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"Humanized" mouse models created by engraftment of immunodeficient mice with human hematolymphoid cells or tissues are an emerging technology with broad appeal across multiple biomedical disciplines. However, investigators wishing to utilize humanized mice with engrafted functional human immune systems are faced with a myriad of variables to consider. In this study, we analyze HSC engraftment methodologies using three immunodeficient mouse strains harboring the IL2rγnull mutation; NOD-scid IL2rγnull, NOD-Rag1null IL2rγnull, and BALB/c-Rag1null IL2rγnull mice. Strategies compared engraftment of human HSC derived from umbilical cord blood following intravenous injection into adult mice and intracardiac and intrahepatic injection into newborn mice. We observed that newborn recipients exhibited enhanced engraftment as compared to adult recipients. Irrespective of the protocol or age of recipient, both immunodeficient NOD strains support enhanced hematopoietic cell engraftment as compared to the BALB/c strain. Our data define key parameters for establishing humanized mouse models to study human immunity.
The discovery of the severe combined immunodeficiency mutation (Prkdcscid, abbreviated as "scid") in a stock of C.B-17 mice in 1983 [1] led to the assessment of their ability to support engraftment with human peripheral blood mononuclear cells [PBMC, termed Hu-PBL-SCID, 2] or fetal [termed SCID-Hu, 3] or adult [termed Hu-SRC-SCID, 4] hematopoietic stem cells (HSC). These human hematolymphoid engrafted mouse models have been used in multiple biomedical disciplines for the study of human immunobiology and hematopoiesis (reviewed in [5–8]). However, the utility of the early strains of immunodeficient mice transplanted with human hematolymphoid cells was hindered by the inability to achieve multi-lineage hematopoietic engraftment leading to the generation of a functional human immune system [5,6]. In the Hu-PBL-SCID model, although all lineages of peripheral blood mononuclear cells (PBMC) are injected, the majority of human cells engrafted are T cells, with few B cells, myeloid cells or natural killer (NK) cells surviving. In the Hu-SRC-SCID model, the major obstacle was the inability of HSC-engrafted mice to develop all components of a functional human immune system, particularly T cells [5].
A major technological breakthrough occurred when investigators created genetic stocks of scid, Rag1null or Rag2null mice that also harbor mutations in the IL2 receptor common gamma chain (IL2rγ) gene [9–12]. IL2rγ is required for high affinity signaling through multiple cytokine receptors, including IL2, IL4, IL7, IL9, IL15 and IL21 [13]. Immunodeficient mice bearing a mutated IL2rγ gene support much higher levels of human hematolymphoid engraftment than all previous immunodeficient stocks. Multiple immune lineages develop following engraftment of human HSC that can lead to the generation of a functional human immune system [5–8].
Confounding widespread establishment of humanized mouse models in multiple laboratories is the diversity of approaches that have been reported for engrafting human HSC. For example IL2rγnull mice are available on a number of strain backgrounds. The ability of immunodeficient mice to support engraftment of human hematolymphoid cells has been shown to be strongly affected by the genetic background of the host [5,14–17]. In addition to strain background, reports differ significantly with regard to engraftment methodologies, which include intravenous (IV) engraftment into adult mice [11], or intrahepatic (IH) [10], intraperitoneal (IP) [12], and IV [9,18] injection into newborn mice.
In the present study, we compared a number of variables of human HSC engraftment, including strain background, age of recipient, and engraftment route. The three strains of immunodeficient mice tested were NOD-scid IL2rγnull, NOD-Rag1null IL2rγnull, and BALB/c-Rag1null IL2rγnull. To provide a consistent comparison, we used human HSC derived from umbilical cord blood (UCB) in all of our experiments. We matched UCB donors within an individual experiment, allowing direct comparison of the engraftment characteristics in each strain, independent of donor-to-donor variability. We observed that in the variables tested, newborn immunodeficient NOD strain mice provided the optimal hosts with respect to their ability to support human hematopoietic cell engraftment.
NOD.Cg-Prkdcscid Il2rgtm1Wjl/Sz (NOD-scid IL2rγnull) and NOD.Cg-Rag1tm1Mom IL2rgtm1Wjl (NOD-Rag1null IL2rγnull) mice have been described previously [11,19]. C.Cg-Rag1tm1Mom Il2rgtm1Wjl (BALB/c-Rag1null IL2rγnull) mice were generated by LDS by crossing C.129S7(B6)-Rag1tm1Mom mice (BALB/c-Rag1null) with C.129S4-Il2rg tm1Wjl (BALB/c-IL2rγnull) mice and then intercrossing the F1 progeny to fix the Rag1null and the IL2rγnull alleles to homozygosity. Mice were housed in a specific pathogen free facility in microisolator cages, given autoclaved food and maintained on acidified autoclaved water and sulfamethoxazole-trimethoprim medicated water (Goldline Laboratories, Ft. Lauderdale, FL), provided on alternate weeks. All animal use was in accordance with the guidelines of the Institutional Animal Care and Use Committee (IACUC) of the University of Massachusetts Medical School and The Jackson Laboratory and conformed to the recommendations in the Guide for the Care and Use of Laboratory Animals (Institute of Laboratory Animal Resources, National Research Council, National Academy of Sciences, 1996).
Umbilical cord blood (UCB) was obtained in accordance with the Committee for the Protection of Human Subjects in Research guidelines of the University of Massachusetts Medical School and was provided by the medical staff of the UMass Memorial Umbilical Cord Blood Donation Program. The program educates and consents mothers regarding UCB collection for research and public banking and performs collections at the time of delivery. Red blood cells were removed from UCB by hetastarch (Baxter HealthCare Corp, Deerfield, IL) to reduce RBC content, followed by double-density Percoll gradient centrifugation (1.05/1.077). The recovered cells were then depleted of T cells using commercially available magnetic bead kits according to the manufacturer's instructions (Miltenyi Biotec, Auburn, CA or Stem Cell Technologies, Vancouver, BC, Canada). Efficiency of T cell depletion and percentage of CD34+ cells were evaluated by flow cytometry prior to injection into recipient mice and in all experiments revealed less than 0.5% contaminating CD3+ cells in the UCB preparation. T cell-depleted cord blood was suspended in PBS in a volume to deliver 3×104 CD34+ HSC per recipient (50μL for newborn recipients; 500μL for adult recipients).
Recipient mice were engrafted with one of the following engraftment protocols, as indicated in the Results section. Within an individual experiment, mice of each strain received CD34+ stem cells from the same cord blood donor. At least 3 experiments, each with a unique UCB donor were performed. In all cases, recipient mice were evaluated for human hematolymphoid engraftment at 12 to 16 weeks post-injection.
Adult (6–12 week old) NOD-scid IL2rγnull, NOD-Rag1null IL2rγnull, and BALB/c-Rag1null IL2rγnull mice were irradiated with either 240 cGy (scid mice) or 550 cGy (Rag1null mice) using a 137Cs source (GammaCell 40, Atomic Energy of Canada, Ottawa, Canada). Four hours after irradiation, 3×104 human CD34+ HSC were injected IV into the lateral tail vein.
NOD-scid IL2rγnull, NOD-Rag1null IL2rγnull, and BALB/c-Rag1null IL2rγnull mice 24 to 48 hours old were irradiated with either 100 cGy (scid mice) or 400 cGy (Rag1null mice). Mice were injected shortly after gamma-irradiation with 3×104 CD34+ HSC via IC injection using a 27 G winged infusion kit attached to a 1cc syringe. Injected pups were returned to their nursing mothers until weaned [20].
This protocol was performed identically to Protocol B, except that mice were injected via IH injection [20].
For analysis of human cell populations in engrafted mice, anti-human CD3, CD4, CD8, CD34, and CD45, anti-mouse CD45 fluorochrome-conjugated monoclonal antibodies (mAbs), and appropriate isotype control Abs were obtained from BD PharMingen (San Diego, CA). Antibodies were conjugated with FITC, PE, PerCP, APC, Alexa 405, Pacific Blue, or Alexa700.
Single-cell suspensions of bone marrow, spleen, and thymus were prepared from nonengrafted and human HSC engrafted mice. Whole blood was collected in EDTA. Red blood cells in bone marrow and spleen were removed by lysis with a hypotonic solution. Cells counts were determined using a Coulter Counter (Beckman, Miami, FL). Single cell suspensions of 1×106 cells in a 50μL volume or 75μL of whole blood were pre-incubated with rat anti-mouse FcR11b (clone 2.4G2) to block Fc binding. Cells were incubated for 30 min at 4°C in appropriate dilutions of antibodies as determined in preliminary experiments. For analysis of engrafted mice, PerCP-conjugated anti-mouse CD45 mAb and APC-conjugated anti-human CD45 mAb were included. All cells that were negative for murine CD45 and positive for human CD45 were analyzed for the presence of multi-lineage differentiation markers. Labeled single cell suspensions were washed in RPMI and fixed in 1% paraformaldehyde in PBS. The staining protocol of whole blood was performed according to manufacturer's instructions (BD Biosciences, San Jose, CA). At least 50,000 events were acquired on BD Biosciences LSRII or FACSCalibur instruments (BD Biosciences). Data analysis was performed with FlowJo (Tree Star, Inc., Ashland, OR) software.
Tissues recovered from non-engrafted and HSC engrafted mice were fixed in 10% buffered formalin. H&E and human CD45 staining of tissue sections were preformed as previously described [11].
Parametric data were compared by one-way ANOVA with Bonferroni post-tests to compare individual pair-wise groupings and nonparametric data were compared by a Kruskal-Wallis test with Dunns post-test to compare individual pair-wise groupings. Significant differences were assumed for p values <0.05. All statistical analyses were performed using GraphPad Prism software (version 4.0c, GraphPad, San Diego, CA).
The optimal mouse strains (described in Table 1) that support engraftment of human HSC harbor mutations in the IL2 receptor common gamma chain (IL2rγnull) and are available on a number of genetic backgrounds, including NOD-scid, NOD-Rag1null and BALB/c-Rag2null [10,11,19]. As the BALB/c-Rag2null IL2rγnull stocks are not commercially available, we generated a BALB/c-Rag1null IL2rγnull stock by crossing the BALB/c-Rag1null strain with the BALB/c-IL2rγnull strain. These parental stocks were obtained from the Repository at The Jackson Laboratory (http://www.jax.org/). Null mutation of either the Rag-1 or the Rag-2 genes results in the inability to generate mouse T or B cells with rearranged antigen receptors [21,22]. As expected, BALB/c-Rag1null IL2rγnull mice are completely devoid of T, B and NK cells (data not shown) and we have shown previously that they can be engrafted with human PBMC, although not as efficiently as NOD-scid IL2rγnull mice [23].
IV injection of human HSC into adult, irradiated mice has been used to establish humanized mice. We have previously shown that this approach is suitable for establishing high levels of human hematolymphoid engraftment into adult NOD-scid IL2rγnull mice and NOD-Rag1null IL2rγnull mice [11,19]. In contrast, it has been reported that reproducible engraftment of adult BALB/c-Rag2null IL2rγnull mice with HSC [24] or PBMC [25] requires macrophage ablation by treatment with chlodronate-containing liposomes.
We first compared directly the ability of the three strains of adult IL2rγnull mice to be engrafted following the IV administration of HSC into irradiated adult recipients. Mice 6–12 weeks of age were sublethally irradiated with 240 cGy (scid mice) or 550 cGy (Rag1null mice) and injected IV with 3×104 CD34+ cells from the same UCB donor. NOD-scid IL2rγnull mice received a lower dose of radiation because the scid mutation results in increased radiosensitivity [19]. Twelve weeks later, engraftment levels were determined by quantifying the percentages of human-CD45+ cells in the peripheral blood. We found that immunodeficient NOD strain background mice engrafted at significantly higher levels as compared to BALB/c-Rag1null IL2rγnull mice (Figure 1). Engraftment levels in the blood of the adult recipients were reflective of the relative engraftment in the spleen, both within an individual recipient and among various strains (data not shown).
As an alternative to adult engraftment with HSC, many investigators have chosen to deliver HSC to newborn mice [10,18]. Using NOD-scid IL2rγnull mice, we compared directly the engraftment levels obtained in peripheral blood of mice engrafted as newborns (IC injection) versus adults (IV injection). While overall human CD45+ engraftment did not differ statistically between these two cohorts of mice, the data trended towards higher engraftment in newborns (Figure 2A). However, the ability to support human T cell development was dramatically improved in newborn NOD-scid IL2rγnull mice as compared to their adult counterparts (Figure 2B). In addition, both newborn and adult mice engrafted at high levels with human B cells (Figure 2C).
These data document that human T cell levels in HSC-engrafted adult mice are reduced as compared to HSC-engrafted newborn mice. We next questioned whether this engraftment deficiency is related to defects within the thymic architecture of adult mice rendering the thymi as unsuitable environments for the development of human prothymocytes. Histological sections of thymic lobes from unmanipulated newborn and 5–7-week old immunodeficient mice were used to evaluate their overall structure. Thymic lobes from all three immunodeficient strains showed an absence of lymphocytes at 1–2 days (Figure 3). At 5–7 weeks of age, all thymi remained alymphoid and contained hypoplastic cysts, consistent with the report that introduction of the IL2rγnull mutation results in the development of hypoplastic cysts that increase with age in mice with impaired immune systems [26]. Overall, it appeared that the hypoplastic cysts were more prevalent in the thymi of adult BALB/c-Rag1null IL2rγnull mice than in the thymi of the adult immunodeficient NOD strains (Figure 3).
We next sought to determine if the strain-dependent differences in engraftment observed in adult recipients would be recapitulated in newborn mice. One to 2-day-old NOD-scid IL2rγnull, NOD-Rag1null IL2rγnull and BALB/c-Rag1null IL2rγnull mice were irradiated with 100 cGy (scid mice) or 400 cGy (Rag1null mice) and engrafted via IC injection with T cell-depleted UCB containing 3×1044 CD34+ stem cells. Twelve to 16 weeks after injection, mice were analyzed for the presence of human hematolymphoid cells (Figure 4). In agreement with data from adult engrafted mice, we observed that overall human CD45+ and human CD3+ cell engraftment was higher in the blood of immunodeficient NOD strains as compared to the immunodeficient BALB/c strain mice (Figure 4A). The percent of human CD45+ cells that were CD20+ B cells was higher in the blood of mice on the BALB/c background reflecting the decreased percentages of CD3+ T cells that develop in these HSC recipients.
Human hematopoietic cell engraftment in the spleen and bone marrow was examined in a subset of HSC-engrafted mice (Figures 4B and 4C). The percentages of human CD45+ cells were significantly higher in the spleens of immunodeficient NOD strains as compared to that observed in the immunodeficient BALB/c strain. Percent CD3+ T cell levels in the spleen trended higher in NOD strain mice but this difference was not significant and there were no differences in the percentages of CD20+ B cells between three strains. However, the total numbers of human CD45+ (9 to 11-fold) CD3+ (12–17-fold) and CD20+ (6 to 9-fold) cells recovered from the spleen were significantly higher in NOD strains as compared to the BALB/c strain (Table 2). In contrast to the spleen, the percentages and total numbers of human CD45+ cells engrafted in the bone marrow were similar between the 3 strains (Figure 4C and Table 2). The percentages and numbers of human CD34+ cells in the bone marrow were also not significantly different between the strains (Figure 4C).
The CD4+ and CD8+ T cell ratios of the CD3+ population of human CD45 cells in the spleen of the 3 strains of mice ranged from 0.3 to 8.1 with the median values being 1.5 for NOD-scid IL2rγnull mice, 2.8 for NOD-Rag1null IL2rγnull mice and 0.65 for BALB/c-Rag1null IL2rγnull mice (Figure 5A). Human CD4 T cells with a regulatory T cell phenotype (CD25+ and Foxp3+) were also detected in the 3 strains of mice as indicated by the representative flow cytometry histograms (Figure 5B).
Because of the large discrepancy of T cell numbers in the spleens of immunodeficient NOD and BALB/c strains of mice, we next investigated the levels of thymic engraftment to determine if the reduced numbers of peripheral T cells in the BALB/c strain was due to reduced levels of intrathymic development of human T cells. Histological analysis revealed similar cell numbers and disorganized structure in the thymi of HSC-engrafted immunodeficient NOD and BALB/c strain mice (Figure 6A). Moreover, the percent human CD45+ engraftment was not significantly different between the 3 strains although there was a trend towards higher numbers of human CD45+ cells in BALB/c-Rag1null IL2rγnull mice (Figure 6B). Representative flow cytometry histograms of thymocyte subsets in the three strains of HSC-engrafted immunodeficient mice are shown in Figure 6C. Similar percentages of each of the major populations of human thymic T cells were detected among the strains with a majority of the cells, as expected, being double-positive for CD4 and CD8 with small populations of single positive cells.
As the original description of engrafting BALB/c-Rag2null IL2rγnull mice with human HSC utilized an alternate newborn engraftment methodology, IH injection [10], we sought to compare the hematopoietic cell engraftment achieved in mice using this route of injection. When NOD-scid IL2rγnull and BALB/c-Rag1null IL2rγnull mice were irradiated and injected with 3×104 CD34+ stem cells via the IH route, a higher percentage of human CD45+ cells were detected in the peripheral blood and spleens of NOD-scid IL2rγnull mice as compared to BALB/c-Rag1null IL2rγnull mice (Figure 7A and 7B). Again, the percent of human CD3+ T cells was significantly higher in the blood of NOD-scid IL2rγnull mice as compared to BALB/c-Rag1null IL2rγnull mice but was not significantly different in the spleens. Human B cell engraftment was similar in both blood and spleen among the strains.
In this study we have compared NOD-scid IL2rγnull, NOD-Rag1null IL2rγnull, and BALB/c-Rag1nullIL2rγnull strains of mice for their ability to support engraftment of human HSC from the same cord blood donor within individual experiments. We observed that newborn mice engrafted at higher levels than adult mice, and that in all parameters tested, NOD strain mice provided the optimal environment for engraftment of human hematopoietic cells. Although all strains of newborn mice engrafted with human HSC generated equivalent numbers and phenotypes of human thymocytes and bone marrow cells, NOD strain mice exhibited significantly higher numbers of human T cells and B cells in their spleens. No differences were observed in newborn NOD-scid IL2rγnull mice when engrafted via the IH route as compared to the IC route. These data suggest that newborn immunodeficient NOD IL2rγnull mice represent the optimal recipients for engraftment of human HSC.
The engraftment of human HSC into mice and subsequent development of a human immune system requires the elimination of murine adaptive immunity and significant reduction of innate immunity (Table 1). Initial efforts to "humanize mice" focused on models in which the adaptive immune response was abrogated, including mice bearing the scid, Rag1null or Rag2null mutation [5]. The CB17-scid mouse, which does not develop T or B cells but has moderate NK cell activity [1], was the first model used extensively to engraft human HSC, but the overall engraftment was extremely low and these mice did not develop a functional human immune system [4]. Further experimentation revealed that strain background can dramatically impact the engraftment of HSC in immunodeficient recipient mice [27]. For example NOD-scid mice show significantly higher levels of engraftment with human HSC as compared to CB17-scid mice [11]. However overall engraftment levels were still low in these models, and this was attributed to the innate immune system, including NK cell function. Thus the next objective was to develop genetic stocks of mice with a weakened innate immune system in addition to absence of adaptive immunity. The focus was to introduce specific mutations to reduce NK cell activity in NOD mice bearing the scid, Rag1 null or Rag2 null mutations. The mutant genes included β2mnull [28], to reduce NK development, perforin [29], to reduce NK cell cytotoxicity and Lsytbg [30], which interferes with NK cell degranulation. While the introduction of these mutations successfully increased the engraftment levels as compared to NOD-scid mice, overall engraftment was variable and still not optimal [5].
The next significant advancement in humanized mouse models was creating stocks of scid, Rag1null or Rag2null mice that expressed targeted mutations within the IL2rγ gene, either by a complete deletion or truncation of the receptor [9,10,12,18]. Immunodeficient mice harboring mutations within the IL2rγ gene show high levels of engraftment with human HSC and develop multiple human immune cell lineages [5]. In addition efforts are underway to improve the IL2rγ-deficient models for the development of functional human immune systems including the transgenic expression of human HLA, both class I and II, and human-specific cytokines and growth factors and by the additional deletion of murine genes to further dampen the innate immune system [31]. Two recent studies have documented the generation of HLA-A2-restricted CD8 T cell responses within A2-transgeneic NOD-scid IL2rγnull mice engrafted with HLA-A2 HSC and then infected with either EBV or Dengue virus [32,33]. However, the creation of humanized mice is still complicated due to the diverse protocols used to engraft HSC, including the growing number of mouse models available and the choices of recipient age, injection route and source of human HSC.
For the data presented in this study, we have used T cell-depleted human UCB, delivered at a dose of 3×104 CD34+ cells per recipient. T cell depletion is required to prevent the development of graft-versus-host disease in recipient mice mediated by the T cells present in unfractionated cord blood. The dose we chose is a stringent test of engraftment, as many studies have published CD34+ doses of 1×105 or greater [5–8]. However, in a practical sense, the fact that our lowered CD34+ cell dose leads to robust human hematolymphoid engraftment means that many mice can potentially be engrafted from a single UCB sample, where 1×106 or more CD34+ cells can often be obtained. We have not specifically tested the engraftment levels supported by the three strains when the cord blood sample is enriched for CD34+ cells by either lineage depletion or CD34+ positive selection. In our laboratory, we have found that more CD34+ cells are required when injecting lineage-depleted cord blood or CD34+ purified cells in order to achieve engraftment levels similar to that achieved following the injection of T cell-depleted cord blood cells (unpublished observations).
HSC can be obtained from many different sources in addition to umbilical cord blood, including G-CSF mobilized peripheral blood [11], bone marrow [4], and fetal liver [3]. A recent report suggested that UCB was equivalent to fetal liver CD34+ cells in their scid repopulating activity in immunodeficient mice [14]. We chose umbilical cord blood to do these comparisons as it is readily available and has suitable numbers of CD34+ HSC to engraft large cohorts of mice. While the human immune systems created from cord blood-derived HSC are extremely useful in studying basic aspects of human immunology, there are advantages for using HSC derived from alternative sources. For example, CD34+ cells can be isolated from the peripheral blood or bone marrow of patients with pre-existing immunologic abnormalities, such as autoimmune diseases, to study the development of clinical disease. However there are no guarantees that HSC derived from a patient would manifest the same disease in the humanized mouse models.
Three strains have emerged as the preferred hosts for human HSC engraftment: NOD.Cg-PrkdcscidIl2rgtm1Wjll (commonly referred to as NSG mice) [11,18], NOD.Cg-PrkdcscidIl2rgtm1Sug (commonly referred to as NOG mice) [9,34] and C.129(Cg)-Rag2tm1FwaIl2rgtm1Sug (abbreviated as BALB/c-Rag2null IL2rγnull) [10] mice. As the BALB/c-Rag2null IL2rγnull strain is not commercially available, we generated our own stock that is similar to the published version, except that the Rag1null mutation is employed instead of Rag2null. However, the phenotype of Rag1null and Rag2null mice are identical [21,22] and our stock of BALB/c-Rag1null IL2rγnull mice has a phenotype that is predicted from the existing BALB/c-Rag2null IL2rγnull strain and from our own NOD-scid IL2rγnull and NOD-Rag1null IL2rγnull mice.
Using this new stock of mice, we compared directly the role of the NOD and BALB/c strain mice bearing the Rag1null mutation for their ability to support human HSC engraftment. In the majority of cases, NOD-Rag1null IL2rγnull and NOD-scid IL2rγnull mice support higher levels of human HSC engraftment in the blood and spleen, and importantly for studies of human immunity, higher levels of T cell development. As previously reported [5–8], we confirmed multilineage development of all hematopoietic cells in all engrafted mice irrespective of the strain or protocol of engraftment (unpublished observations). The major differences in lineage-specific engraftment that were observed were in the T cell population between NOD and BALB/c strain mice. In contrast, human CD45+ engraftment in the bone marrow and thymus between the strains was similar. The peripheral T cell lymphopenia in HSC engrafted BALB/c-Rag1null IL2rγnull mice could result from a survival defect of human cells or a lower proliferation rate of human cells compared with the immunodeficient NOD mice. We are currently investigating these possibilities. The direct comparison of newborn and adult BALB/c-Rag1null IL2rγnull mice as was done for NOD-scid IL2rγnull mice was not performed as neither strain exhibited human HSC engraftment at levels observed in the NOD strains. However, we must caution that there are multiple substrains of BALB/c mice, and the BALB/c-Rag1null IL2rγnull stock we have generated is not identical to that developed in the laboratory of Manz [10], and may display different engraftment characteristics.
The reasons for differences in strain background are not well understood, but it is clear that modifying genes are contributing to the ability of the host to support engraftment. Although NK cells are important factors in determining human HSC engraftment and vary considerably between background strains, no functional NK cells develop in mice with a mutated IL2rγ gene [5–8]. An important advance in this regard is the recent report documenting the role of the signal regulatory protein α (Sirpα) gene in human HSC engraftment [35]. Polymorphisms in this gene product alter its ability to bind human CD47 and promote human HSC engraftment in immunodeficient mice. Continued screening for host genetic pathways will likely lead to further insights on the role of mouse genetic polymorphisms on their ability to support human hematopoietic engraftment.
We have found that engraftment of newborn rather than adult mice with HSC is optimal for several reasons. First, for studies of immune function where a robust T cell population is required, newborn injection is clearly superior to adults as hosts in their ability to support T cell development. While the mechanism underlying the superiority of the newborn host in generating human T cells is unclear, it is likely due to thymic organogenesis in these mice. In the absence of developing T cells, untreated scid mice rapidly undergo thymic hypoplasia, so that by the time HSC are administered to adult mice, thymic architecture is severely altered. It has been reported that introduction of the IL2rγnull gene results in the development of hypoplastic cysts that increase with age in immunocompetent mice [26]. We extended this observation to IL2rγnull scid and IL2rγnull Rag1null mice, and did not observe large differences in the appearance of these hypoplastic cysts with age between strains. In contrast, in newborn mice, HSC can be delivered before thymic dysplasia occurs and the HSC inoculum can potentially contribute thymic organizing cells to thymus organogenesis. This is suggested by the apparent lower appearance of hypoplastic cysts we observed in human HSC-engrafted mice. It is also possible that the higher ratio of HSC number to body mass achievable with the same number of cells in newborn mice as compared to adult mice supports enhanced human T cell development. Previous studies with umbilical cord blood derived HSC have shown that increasing the cell dose (cells/kg) results in superior engraftment and survival for human recipients [36], and this may benefit T cell development in newborn mice.
Several approaches have been used to engraft newborn immunodeficient mice with human HSC, including IH [10], IV (facial vein) [37] and IC (this report). We believe that engraftment success of IV injection through the facial vein and IC injection are essentially identical, as they both introduce the HSC directly into the circulation. However, we favor IC injection as it is a rapid, relatively simple technique and does not require microscopic visualization of the injection site. We compared engraftment in newborn NOD-scid IL2rγnull and BALB/c-Rag1null IL2rγnull mice established by IH versus IC injection routes to determine if either route promoted higher levels of engraftment. Interestingly, the route of injection did not influence the results in either strain, as both methodologies readily led to equivalent human HSC engraftment within an immunodeficient mouse strain. Our laboratory favors the IC injection, as the "flash" of blood that is often observed during injection provides confirmation of delivery of the HSC into the intended anatomical site [20].
In summary, we have compared directly human HSC engraftment in NOD-scid IL2rγnull, NOD-Rag1null IL2rγnull, and BALB/c-Rag1null IL2rγnull mice. We observed that human HSC engraft more efficiently in newborn mice as compared to adult mice, and that both immunodeficient NOD strains tested displayed enhanced hematopoietic cell engraftment as compared to the immunodeficient BALB/c strain. Although all strains permitted the engraftment of multi-lineage cell populations, significantly fewer human T and B cells were present in the spleen of BALB/c mice as compared to both NOD strains. We conclude that NOD-scid IL2rγnull and NOD-Rag1null IL2rγnull strains engraft at higher levels and generate more human T cells following HSC engraftment than do the BALB/c-Rag1null IL2rγnull mice. These data identify key variables for establishing humanized mouse models for the study of human immunity.
We thank Jean Leaf, Linda Paquin, and Allison Ingalls for technical assistance. This work was supported by National Institutes of Health research grants AI46629, DK53006, HL077642, CA34196, an institutional Diabetes Endocrinology Research Center (DERC) grant DK32520 and grants from the Beta Cell Biology Consortium of NIDDK, grants from the Juvenile Diabetes Foundation, International and a grant from the University of Massachusetts Center for AIDS Research, P30 AI042845. The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of the National Institutes of Health.
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Winter batters foreclosed Schenectady homes
Winter is taking a toll on the houses Schenectady seized through foreclosure last year. There's so much damage that city workers can't keep up with it all.
Wind has pulled shingles off roofs, leaving leaks to drip unnoticed inside vacant rooms.
Heavy snow has left cracked and broken sheds in its wake.
At one house, the siding is stripping off in chunks. It has lost about an eighth of its siding already, and a full-story swath is missing on one side, leaving the house frame vulnerable to the elements.
In one of the worst cases, part of a roof piece fell off the vacant house at 1352 Albany St., landing in a tree nearby. Electrical wires that had gone through that piece of the roof to the house are dangling in the air.
City Council members had worried last summer that the city might get stuck with expensive repairs if it held foreclosed houses over the winter. At the time, they hoped to quickly sell the houses.
But many were in serious disrepair and the city's contractor-rehab program only just started this winter. Three houses have been handed over for renovations, leaving more than 200 to the elements.
Building Inspector Eric Shilling said his office is overwhelmed by the amount of repairs needed on those houses.
"We're trying. We're doing the best we can," he said. "There's no way we are going to be able to keep them up to pristine condition."
Mayor Gary McCarthy did not return a call seeking comment.
City workers did perform emergency repairs soon after the foreclosures last August. Shilling inspected all of the houses and ordered the necessary repairs. Then workers boarded up windows with special techniques and tamper resistant screws that appear to have kept vandals out.
But they only did emergency work, such as roof patches.
Some residents, who were cited for peeling paint on their properties, complained to Shilling recently because some of the foreclosed houses also had peeling paint.
"They said, 'Are you going to cite yourself?' No, because I can't paint 300 houses. You can paint one," Shilling said. "They moved in to a house they chose. We got these because we were dealt this hand."
And he doesn't have much help dealing with it.
This winter, he has just three property maintenance workers. The rest of the crew is made up of seasonal workers who are only hired for the summer.
He wishes he had them now.
"This is the first winter in our foreclosures. We can't anticipate when to keep people on," he said. "When they come on, we'll start ramping things up."
But for now, he has to triage. The house with the dangling wires got priority when it was reported Friday afternoon.
"Anything with this kind of concern we can get to," he said, adding that the wires could be a serious danger.
"That may still be energized," he said.
The utilities to the house were turned off months ago, but National Grid usually only cuts the power at the house, not at the pole, he said. That would leave the dangling wires live.
City workers will ask National Grid to cut power to those wires. Then they will remove the piece of roofing and make emergency repairs, he said.
He added that residents should not hesitate to call 911 or National Grid if they see similar situations elsewhere.

subset: commoncrawl


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Shelby Hotels
Hotel bookings and reviews
Hotels in Shelby
Hotel Ratings and Locations
Comfort Inn Gateway to Glacier
Shelby is the perfect destination for outdoor and sports enthusiasts. The part of Montana around She...
Best Western Shelby Inn & Suites
Welcome to BEST WESTERN Shelby Inn & Suites! A meeting space, indoor pool and spa, and a generous co...
+44 1273 728862
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And I mean genuine anger.
I've just had a clown threaten to knock my teeth out, calls me a dog, before I could reply... he blocks me!?!? Why threaten someone and then block them?!?!?! I'm fuming
Most Helpful Guy
Yeah I get irritated by peoples comments and answers on this site at times. I don't care for any kind of disrespect, ignorance or arrogance. Some have absolutely no idea what they are talking about. Admit some humility. I try not to get involved in meaningless arguments or trolling.
I myself know that I do not know everything, and do not answer questions that I have no experience or knowledge of. There is a big difference between "experiencing" something in oppose to being told, or reading about it. But... I try to steer clear of commenting or correcting. If you want to sound like an idiot that is your business. As for opinion, It is just the preference or view of the individual.
This site is good in a way for provocative and progressive debate. I Have learned some things about behavior myself.00|00|0


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Lacrosse Arcade
Carlo Sunseri, LLC
- April 4, 2014
( 385)
Dodge, pass, shoot, and defend like never before on your mobile device. Lacrosse Arcade is a simulation lacrosse video game featuring five lacrosse game modes, worldwide leaderboards, and intense lacrosse action! Download Lacrosse Arcade today and be part of the lacrosse revolution! #laxvideogame #growthegame
Worth paying for Good game. Decent graphics, good controls and good camera angles. I like the 4v4 and practice mode but this game needs like a season mode or team mode were you can create a team. Also this game needs to be multi-player like bluetooth with devices around you. So yea overall everything is good but needs more game modes. Gets boring playing alone
Has great potential BOTTOM LINE: Currently not worth the $4 unless you've got cash to throw. I see $1 as a fair price. First, this app is known to not work on some devices. Works on my Samsung GS4 though. Glitchy... ball carrier AI sometimes falls and rolls around all twitchy. Also, my goalie re-appeared with the ball in the stands. Animations could use work. Needs season/career mode, create-a-player, and create-a-team
Good game but can improve I like the game but it needs to have a season mode or something along those lines because playing meaningless games get very boring. Be sides that I like that game, graphics are decent and controls are good
Almost broke my phone I want my money backDoes not pick up touches easily and consistantly doesnt pass when i want. Should be a free app. Still kinda fun cannot take seriously- reveiw from laxbro
Great This game is amazing but is disappointing that the actually lacrosse game mode is separate. Worth to buy even if your not gonna play because it funds the ps4,xb1, and steam version of lacrosse 16
Please update this game it is one of my favorite games on the app store, please add 10v10, mll,nll, and/or college teams, have the defenders have long sticks, team colors, create a team, create a player, regular season, tournament, logos and some good graphics. Another than that please update the game. The game is growing fast and we need more games like this so please. The best game on the app store
What's New
-Fixed issue with leaderboards not posting scores.
-Improved reactions when player has ball and is knocked down.
-Resolved crash in practice mode.
-Fixed issue where practice mode options were not displaying on some devices.
-Pause button no longer shows on main menu if user quits the game.
Keep Posting feedback for us to continue improving the game!
Lacrosse Faceoff Practice: Drills and Workouts to Improve Face Off Reaction Time


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I wear these only on the weekends (only Saturday basically) and all the other days, I just don't wear any. But just focusing on the amount of makeup itself, do I wear too much?
I wear:
- bb cream
-eyeliner (sometimes)
-lip gloss
-fill in my eyebrows (sometimes)
- yessssVote A
- nooooVote B
Most Helpful Guy
honestly, I don't think that's much makeup at all. many people wear more on a daily basis.
but makeup is just makeup and there really isn't a "too much" point. as long as you're comfortable with what you're doing, then i don't really care either way.00|00|0
I love you
haha, thanks! I'm just saying what I think
Most Helpful Girl
I don't know. Do you lightly put some on? Or make it really obvious that it looks like you applied it on with a paint brush? Because if it's just subtle amounts I don't think it's much at all. Plus it's only on Saturdays00|00|0


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The Boudicca will sail from Southampton, England, on January 6, 2009, on a 97-day westbound voyage through the Caribbean, the South Pacific, Australia (where she will visit six ports), Tasmania, South Africa and up the west coast of Africa back to Southampton. Among the features of the cruise are a transit of the Panama Canal and three-day calls at Sydney, Australia and Cape Town, South Africa. Fares with early booking discounts for inside single cabins start at $29,995. single outside cabins start at $25,405, and single balcony cabins start at $42,410. Per person double occupancy fares with early booking discounts start at $21,715 for outside cabins, $27,505 for superior outside cabins and suites from $35,405.
The Black Watch will depart Dover, England on February 15, 2009, on a 104-day eastbound route through the Mediterranean, Middle East, India, Vietnam, Hong Kong, China, Japan, Petropavlosk, Russia, the Aleutian Islands, Alaska (where she will visit three ports), Vancouver, three western U.S. ports, Mexico, the Caribbean and the Azores Islands before returning to Dover. The cruise features transits of both the Suez Canal and the Panama Canal. Fares with early booking discounts for inside single cabins start at $32,120 single outside cabins start at $37,960 and single balcony cabins start at $45,260. Per person double occupancy fares with early booking discounts start at $23,285 for outside cabins $29,945 for superior outside cabins and $37,960 for suites.
All passengers making a deposit on one of the entire cruises by December 31, 2007 will receive a $950 per person onboard credit, private roundtrip car transfers between London airports and the ship, and an airfare allowance of $300 to $500 per person, depending on cabin category.
The two ships are were both originally built for Royal Viking Lines. The 839-passenger Boudicca is the former Royal Viking Sky and features primarily outside cabins. A total of 61 cabins and suites have balconies. The 807-passenger Black Watch is the former Royal Viking Star with 90 percent of the cabins having portholes or windows and 66 suites and cabins having private balconies. Four cabins on each ship are fully accessible for handicapped passengers.
Booking for Fred. Olsen Cruises in the United States and Canada is handled by their GSA, Borton Overseas, at 800.843.0602. ext. 6. The cruise line's website is www.fredolsencruises.com.


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Queen of Broken Hearts Costume
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Order within 00 days 00 hours 00 minutes and 00 seconds and choose UPS Next Day Shipping. See Details.
Product Description
Mend that broken heart with a Queen of Broken Hearts costume. This versatile outfit will give you all the power in the bedroom but can still turn heads at your neighborhood celebration. The complete looks comes with the mini dress, puff sleeves, petticoat and wide fishnet stockings. It also includes a stand-up collar, jeweled choker and broken hearts crown. Let everyone know who you are this season. And for an extra kick, accessorize with a sleek pair of high heel shoes.
*Queen of Broken Hearts Costume for adult women includes:
*Red and Black Mini Dress
*with Sheer Stand-up Collar and Attached Broken Hearts Panels
*Puff Sleevelets
*Wide Mesh Fishnet Stockings
*Jeweled Choker
*Broken Heart Crown


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eek (Craig T. Nelson) and Camille (Bonnie Bedalia) finally agree on what to do with the house. Julia (Erika Christensen) jumps back into work helping out Kristina (Monica Potter) and Adam (Peter Krause), but is struggling on how to proceed in her personal life. Sarah (Lauren Graham) discovers what her children have been up to and puts her foot down with Drew (Miles Heizer).Resumen IMDb.com
|Nombre del documento
Craig T. Nelson
Zeek Braverman
Monica Potter
Kristina Braverman
Bonnie Bedelia
Camille Braverman
Savannah Paige Rae
Sydney Graham


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Diminish the signs of aging with this best-selling tinted moisturizer that gives skin a touch of sheer, healthy color. Formulated with an advanced peptide and broad-spectrum UVA and UVB protection, it's the smart way to care for your skin.
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Product Details
GReddy 14023011N GREX Front Strut Tower Bar OS Nissan 350z
**Item has been DISCONTINUED**
GReddy is highly known for it's fair share of suspension parts. The GREX Strut Tower Bars help improve the chassis stability and decreases the flex when being driven hard. With it's monocoque aluminum triple tube shaft, these GREX bars are very simple to install, durable for years of abuse and barely add any weight to the car.
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Kratie – Authorities respond to PLCN reports of illegal logging
December 13, 2013 | Posted in: News
Following a large-scale PLCN patrol at the end of November, authorities responded to the group's reports of illegal logging activity. On December 6 and 7, more than twenty representatives of provincial authorities, along with commune chiefs from Boeung Char, Chroy Banteay and Vatanak, found and confiscated three cartloads of illegally felled timber from loggers inside Prey Lang's protected Trapeang Tea and Phnom Krahorm areas. Thirty chainsaws were included in the haul.
Also in response to PLCN's November patrol, on 12 December three provincial forestry administration representatives, along with a police officer, two commune councilors and a village chief, visited Domrei Phong to look for evidence of illegal logging and document the number of migrant families living in the area. The group discovered an estimated 100 hectares of illegally cleared land and found that 50 migrant families were living there illegally.
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|Mr Stark
|Mrs Stark
|Jean Stark
|First appearance
|22nd May 1961
|Number of appearances
|Played by
|John Barrett
Mr Stark, and his wife Mrs Stark, were the overprotective parents of Jean Stark. In 1961, Jean lodged with Christine Hardman at 13 Coronation Street to rebel against them. The Starks went to see Jean and assured her that she was free to make her own decisions.


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Visit the
Management MSc (with Specialist Pathways in Strategy, Operations, Entrepreneurship & Innovation and Organisational Leadership & Change)
page on the University of Birmingham website for more details!
After completing her undergraduate degree in Business Administration in Amsterdam, Rozemarijn decided it was time for a change. She made the decision to move to the UK and start her postgraduate course in MSc Management at the University of Birmingham.Rozemarijn is a postgraduate mentor, you can contact her online with questions or read further about her experience at the University of Birmingham on her mentor page: http://www.pg.bham.ac.uk/mentor/rozemarijn-stolvoort/
Timo is Masters student and passionate athlete interested in the fields of urban development and destination branding. Growing up in sunny South West Germany he set off to explore South America and South East Asia before commencing a Bachelor degree in Tourism Consultancy in the Netherlands. Having arrived in Birmingham he embraces another year of study at a great university with new people and opportunities.Timo is a postgraduate mentor, you can contact him online with questions or read further about his experience at the University of Birmingham on his mentor page: http://www.pg.bham.ac.uk/mentor/timo-bornkessel/
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Grand cru (French for 'great growth' or 'great classed growth') is a regional wine classification that designates a vineyard known for its historically favourable reputation in producing exceptional wine.
Although often used to describe grapes, wine and cognac, the term is not technically a classification of wine quality per se, but is intended to indicate the potential of the vineyard or terroir. It is the highest level of classification of AOC wines from Burgundy and Alsace. Plus the same term is applied to Chateaux in Saint-Emilion (right bank of Bordeaux), although in this region it has a different meaning and does not represent the top tier of classification. In Burgundy the level immediately below grand cru is known as premier cru, sometimes written as 1er cru.
Early Burgundian wine history is distinctly marked by the work of the Cistercians with the Catholic Church being the principal vineyard owner for most of the Middle Ages. Receiving land and vineyards as tithes, endowments and as exchanges for indulgences the monks were able to studiously observe the quality of wines from individual plots and over time began to isolate those areas that would consistently produce wine of similar aroma, body, colour and vigour and designate them as crus.
Following the success of the Bordeaux Wine Official Classification of 1855, Jules Lavalle developed an informal classification of vineyards of the Cote d'Or in his book 'History and Statistics of the Cote d'Or'. In 1861, Lavalle's classification was formalized by the Beaune Committee of Agriculture. The designations of grand cru and premier cru were later developed and expanded on in the 1930s with the creation of the 'Appellation d'Origine Controlee' (or AOC) system.


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How does everyone store their leftover MMF? I tried keeping mine the fridge and it seems to dry out. I have each other in a zip baggie and then all of the little baggies in a large zip baggie. I squish out as much air as possible. When I get it out the next time to use it, it seems very dry. I have a very hard time rolling it into a crack free ball.
Are you covering each ball of MMF with a little film of Crisco before placing in the bag and storing in the fridge? I give each ball of my leftover MMF a light coating of Crisco before storing and they are fine even after 2 weeks. Hope this helps!!
After you put the Crisco on, instead of putting it in a bag, wrap it tightly in plastic before putting them all in the bigger bag. That will also help.
Same here. Coat it with shortening before putting it in a storage bag. Welcome Newbies.
Thanks everyone. I'll give the shortening a try. After rubbing the shortening on should I then wrap in some plastic wrap?
Yep, wrap it in plastic wrap and then in a ziplock bag. You want to keep it from getting air in.


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NYC Parks Events Calendar
Get up, get out, and do something! Just about any day of the year, you can find something fun, informative, and healthy to enjoy at an area park. Feel free to browse the listings below, or use the search tools to find an event that matches your interests and location.
Viewing 1–10 out of 15,385 events
from February 3, 2013 (x) to January 20, 2018
Monday, July 18, 2016
Morningside Park Green Gym Program
at Morningside Field House (in Morningside Park), Manhattan
12:00 a.m.–12:00 a.m.
Learn how to use Morningside Park's Green Gym program of 12 instructional banners installed in the park—and make it your own.
Category: Fitness, Outdoor Fitness
Kids Workshop: How Does Soil Impact Plant Growth?
at East 43rd Street Block Association Community Garden Garden, Brooklyn
2:00 a.m.–2:45 a.m.
At this workshop, we will learn how soil impacts plant growth. We will use hands-on explanations to explore soil health, while using our five senses.
Category: Nature, Best for Kids
Sunrise: Abhaya Yoga
at Pier 6 (in Brooklyn Bridge Park), Brooklyn
7:30 a.m.–8:30 a.m.
Join Sunrise Abhaya Yoga for a yoga class that integrates postures, breathing exercises, relaxation, and meditation to stretch, strengthen, and condition.
Category: Fitness, Outdoor Fitness
Exhibition - SPF16: NYC Pools and Beaches in Contemporary Photography
at Arsenal (in Central Park), Manhattan
9:00 a.m.–5:00 p.m.
Celebrate the 2016 summer season with an exhibition of contemporary photography that highlights the city's most popular warm weather amenities—its public pools and beaches.
Category: Art
Summer Sports Experience at Sunset Park
at Sunset Park Recreation Center (in Sunset Park), Brooklyn , Sunset Park, Brooklyn
9:15 a.m.–4:45 p.m.
The Summer Sports Experience provides a friendly introduction to sports for kids!
Category: Best for Kids, Sports, Summer Sports Experience
It's My Park at Gateway Triangle
at Gateway Triangle, Brooklyn
9:30 a.m.–2:00 p.m.
This It's My Park season, volunteer with League Education & Treatment Center to help care for Gateway Triangle through weeding, litter removal, and leaf raking.
Category: Volunteer, It's My Park
Prospect Park Alliance Monday Eastside Revival
at Prospect Park, Brooklyn
10:00 a.m.–2:00 p.m.
Tasks may include raking, sweeping, path edging, and litter removal.
Category: Volunteer
Landmark at Socrates Sculpture Park
at Socrates Sculpture Park, Queens
10:00 a.m.–8:00 p.m.
Marking Socrates Sculpture Park's 30th anniversary, the park presents Landmark, a series of artist commissions and projects that transforms the land both physically and symbolically.
Category: Accessible, Art, Waterfront
Flora/Fauna/Feathers of Richmond County
at Conference House Park Visitor Center (in Conference House Park), Staten Island
10:00 a.m.–4:00 p.m.
Come view original images of nature celebrating the natural environment of Staten Island.
Category: Art
Summer Sports Experience at Playground for all Children, Queens
at Playground for All Children (in Flushing Meadows Corona Park), Queens
10:00 a.m.–6:00 p.m.
Join NYC Parks at the Playground for all Children Mondays- Fridays this summer to learn sports such as Wiffleball, Kickball, Ultimate Frisbee and Flag Football. All Levels appropriate.
Category: Best for Kids, Sports, Summer Sports Experience
Related Information
- Get information on upcoming events directly in your inbox by signing up for our Email Newsletters.
- Know about an event that's not on the calendar? Send us a tip!
- Planning an event and considering hosting it in a park? Make sure to read our Planning an Event page.


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It was Valentine's day and yeah my bf bought me a flower when a men was selling them at the city we had a nice day at the cinema, but he bought his bestfriend a bouquet of flowers! and she kissed him on the cheek as well when he gave her the flowers... i was furious i mean he's my bf not hers and she already has a boyfriend who bought her flowers why did he buy her flowers? Is it wrong to be angry and jealous? He said I'm overreacting and that he was always buying her flowers But i don't give any guys presents and he'd kill any guy who'd buy me presents, I'm just pissed because I'm jealous, she got a bouquet and i got a single flower.. And that kiss was just too much he's angry at me now and said that i should stop being jealous and accept her because he's not going to give her up, i don't want their friendship to end but he shouldn't do this to me right?
Most Helpful Girl
So wait, you got ONE flower? And she got a whole bouquet? Yea my boyfriend would've been eating that damn flower! Now I wouldn't mind if he got HER the one flower and me a bouquet. But if she has a boyfriend why get her anything at all? I could see if she was single and maybe feeling lonely. But to buy her a whole bouquet? Yea I would have a SERIOUS problem with that.11|00|0
I know! And now he's the one mad at me! -_-
- Show AllShow Less
So it's okay she got a huge bouquet and i just got a single flower? That's ridiculous sorry it'd be okay if he didn't give her anything or if it was the other way around that i get the bouquet and she the single flower
Thanks for MHO. And it's not just be glad. He shouldn't be buying other girls whole bouquets and getting his GIRLFRIEND one flower.


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|Games & Hobbies
|Video Games
The Angry Chicken is a podcast that cracks 40 packs, throws down a 1/1 chicken w/ a +5 Enrage, all the while keeping you up to date on everything going on in the world of Hearthstone . Each week the latest news, strategies, crazy game stories and your emails are covered. Garrett Weinzierl, William "Dills" Gregory, and Jocelyn Moffett together host The Angry Chicken.
December 23rd, 2014
Episode 35 of 201 episodes
On The Angry Chicken this week, Brian Kibler of BMKgaming.com guest hosts with Garrett and Dills. Brian talked about his experience so far with Goblins vs Gnomes, his games against Darkwonyx in the ESL Legendary Series, Fel Reavers, Mech Mage, and more! All of the GvG Priest cards were also reviewed this week, spoiler alert: everyone loves Vol'jin. There were also many emails concerning the Good Gaming Tournament that turned out to be one of the biggest messes in tournament management the Hearthstone world has seen in awhile.Vote for thenextGvG class-cards to be reviewed on episode 60here:tinyurl.com/tac60You can support The Angry Chicken by going tohttp://www.patreon.com/tac.Follow us on http://Twitch.tv/amovetvand subscribe to the Amove Youtube athttp://Youtube.com/starcastshow. You can email the show at [email protected]:https://itunes.apple.com/us/podcast/angry-chicken-blog-amove.tv/id724741688RSS:http://feeds.feedburner.com/theangrychickenThe Angry Chicken #59Show NotesDownload TAC #59Permalink
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Welcome to the Brain Training Podcast, the daily audio workout for your head. In this podcast we have two games for you, each with three rounds which get progressively harder. To enjoy the full experience, relax, and avoid distractions whilst you listen.


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The 28th and 27th Centuries are the third section of the Bronze Age of history.
|Bronze Age Pt. 2:
|Bronze Age Pt. 3
|Bronze Age Pt. 4:
With Hotepsekhemwy taking hold of the Pharaoh's throne, records show that his first issues was settling down and stabilizing the kingdom that he had just taken control of. Hotepsekhemwy's biggest accomplishment was establishing the Netjer-Achty, or the "Chapel of the White Crown", in Buto. The chapel's main purpose was to affirm his power as the ruler of Egypt and cementing Hotepsekhemwy's heritage from the Upper Kingdom of Egypt. With Raneb, Hotepsekhemwy's son, he secured and solidified the throne for the Second Dynasty. Most records date that most of Raneb's reign was spent helping rebuild the Egyptian economy after years of rebuilding the army and the destroyed cities during his father's uprising and reign.
Egypt would meet another troubling moment in time. When Nynetjer took the throne after succeeding his father, he created a heavy system of integrated administration. Putting on a heavy tax and toll on the peasants and nobles of the kingdom. It would not help that during his reign Egypt would suffer one of its severest droughts in history after mismanagement with the irrigation technique of the Nile. Nynetjer would leave Egypt in a troubling time, he would crown his first son Wadjenes the next Pharaoh and the King of Lower Egypt, while his second born, Weneg, would be crowned King of Upper Egypt. Nynetjer's thought was that dividing up the kingdom would allow his son's the ability to effectively govern the kingdom and fix the problems that he had built. Within the first decade Weneg would declare war against Lower Egypt and pronounced himself the true. After a 5 Year Civil War, Weneg would die in battle, leaving Wadjenes the true and only Pharaoh, allowing him to reunite the kingdom under his crown. With Wadjenes reuniting his kingdom, he spent the rest of his reign rebuilding it, as it was another civil war that had occurred in the past 200 years of Egypt's history.
When Senedj took to the throne no advisors would expect his reign to be great. Though Senedj would be known as The Great Architect, with the constructions of the Pyramids of Giza and the Great Sphinx. These statues reaffirmed his kingdoms greatness for all to see, scripts tell of traders coming into the kingdom and seeing these great tombs of stone and being awed and amazed by their greatness. Senedj's move to build the Pyramids and Sphinx proved risky, but it promoted the need for better schooling, better tools, better measurement systems, and better farming for the workers. He would never see all of his creations fully built, as scripts from his advisors tell, he would live until those tombs were built. He unfortunately died 3 years after construction began."I will not pass on to meet my former kings, until they find a resting place that they can call their own."
Indus Valley
In what is now the Persian province of Hindustan, the population of the region flourished as many more cities were formed. In what was known as the Indus Valley Civilization, over 2500 cities were formed. There was no main government that ruled at the time, but each city had their own similar system of worship. Around this time, there are myths of small Mesopotamian trade ships reaching these cities to trade around this time. Historians believe it could have been motivation to explore because of Egyptian expansion, but whatever else was founded quickly fell after contact.
The main cities in the region bustled, and were estimated to have tens of thousands of people in each. These cities included Harappa, Ganeriwala, and Mohenjo-Daro, which all traded with one another through the invention of the wheel. This civilization relied heavily on trade. Barter communities formed, and the Indus Valley was the biggest it would get until it's demise in then coming millennium.
The first real records recorded of Chinese history began in 2697 BCE, when the Yellow Emperor, Huangdi, took power over most of China. Huangdi was the first true ruler of all of China after many centuries of being divided between what was known as the Three Sovereign Kingdoms. The few records on Huangdi portray him as the great unifier and thinker, able to solve any problem with a new idea or invention. Hunagdi was also known as the one who helped create the knowledge on how to make tea using the right plants. Then helped unlock China's greatest export in history, Silk.
However, after Huangdi's death, many of the Chinese cities and nobles would fall back into warring states, his teachings and ideals would be passed on to the various Dynasties that followed after his own.
Rest of the World
Cities all over the world grow, with Caral becoming the center of trade for the Pre-Inca peoples. In Sumer, the events that the Epic of Gilgamesh is based off of occur, and the cities in the region grow and begin to trade with Egypt. Some ships go off on exploration missions in search of Egyptian ports, leading to a few small communities on the Arabian peninsula.


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text: HALL Corsa 1.3 CDTi SXi, 5 Doors, Manual, Hatchback, Diesel, 2004 04 Reg, Blue. ABS, Air conditioning, Alloy wheels, Central locking, Folding rear seats, Immobiliser, Passenger airbag, Radio/Cassette, Radio/CD, Rear headrests, Sports seats, Audio remote control, Body coloured bumpers, Drivers airbag, Electric door mirrors, Front electric windows, Front fog lights, Heated door mirrors, Isofix child seat anchor points, Remote central locking, Steering wheel rake adjustment, Trip computer, 3x3 point rear seat belts, Cloth seat trim, Height adjustable drivers seat, PAS, Rear wiper, Service indicator, Solid Paint, Front head restraints, Full size spare wheel. Insurance Group:3


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Status: Open
Know more about this business than we do? Cool! Please submit any corrections or missing details you may have.Help us make it right
We offer a large selection of souvenirs, beachwear, sportswear, footwear, sun glasses, Gulf Shores t-shirts and more...everything you need for a great day at the beach! We provide outstanding customer service and an enjoyable shopping experience in a family friendly atmosphere. We also rent bicycles, beach umbrellas and chairs.
Be the first AL.com user to add photos or videos of Island Outfitters Store
Since 2008, Island Outfitters Store has been providing Miscellaneous Retail Stores, Nec from Gulf Shores. Island Outfitters Store has estimated annual revenues of $330,000.00 and also employs an estimated 5 employees.
Miscellaneous retail stores, nec, nsk


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Bring Happiness and Comfort To Our Patients and Families
40.5825, -75.61499)
Cause Areas
WhenIt's flexible! We'll work with your schedule.
WhereLehigh ValleyAllentown, PA 18106
AseraCare Hospice is currently seeking kind and compassionate individuals to spend some time comforting patients that are critically ill. We ask that you simply read, play an instrument, or hold the hand of someone near the end of their life. While our office is in Allentown, Pa, we serve patients in Lehigh,Berks,Carbon,Schuylkill,Northampton counties.
Good Match For
Teens People 55+
Requirements & Commitment
- Driver's License Needed
- Background Check
- Must be at least 18
- Orientation or Training
- Flexible


text: I've been through the benchmark post and began looking for items and doing a lot of research on updating my laptop. I know its generally hard to do but I have found a processor that is compatible with my laptop and I am able to replace it.. now I need to find a compatible GPU. Currently I have 7950 Ge Force Go, which is no where near meeting specifications. I can run FFXIV but its pretty laggy and almost impossible to move. Can anyone point me in the right direction?


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mission is to unify and represent the
graduate student body at the University of Kentucky
in matters affecting the quality of graduate student
life and work and to facilitate interdisciplinary
collaboration and promote professional development
for graduate students through seminars, forums, and
social functions.
Visit us anytime to find updated
information about events, resources, and to keep
up with what GSC is doing for you!
Thank you everyone for filling out our assistantship survey!
Find the analysis of the results in our Report
Spring 2017 Meeting Dates
All graduate students are welcome to attend!
Meetings will be held on the following days in
Little Fine Arts Library:
• 4th GSC general meeting - Thursday, April 13 at 5:30pm at at Lucille Little Fine Arts, Rm. 311
©2015 Graduate Student Congress | The University of Kentucky is an Equal Opportunity University
Stay connected! Follow us on Facebook
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Boost your child's performance. Get them ready for tomorrow's careers with Zaniac's fun learning style. Test drive our many programs in math, reading, coding, LEGO robotics, 3D printing, and more. We can teach your child Newton's laws, electromagnetism, rocket design and more with MINECRAFT. Find out which after school program works for you and your child. Do you need occasional childcare? Choose our Bronze Edison Club! Do you need a higher quality after school program with STEAM Enrichment or homework help? Choose Edison Club with transportation. Do you want full STEAM enrichment with reading, math and technology programs? Choose...Continue reading
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Designed for Baby's Comfort and Oral Health, Recent Clinical Studies Prove MAM Perfect Supports Healthy Teeth and Jaw Development Purchase, NY, July 2017 -- MAM, the leading designer brand of award-winning, medically sound and innovative baby products debuts new designs for its signature MAM Perfect Pacifier which has now been clinically proven to support healthy jaw and tooth development. The MAM Perfect Night, a glow in the dark pacifier is now offered as a new design. A recent independent study by Dr. Yvonne Wagner, a pediatric dentist at the Jena University Hospital (GER)* examined the effects of the MAM Perfect...Continue reading
I attended the open night performance of The Music Man, which is the new summer musical at Derby Dinner Playhouse and I was in pure Bliss for the entire 2.5 hour show. This by far was one of the best musicals I have seen at the playhouse ever! They can toot their own horn, trombone, brass band, and more because they surely deserve three cheers for a performance well done! The Music Man is a classic musical, in which, fast-talking traveling salesman Harold Hill, comes to River City, Iowa with a plan to sell the innocent town folk on the idea...Continue reading
I hate to admit it with the summer sun providing so much fun lately, but Back to School is just around the corner and now is the time to start thinking about sending the kids back in great footwear! KidsShoes.com is #1 place to shop for Kids Shoes! They are your destination for the best in children's footwear offering a variety of designer brands you know and love such as •Michael Kors •Sam Edelman •Kenneth Cole •Tommy Hilfiger •Ash Sneakers •Stuart Weitzman •Badgley Mishka •Sydney Jordyn They the only retailer to get styles in these brands...Continue reading
Graeter's Ice Cream Reveals a Month of Surprises for Ice Cream Lovers and Loyal Consumers Graeter's Ice Cream, a 147-year-old, family owned craft ice cream company, will celebrate National Ice Cream Month this July with a calendar of events meant to surprise and delight Graeter's fans nationwide. The company begins the summer ice cream season by celebrating its 147th birthday in a variety of delicious ways, including birthday scoops for just $1.47, surprise bonus flavors, and a new flavor to be unveiled to support the Cincinnati Zoo. Graeter's Ice Cream will take advantage of National Ice Cream Day on...Continue reading


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16 solid senders in a variety of rockin' styles - this is Marti Brom's most diverse release so far.Using only the best musicians, with guests including Nick Curran and Ted Roddy, Wise To You is a stunning release from Texas' Queen of Real Rockin' Music.
Raucous Records is the longest establisted 1950s Rock 'n' Roll, Rockabilly and Psychobilly CD, DVD and vinyl specialist on the internet.A world of Rockabilly, Psychobilly, Surf, Swing and 1950s Rock 'n' Roll delivered to your door.
© Raucous Records 2017 All Rights Reserved


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Beachside Weddings – Private Wedding Venue's South Australia.
Choose Life Time Private Retreats on Kangaroo Island as your exclusive beach side venue to get married or propose.
Life Time has created small and intimate weddings for ten – twelve guests including larger receptions for up to over one hundred guests set up in a spectacular marquee on the beach, or in our orchard by the sea.
With our range of exclusive ceremony and reception locations around Snelling's beach on Kangaroo Island, Life Time can offer full wedding management and planning, or just provide the venue and catering for your unforgettable day.
Email us below for exclusive luxury accommodation wedding packages as well as hand crafted menus and dining feast options and locations.
The owners themselves bring an abundance of skills and expertise to the event management team.
Email: Sasha Sachs from Hannaford and Sachs Catering here for information or quote.


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Listen to free music played by point5
Blip.fm is internet radio made social. It's easy to search for, play, and discover free music recommended by real people. Join today to create your own free station and start listening to @point5.
point5 I'll be damned if this year isn't a strong turning point … @zackradio, can you relate?!!
50 Cent – I Get Money| play
Apr 22, 2010 at 4:31 am via YouTube | AMAZON | ITUNES | get ringtone
point5 That's you up in dat Twittah haaa …
Apr 15, 2010 at 6:31 pm via YouTube | AMAZON | ITUNES | get ringtone
point5 never too early for Bassnectar
Bassnectar – Bassnectar - Bass Head| play
point5 one of the songs that got me into drum n bass
Mar 30, 2010 at 7:46 pm via YouTube | AMAZON | ITUNES
point5 Mos kills it as usual, and I actually like Jim Jones on this flow
point5 La Roux … it's like brand new hip 80s oldness. cop it …
La Roux – Colourless Colour [[WITH LYRICS]]| play
point5 whoa … one of the only east coast anthems I liked back in the day. DANGER!
Blahzay Blahzay – Danger| play
point5 "It's like that now…" Outkast ft. Slick Rick "Da Art Of Storytellin' (Part I)"
Outkast – Da Art Of Storytellin' (Part I)| play
point5 burn em up with some "Ether"; Nas might need to drop a diss track on Kelis soon
point5 does it get better than Red and Mef?! … "A-YO!"
Method Man & Redman – A Yo (Clean)| play
point5 it's the point5 afternoon wake up hour…
Mint Condition – U Send Me Swingin'| play
point5 Mef & MJB did this song right, but the O.G. version is just so correct…
Marvin Gaye – You're All I Need to Get By| play
point5 I always fell like…
Rockwell – : Somebody's Watching Me| play
point5 Uptown Baby? Nah, Black Cow…
Steely Dan – Black Cow| play
point5 real beats, real rhymes, don't sleep on the 'R'
Eric B. & Rakim – My Melody| play
point5 In case some of ya'll forgot… legendary Paid In Full


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TuffCare S800 Commode Capable Shower Chair with padded seat
800 Aluminum Transport Shower chair
Discount Commode Showerchair
Anodized aluminum frame.
Fits over most toilets.
Padded vinyl seat for comfort and easy cleaning with commode pail.
Waterfall plastic arm rests.
4" swivel caster wheels with rear wheel locks.
Dimensions for the showerchair are 23" x 24"
Assembly of this product is required.
Weight limit: 250 lbs.
Commodes, due to health regulations, may not be returned. All sales are final.
1stSeniorCare works to ensure your satisfaction with every purchase. 1stSeniorCare strives to make every purchase the right purchase for our customers.


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In anticipation for the RAW: Seattle show next month, I've decided to focus significantly more on larger skull renderings, specifically on wood/board.
Below is "Harbingers", a 12"x12" pen and ink with gold enamel on board. I'm planning on also working on a small series of gold beetles and smaller skulls on smaller pieces of maple, which I will be posting regular on my social media, so feel free to follow me on these platforms:
Twitter: @designbyzoe
Facebook: DesignByZoe
Instagram: Zee_Bees


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Glass Eiffel Tower Vase 16" Tall
Get tall glass vases for your next event like this gorgeous and modern glass Eiffel Tower vase in clear. Simple but super elegant when filled with flowers, this glass vase will give your DIY wedding centerpieces stunning height and beauty!
- 16" Tall
Shipping & Returns
- Economy Shipping: 5-10 business days
- Ground Shipping: 1-5 business days
- Expedited, International, and SmartPost options available at checkout.
- More Shipping Information »
Return this item within 30 days for a full product refund. Return Policy »


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Senior censor/education and training vice president
Gerrard Phillips was elected senior censor/education and training vice president in July 2014 and leads for the RCP on all matters relating to education and training.
He is a member of the RCP's Board of Trustees, is medical director of the RCP's Education Department, and is also medical director for CPD.
He qualified from Oxford and Barts and after initial training as a surgeon, 'saw the light' and trained in respiratory and general internal medicine in Southampton (research), and as a senior registrar at St Georges' and the Royal Brompton Hospitals, London. He has published over 40 research and other articles.
Gerrard has been a consultant physician at Dorset County Hospital, Dorchester, since 1994. He is a 'jobbing' clinician, still very active in both the acute medical take and his specialty. He has been involved in medical education for more than 15 years. He has been chair of the British Thoracic Society Education and Training Committee, Wessex regional training programme director for respiratory medicine and SAC chair for respiratory medicine (8 years).
He wrote the 2007 and 2010 UK RM curricula and co-wrote the European RM curriculum. He has co-written other national training programmes and advised on the consultant scientist curriculum. Recently, he has been a member of the GMC Curriculum Advisory Group and an RCP Censor. He is currently deputy head of the Wessex School of Medicine.
Gerrard's current RCP role involves consulting on RCP policy, particularly with regard to education and training, and liaising with stakeholders. He has represented the RCP at political party conferences and has been extensively involved in discussions over the implementation of Shape of Training and the new internal medicine curriculum. As senior censor, he coordinates the activities of the six college censors. He is also responsible for overseeing the diplomas in tropical medicine and in geriatric medicine.
Gerrard was involved in developing the recent RCP strategy, especially 'theme 2' which concerns education and training. He chairs the RCP Education Board. He is a member of the Academy of Medical Royal Colleges (AOMRC) Joint (Deans') Training Forum, the HEE Medical Advisory Board, the HEE Improving Junior Doctors' Working Lives Group, and the AOMRC Revalidation and Professional Development Committee. He also represents the RCP on the NHSE Cross System Sepsis Board.
Particular achievements have included the delivery of the chief registrar project, establishing a committee for coordinating college policy for sepsis and antimicrobial resistance/stewardship, a roundtable event on the challenges faced by remote and rural hospitals, working towards a post-CCT credential in acute medicine for GPs with the RCGP, and helping to open a dialogue with the RCPsych on cooperative working between the two colleges.
Gerrard is keen to increase the research profile of the education department and its contribution to the development of medical leadership, and to translate the learning from the Care Quality Improvement Department (CQID) and the Future Hospital Programme into education and training products. He is also keen to increase the diverse education outputs from the department, including streaming and e-learning.
A particular pleasure has been to oversee the development of an inaugural annual RCP Medical Education Conference, to take place on 7 June 2017.


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A murder suspect who has been on the run for nearly six months is the latest celebrity on Facebook with over 1,500 friends accompanied with a fan page of his own. Known as John Anslow, the suspect broke out of prison van by a gang of balaclava clad men while he was on his way to court in January. This made him the first category A prisoner to escape in 17 years.
The 32-year-old suspect who is believed to have fled to Spain or North Africa broke cover from his concealment last month to read messages sent to him by his loved-ones and at the same time accept invitations sent to him by 400 of his friends. With the recent addition he has more than 1,500 friends on his personal Facebook page at present. Just after three days of his prison escape, a Facebook page was made in his honor by the name of 'John Anslow is A Innocent MAN Being SET UP'.
The page has over 300 likes and messages of encouragement from well wishers. Both his pages feature photos of Anslow posing with guns and swimming with dolphins.
On the flip side, police officials believe that such an attitude from people is inappropriate. 'This is embarrassing for the police. He thinks he's untouchable now and it's like he's playing to crowd.'
A reward of 10,000 pounds has already been offered to anyone who can help bring Anslow to justice.


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HEIDI KLUM INTIMATES Mon Coeur Thong - Black
Heidi Klum Intimates Mon Coeur Thong – Black
Inject some supermodel glamour into your lingerie edit with this 'Mon Coeur' thong by Heidi Klum Intimates. Crafted in fine, floral lace with contrasting scalloped trims and a stretch finish for comfort, twin bow embellishments complete the design with a playful twist.
Model Sizing
Model is 5'10", standard UK size 8 and wears a size S. Wear true to size.Colour: Black. Hand wash.


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The following infographics provide a global and regional overview of key findings uncovered from Venture Pulse Q2'16, created by KPMG and CB Insights.
According to the Venture Pulse report, following two consecutive quarters of declines, global funding to VC-backed companies edged up 3 percent to $27.4 billion in Q2'16.
The following infographics provide a global and regional overview of these findings and other key developments uncovered from the report created by KPMG and CB Insights.
Quarterly global report on VC trends published by KPMG Enterprise.
Blog series covering topics related to venture capital investment globally.
KPMG International has created a state of the art digital platform that enhances your experience, optimized to discover new and related content.


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Each month, Rated People connects over 50,000 homeowners with quality tradesmen - simply enter your job details in the form above to get started.
We'll match your job to the relevant tradesmen in your area - those interested will pay to see your contact details and call you to quote.
As each tradesman expresses interest in quoting on your job, we'll send you their profile and ratings to help you choose the right one for you.
After the work is complete, return to the site and rate your tradesman based on their quality of work, reliability and value for money.
Tree Surgeon
Added 02/10/2016
We have a very old apple tree in our garden. Approx width of canopy 20ft hight 25ft. We woild...
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Tree Surgeon
Added 02/08/2015
Trim two trees, need side branches and tops
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Tree Surgeon
Added 21/07/2015
Removal of one tree at rear entrance, and reduce crown areas plus and existing limbs over hanging...
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Managing your job is easier with the Rated People Homes App, download it today.


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The Old Bakery B&b
Escape to the peace and tranquility of The Old Bakery, in the heart of the countryside and very close to Holt and the beautiful North Norfolk coast. In a great location for bird-watching, walking,cycling, canoeing, kayaking or just relaxing. The Old Bakery dates back to the 1700s and retains lots of character but is coupled with contemporary comfort in the guest rooms. Winner of an AA Gold Award for hospitality. Large suite with comfortable seating area overlooking our walled garden, super-king-size bed with feather duvet and topper that feels like sleeping on a cloud. Ensuite bathroom with bath and shower. Tea and coffee making facilities in room, flatscreen TV, with freeview and DVD player, i-pod docking station and free Wi-Fi. Silent fridge for your own drinks. Second large bedroom with king-size bed, feather duvet and topper for an ultra comfortable night s sleep. Ensuite bathroom with shower. Tea and coffee making facilities in room, armchairs, flatscreen TV with freeview and DVD player and free Wi-Fi. Silent fridge for your own drinks. Guests lounge with squashy leather sofas, tv and books to read. Dining hall with patio doors overlooking garden. Breakfast can be served on the patio, if desired, on sunny mornings. Breakfasts are a speciality and we are very proud that we ve been awarded the AA Breakfast Award we offer choices of starters and main courses all using locally sourced produce wherever possible with fruit from our garden and home-made jams. Tea and home-made cake on arrival. Evening meals available by prior arrangement. We also offer coaching retreats to help you with your work and life issues and quilting retreats. Relax in our walled garden or summerhouse surrounded by flowers and fruit trees, garden games including croquet, boules and badminton for the more energetic. Some off street parking and secure storage for bikes, canoes and kayaks.
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About us
Contact Us
We've already taken care of the advertising, interviewing, police and reference checking for you.
Welcome to QTKids
where we know how important
finding a qualified carer for your kids is. We can help you
by providing temporary or short-term experienced
nannies, or babysitters to suit your childcare needs while
holidaying or living in Queenstown.
To ensure the highest quality care for
your child, all of our carers are experienced, police,
reference and credential checked, and we have available
carers with current first-aid certificates.


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Get ready to undertake your literature review using your own research questions to build a successful search and apply it to a range of library resources.
Objectives - By the end of the session you will be able to:
Intended Audience: All DPhil students and research staff in the Social Sciences
Dates and booking
|Thursday 27 April 10.00-12.00
|Social Science Library, Information Skills Training Room
|Booking now closed
Presenters: Craig Finlay and Helen Worrell
Format: Presentation with practical exercises
What you will need? A computer will be provided. If you prefer to bring your own laptop, please read our guidance notes.
Related courses


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Wicketkeeper Brad Haddin has been rested from a near full-strength Australia squad to take on South Africa in their pre-World Cup series with the No.1 one-day international ranking at stake.
The selectors said veteran Haddin had been granted a break to freshen up ahead of the home summer Tests beginning in December, giving fellow gloveman Matthew Wade a return to international duty.
They have included a battery of six pace bowlers and no specialist spinners in the 13-man squad for first two games of next month's five-match home ODI series against the Proteas, with those matches on the pace-friendly WACA pitch in Perth on November 14 and 16.
Allrounder James Faulkner and paceman Mitchell Starc have been released to play in Sheffield Shield while 23-year-old NSW Josh Hazlewood has been called into the national ODI squad after overcoming back injury woes.
Hazlewood, who has played two ODIs for Australia, one each on the 2010 and 2013 tours of England, joins fellow pacemen Mitchell Johnson, Kane Richardson and Nathan Coulter-Nile as well as all-rounders Mitchell Marsh and Shane Watson.
"We are pleased with the batting line-up that we have at our disposal," said national selector Rod Marsh.
"It has great balance with the likes of David Warner, Aaron Finch and Shane Watson at the top of the order complemented by players such as Michael Clarke, George Bailey and Steve Smith.
"At this stage the squad is only for the first two matches of the series and balances out player preparation for long and short form cricket as we build into a big summer."
South Africa claimed the ODI No.1 top ranking from Australia this week.
They begin their month-long Australian tour with three T20 internationals against Australia in Adelaide, Melbourne and Sydney before the ODI series begins.
Australia: Michael Clarke (capt), George Bailey (v-capt), Nathan Coulter-Nile, Aaron Finch, Josh Hazlewood, Mitchell Johnson, Mitchell Marsh, Glenn Maxwell, Kane Richardson, Steve Smith, Matthew Wade, David Warner, Shane Watson.


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- Futurama Comics Issue #11
In this issue there is story titled The Cure for the Common Clod in which Fry is exposed to the common cold virus while on a gangster planet. He then brings it back to earth where it wreaks havoc on the populace of New New York. While the effects of the cold are different in both stories the basic premise is essentially the same.
- Obama appeared to work at Panucci's Pizza before Fry.
- It is revealed that Zoidberg is allergic to Amy Wong's sweaty feet and that his nasal system is located in his four tentacles, which are covering his mouth.
- The scene where Manhattan is covered in a dome, cut out and removed from Earth is reminescent of a book "Manhattan Transfer" by John Stith.


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Education for Tomorrow, Faith for a Lifetime.
Hello! My name is Caitlin Bruni and I have been an active teacher at St. Paul School since April 2016. I am a teacher who believes that children learn best when they feel safe and supported in a classroom environment. It has been an extreme pleasure being a part of this educational family that provides a faith, family based, and loving environment. I graduated from Gwynedd Mercy University with a Bachelor's Degree in Special Education and Cabrini University with a Master's Degree + 30 in Elementary Education/ Reading Specialist. I have been a teacher, in a classroom, since 2009 and a tutor since I was in High School. As an enthusiast, I am very excited to work with the students and help them build a strong foundation for success in their futures.
In my spare time, I enjoy spending quality time with my family and friends, watching sporting events, being involved in Community Service, and playing with my dog Oscar.


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Sacred Truth Home was a holding of the small Mantis Hogosha family located in the Noble Quarter at Ryoko Owari Toshi. [1]
- ↑ Secrets of the Mantis, p. 28
|This Mantis Clan related article is a stub. That means that it has been started, but is incomplete. You can help by adding to the information here.


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Berthon is represented at London, Southampton, Dusseldorf, Cannes, Newport, Annapolis and Paris Boat Shows. In addition we host our own brokerage show – the Berthon International Collection, an annual event originally started in 1995 which runs concurrent with the Southampton Boat Show each September. We also attend annual brokerage shows at Antibes and Newport Rhode Island. These events are specifically targeted to market our brokerage listings.
All of these can be downloaded from our website which also offers general access to information about all the brokerage yachts that we have for sale. The website is updated and optimised on a monthly basis.
Berthon advertises widely in the international yachting press including Boat International, Yachting World, Motor Boat & Yachting, Seahorse, Yachting Monthly and various lifestyle publications as appropriate.
Berthon UK – our HQ – open 7 days a week and on average 100 yachts for sale on site either afloat or on our 'land marina' of 3 custom built gantries, each of around 220' and of varying heights to suit all our listings. It is also home to Berthon Boat Company and Lymington Marina, our sister companies. We have an experienced selling team and support staff, and where we also sell new Windy Boats of Norway, and act for Baltic Yachts, Dashew Offshore in the sale of the FPB64 project, and are preferred brokers for Discovery Yachts with whom we work closely, as well as exclusive brokers for the World Cruising Club and the Ocean Cruising Club.
Berthon France – in Golfe Juan in the yachting centre of the Cote d'Azur. We offer a professional brokerage service and have excellent local knowledge enabling you to get the best out of this area. The office is headed up by Bruno Kairet who will be delighted to help you. In France we also distribute Windy Boats of Norway.
Berthon USA – the office is in a centre of yachting on the East Coast of the USA at Newport Rhode Island. Alan Baines, Jennifer Stewart and Randy Altemus, bring a wealth of knowledge of the market and years of brokerage experience. Jennifer was the Nautor Swan agent in the USA for over 12 years. In the USA we distribute Najad and Rustler yachts.
Whichever office you are co-ordinating with, we will keep you in touch with the market, values and the presentation of your yacht. The market changes all the time so you need to know this.
Once we have found a buyer we conduct the sale on your behalf. We are members of the YBDSA (Yacht Brokers, Designers & Surveyors Association) and MYBA (Mediterranean Yacht Brokers Association). All Berthon documentation has been drafted for us by Hill Dickinson in London. The process works this way:
You will see that you have the option of entering into a Central Listing arrangement with Berthon. This is our preferred route. Central Listings receive priority treatment as we control all aspects of the marketing. We are delighted to work on an Open Listing arrangement but by the nature of the contract we cannot exercise the same level of control over the marketing.
At Berthon, our aim is to find a purchaser for your yacht promptly at market value. We work for you, using our unrivalled experience within the brokerage market.
Call Berthon on 01590 679222 to discuss the purchase or sale of your yacht.