community
- The home site building envelopes have been selected to insure the most inviting and unobstructed views while maintaining privacy
- Buried utilities including propane
- Two Dogs offers the opportunity for membership in The Glacier Club and The Dalton Ranch Golf Club
Property Grounds and External Features
- Two premier lots combine to make these 9.42 acres that capture the most dramatic views of Engineer Mountain, Spud Mountain, the West Needles Mountains (Twilight Peak), Needles Mountains and the Hermosa Cliffs
- The homesite enjoys direct access and rights to a private lake on its western boundary and borders the San Juan National Forest on its northern property line
Several short trails have been established connecting the residence to adjacent private lake and to an historic wagon trail directly accessible on the north property line in the San Juan National Forest
- Acreage is open and forested with large ponderosa pine, aspen, juniper, gambel oak
- Landscaped yard featuring native ground cover surrounds the home with meandering stone walkways and retaining walls
- Lawn sprinkler systems on timers
- Asphalt circular driveway spanning the two lots
- Gas fire-pit in backyard
- Quartzite flagstone patio on lower level, partially covered, allows for expanded views of mountain ranges
- Exterior has been freshly stained; all logs in very good condition
- 12" cold roof system of .5" cedar shake shingles; 3.5" extra layer to vent cold air and maintain temperature, alleviating ice build-up
- Full gutters surround exterior of home
- 3' x 3' stone columns with large stone caps; log 4.5" x 12" window bucks
- Outside back decks of Brazilian ironwood (Epay); very large overhang; hand-forged exterior hand railings
- Built-in stainless steel propane BBQ
- Amazing views of Engineer Mountain, Needles Mountains, Spud Mountain and Tank Mesa along Missionary Ridge
Residence Features Include:
- 10,215 SF gracious living space in 3 levels with loft and separate guest wing; full of large 360 degree panoramic windows allowing for a very light and bright interior
- 5-bedrooms; 5-full and 2-half baths
- 20" and up logs create multiple points of interest throughout home
- Elevator from lower level services second floor
- Security system
- Sound system inside and out
- Cathedral and 9', 10', 12', and 16' ceilings
- Kolby casement windows
- Ceiling fans in most rooms
- Radiant in-floor heat; propane heating
- On-site hand-hewn support beams throughout
- Hand-forged custom metal work by Rod Pickett, notable local artisan
- Radius corners throughout
- 7'6" solid wormhole and distressed knotty alder doors throughout the house, featuring oil-rubbed bronze Rocky Mountain Hardware
- Custom wormhole and hand-distressed knotty alder, raised panel, full-overlay cabinets with Rocky Mountain Hardware
- Wormhole and distressed knotty alder 6" baseboard throughout
- Combination of hand-scraped hickory hardwood with wood plug detail, wool carpet, stone, and cement floors
- 2 separate front and back stairways – one to guest wing, second to owner's wing
- Custom plumbing and light fixtures; lighting dimmer controlled
- Recessed and up lighting; chandelier; lighting sconces
- Heavy insulation and sealing provides a "no draft" home
- 3-car+ oversized heated garage with concrete floors; cedar garage doors; extensive storage in built-in maple veneer cabinets for ample storage; built-in workbench & pegboard; doggie shower; stainless steel sink basin; Whirlpool Duet washer/dryer; ski and snowboard racks; large storage closet underneath stairs
- Mechanical Room: Culligan water softener; central vacuum; HEPA air filtration/heat recovery system; 150-gallon water heater; air exchange and handling unit; full humidification system; state-of-the-art fire suppression system; multiple heating zones clearly marked
- 600 AMP electrical to service home
Main Foyer
- 9' double 2¼" doors with flanking 3' x 5' side-light casement windows
- Random-set slate flooring with hand-scraped 10" hickory planks
- Foyer flanked by 2 stone walls
- Very large stone columns outside main entry
- Flagstone porch with overhang
Great Room
- 4' hand-distressed wet bar with bronze faucet and under-mount sink
- 35' ceiling from floor ridgeline with large prow
- Very large sitting area with walls of oversized and trapezoid windows take in spectacular views of Engineer Mountain, Needles and Twilight Mountains, Spud Mountain
- Floor-to-ceiling stone wall with 4' x 4' Rumsford-type fireplace; wood storage underneath
- 6' radius custom Mexican hand-made chandelier
- 6" pine tongue and groove ceilings walk-out to large Brazilian ironwood (Epay) decks with hand-forged exterior railings
- 10' ceilings
- Coppers Mine slate flooring with hand-scraped 10"-wide hickory planks with steel hand-forged medallions on a grid pattern
- Hand-distressed Ogee alder cabinets with open top shelves for decorations; glass inserts upper cabinets; sand-cast nickel hardware
- 48" Viking cook top with 6-burners and center griddle
- Custom plaster hood with metal tile inlays; commercial exhaust fan with heat lamps
- Black 3CM double-layer granite counters
- Large radius center island with copper vegetable sink; double-layer black 3CM chiseled-edge granite; 6 turned wooden legs supporting to create a furniture-like piece with granite top
- 4' x 7' 3-panel casement windows looking out from sink area to Hermosa Cliffs
- Two Bosch dishwashers
- Upper and lower Viking convection and conventional ovens
- Sub-Zero side-by-side fridge/freezer
- Kitchenaide trash compactor
- Double stainless-steel sinks
- Travertine tile backsplash
- Walk-in pantry with built-in and open shelving; power supply for appliances
Dining Room
- 10' ceilings
- Two custom light chandeliers over dining room table seating for 10-12
- Coppers Mine slate flooring with hand-scraped 10"-wide hickory planks with steel hand-forged medallions on a grid pattern
Owner's Master Bedroom and Bath Suite
- 18' vaulted ceiling
- Large windows with northern views taking in Needles and Twilight Mountains; fully lined floor-to-ceiling double-lined linen drapes with accent trim
- SW style Kiva fireplace
- 8' door goes out to private Brazilian ironwood (Epay) deck
- Walk-in closet with 13' ceiling; built-in adjustable shelving; his and hers sides with center island; various shelves, cubbies and drawers for jewelry, belts, shoes, built-in clothes hamper, etc.; large 5' x 7' opening windows facing south with lined linen window coverings
- Secondary master closet is 5' x 6' has separate door; cedar lined with adjustable shelves
- Custom knotty alder distressed cabinets; pulls sand-cast brushed and pitted nickel
- Large master shower with two shower heads, steam unit; long sitting bench and two soap niches
- Rainforest granite countertops; separate make-up vanity with knee hole
- Kohler porcelain dual sinks with separate mirrors; granite accent tile at corners
- Travertine tile floors with granite accent tiles
- Oversized serpentine tub deck with 12" x 12" travertine tiles and ½" mosaic facing; large 2-seater Kohler Jacuzzi jet tub
- Large windows facing west
- Separate large water closet with built-in bookshelf and phone jack
Master Office/Study
- 16' vaulted ceiling
- Pocket half-light doors with seeded glass, wood panels below; clavos accent inlays
- 15'W x 7'H built-in bookcase and desk with slide-out keyboard tray
- 8' tall bay windows with custom cushions; incredible views to Engineer Mountain, Twilight Peaks, Needles Mountains, and Spud Mountain
Powder Room
- 5' x 9'
- 24" French Quarter setting in travertine flooring
- Distressed wormwood vanity with copper sink
- Oil rubbed bronze mirror, faucet and wall sconces
Gallery Foyer
- 2 entrances; one from south of home enters from outside into guest wing; other door opens to back deck; both large doors with half glass to capture mountain views
- 12' ceilings
- Pine tongue and groove ceilings
- Custom iron chandelier
- Elevator access
Guest Wing
- Built-in desk separates guest room suites
- Large utility closet with built-in cabinetry stores cleaning supplies and central vac parts
Main Laundry Room
- 12' x 12'
- Built-in desk "bill-pay" center with under-mount keyboard tray; closed cabinets on top
- Separate wall-to-wall cabinets and drawers for storage; ample counter space for folding clothes
- Under-mount large stainless steel basin
- Whirlpool high-end washer/dryer
- Central vac "dust-pan toe-kick" allows for quick easy disposal of dust and dirt
Two Guest Bedroom and Bath Suites
- 16' vaulted ceilings with 30" ridge beam
- Wool carpet
- Window coverings are full length, double-lined linen with accent trim; wood rods
- Built-in drawers
- Walk-in closets, fully adjustable hanging, shelving and cubbies; slide-out drawers
- Honed black slate tile floors
- Showers porcelain tiles with black slate deco bands; brushed nickel fixtures
- Vanities of porcelain tiles with Kohler under-mount sink; rusted metal wall sconces; Western mirror
Owner's Entry – Mud Room
- Large mud room with concrete floors; built in seating bench with floor-to-ceiling wall cubbies and hanging spaces for outdoor gear
- Elevator to second floor
- Large walk-in coat closet
- Double staircases to main level with stairs of 5" x 13" Douglas fir timber steps with alder risers and skirting
- Long gallery hallway with 12' barrel ceiling of corrugated rusted metal
TV Room, Billiard and Wet Bar Area
- Five 10" x 14" hand-hewn on-site, hand-distressed with burnt edging ceiling beams with hand-forged plates, replicating metal detail on handrails throughout
- Wood-burning 4' x 3' Rumsford-type fireplace set in floor-to-ceiling wall of Telluride fieldstone and lead stone with brick accents; wood storage underneath; serpentine floor hearth of quartzite flagstone
- Copper radius wet bar; black granite counter encloses icemaker/mini-fridge combo; copper sink with oil-rubbed bronze fixtures; upper cabinets combination of fluted glass and open-shelves
- Billiard area has full slide-out A/V cabinets; separate built-in computer desk
- Glass block wall separating workout room
- Floor is integral-colored, stained concrete
- Large sitting area with doors out to quartzite flagstone patio
Wine Cellar and Card Room
- Mounted ½ wine casks on wall of entry area
- Separate 8' x 12' Poker Room with large windows; hand-hewn ceiling beams; rusted tin walls sconces; quartzite flagstone floor; German smears stone wall; ¾ light door with fluted glass and rusted clavos nails
- Separate fresh-air intake to exhaust cigar smoke
- Separate climate controlled, air-tight wine cellar with Speak-Easy door
- Secondary owner's deadbolt closet off main wine room
Work-out Room
- Climate controlled with separate fresh-air intake
- Full floor-to-ceiling tempered glass full-wall entry
- Full length floor-to-ceiling mirrored wall; trim also mirrored
- Glass block wall separates view from Billiard Room
- Enclosed 6-8 person, cedar-lined dry/wet sauna with tile floor and drain; Finnleo state-of-the-art sauna control system
'Boys Dorm and Bath' – Guest Room
- Bunk room to sleep 5 people; 2-up/down hand-hewn log bunks; built-in hand-hewn shelving on wall with clavos detail
- Large window facing the range of mountains
- Walk-in closet with built-in fully adjustable hanging, shelving and cubbies, slide-out drawers; built-in dressers and clothes hamper
- Wool carpet
- 8' porcelain tile vanity with double Kohler under mount sinks; accent trout backsplash tiles; oil rubbed bronze fixtures
- 16" x 16" copper slate floor
- 2 full showers; oil-rubbed bronze fixtures; soap/shower niches; matching porcelain tiles, slate; trout tile deco bands
'Outhouse' – Powder Room
- Replicated outhouse using old, reclaimed lumber on walls, counter and Kohler toilet platform
- Original turn-of-the-century copper farmhouse water pump
- Copper basin
'Girls Dorm' – Guest Suite
- Landing area has storage room underneath roof
- Alder hand-distressed, multi-layered wood cap going upstairs
- Stairs slab 5" x 13" Douglas fir timber steps with alder risers and skirting
- Spacious, spit-level room with two separate sitting areas
- Vaulted ceiling – 20' to peak
- Carpeted
- Radius cushioned sitting bench looking over Great Room
- Built-in floor plug locations across length of loft
- Built-in writing/computer desk on west side; built-in linen storage on east side of upper level sitting area
- Large windows taking in spectacular views of above said mountain ranges
- 6" pine tongue and groove ceilings
- Bedroom area has access to separate loft area with hand-hewn log ladder and handrail
- 6' x 9' panel casement windows looking out to mountain ranges
- Fabric Roman shades with lining
- Walk-in closet with built-in closet with built-in fully adjustable hanging, shelving and cubbies
- Travertine tile floors with 3" Mexican tile inlays
- Porcelain 6' vanity with double Kohler under-mount sinks; brushed nickel hardware and fixtures
- Separate make-up vanity with knee hole; magnifying mirror
- Separate room for toilet and tub/shower combo
Wildlife and Recreation
The property has an ideal combination of wildlife habitat, manicured lawns and gardens in a very private setting. There is abundant wildlife, including: mule deer, elk, black bear, turkey and soaring raptors. There is direct access to the National forest for a myriad of recreational opportunities. For downhill skiing, Durango Mountain Resort is a few minutes from the estate, while Wolf Creek and Silverton Mountain Ski Areas are hour-and-forty minutes away. Cross-country skiing, snowshoeing and snow-mobiling are also available throughout the area. Summertime is filled with opportunities for biking or horseback riding, hiking, rock climbing, fly-fishing, boating or canoeing in the area. Also, the Animas River provides great fly-fishing, kayaking, canoeing and rafting. There are many other things to do in the area, like visiting the local Durango to Silverton Narrow Gauge Railroad, Mesa Verde National Park and the Four Corners National Monument.
Excellent golf can be found at Dalton Ranch Golf Club, Hillcrest Community Club, the private Glacier Club (memberships available), and a top-rated public course at Pinion Hills in Farmington, NM (approximately 60 miles).
Now $5,545,000 – Significant Price Adjustment
By Appointment only
This property is offered by Legacy Properties West Sotheby's International Realty, LLC, Tom and Zach Morse, Brokers. All information provided by Legacy Properties West Sotheby's International Realty, LLC or its sales representatives in connection with this property was acquired from sources deemed reliable, however, we do not warrant its accuracy or completeness and any users of such information waive any claims related thereto. Legacy Properties West Sotheby's International Realty, LLC recommends that every item of interest to the purchaser (i.e. issues regarding title, water, minerals, survey, zoning, permits, land use regulations and other applicable laws, and any other issues) be independently verified by the purchaser's attorney.


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Joint BSM and MBA
Admission Information
Interested students with a 3.4 grade point average in the Pepperdine Bachelor of Science
in Management (BSM) program who have all general education requirements completed
should submit applications during their fourth term of enrollment, or upon completion
of 40 BSM units. This permits eligible students to be provisionally accepted for the
MBAJ and allows them to register for their last two terms in a modified curriculum
(transition track) that includes transition classes to assist in preparing for the
MBA program.
Students who complete the modified program are then awarded their BSM degrees and, providing eligibility is maintained, are fully admitted to the MBAJ program at the end of their sixth term.
Spring 2018 Term
Classes Begin
December 5, 2017
January 5, 2018


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At Gemvara, we believe that the jewelry you wear should be one of a kind, like you. That's why we make each piece to order, one at a time, just the way you want it. Choose black onyx and palladium or emerald and sterling silver: it's your choice. Mix and match gems and precious metals for a look that's exactly you. It's jewelry for all life's occasions, from a birthday or anniversary to your own personal milestones and celebrations. Our inspired gift ideas with a personal touch will make this Hanukkah memorable. Gemvara uses only the highest quality materials to craft your july birthstone bold jewelry design with meticulous attention to detail. Our precious metals are recycled or responsibly mined. The July birthstone, ruby, is the essence of red, adding a bold dash of drama to everything you wear. Our wide selection of july birthstone bold jewelry by Carl Ansley, Ruzanna Davtian, David West, and other talented designers means you can find jewelry that's exactly your style. You can also shop by style, choosing our solr.product_tag_style.1483, Delicate, or Customer Favorite Collections. Gemvara's fine quality july birthstone bold jewelry is designed to be passed down for generations to come, crafted by hand in the United States.
We can help right away. Chat with a jewelry expert now!


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Become an Animal Reiki Ambassador and raise awareness for Animal Reiki in your community. Help us spread compassion and healing for animals and their caregivers through Reiki practice. Recognize and honor animals as spiritual teachers and healers.
Our Mission
To raise awareness around the globe of the healing benefits of Reiki for animals as well as honor all animals as our spiritual teachers.
To achieve this, we encourage shelters, sanctuaries and other animal welfare organizations, veterinary clinics, animal groups, animal-related businesses and individuals who love animals to organize Reiki events in celebration of World Animal Reiki Day.
In addition to larger, organized events, celebrating can also be as simple as lighting a candle to honor your animal teachers, sitting in silent Reiki meditation with your animals, or doing a compassionate act for your local shelter or sanctuary.
Unite the world Animal Reiki community by sharing your thoughts, messages and pictures of your celebration of World Animal Reiki Day on our Facebook page. For Twitter:#WorldAnimalReikiDay and tag us @SARA_news.
Who Is Involved
World Animal Reiki Day is organized by the volunteer members of The Shelter Animal Reiki Association (SARA), whose mission is to bring the healing benefits of Reiki meditation to rescued animals and their caregivers worldwide. We welcome all Animal Reiki practitioners from all lineages and forms of practice to join us to celebrate this special day!
Why February 5th?
February 5 was the birthday of SARA President Kathleen Prasad's first Animal Reiki teacher, Dakota. An Australian shepherd mix, Dakota was rescued from Sacramento Animal Control as a puppy and spent over sixteen years by Kathleen's side. In addition to awakening Kathleen to the benefits of Reiki for animals, Dakota inspired Kathleen to create the Shelter Animal Reiki Association (SARA). To view a memorial tribute video to Dakota's life click here.
Becoming an Ambassador
- Thursday, February 2, 4-6pm in Mill Valley, CA: Book Launch Party and SARA Fundraiser! CLICK HERE to RSVP
- Sunday, February 5, Noon – 3pm, Animal Reiki Share at BrightHaven Sanctuary in Santa Rosa. Email Kathleen to attend this special event. Space is limited. $25 love donation to BrightHaven.
- Tune into Kathleen's YouTube Message for the day! Find the link on our newsletter, FB and Twitter, #WorldAnimalReikiDay.
- Spread the Word! Add the World Animal Reiki Day logo to your website and social media graphics. Click Here for the LOGO and more information on how to create your own personalized World Animal Reiki Day photo!
Facebook Photo Contest
February 1st to the 4th
Help us celebrate World Animal Reiki Day on February 5 by sharing a photo of the cutest animal you have ever offered Reiki to.
If your picture gets the most votes, you will win:
- Autographed copies Reiki For Dogs and Kathleen's newest book, Heart to Heart with Horses: The Equine Lover's Guide to Reiki.
Organizing an Event
• Awareness & educational events at shelters, sanctuaries and other animal organizations.
• Conferences & workshops.
• Animal Reiki sessions or chair sessions for animal caregivers.
• School Animal Reiki events to educate the younger generation.
• Radio & TV interviews to raise awareness of Animal Reiki & World Animal Reiki Day and its mission.
World Animal Reiki Day Swag
Stick-It. Pin-It. Hang-It.


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KaBa Hostel offers precisely that what the lonely backpacker or the smart budget tourist is looking for. A cosy atmosphere in the green garden and arty lounge, pleasant comfort in the dorms or private rooms, a fully equipped kitchen and an internet bar. Located within walking distance from the historical centre but just outside of the busiest city area. And very nice hosts at the reception desk who you can always ask anything.
With a total of 37 beds and 9 rooms, KaBa Hostel is a rather small hostel, perfect for tourists travelling the globe or fun families looking for a nice accommodation alternative. Less suited for big groups and totally not suited for bachelor parties and such. We are not a party hostel!
Incl. breakfast, bed linen, internet and Wifi, selfservice kitchen, personal locker. Excl. city tourist tax (3 euro pp/night).
Incl. breakfast, bed linen, internet and Wifi, selfservice kitchen, personal locker. Excl. city tourist tax (3 euro pp/night).
KaBa Hostel, a charming and authentic hostel in the center of Ghent. We offer a soft bed, with private or shared bathroom, a yummie breakfast, a nice garden, a fancy kitchen, and of course a warm welcome with heaps of advice on tours, activities, shops, bars and restaurants...
KaBa Hostel is raised and loved by Bart and Katrijn, both of them fully experienced in the art of budget travelling and seeking out the nicest spots to sleep all over the world.


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|After an incredible engineering effort, the O2 arena was transformed into a Super Cross track by adding hundreds of tons of earth where normally there would be a mush pit or rows of seated concert goers.
The riders provided as big a spectacular as many of the other performers I've seen at the O2. The height of the jumps and the speed between the bumps was amazing. How the evening finished without at least one person in an ambulance I'm not sure. Every age group fearlessly charged round the track on ever large and more powerful bikes. The larger struggled in the confined space to over take each other – leading to a number of spectacular collisions!
The only downside was the requirement for one end of the arena to be open to allow the exhaust fumes out and the bikes in and out resulted in the normally warm location to take on the near freezing outside temperatures!
Saturday, January 30, 2010
Motorbikes replace the musicians
Posted by James Hammick at 1/30/2010 10:04:00 pm 0 comments
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Im Melting Im Melting.....
Discussion in 'Lipotrim Forum' started by thefutureisbright, 9 June 2008 Social URL.
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"Whole foods are those that are unprocessed and unrefined, or processed and refined as little as possible before being consumed. They typically do not contain added ingredients, such as sugar, salt, or fat."
Monday, January 23, 2017
Raspberry Clementine Muffins with Roasted Cashews
Posted by WholeFoodVegan at 10:14 AM
Labels: breakfast, fruit, nuts, whole grains


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If you were wondering what happened to the expectation of a Sainsbury's own brand virtual network, don't ask me. The promise was great, and it has been 'imminent' for over a year, but it has failed to materialise. Sainsbury's own brand mobile network appears to have been stifled amid finding a proposition that chimes with its customers, bigger priorities at the supermarket and 'the economy'.Interest from operators to act as a wholesale partner is certainly not one of the obstacles. Relentless growth for Tesco at one end, and Lebara Mobile and Lyca at the other has whipped up considerable appetite from the networks to sign up more wholesale deals. It marks a complete reversal of the suspicion towards MVNOs from the main operators a few years ago.Orange and Vodafone have seen MVNO wholesale represent growth engines over the last year, and 3 now wants a slice of the cake too. Lebara and Lyca are ripping lumps out of each other in the booming ethnic/overseas calling market, which is still growing. The search for the next MVNO success is the holy grail for operators as their core businesses are flattening or declining.The supermarket sector is undoubtedly one area. Taking the Lebara/Lyca format with more precise targeting of a specific ethnic group, say the Polish market, is also being talked about. There is also a possibility of reviving the EasyMobile low-cost, web-based format, in the belief that it was the right model but at the wrong time. Despite the saturation in the market, there is belief that there is room for more good MVNO ideas, executed well and with strong distribution routes. Over to you, Sainsbury's.
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Because of privacy laws, lenders are not allowed to call you when a loan goes into default or might be foreclosed.
The best you can expect is that a lender might give your card to a borrower in trouble, as someone who might help. This is certainly a good strategy, and you should pursue it with local real estate lenders.
After foreclosure, lenders and servicing companies generally rely on real estate agents to market their properties. Learn which companies in town are the preferred brokers for foreclosure properties.
Get to know the agents. Once you establish your credibility as a serious buyer, ask them to notify you as soon as something new comes into their inventory. This gives you a head start before the other buyers see the ads.
It also saves the agent some advertising and marketing money if you buy before those expenses are incurred.


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Register on; www.triathlonireland.com
Fingal Tri is affiliated with Triathlon Ireland. Triathlon Ireland affiliation provides 3rd Party insurance cover when training with the club. Once you join the club you are bound by our club constitution.
If you have any issues or questions or would like a copy of the club constitution please click HERE contact me on 0879974781 or email firstname.lastname@example.com
* For members who marshal at the Club Race, they will be refunded €25.
Please make sure your name appears as the transaction reference so we know who its from
Bank of Ireland
Account number: 40698271
Sort Code: 90-07-46
© Fingal Triathlon Club


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Hello Love
- reviewed by Russ Breimeier Copyright Christianity Today International
- 2008 1 Sep
- Sing, Sing, Sing
- Jesus Messiah
- You Lifted Me Out
- God of This City
- I Will Rise
- Love
- Praise the Father, Praise the Son
- God Almighty
- My Deliverer
- With Me
- Exalted (Yahweh)
- All the Way My Savior Leads Me
He's one of Christian music's best-selling recording artists with several radio hits and Dove awards to his credit. He's also one of the world's most revered worship artists, excelling at writing songs for the church that are easy to pick up yet hard to forget. When you put it all together, it's no wonder that Chris Tomlin has become one of the biggest artists in Christian music, but such success makes it easy for fans of worship and Christian pop to lose perspective through raised expectations.
Tomlin's fifth studio recording
Truth be told, there's validity to both sides of that debate. Even the first radio single "Jesus Messiah" has drawn mixed reactions, some calling it another catchy Tomlin classic, with others saying it sounds exactly like his past work, little more than an excuse to build a song around the word "Messiah." Personally, I see it as overly familiar, yet an accessible ballad that beautifully explains the Son of God's sacrifice.
Other songs are similarly mixed. "Sing, Sing, Sing" is a song of praise similar to plenty other driving rockers from modern worship in the last decade, particularly Tomlin's own cover of Jon Abel's "Awesome Is the Lord Most High." Yet lower it several keys (like most Tomlin songs) and the average contemporary worship service will readily embrace it. The same could be said of "You Lifted Me Out," a catchy rocker based on
For those wanting something sonically different from this worship leader, Tomlin and longtime producer Ed Cash do kick the sound up a notch in some spots. "Love" is a musical highpoint with an African Children's Choir bringing something different to the mix, adding up to a joyous anthem with some meaty verses about the nature of love: "It heals the sick, comforts the weak/Breaks the proud, raises the meek/In this life no guarantees, but there is love." There's also "God Almighty," which relies on a cool rhythmic hook carried by drums, piano, and strings—maybe not the sort of thing the average church will replicate, but the song offers one of the Tomlin's strongest choruses to date with lyrics borrowed from "Holy, Holy, Holy."
Speaking of which, Tomlin seems at his best on
I'd be remiss in failing to mention "God of This City" as another highlight, a hopeful and timely anthem by Irish worship band Bluetree, previously heard on the Passion album of the same name. Stylistically it resembles the writing of Redman, grounding worship in realities of the world we live in. But that said,
If I sound like I'm on the fence, it's because I am. This album is not as good as Arriving or See the Morning, but perhaps things would be different if


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1st Race: STORMFIELD (#6) showed improvement in his second race back after an 18-month layoff. He gets the nod for trainer Buff Bradley with James Graham picking up the mount. JEMROSE (#3) drops a notch in class and looks like the one to beat. TIZ GIG GEE (#1) is in for a tag for the first time and fits with these.
2nd Race: ULTIMA D (#8) is a $300,000 Keeneland sales grad who debuts for trainer Wesley Ward. She worked head to head with Fairyland, who won the baby race her on opening day. CLASSY MUSIC (#5) is training sharply for Kenny McPeek and has to be respected. BUY SELL HOLD (#2) is trained by Steve Asmussen and is a contender.
3rd Race: PUCA (#2) is graded stakes-placed on the dirt but may be better with the switch to the turf. She was fourth behind Coolmore Jenny Wiley (G1) winner Dickinson last out and rates the edge with Joel Rosario picking up the mount for trainer Bill Mott. SASSY LITTLE LILA (#5) had a solid 3-year-old campaign capped by a runner-up finish in the American Oaks (G1). She is training well for Brad Cox, who is very good with returning runners. SHAAN (#3) comes off a couple of solid efforts for Todd Pletcher and has to be respected.
4th Race: SHE STAYS ROLLIN (#5) is a first timer by War Front who has several solid works over the Keeneland strip. She looks like another solid win candidate for trainer Wesley Ward. WAKI PATRIOT (#3) was no match for Fairyland on opening day but ran well to finish in a dead heat for second. TRUE DREAM (#9) is another one training well for Kenny McPeek.
5th Race: ANCIENT WARFARE (#8) closed well to finish second in his 3-year-old debut and figures to move forward off that one. Give him the slight edge for trainer Mark Casse with Julien Leparoux aboard. BOUND FOR NOWHERE (#12) didn't beat much at Turfway but he did it very easily. He had a good work on April 9 in company with European champion Lady Aurelia, winner of Saturday's Giant's Causeway here, and is a solid contender in a wide-open race. KISS THE TOAD (#11) has shown improvement in his last two and has to be respected.
6th Race: FACTION CAT (#3) came back after a long layoff and ran a decent race on the grass in Tampa. He closed his 3-year-old campaign with two good races here and at Churchill. His best would make him a winner today. COAL FRONT (#9) was a high-priced 2-year-old in training purchase last April and debuts for Todd Pletcher off a series of solid works. He is the one to beat. ONE RIVER PLACE (#7) makes his first start for trainer Al Stall and posted a recent bullet work at Fair Grounds.
7th Race: CONQUEST BABAYAGA (#2) has won three of her five races and comes off a game score at Gulfstream for trainer Chad Brown. She rates a slight edge in what appears to be a very well-matched turf race. SCATTER GUN (#1) has also been very good on the grass and has the speed to be on or near the lead from the inside post. She could be tough to catch with Florent Geroux aboard for trainer Neil Pessin. CROOKED STICK (#5) is capable of running well at a nice price.
8th Race: PABLO DEL MONTE (#2) is a graded stakes-placed runner who was claimed from Wesley Ward in 2015. He showed nothing in his four starts last year but Ward claimed him back in June 2016 and he has been training well for his return. He should be ready to go today. RAY'SWARRIOR (#5) would win this one with his best performance but seems to have lost a step or two this year and the class drop isn't a good sign. CIRCLE UNBROKEN (#4) needed his race at Oaklawn and rates a chance for trainer Garry Simms.
9th Race:KAHRUMANA (#9) just missed in her debut on the Gulfstream and appears to be in a good spot to get her maiden win for Chad Brown and Joel Rosario. HOW (#12) had two rough trips in New Orleans and figures to be tough with Florent Geroux picking up the mount for trainer Tom Proctor. POUNDS TO PENNIES (#7) is another one who ran very well in her debut for Bill Mott and is a solid contender.


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Bold and bare
Rome, Dec. 3 (Reuters): Calendars in Italy use bare breasts to sell everything from cars to coffins.
Now, a different calendar uses photos of nude women who have had breast reconstruction to sell a very simple but vital message that there is life for women after breast cancer.
Calendars in Italy are usually associated with female beauty and we wanted to send a message that courage is beautiful too, Dr Simonetta Franchelli told Reuters yesterday.
Franchelli and her colleague Marisa Muggiano, both of whom work at the Genoa Tumour Institute, came up with the idea to encourage women who have undergone radical mastectomy and reconstructive surgery to lead normal lives.
Film fever
Hanoi (Reuters): Cable television viewers in Vietnams capital, frustrated by the governments blackout of Apocalypse Now Redux, can just run out to their local bootleg DVD store and buy a copy of the Vietnam War classic. Local cable operator Vietnam Cable Television Co, bowing to the governments ban on sex, violence and politically sensitive issues, stopped the screening of Apocalypse Now Redux on Sunday night about a half-an-hour after it started. We cannot relay (the film) when we see scenes of shooting and killing our people in the film, Nguyen Huu Loc, director of the company, said. Yet copies of the critically acclaimed classic by Francis Ford Coppola are easily available in stores in downtown Hanoi. A visitor to a shop on Tuesday bought a pirated DVD copy of the movie for $1.30.
Hamburg (Reuters): Extramarital sex can increase your risk of having a heart attack, a British cardiologist said. Graham Jackson, a heart specialist at St Thomas Hospital in London, found couples in long-term relationships were far less likely to have heart attacks while having sex than those having affairs or one-night stands. We found that 75 per cent of the cases of sudden death during sexual activity involved people who were taking part in extramarital sexual intercourse, Jackson said. He added the danger of having heart failure during extramarital sex was exacerbated even further when there was a significant difference in age between the partners. Jackson said heart rates accelerate considerably during all sexual activity. He said measurements on blood pressure and heart rates found sex is generally comparable to a brisk 20-minute walk, with an orgasm putting a strain on the heart similar to an ensuing walk up a flight of stairs.


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Wessex FM News
Tributes Paid To Former Fire Authority Chairman
12:38pm 7th May 2014
(Updated 12:55pm 7th May 2014)
The flag at Dorset Fire and Rescue's headquarters in Dorchester is flying at half- mast today.
It's in tribute to ex-Chairman and former councillor David Fox who died yesterday morning.
David worked with Dorset Fire Authority for 30 years and oversaw Dorset Fire Brigade move to become Dorset Fire and Rescue.
He leaves behind his wife and two sons.
Chief Fire Officer Darran Gunter said: "I am extremely saddened to learn of David's passing. For over 30 years David was hugely committed to the Fire and Rescue community both within Dorset, Bournemouth and Poole and also involved in wider national leadership issues. He was a passionate and caring man and will be missed by us all."
Chairman of Dorset Fire Authority, Councillor Rebecca Knox said: "David was one of the first county councillors I met when I was elected in 2009. His passion for the fire and rescue service was evident and he immediately inspired me to accept his invitation to become part of the Dorset Fire Authority. His dedication had been evident as I met with many councillors and officers both in Dorset and nationally who wished him well. On behalf of the Members of Dorset Fire Authority, our thoughts are with his family and friends, David's contribution will be remembered."
(Picture supplied by Dorset Fire and Rescue)
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Yesterday was World Water Day…..Birdelini is showing her support!
"Water pollution is a main cause of reduced water availability and can have serious impacts on the environment and on human health. Pollution stems for inappropriate industrial and agricultural practices as well as from urban waste production. In agriculture, overuse of chemical fertilizers and of pesticides leads to contamination of water bodies (rivers, lakes, underground aquifers). Intensive livestock production can also impact the quality of water resources if measures are not taken. New, more integrated approaches to food production can substantially improve the situation and limit pollution.
Protection also means conservation. A good soil, well maintained, can capture much of the rainwater, and avoid surface runoff which causes erosion and the loss of soil nutrients. Conservation agriculture is a farming practice that makes best use of available water, increases the resistance of plants to droughts and at the same time contributes to improving both the quantity and quality of groundwater and rivers.
Management of watersheds and the protection of water sources are also important. Forests can play an important role in protecting water resources."


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Right now, I am down to the last few days before my second marathon which will be the ING Miami Marathon that takes place on January 27, 2013. A lot of time was put into the training for this race, but it was required if I wanted give myself every chance possible to meeting my goal.
For a race, typically you download a plan and you follow it as much as possible to ensure you are prepared to reach your goal for that race. However, what is more important is during the training weeks is that you learn how much you truly are capable of if you have a goal and a plan in place.
· You realize how much you can handle on a daily basis.
oYou make time for the training that is required for you to be prepared for race day.
· You learn to make your goal an important part of your daily life.
oYou make daily choices that will affect how you will be ready for the big day.
· You stop making excuses.
oIf race day is the big goal, you will make sure you no longer make excuses and get the necessary exercise in.
· You eliminate out of your life (at least during the training) what is not important.
oTo ensure you get your training in for the day or week, you no longer spend time on consuming tasks that do not contribute to your goal.
Are there some tough times during the training? You better believe it. You become irritated at times for a variety of reasons:
· Tired from the training you put in on a daily basis.
oSometimes you get up earlier than you want, other times you stay up later than you prefer.
· Tired from passing up some of the food items you are used to having.
oIf you are trying to lose weight along the way or at least not gain weight, you have to consistently make great food choices.
· Frustrated on not having time for some of the activities your family or friends are attending.
oThere will be some gatherings that you may need to pass on because you have to get up early in the morning for a run. It can become an issue, but if you share your goal with others they will understand why you are making your choices.
Does this sound difficult? No doubt, it is a bit overwhelming at first. However, like the first four bullet points state, you learn so much during this journey and more importantly you become a stronger person. Typical training plan is only 16 weeks. Only four months away from reaching a goal that at one time you only dreamed of accomplishing. If you decide to download a plan, you will have the outline of what needs to be done. Now the only action to take is to commit to it.
What truly has stopped you from doing a "race"? Fear? As Steven Pressfield said in his The War of Art book, "The more scared we are of a work or calling, the more sure we can be that we have to do it."
I know I have stated this before, but I challenge you to go to a local race so you can see all the different types of people that finish them. You will see young and old, fit and out of shape. Age, color and shape do not matter at this type of event. What is special about these people though was that they signed up and showed up for the race, even if they knew it was going to be difficult. They conquered their fear.
I know I have stated this before, but I challenge you to go to a local race so you can see all the different types of people that finish them. You will see young and old, fit and out of shape. Age, color and shape do not matter at this type of event. What is special about these people though was that they signed up and showed up for the race, even if they knew it was going to be difficult. They conquered their fear.
Have you been doing 5Ks, but have not stepped it up to 10Ks? Why not? If you can run 3.1 miles, you can run 6.2. What really scares you from signing up for your first or next race? Why continue to take the easy road when you can learn to travel along the difficult path that gets you closer to the person you are capable of being. There will be some bumps along the way, but it will make you a better person at the end of the journey.
You can also develop some type of plan to reach non-running goals and it will assist you in getting to your ultimate dream in your career and life. However, to get where you want to go you need a map or plan to ensure you get to your ultimate destination.
By: Fred Lechuga
By: Fred Lechuga


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Miami Marlins starting pitcher Kevin Slowey has been outstanding in four starts this season. Unfortunately, he has nothing to show for it as Miami's offense has yet to give him any significant run support in his outings. Will Slowey be able to record his first win of the 2013 Major League Baseball season when the Marlins take on the Chicago Cubs Thursday night?
The 28-year-old is 0-2 with a 1.90 earned run average, 15 strikeouts, six walks and a .261 opponent batting average in 23 2/3 innings this season. His ERA is the best on the team among pitchers who have tossed at least four innings this year. Only rookie starter Jose Fernandez can rival Slowey's overall numbers, with the exception of ERA, and he has not received any run support either.
Slowey has only faced the Cubs one time in 104 career games. That contest occurred on June 12, 2009 when he was making starts for the Minnesota Twins. Slowey allowed three earned runs on four hits while striking out a whopping 10 batters in six innings. He walked away with the win, something the Marlins are hoping he can do again in his second time around against Chicago.
The right-hander has only given up more than one earned run in one game in his first four starts. That came against the Atlanta Braves on April 8 in which he surrendered two earned runs on six hits in seven innings. If Slowey can keep the Washington Nationals, Philadelphia Phillies and Cincinnati Reds sluggers at bay then he should have no problem keeping the Cubs quiet.
Hopefully, the Marlins will be able get some runs off of Chicago starter Edwin Jackson in order for Slowey to be eligible for the win.
Michael Terrill is a Senior Writer for Rant Sports. Follow him on Twitter @MichaelTerrill, "Like" him on Facebook or add him to your network on Google.


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3 and a half days ago, my 20 month old daughter had redness on her top lip. The next day it turned into what looked like a blister and looked a little infected. When we first noticed the redness, we checked the inside of her lip which looked like she had bruising/blood spotting. I took her to three pharmacies.. One said no to cold sore and the other two weren't sure. I took her to one doctor who said 99% cold sore.. I took her to another doctor who said no to cold sore and more like a school sore.. A sore which has become infected and bacterial rather than viral. There has only been one big blister - not little multiple blisters. It also started to clear quite quickly and today is only a little red mark with a little bit of skin. There was never any crust.. Moreso just a little scab. It hasn't seemed to cause her any pain whatsoever and she hasn't been trying to touch it or anything. Is it likely that this is a cold sore? As mentioned it has been 3 and a half days and it practically cleared up - I thought cold sores last quote some time. My daughter has never been in any type of care.. My mum gets cold sores but is always super careful. I therefore don't know where she would have caught the virus. The second doctor also took a swan yesterday but said that because the sore is already quite healed, it would likely give a negative regardless of whether it is or isn't a cold sore. Thank you.
Does this sound like a cold sore?
- Asked
- 3 May 2016 by worriedmummy123
- Topics
- cold sores, blister, lips, sores
Added 3 May 2016:
Ps it didn't get any bigger at all.. She often puts her fingers in her mouth and uses a dummy - which I thought, if it was a cold sore, would irritate and cause spreading?
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text: by Clare Plested on 23rd August 2016 Clare Plested is spending her Fringe in bars drinking cocktails - just so you, dear reader, can sample a new one every day. Clare is also a borderline alcoholic."Salty Margarita at the Abattoir... Definitely not one of my five a day."Clare Plested: Flock UpVenue: PBH Free Fringe – Ciao Roma (Venue 283)Dates: 6th to 27th August 2015 (not 17th) Time: 5.50pm (6.50pm)


text: Does anybody know of a way to stop status bar notifications for email WITHOUT stopping the ringtone or vibration that goes with the email? I am using K9 mail on Droid X 2.2. My single biggest nag about this phone is that when I go directly to my email app, bypassing the status bar, the status bar notification doesn't go away unless I go back and re-read the email from the status bar. This is one of those crazy things that is just driving me nuts. I do not want to use the status bar for email. Oddly, texting (via handcent) works exactly like I would like for it to work. Once I check the text directly from the app, the status icon goes away. Thanks,


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|Submission Period
|Winter 2018
|October 16, 2017 to December 30, 2017
|Spring 2018
|Poetry: Formal Poetry
Any form – specify the form with your submission.
Fiction: Short Story 1000-500 words or Flash Fiction 500-100 words.
|January 16, 2018 to March 30, 2018
We want your best original, unpublished literary work on the theme for the current submission cycle. You are free to interpret the theme in your own way. For our tastes in fiction, feel free to read this article by our editor Pratibha Kelapure.
By submitting, you declare that your work:
- is unpublished,
- is your own work,
- and you own the rights to the work.
Formatting Your Manuscript:
- Type the manuscript in 12-point Times New Roman font, with one-inch margins, and sequentially numbered pages.
- Use double-spacing for fiction and single-spacing for poetry.
- Include the author's name, address, telephone number, and email address at the top of the first page.
- Include a 75-word third-person biography at the end.
- Send only ONE document containing everything. We accept.doc, .docx, or .rtf files.
Simultaneous Submissions: If this is a simultaneous submission, please indicate in your submission and if it is accepted elsewhere before we send an acceptance, please let us know immediately. Once accepted by us, you can not withdraw it later.
No revisions to the submitted work are allowed. The editors reserve the right to do minor edits for clarity.
Word Count Guidelines:
Fiction: 1000-5000 words.
Poetry: Up to five poems. Each poem must be 50 lines or fewer. We love all forms of poetry: all forms of metered, free-verse, form, and prose poetry.
How to Send Us Your Work: We accept submissions only through email at this time. You may send us manuscripts for the following categories: Short Story and Poetry. Your manuscript must be in one of the following file forms: .doc, .docx, or.rtf.
Before you submit, follow us in at least one of the following ways.
Please send only ONE document to email@example.com with "Submission" in the subject header.
Please wait at least three months between submissions.
Submissions not adhering to these guidelines will be deleted unread.
Publication Rights: The Literary Nest acquires First North American Serial Rights to published works, which means that copyrights for any works published in The Literary Nest revert to the author upon publication. We reserve the right to feature published works or excerpts on our website and in its promotions. We also reserve the right to use your work in any future print anthology.
Every submission is acknowledged upon receipt. We send acceptance/rejection letters before the issue is published. If you didn't hear from us, feel free to contact us.
Please direct any questions to email@example.com.


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Damian Burgess
Phone: 0428 558 590
Wollermann & Associates
415 Canterbury Road,
Surrey Hills, VIC 3127
Phone: 03 9888 6488
Fax: 03 9888 4040
Mildura, MILDURA 3500
Retail for sale
Business Details
Mildura - Cappy's Corner Store and Post Office (DB4416)
Post Office and Mixed Business located in Mildura, regional city in northwestern Victoria. It is located in the Sunraysia region and is on the banks of the Murray River. The population is just over 51,000. Life in Mildura revolves around the Murray River, with fishing, water sports and the desert environment of Mildura-Sunset National Park.
Products & Services :
Australia Post Banking facilities (large banks and credit unions), bill payments, newspapers, magazines, post office supplies, mobile phones & recharge vouchers, stationary, gift lines, groceries, confectionary including drinks, tobacco, cards for all occasions. Eftpos accepted.
Ideal Location:
The business is situated on a main arterial road connecting NSW and Victoria and linking with South Australia, on a busy corner with ample parking directly outside. Bright sunny shop that utilizes all available space.
Well presented premises with proper presentation of stock.
Trading Days:
This business trades seven days per week.
Owners Role: Hands on
This business requires 2 part time employees when operated as a partnership.
Australia Post Epos Terminals: Two
Post Boxes:
There are 80 boxes on site, of which 56 are currently leased.
Price: $250,000 + SAV (approximately $90,000)
Mildura also has a strong arts culture with the Art Gallery adjacent to the Murray River. The centre includes a 384 seat theatre, the Mildura Art Gallery and the original Caffey House.
Mildura presents significant economic opportunities as the primary activity centre for the region.
Mildura connects Victoria, South Australia and New South Wales by road, rail and air. The Mildura Airport is strategically vital to the Sunraysia region. It is the busiest regional airport in Victoria for passenger services.
The central business district features a pedestrian mall and shopping centre. However, this shopping precinct competes with the Mildura Central Shopping Centre, located at the opposite end of the urban area which is also the main strip of big box stores and other commercial enterprises.
The Mildura Senior College (AKA Mildura Secondary College), Chaffey Secondary, Sunraysia Institute of TAFE and the La Trobe University operates a regional campus in Mildura. There are several primary and secondary public and private schools to cater for all students in the district and beyond.
Excellent sporting facilities are available to cater for everyone including AFL football, cricket, horse racing, bowls, golf, darts, tennis, motor sports and swimming.
For further information please contact Damian Burgess on
Click to call or Click to call or email
Click to email
Summary »
- Business Category: Retail
- Business Sub Category: Post Offices


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November 6, 2009 – Today representatives of the Animal Welfare Network Nepal and Anti-Animal Sacrifice Alliance handed over a petition signed by 3200 individuals from all over the world to Karima Bagam, State Minister of Agriculture and Cooperatives. Pramada Shah, Manoj Gautam and Lucia de Vries urged the minister to ensure that all animals brought to Gadhimai to be sacrificed carry a certificate from an Indian or Nepali quarantine office.
Ms Begam promised to discuss the suggestion with her officers. She explained that Gadhimai is a very sensitive issue and that due to a limited number of quarantine offices and resources ensuring the wellbeing and health of the animals is difficult. She invited the representatives to join a meeting to finalise the long awaited Animal Welfare Act at her Ministry.
According to the campaigners the festival is not only cruel and inhumane but also dangerous. 'Due to the fact that the transport, killing, disposal and preparation of the animals and their meat goes unmonitored and the close proximity between humans and animals, Gadhimai can result in widespread food poisoning (Salmonella, E coli, Campylobacter, etc), Tuberculosis, as well as bird and swine flu,' the campaigners argue.
Friday, November 6, 2009
3200 signatures handed over to State Minister
Posted by Animal Welfare Network Nepal at 9:30 AM
Labels: awnn, Karima Begam, State Minister
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Stay with us for your next corporate event in Boston
Travelers attending corporate events in Boston can improve their business trip by staying at John Jeffries House. Conveniently located in downtown's Beacon Hill neighborhood, the intimate hotel is only minutes away from conference hubs like the Boston Convention and Exhibition Center and the Hynes Convention Center. Guests can walk to meetings and events at the State House, Government Center, Massachusetts General Hospital and much of the Financial District in under a fifteen minute walk.
John Jeffries House provides complimentary Wi-Fi throughout the hotel, letting business travelers stay connected as they work in their rooms, common areas and garden. Guests can enjoy complimentary Continental breakfast before attending long meetings at any of the nearby corporate event venues in Boston. Tastefully-designed apartment-style guestrooms, bookable in room-blocks, make John Jeffries House Hotel perfect for companies attending conferences as a group.


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Dom Pedro Portobelo, Vilamoura's most popular private apartments provides the ideal location for visitors seeking a family holiday that offers the privacy and comfort of your own apartment complete with private balcony and kitchenette, with the benefits of four star aparthotel services. Dom Pedro Portobelo offers a privileged location in the heart of Vilamoura, facing the Casino and a short walk from the International Yacht Marina and the Atlantic Ocean. Dom Pedro Portobelo - Vilamoura at it's finest.
Cancel by 4pm hotel time, day of arrival to avoid a penalty of one night\'s room and tax.
Avenida Tivoli, Lote H5CP:8125410,Faro,Portugal


text:
Game Four: Cubs (1-2) vs. Brewers (1-2)
Game Time – 6:05pm CDT
Coverage – TV: ESPN Monday Night Baseball/WCIU – Radio: 720 WGN, XM Radio Channel 185/901
Location – Wrigley Field, Chicago
Chris Volstad (0-0, -.–/-.–) vs. Shaun Marcum (0-0, -.–/-.–)
The first series of the year is in the books and the season gets into full swing tonight with the first of four against the defending NL Central Champions. The Cubs and Brewers have created quite a rivalry in recent years and tonight's opener will feature two firsts … the first time Dale Sveum has faced his former organization and the first time Aramis Ramirez has been in the visitor's dugout at Wrigley Field since 2003.
Chris Volstad gets the tall task in his Cubs' debut of not only facing a very good offense but trying to follow the performances of Ryan Dempster, Matt Garza and Jeff Samardzija with a quality start of his own. The Brewers will also be looking to put their rough weekend behind them … Milwaukee dropped two of three to the Cardinals.
Starlin Castro begins play tonight riding a career-high 14-game hitting streak. Dating back to last season, Castro has reached base safely in his last 43 games. Castro is close to tying the Cubs all-time mark of 44 set by Riggs Stephenson in 1928.
Blake DeWitt was a late scratch Sunday and is listed as day-to-day with back spasms.
Last year's group of players posted a 6-10 record against the Brewers and the top three teams in the division (Brewers, Cardinals and Reds) are looking at the Cubs again as a way to add to their season-win total.
The Cubs first night game of the year all begins with Chris Volstad …
The Cubs were very happy with the adjustments Volstad made in the spring. Volstad worked with Rick Sutcliffe throughout the Cactus League and the Red Baron thinks Volstad will have a good year. Tonight is the first step in the process.
Volstad will make just his fifth career start at Wrigley tonight … and he's not posted a win in the first four (0-1 with a 4.91 ERA and a 1.27 WHIP).
Chris Volstad is 1-1 in four career starts against the Brewers with a 1.71 ERA and a 0.99 WHIP … last season, 0-1 in two starts with a 2.57 ERA and a 0.93 WHIP.
Brewers Career Numbers against Chris Volstad
- Ryan Braun – 2-for-12
- Carlos Gomez – 1-for-3
- Alex Gonzalez – 3-for-12 with two doubles
- Corey Hart – 2-for-10 with two walks
- Travis Ishikawa – 1-for-3
- Cesar Izturis – 0-for-2
- Jonathan Lucroy – 2-for-6 with a double
- Nyjer Morgan – 3-for-17 with a home run
- Aramis Ramirez – 6-for-12 with two doubles, a home run and a walk
- Rickie Weeks – 2-for-12
The Cubs will get their first look at Shaun Marcum after he dominated a much different lineup a year ago. Marcum is one of the keys to the Brewers success this season. He was solid a year ago (13-7 in 33 starts with a 3.54 ERA and a 1.16 WHIP) but faltered down the stretch and was horrible in the playoffs.
Marcum shutdown the Cubs in his first season with Milwaukee … Marcum won both starts (2-0) and posted a 1.93 ERA and a 0.93 WHIP.
Cubs Career Numbers against Shaun Marcum
- Darwin Barney – 3-for-7 with three doubles
- Marlon Byrd – 1-for-10 with a walk
- Starlin Castro – 1-for-7 with a home run
- David DeJesus – 0-for-16 with three walks
- Reed Johnson – 1-for-1
- Bryan LaHair – 0-for-5
- Alfonso Soriano – 1-for-6 with a double
- Geovany Soto – 1-for-6
- Ian Stewart – 0-for-2
Let's Talk Cubs Baseball …
Follow ChicagoCubsOnline on Twitter: @TheCCO