text:
Key Points
ICOS-based CARs program bipolar TH17/TH1 cells with augmented effector function and in vivo persistence.
The expression of selected CAR endodomains can program T cells for their subsequent differentiation fates and effector functions.
With the notable exception of B-cell malignancies, the efficacy of chimeric antigen receptor (CAR) T cells has been limited, and CAR T cells have not been shown to expand and persist in patients with nonlymphoid tumors. Here we demonstrate that redirection of primary human T cells with a CAR containing the inducible costimulator (ICOS) intracellular domain generates tumor-specific IL-17-producing effector cells that show enhanced persistence. Compared with CARs containing the CD3ζ chain alone, or in tandem with the CD28 or the 4-1BB intracellular domains, ICOS signaling increased IL-17A, IL-17F, and IL-22 following antigen recognition. In addition, T cells redirected with an ICOS-based CAR maintained a core molecular signature characteristic of TH17 cells and expressed higher levels of RORC, CD161, IL1R-1, and NCS1. Of note, ICOS signaling also induced the expression of IFN-γ and T-bet, consistent with a TH17/TH1 bipolarization. When transferred into mice with established tumors, TH17 cells that were redirected with ICOS-based CARs mediated efficient antitumor responses and showed enhanced persistence compared with CD28- or 4-1BB-based CAR T cells. Thus, redirection of TH17 cells with a CAR encoding the ICOS intracellular domain is a promising approach to augment the function and persistence of CAR T cells in hematologic malignancies.
Significant progress has been achieved during the past few years demonstrating the potential for adoptive T-cell transfer to treat cancer. One of the most promising approaches is the introduction of chimeric antigen receptors (CARs) to redirect T-cell specificity with high affinity antibody-based recognition units.1 CARs are synthetic molecules containing 3 distinct modules: an extracellular target binding module, a transmembrane module that anchors the molecule into the cell membrane, and an intracellular signaling module that transmits activation signals.2 Transmembrane modules are most commonly derived from molecules involved in T-cell function such as CD8 and CD28. The intracellular module almost always contains the CD3ζ chain and other costimulatory domains linked in cis. After initial reports demonstrating safety, with disappointing clinical results,3-5 the most recent clinical results with CAR-redirected T cells show remarkable antitumor effects in patients with neuroblastoma, chronic lymphocytic leukemia, non-Hodgkin lymphoma, and acute lymphoid leukemia.6-10
Since the mid-2000s, a new effector CD4+ T helper cell subset that secretes IL-17 was discovered,11,12 and it has become clear that TH17 cells represent an independent subset of T helper cells. TH17 cells regulate host defense and exacerbate autoimmune diseases. Naturally arising endogenous TH17 cells have been found in various human tumors, however their function in cancer immunity is unclear. When adoptively transferred into tumor-bearing mice, TH17 cells have been found to be more potent at eradicating melanoma than TH1 or nonpolarized (TH0) T cells.13-15 Importantly, TH17 cells have considerable plasticity and can acquire certain type 1 characteristics (such as IFN-γ production) depending on the inflammatory conditions. The ability of TH17 cells to acquire TH1 cell-like features appears to be a prerequisite for potent antitumor activity.13 One obstacle to the use of TH17 cells for adoptive cell transfer is the identification of robust culture conditions that limit the inherent plasticity of this subset.16-18
Two properties of CAR T cells that correlate with potency are the specific lymphocyte subsets that are infused and the signaling domains of the CAR. Preclinical studies indicate that cells with extensive proliferative capacity are more potent.19-21 Adoptive transfer experiments in mice indicate that TH17 cells have higher in vivo survival and self-renewal capacity than TH1 polarized cells.14 In studies using CAR T cells, incorporation of signaling domains from CD28 or from tumor necrosis factor (TNF) family members CD137 (4-1BB) or CD134 (OX40) has been shown to prevent anergy and to enhance antitumor effects.2,22
Inducible costimulator (ICOS, also called CD278) is a member of the CD28 family. We have previously shown that ICOS, but not CD28, is necessary for optimal expansion and function of human TH17 cells.15 ICOS is constitutively expressed on TH17 cells and anti-CD3/ICOS stimulation induced RORγt and T-bet expression in these cells, leading to increased secretion of IL-17A, IL-21, and IFN-γ compared with CD3/CD28 stimulation. We herein report that TH17 cells expressing CARs bearing ICOS signaling domains exhibit enhanced stability as TH17/TH1 cells and increased persistence after transfer into tumor-bearing mice.
Materials and methods
Generation of SS1-CARs and lentivirus production
Mesothelin-specific SS1-based CARs containing the TCR-ζ signal-transduction domain alone (ζ) or in combination with the CD28 (28ζ) or the 4-1BB (BBζ) intracellular domains were generated as previously described.23 The third generation self-inactivating lentival expression vector containing the SS1-ICOSz CAR was generated as described in the supporting information text. High-titer replication-defective lentiviral vectors were produced and concentrated as previously described.24
Isolation, polarization, transduction, and expansion of TH17 and TC17 cells
Blood samples were obtained from the Human Immunology Core of the University of Pennsylvania. The study was conducted in accordance with the Declaration of Helsinki. Peripheral blood CD4+ and CD8+ T cells were negatively isolated using RosetteSep Kits (Stemcell Technologies) and cultured under TH17 conditions as previously described15 (Figure 1B). For stimulation, CD4+ and CD8+ T cells were cultured with activating beads coated with antibodies to CD3 (eBioscience, San Diego, CA) and ICOS (Japan Tobacco Inc., Japan, and eBioscience) at a 1:3 cell-to-bead ratio.
Cytokine production of restimulated T cells
T cells (4 × 105) were cocultured with 2 × 105 K562, K562.meso, or mesothelin-expressing tumor cells, and supernatants were harvested 24 hours later. Concentrations of IL-17A, IL17-F, IL-22, IL-2, IFN-γ, TNF-α, IL-10, IL-4, and CCL20 were determined using the DuoSet enzyme-linked immunosorbent assay (ELISA) Development Systems (R&D Systems, Minneapolis, MN). Concentrations of IL-21 were determined using Human IL-21 ELISA Ready-SET-Go! (eBioscience).
Flow cytometry
The following conjugated antibodies were purchased from eBioscience: anti-CD161 phycoerythrin (PE), Ig (PE), anti-CD8 allophycocyanin (APC), anti-CD4 PerCp-Cy5.5, anti-CD4-PE, anti-CD45 PerCp-Cy5.5, and anti-CD45 APC. Expression of the various SS1 scFv fusion proteins on T cells was detected using biotinylated goat anti-mouse IgG (specific for scFvs of murine origin) from Jackson ImmunoResearch (West Grove, PA). Streptavidin (PE) and streptavidin (APC) were purchased from BD Biosciences. Cells were stained for viability with aqua amine-reactive viability dye (Invitrogen, Frederick, MD) for 30 minutes at room temperature. Surface staining was performed at 4°C for 30 minutes in phosphate-buffered saline (PBS) supplemented with 3% fetal bovine serum. Samples were analyzed in the LSRII flow cytometer using the DiVa software (BD Biosciences, San Jose, CA), and results were evaluated using the FlowJo software (TreeStar, Eugene, OR).
Microarray studies
Redirected T cells from 3 different normal human donors were stimulated with immobilized yeast-derived recombinant mesothelin.25 Cell pellets were collected and frozen at indicated time points on antigen recognition. Gene expression levels were determined with GeneChip Human Gene 1.0 ST arrays according to manufacturer's protocols (Affymetrix, Santa Clara, CA). Description of methods and statistical analysis is included in supplemental data. (Gene Expression Omnibus accession number: GSE58867.)
Gene set enrichment analysis
Gene set enrichment analysis (GSEA) for gene lists that were ranked by log2-fold changes were constructed, representing the expression patterns of TH17 cells expressing 3 different CARs. Analysis was performed using a modified R version of the GSEA code provided by the Broad Institute. In all cases we used a list of 74 TH17-relevant genes that was previously curated from the literature.26
Multidimensional scaling analysis
We performed multidimensional scaling (MDS) analysis on 886 differentially expressed genes among 3 CARs (P < .05, Student t test) using Euclidean distances, and the R function cmdscale() with parameters set as k = 2 dimensions, and eigen values = TRUE.
Quantitative gene expression analysis by RT-PCR
Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was used to validate select microarray results, using the same RNA samples. cDNA was synthesized with the High Capacity RNA-to-cDNA Kit (Applied Biosystems). qRT-PCR was performed in duplicate using TaqMan Universal PCR Master Mix, in a ViiA 7 Real Time PCR system following manufacturer protocols. All probes used are commercially available (Applied Biosystems).
Flow cytometry-based assay to quantify cell-mediated cytolysis
Target cells (M108) were stained with CFSE and seeded at 150 000 cells/well in 48-well plates. After 16 hours, effector T cells and CFSE-labeled target cells were cocultured at multiple Effector:Target ratios for 18 hours. Total cells were trypsinized and stained for viability with aqua amine-reactive viability dye for 30 minutes. After washing, cells were stained with annexin V, and T cells were stained with an anti-CD45 antibody. Finally, cells were resuspended in 1% HuSA PBS and a known number of counting beads. Data were collected on an LSRII flow cytometer. 4000 beads were collected for each sample.
The University of Pennsylvania Institutional Animal Care and Use Committee approved all animal experiments. NOD scid gamma (NSG) mice were purchased from The Jackson Laboratory and were bred in the vivarium at the University of Pennsylvania. The mice were housed under specific pathogen-free conditions in microisolator cages and were provided ad libitum access to autoclaved food and acidified water.
In vivo assessment of anti-mesothelin CAR T cells
Xenograft tumors were established by subcutaneous injection of 5 × 106 M108 mesothelioma cells or 5 × 106 L55 nonsmall cell lung cancer cells in the presence of a 50% Matrigel solution (BD Biosciences) in PBS. Tumor dimensions were measured with calipers, and tumor volumes were calculated using the formula V = 1∕2 × L × W × W, where L is length (longest dimension) and W is width (shortest dimension). Peripheral blood was obtained from retro-orbital bleeding or intracardiac puncture and was stained for the presence of human CD45+, CD4+, and CD8+ T cells. After gating on the human CD45+ population, the CD4+ and CD8+ subsets were quantified using TruCount tubes (BD Biosciences). All experiments were performed in a blinded, randomized fashion.
Statistical analysis
All results were expressed as means ±standard deviation (SD) or standard error of the mean (SEM), as indicated. Data were analyzed with a 2-tailed Student t test using Prism (GraphPad Software).
Construction and expression of ICOS-based CARs
The CAR constructs used in this study contain the SS1 scFv that recognizes human mesothelin (Figure 1A). The scFv was fused to the TCR-ζ signal transduction domain with the CD28, 4-1BB, or ICOS signaling domains in tandem. As negative control for signal transduction, we used a chimeric receptor containing a truncated form of the TCR-ζ intracellular domain (delζ). CD4+ T cells were activated with beads coated with anti-CD3 and anti-ICOS antibodies in the presence of TH17 polarizing cytokines, transduced with lentiviral vectors encoding SS1-based chimeric receptors, and expanded (Figure 1B). For all functional assays, T-cell cultures were normalized for equivalent CAR expression by the addition of mock transduced T cells as required. The surface expression of all CAR constructs was similar (Figure 1C).
In vitro function of ICOS-based CAR cells
One of the functional hallmarks of TH17 cells is the production of large amounts of IL-17A, IL-17F, and IL-22. To test the cytokine response of the mesothelin redirected T cells, we cocultured them with tumor cells that expressed mesothelin and quantified the concentrations of cytokines in the supernatants by ELISA (Figure 2A). We observed a significant enhancement of IL-17A, IL17-F, and IL-22 secretion by the ICOSζ cells, consistent with a predominantly TH17 phenotype. The secretion of IL-17 required surrogate antigen signaling because cells expressing a CAR with a truncated ζ chain did not secrete IL-17 (data not shown). Importantly, the addition of the CD28 or the 4-1BB costimulatory domains to CD3ζ domain impaired expression of TH17 cytokines, whereas the addition of the ICOS domain significantly increased IL-17A production. In contrast, no differences in IL-17A secretion were observed when T cells were stimulated through their native TCR receptor (Figure 2B), confirming that differences observed in IL-17A secretion following antigen encounter require signaling through CAR costimulatory domains. The enhancement of IL-17A secretion following ligation of the ICOSζ CAR was consistently observed on T-cell exposure to cancer cells derived from various tissues including lung, mesothelioma, pancreas, ovary, and breast (Figure 2C), and when using ICOSζ−based CARs with distinct antigen specificities (supplemental Figure 1; available on the Blood Web site). In addition, ICOSζ CART cells cultured under TH17 polarizing conditions, but stimulated with anti-CD3/CD28 beads rather than anti-CD3/ICOS beads, showed enhanced IL-17A secretion when compared with ζ alone, 28ζ-, and BBζ-redirected T cells, although the amount of IL-17A secreted was 20-fold lower (Figure 2D). These TH17 cells expressed similar levels of IFN-γ compared with cells preexpanded with anti-CD3/ICOS beads.
In contrast to the enhanced secretion of TH17 cytokines by the ICOSζ cells, we found that only 28ζ CAR T cells secreted robust amounts of IL-2 after coculture with mesothelin-expressing targets (Figure 2A,C). 28ζ cells secreted significantly more TNF-α and consistently more IL-21 (although significance was not reached because of donor variability) than cells transduced with the other CARs. This is consistent with studies showing that stimulation by natural ligand or agonistic antibodies to CD28 induces IL-2 and IL-21 secretion, whereas ligation of the endogenous ICOS molecule does not.27 Cells expressing either ICOSζ or 28ζ CARs secreted similar amounts of CCL20, IFN-γ, and IL−10, whereas cells expressing the BBζ CAR produced low-level secretion of all cytokines. IL-4 secretion could not be detected in any of the conditions tested.
Surface phenotype of ICOS-based CAR T cells
Human TH17 cells can be identified by their surface expression of the lectin-like receptor CD161/NKRP1A, a marker of all human IL-17-producing T-cell subsets.34 Therefore, we measured the frequency of CD161+ T cells after surrogate antigen recognition. We observed a progressive enrichment of CD161+ cells in the ζ and ICOSζ CAR cultures. In contrast, cells expressing CD161 were not enriched in the cultures containing BBζ or 28ζ CAR T cells (Figure 3A). This enrichment was reproducible, as it was observed in ICOSζ CAR expressing T cells from 12 healthy donors (Figure 3B). In contrast to TH17 cytokine production, no differences in CD161 expression were observed between cells redirected with ζ or ICOSζ CARs (Figure 3C), indicating that signaling through 4-1BB and CD28 are detrimental for the expansion of CD161+ cells, whereas ICOS is not necessary but not detrimental. Enrichment required initial exposure to TH17 polarizing conditions, as T cells transduced with the panel of CARs under nonpolarizing conditions did not exhibit subsequent enrichment for CD161 expressing cells when cultured with mesothelin-expressing tumor cells (Figure 3D).
Transcriptional signature of T cells expressing ICOS-based CARs
The enhanced expression of CD161 together with the selective secretion of IL-17 suggests that the ICOSζ CAR can be used to maintain a TH17 cell phenotype. To characterize fully the spectrum of Th17-related markers in our cells, we next used gene expression analysis to perform an unbiased characterization of the effects of the CAR signaling domains on T-cell activation after antigen recognition. Redirected CD4+ T cells from 3 healthy donors were restimulated with surrogate antigen using immobilized recombinant mesothelin-Fc and gene expression levels determined before activation and at multiple time points after stimulation. Signaling through CARs induced the differential expression of 886 genes compared with resting T cells at 4 hours on antigen recognition (false discovery rate [FDR] <0.05, >twofold change [FC] in expression). MDS analysis of these 886 genes showed that T cells redirected with ICOSζ and 28ζ signaling domains presented a closely related gene expression profile, with only 15 differentially expressed genes (Figure 4A). By contrast, BBζ T cells exhibited a distinct gene expression profile, with 163 and 140 differentially expressed genes when compared with ICOSζ and 28ζ T cells, respectively. Supplemental Tables I-VIII display all genes differentially expressed in ICOSζ CAR CD4+ T cells vs 28ζ and BBζ CAR T cells at different time points following stimulation. Supplemental Figures 2-4 show the expression of selected genes.
Our results in stimulating T cells through the CAR signaling domains are similar to previous studies after ICOS and CD28 costimulation through the endogenous receptors.27 We identified only 2 genes that were markedly upregulated by CD28ζ-expressing T cells, compared with both ICOSζ and BBζ CAR T cells: IL-2 and IL-8 (supplemental Figure 2). In contrast, we found 11 genes that were differentially upregulated in ICOSζ cells compared with both 28ζ and BBζ T cells. Of note, 5 of the 11 genes upregulated in the ICOSζ cells were TH17-associated transcripts, such as Il17a, Il17f, Ccl20, Il1r1, and Klrb1 (which encodes CD161) (Figure 4B-C and supplemental Figure 3). As predicted by the cytokine secretion data, Il17a was the most differentially expressed transcript in ICOSζ-redirected cells, and was induced 16-fold (4 hours vs 0 hours) (Figure 4B). Other genes that were preferentially activated by ICOS include Thbs1, Xcl2, Fam184a, Cldn1, and klrk1, although their specific roles for TH17 biology are currently unknown. Differences in gene expression observed by microarray analysis were validated independently using ELISA or RT-PCR analysis (Figure 4B-C).
The TH17 developmental regulatory network in the mouse has been characterized,26 and it was shown that a gene's involvement in TH17 function can be predicted using a network-based "KCRI" score.26 This score quantifies the extent to which each gene is regulated by 5 TH17 core transcription factors (TFs): Stat3, Batf, Irf4, Maf, and Rorc. Using this network resource, we tested whether the 6 uncharacterized genes we identified were regulated by the 5 core TH17 TFs. We found that Ncs1 was significantly upregulated in TH17 cells compared with TH0 cells, and was ranked 46 out of more than 22 000 tested genes for TH17 function based on the "KCRI" scoring scheme, similar to well-established TH17 genes including: Il17a (ranks in parentheses; 6), Il23r (7), Il17f (8), Rorc (11), Il1r1 (19), Ahr (107), and Ccl20 (143). Figure 4D shows the significant regulatory inputs for Ncs1 identified in the TH17 differentiation network model.
CD4+ T cells redirected with ICOSζ showed a TH17 core molecular signature with a TH1 bias
We next performed GSEA of the microarray data obtained using cells at 4 hours following antigen recognition (Figure 5A). We observed a nonrandom distribution of genes associated with TH17 cells at the top of the ranked gene list for all groups. Cells expressing the ICOSζ CAR had the highest maximal enrichment score (ES) of 0.784, followed by 28ζ and BBζ, confirming that redirection of TH17 with an ICOSζ-based CAR can be used to sustain a TH17 core molecular signature.
Because it has been suggested that TH17 cells must acquire TH1 cell-related features to eradicate tumors in mice,13 we next analyzed whether signaling through ICOS could induce the expression of TH1-related genes, as well as TH2- and Treg-related genes. Heat map representation of log2-fold changes of T helper signature gene expression at 4 hours after stimulation relative to nonstimulated cells demonstrated that TH17 cells redirected with ICOSζ highly overexpressed TH17-related genes, including Rorc, Il22, and Il26 (Figure 5B-C). Notably, they also overexpressed some TH1-related genes, including Ifnγ, Tnfα, and Tbx21 (T-bet) (Figure 5B-D). Importantly, ICOS signaling in TH17 cells did not induce the expression of Treg- or TH2-related genes, with the exception of Il13 (Figure 5B). Also, contrary to cytokine and transcription-factor expression, signaling through CAR did not induce significant changes in expression of most cell surface markers, including Ccr6, Ccr4, and Ccr10 (supplemental Figure 4A). Collectively, these results suggest that ICOS-based CARs can sustain TH17 function with a TH1 bias.
Comparison of antitumor efficacy of TH17/TH1 polarized T cells expressing CD28, 4-1BB, and ICOS-based CARs
We next analyzed the antitumor activity of retargeted TH17 cells. For this experiment, in addition to redirected TH17 cells, we also redirected TC17 cells (CD8+ T cells cultured under TH17 polarizing conditions).28 Both TH17 and TC17 redirected cells efficiently lysed mesothelioma (M108) tumor cells, and no differences in cytotoxicity were observed between CAR signaling domains (Figure 6A). TC17 cells secreted robust amounts of IFN-γ following antigen recognition, although IL-17 secretion was not detected (supplemental Figure 5). Next, the antitumor activity of mixed redirected TH17 and TC17 cells was analyzed in NSG mice bearing M108 tumors (Figure 6B-C). A potent antitumor effect was observed in all groups treated with mesothelin-specific CAR T cells. T cells redirected with BBζ showed a relatively slow tumor decline, with all mice showing significant tumor shrinkage after treatment. 28ζ and ICOSζ treatments showed a more rapid tumor decline, leading to tumor eradication in most mice (Figure 6B and supplemental Figure 6). Treated mice were sacrificed on day 51 after T-cell injection as they developed xenogeneic graft-versus-host disease (GVHD), and mice treated with the ICOSζ CAR T cells had the most severe GVHD (supplemental Figure 7). We have previously shown that this tumor does not regress following injection of nongene-modified allogeneic T cells.23
CAR TH17 cells reprogramed with ICOS endodomains have long-lived persistence in vivo
Long-term persistence of adoptively transferred T cells correlates with antitumor effects in clinical trials.29,30 Because it has been shown that natural TH17 cells are long lived in vivo,14 we next sought to assess the in vivo persistence of CAR-modified TH17 and TC17 cells in the peripheral blood of treated mice (Figure 7A-B). The average number of CD4+ T cells per µL of blood and per spleen at the time of sacrifice (day 51) was higher in mice treated with ICOSζ-based CARs than when compared with 28ζ and BBζ groups. Of note, TH17 cells redirected with ICOS also showed a higher frequency of CD161+ cells, correlating with in vitro results (Figure 7C). CD8+ T cells redirected with BBζ persisted better, although differences were not significant (Figure 7A).
To extend these results, we performed a similar experiment with NSG mice bearing non–small cell lung tumors. To confirm that T-cell persistence was a result of tumor recognition, and not a result of recognition of xenoantigens by the endogenous TCR of these cells, this experiment was performed in duplicate using mice with or without tumors (empty mice). TH17 cells redirected with ICOSζ showed an increased in vivo expansion and persistence that was significantly greater than the 28ζ and BBζ groups. Importantly, ICOSζ T-cell expansion was dependent on tumor antigen encounter, as empty mice showed significantly fewer ICOSζ cells compared with mice with tumors (Figure 7D). In conclusion, these results suggest that reprogramming CD4+ T cells with an ICOS-based CAR signaling domain promotes TH17 cell function and phenotype and enhances T-cell persistence in tumor-bearing mice.
A central issue in the field of adoptive cell therapy is optimizing the proliferative capacity of the infused T cells while stabilizing the phenotype and limiting the plasticity of the infused cells so that they are resistant to the induction of regulatory or exhausted phenotypes that commonly emerge during cancer therapy.31,32 Here we find that CAR T cells expressing an ICOS signaling domain in TH17 cells are preferable to CAR T cells expressing CD28 or 4-1BB domains resulting from a stabilized TH17 core molecular signature and an enhanced persistence of the CAR T cells in tumor-bearing mice. CAR T cells with ICOS domains retain a "bipolar" bias toward a TH1 and TH17 signature, which is a plasticity that appears to be a prerequisite for potent antitumor activity in several preclinical models.17 Shen et al constructed an ICOS-based CAR that recognizes EGFRvIII,33 and demonstrated that TH1 cells redirected with this CAR could release IFN-γ and destroy glioma cells in an antigen-dependent manner. Our results confirm and extend this work by performing a comprehensive analysis of CARs expressing ICOS signaling domains. We show for the first time that ICOS based CARs have better persistence than the signaling domains currently being tested in the field, principally CD28 and the TNF family member 4-1BB.
We cultured CAR T cells under conditions that are used for clinical trials using antimesothelin redirected CAR T cells. When the cells were cultured in TH17 polarizing conditions, only ICOS-based CARs secreted high amounts of IL-17A, IL17-F, IL-22, and expressed CD161 after encounter with the redirected antigen. In humans, interleukin 17-producing cells have been shown to originate from a CD161+CD4+ T-cell precursor.34 Consistent with TH17 polarization, CARs with ICOS domains did not secrete IL-2 in contrast to CAR T cells with CD28 domains. IL-2 is generally considered to be essential in adoptive therapy protocols that involved transfer of CD8+ T cells; however, IL-2 can also promote the outgrowth of Tregs, so that competing agonistic and antagonistic effects might be observed in vivo. In this regard, Kofler et al showed that deletion of the lck-binding moiety in the CD28 endodomain of a CD28-ζ CAR abrogated IL-2 secretion on CAR engagement, and improved antitumor activity in tumors infiltrated by Treg cells.35 Our finding that ICOS-based CAR T cells don't secrete IL-2 may provide a strategy for the treatment of hematologic and solid tumors heavily infiltrated by Tregs.
Related to our findings are reports that treatment of cancer patients with antagonistic CTLA-4 antibodies results in an enrichment of CD4+ T cells that express ICOS and produce augmented levels of IFN-γ.36 Of note, a report showed that concomitant CTLA-4 blockade and ICOS engagement by tumor cell vaccines engineered to express ICOS ligand significantly improved rejection of established melanoma and prostate cancer in mice.37 Our findings are consistent with reports suggesting that ICOS expression on CD4+ T cells is a predictive biomarker for CD4+ effector T-cell responses generated after CTLA-4 blockade.38
Cioani et al created a validated regulatory network that describes the master transcription factors and genes expressed in TH17 cells.26 We have used those results as a framework to characterize the human orthologs expressed in CAR TH17 cells expressing various signaling domains. We found that the gene expression analysis of human ICOS CAR bearing T cells was closely related to the orthologous set of genes expressed in mouse TH17 cells.26 CAR cells bearing ICOS domains specifically retained a genotype of TH17 cells with the expression of Rorc and Il1r1 in the absence of Foxp3 after surrogate antigen stimulation. IL-1 signaling is reported to promote proliferation and survival of antigen-stimulated TH17 cells. This process involves activation of the serine-threonine protein kinase Akt-mammalian target of rapamycin (Akt-mTOR) pathway.39 In addition, the ICOS CAR cells also retained selective expression of Tbx21 and IFN-γ, consistent with the maintenance of a TH17/TH1 bipolar phenotype. We also found that stimulation of ICOS CAR cells with surrogate antigen promoted the expression of Ncs1. NCS1 is a protein that belongs to a large Ca2+-binding protein family, implicated in the regulation of Ca2+-dependent exocytosis, with a previously unknown function in T-cell biology. However, based in our results, NCS1 is likely to carry an important function in the development and function of TH17 cells, as it was also implicated in the mouse TH17 cell network.26
Enhanced in vivo persistence is a desirable effect for long-term antitumor effects. Our studies suggest that the ex vivo manipulation of CAR T cells with an ICOS signaling domain can stabilize TH17 function and promote CAR T-cell persistence in mice bearing human tumor xenografts. This enhanced persistence required stimulation through the CAR receptor. In our microarray data we observed similar expressions of antiapoptotic genes in all of the CAR T-cell signaling domains tested. However, it is likely that ICOS-CAR TH17 cells had a proliferation advantage because they were consistently recovered in larger numbers from the spleens of humanized mice. This may have important translational implications, especially in patients with nonlymphoid tumors, where the functional persistence of CAR T cells has not yet been observed. A limitation of the present studies is that we do not yet know whether this will be predictive of the results after CAR T-cell transfer to patients, and therefore a future clinical trial will be required to test these principles.
To our knowledge, our findings are the first to demonstrate that the expression of selected CAR endodomains can program T cells for their subsequent differentiation fates and effector functions. The balance of Tregs to effector T cells is an important biomarker for the outcome of many immunotherapies,40 and the ability of ICOS to limit IL-2 secretion may be an important feature in improving this ratio. The observation that CAR T cells bearing ICOS signaling domains have enhanced survival after adoptive transfer provides the rationale to test this concept in human trials in the setting of solid tumors and nonlymphoid hematologic malignancies, where improved persistence of CAR T cells remains as an unmet medical need.
The online version of this article contains a data supplement.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 USC section 1734.
The authors thank Chrystal Paulos for helpful discussions, Dr Katsunari Tezuka (Japan Tobacco, Inc.) for the ICOS mAb, Daniel Powell for the generous gift of Mov19 vectors, Brian Keith for helpful comments on the manuscript, and Ronghua Liu, Anthony Secreto, Omkar Kawalekar, Kathleen Haines, and Shree Joshi for expert technical assistance.
This work was supported by the National Institutes of Health Common Fund Nanomedicine program (PN2 EY016586, RO1CA120409) and the PA DOH award 410005172. This work was supported by a Marie Curie International Outgoing Fellowship within the 7th Framework Programme of the European Union (IOF-GA-2010-275139) (S.G.).
Contribution: S.G. and C.H.J. designed the study and wrote the manuscript; S.G., C.C., S.E.M., M.J.F., J.L., A.D.P.Jr, and J.S. performed experiments; X.C., A.M., and R.B. performed bioinformatics analysis; and N.S. provided vital new reagents.
Conflict-of-interest disclosure: S.G., J.S., and C.H.J. are inventors of intellectual property licensed by the University of Pennsylvania to Novartis. The remaining authors declare no competing financial interests.
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GIFT: Holiday Workshop (2014)
Holiday Workshop is all about making gifts for the Holidays! Come join us to make gifts for family, friends, and the special people in your life! We will serve cookies and hot cocoa to enjoy as you sew.
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The Possibility Project with Mending As Metaphor presents Landfill Harmonic
'The world sends us garbage, we send back music'
We live in an unprecedented time of social and environmental upheaval, so what might the possibilities look like when there is harmony between the two? You are warmly invited to take part in a special screening of Landfill Harmonic, a feature length documentary that follows the journey of a Paraguayan Youth Orchestra that makes its instruments entirely out of waste. The film masterfully demonstrates how ordinary people can make extraordinary choices to rise above conflict and create so much hope in the process.
The Possibility Project is an Australian based social enterprise, they collaborate with slum communities in India to create a slow clothing label called slumwear108, they will share their own insights on how people around the world are collaborating to enact peaceful social and environmental cohesion.
Join us for a wine and cheese gathering, a viewing of the film and a Q+A with The Possibility Project's founders, Kim Pearce and Kath Davis. There will also be a pop-up of clothing from the slumwear108 line.
Note: There is no charge for this program but please RSVP as space is limited
The film is sub-titled – so although very family friendly – reading ability 10+ ??


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I wrote one post this week revealing my asset allocation and my reasoning behind it:
My Asset Allocation
Here are some short takes and some weekend reading:
The Blunt Bean Counter looks into the tax implications of crowdfunding.
Potato looks at a wide range of choices for implementing a passive index investing plan. He rates them by simplicity and costs.
Colleen Jaconetti at Vanguard coins the term "rebalaphobia," the fear of rebalancing your portfolio. I think this is quite common.
Big Cajun Man reports that TD Waterhouse has made it easier to make transactions in their RDSP accounts.
My Own Advisor reviews the book The Empowered Investor.
Million Dollar Journey shows how to figure out where your hydro dollars are going.
Friday, October 31, 2014
Short Takes: Crowdfunding Tax Issues, Index Investing Choices, and more
Posted by Michael James at 6:30 AM
Labels: Short Takes
3 comments:
Easier to move assets around in the account, but still a pain to get money into the actual account. Hopefully fixing transfers into the account is the next step.ReplyDelete
Thanks for the mention, Michael!ReplyDelete
Thanks for the mention Michael, enjoy the rest of your weekend. Interesting to read your asset allocation and reasons behind it, including your comments.ReplyDelete
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The Orichalcos God, or the Great Leviathan (4Kids dub), also known as the "Great Beast", is a gigantic serpent creature who is behind all the events in Season 4 of Yu-Gi-Oh!, the Waking the Dragons arc and thus is the overarching antagonist and final boss of the arc.
He was empowered by the dreaded Orichalcos magic which he is the sole master of and he employed the Orichalcos Stones to take over the world. It was these stones that made Dartz of Atlantis turn evil and use the Orichalcos Stones and cards to seek out victims to steal souls from to feed the Orichalcos God.
The souls of Gurimo, Pegasus J. Crawford, Yugi, Insector Haga, Dinosaur Ryuzaki, Amelda, Valon, Katsuya Jonouchi, Mai Kujaku, Rafael, and Seto Kaiba were among the many fed to the Orichalcos God. The final soul was that of Dartz himself, after he lost a duel to Yami.
Later, the remaining duelists' Duel monsters battled the Orichalcos God, but it was completely ineffective; the Orichalcos God sent numerous shards which easily dispatched all of them, while all the duelists were captured by him, except Yami. Yami, however, converted the darkness inside people and Duel Spirits' souls into light, allowing the three Egyptian gods to be released. It also greatly weakened the Orichalcos God, while the Egyptian gods' powers increased immensely. It required the souls and powers of millions to neutralize him; Yami finally caused him to disappear by removing all of his remaining darkness within his soul.
After the Orichalcos God's death, Dartz was released from his control, and the souls of all his victims were returned during the battle.
- He was also dubbed the "Grand Dragon Leviathan" in English advertisement media.
- On the Yugioh.com site, the artwork displayed for this monster is mistakenly that of "Cave Dragon".
- It is considering to be one of the largest monster that ever seen in the anime, being larger than even all three Egyptian Gods.


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I've been meaning to get back onto regular blogging since I set this new website up, but all the stuff's been kind of running into the other stuff and there hasn't been a window of opportunity! But, here we are, the first weekend of 2017 that I've had a chance to sit down and rattle off some words. The last few months have thrown some amazing amazing photographic challenges at me; these have included band photography for Leicester's own Great Central Magazine; an amazing performance of Outlaw by The Point Youth Theatre (which also tested my ability to do photography with both sinusitis and conjunctivitis!); promotional photography for Gallops and Rich List (the Gallops photo session requiring 2 days of mask making courtesy of Wintercroft's wonderful low-poly mask templates www.wintercroft.com); and Tetrad Collective's multi disciplinary artist takeover of Attenborough Arts Centre.
Running alongside these and other projects has been the longer form documentation of Sleuth, a play by Anthony Shaffer, that opened at the Little Theatre in Leicester on the 6th February. I was asked by director Edward Spence to document the production process from the first reading through to the final dress rehearsal. Having previously worked with Ed to document his productions Butley (again at the Little Theatre) and The Finest Ruin (performed at Leicester's Y Theatre), I absolutely leapt at the opportunity.
The process kicked off in September 2016 with the first cast reading at Leicester's Phoenix, it was apparent that the darkly comic and cynically twisted source material was really going to give the cast of Jaz Cox and Kenton Hall something to get their teeth into; therefore making for an excellent documentary photography subject. So it progressed through the Christmas period, with each visit to the rehearsal studio yielding excellent opportunities to capture images of the play throughout it's development. The trust shown in me by the director, actors and wider production team was both invaluable and hugely appreciated; as it gave me the chance to build up a genuine document of the work in progress.
By late January the production moved to the main house, where a wonderful set had been designed and built by Lynsey Brecknell and Anna Field; having spoken with Ed about the set late last year, it was fantastic to see such grand designs come to fruition. The final set featured a prominent photo of the character Andrew Wyke over the mantelpiece, this was a photograph shot during the rehearsal process, it was great to have physically contributed something to the production itself. Once the production hit the main house I was able to photograph the two tech rehearsals and the final dress rehearsal, as well as being a rare chance to work with a full main house production it was fascinating to see how the work had evolved from the first reading in the Phoenix's basement studio into such an uncompromising tour de force of a production that really was a huge credit to the whole production team and a testament to the tireless hard work and dedication to everyone involved.
From a developmental point of view it really reinforced that this is the sort of documentary photography work that I'd love to do more of; being able to follow projects as they grow and evolve through the developmental process; this method of working really seems to suit my unobtrusive style of photography. So a huge thank you to everyone involved, it was a pleasure to be a small part of such an excellent production; onwards to the next project!


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Tuesday, March 15, 2011
diy: birdie mobile
I came across an adorable little birdie mobile tutorial on Blessed Little Nest. I used it to whip this one up. It is my mock-up. I want to use some patterned scrapbook paper, so I tested it out first.
I used a couple of sticks tied together to create a mobile structure since I am not planning on hanging it from a chandelier.
I can't decide if I will hang it in my studio space, the kitchen, or the laundry room... my laundry room could use some love! Or maybe I'll make several. :)
This look me about 20 minutes from start to finish, and it adds a great pop of color to my space. I love it! You should try one!
xoxo, christa
11 comments:
this is so cute!ReplyDelete
How cute! I love the color.ReplyDelete
Oh, as a total fan of all things birds, I adore this idea!ReplyDelete
ca-uuttee! great colors too luv:)ReplyDelete
That is totally darling.ReplyDelete
That is really cool! Looks so easy, yet so stylish!! Great idea!!ReplyDelete
Btw, you've been tagged - by me! If you want to participate in the "Handwriting Game" - look at my blog. :)
♡ Nadine
Love mobiles~ I made one of beads on fish line last summer...Also a santa one when Tracy and Troy were "young" wondere where it went????? I loved it!ReplyDelete
Hey sis,ReplyDelete
I really decided to do this mobile as a crafting project for kids next week. Where did you find the patterns for the birds?? The link doesn't work...
XO. Nadine
Really inspired to see this awesome blog, keep the good work.ReplyDelete
Now you can win free stuff online, free giveaways, enter every day to win latest iphones, smartphones, macbooks, laptops, accessories, technology and much more.ReplyDelete
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Sickbits is a library of unlimited hacks for android, ios smartphones. Get unlimited amount of hacks and modes for mac & pc. sickbits.netReplyDelete


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The Body Shop has just launched two new products to the Tea Tree range – Tea Tree Cool & Creamy Wash & Tea Tree Squeaky-Clean Scrub.
Together with the all-time popular Tea Tree Oil, the three products form an informidable Power Trio!
In conjunction with the launch, The Body Shop are looking for Super Power Trios who are superheroes in their own way. To participate simply, take a photo of yourself and your two other friends in your superhero pose, and comment on why the three of you are a super power trio. The most creative entry each week gets to walk away with a Tea Tree hamper worth $200 weekly.
Do also check out http://on.fb.me/10VqJit where you can get information on the 3 products and Instagram contest details, as well as redeem free samples and small sized Tea Tree Skin Clearing Facial Wash.


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Many people put their full trust in the American health care system, and accept the diagnoses given to them by knowledgeable physicians. According to the National Center for Policy Analysis, however, between 10 and 20 percent of medical diagnoses in America are inaccurate, costing patients unnecessary medical expenses and potentially their lives. These staggering statistics show the prevalence of … [Read more...]
Seeking a medical second opinion is a good idea
Taking an active role in one's health can prevent medical errors
Although many Nevada residents rely on the medical expertise of health care professionals to provide adequate care in their time of need, medical errors are one of the leading causes of death in the U.S. According to the Journal of Patient Safety, experts estimate that up to 425,000 people die each year as a result of medical errors. Rather than complacently accepting medical treatment and advice, … [Read more...]
Medical mistakes determined third largest cause of death, study says
When patients enter a Las Vegas hospital, they do so with reasonable expectations that they will receive the proper medical care for their conditions. However, new research indicates that a significant number of patients fail to receive even the minimum standard of care in U.S. hospitals, and that lack of care is leading to an epidemic of preventable deaths. According to A New, Evidence-based … [Read more...]
Central line-associated bloodstream infections are a serious problem
While receiving medical care of any kind has its associated risks, patients who are admitted to hospitals or other care facilities in Nevada have the right to expect that they will receive the best possible level of care. However, many individuals and their families must deal with the consequences of receiving sub-par care that results in extra illness or infection, such as seen with central … [Read more...]
Study: Many robotic surgery errors underreported
The number of reported injuries directly linked to robotic surgery errors is rising, according to data collected from the Food and Drug Administration. In 2004, 13.3 adverse events per 100,000 procedures were reported, and this number increased to 50 events per 100,000 procedures in 2012. Many experts believe that the reports only show a small piece of the picture. FDA warning letter lists … [Read more...]
Reducing your risk of a medical misdiagnosis
When patients in Las Vegas seek medical care from a doctor, they expect to receive an accurate diagnosis that they can rely upon. However, this is often not what patients actually receive. When a doctor mistakenly says that a patient suffers from one disease while the patient's symptoms are really caused by another disease, a misdiagnosis has occurred. Misdiagnosis is also a national problem. … [Read more...]
Defective surgical mesh implants pose several risks
As women in Las Vegas grow older, surgeries, childbirth and weight issues can cause muscles in the pelvic area to weaken. When this occurs, additional problems such as urinary stress incontinence and pelvic organ prolapse can develop. Often, the recommended treatment is to reinforce the tissue by inserting a vaginal mesh implant. The risks According to the U.S. Food and Drug Administration, … [Read more...]
Up to 4,000 times a year, people are victims of surgical "never events"
Going through a surgical procedure in Las Vegas is always associated with some risk. However there are some mistakes that just should not happen. These surgical errors are called "never events" and according to a study from Johns Hopkins Medicine, they happen to people in the U.S. on average 4,000 times a year. The estimate comes from a review of medical malpractice claims but researchers … [Read more...]
How You Can Get Injured in Las Vegas
While enjoying the sights and sounds of Las Vegas nightlife for a weekend visit, no one plans to be involved in an auto accident in their rental car, slip and fall at a hotel as a result of someone else's negligence, or visit a popular eatery only to then become the sick with food poisoning. While no weekend vacationer to Las Vegas plans these types of misfortunes, they do sometimes happen. Auto … [Read more...]


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Shredding Coventry
How does it work?
Branch information
Box-it Central is perfectly located for document management services in Coventry and Warwickshire. We are proud to be your local branch of the nationwide Box-it group, operating to exceptionally high standards. Based in Bishops Tachbrook, just off the A46 Warwick / Coventry bypass, we provide a full range of document management services for paper and digital records.
These include document shredding (which is available as a mobile shredding service operating in Coventry City Centre), archive storage, cataloguing and scanning services, as well as our own Box-it Omnidox platform for Cloud based document management solutions.
Opening Hours
- Monday – Friday: 8.30am – 5.00pm
Shredding Coventry
We provide reliable, highly secure document shredding services to organisations in Coventry and neighbouring districts, such as Tile Hill, Allesley, Baginton and Ryton-on-Dunsmore, to name but a few. Our liveried vehicles are fully tracked (as is the whole shredding process) and we only use our own Box-it Central personnel, all of whom are uniformed and DBS screened.
We provide two types of document shredding services:
Onsite shredding Book an appointment for our mobile shredding lorry to come to your Coventry premises for either a one-off shredding purge, or as part of a regular scheduled document destruction service. We then securely shred your documents onsite and you can, if you wish, supervise the whole process for added peace of mind. This is ideal for the safe destruction of confidential paperwork. As long as there is sufficient parking for our lorry, we are happy to bring it to any address in Coventry City Centre, or to the business parks, such as Warwick Science Park or Coventry University Technology Park.
Offsite shreddingWhen you use Box-it Central's offsite shredding we collect your sacks from your premises and return them to our own shredding facility just off the A46 Coventry bypass where they are destroyed in highly secure conditions. The sacks have security fastenings with barcodes which allows us to track them throughout their journey from your Coventry offices to our shredding facility, right through to their destruction. We issue you with Certificates of Destruction to confirm that your shredding has been completed. We offer a collection service to all Coventry addresses.


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Awarded by
Foundation Education (RTO 22557)
Certificate IV
Study Mode
Payment Options
Payment planUpfront payment
12 months
Start Date
Anytime – start today!
Course Fee
Career Outcomes
Dental Assistant | Dental Hygiene Assistant | Oral Health Advisor | Dental Radiographer | View all
This course meets the Nationally Recognised Training standards. It is listed on the Australian government's National Register for Vocational Education and Training (VET). Studying this course leads to a qualification that's recognised throughout Australia, and lets employers know that you have the right skills for the job.
Give your career as a dental assistant the best possible start with this HLT45015 Certificate IV in Dental Assisting (Radiography and Oral Health Promotion Specialisation). The course will equip you with knowledge needed to pursue work in the specialisations of radiography and oral health promotion.
You will complete 250 hours of workplace training in addition to the online coursework to ensure you have a firm grasp on the practicalities and skills required for both the radiography and oral health promotion specialisations, in addition to general duties and administrative work required in a dental assistant role.
Foundation Education offers you the flexibility to make your studies work around your schedule, allowing you to balance coursework with your other commitments.
As a part of this course you will be required to complete a minimum of 250 hours of practical placement in a dental clinic.
You will need to have completed year 12, or equivalent. Equivalency can be demonstrated by one of the following:
Minimum Age
You will need to be at least 16 years old. If you are under 18 your enrolment form will need to be signed by a parent or guardian.
Residency Status
You will need to be a permanent Australian resident or equivalent.
Computer Requirements
In order to complete the on-line part of the course you will need regular access to the following:
Please note that legislative radiation licensing requirements may apply to some elective units.
In this dental assisting course, the units of competency you may study include:
*This course contains a core unit, HLTAID003 Provide first aid, which must be completed through an external RTO. Foundation Education has an agreement in place with Red Cross Australia (RTOID 3605) which allows students to receive training for this unit at a reduced price. Red Cross Australia is a third party training provider who conduct training and assessment services on behalf of Foundation Education.
Foundation Education is one of the few Registered Training Organisations (RTO) in Australia who endeavour to deliver flexible online training solutions designed to fit our students' lifestyle. Since 1999, we've been focused on providing excellence in Vocational Education and Training and building partnership networks that drives the quality of graduates across each industry, with our partner RTO, the Australian Institute of Personal Trainers (AIPT). With more than 30 nationally recognised certificate and diploma level courses on offer, the Foundation Education difference means:
Flexible Online Study – Courses that don't require a practical placement are delivered 100% online so you can study whenever and wherever suits you. This also gives you the flexibility to choose your course length – whether that be the full term of your course or completing your study in as little as six months.
Dedicated Student Support – Our industry experienced Subject Advisors, Assessors and Support Officers will support you from enrolment through to graduation. You can use them as often or as little as you like either via email or the telephone.
Industry Relevant Courses – Course material is developed in consultation with industry experts to equip you with the job-ready skills and up-to-date knowledge required to support your career goals.
Foundation Education provides a dedicated student support team throughout your studies. Whether it is access to experienced Subject Advisors, Assessors or general Support Officers, we will support you from enrolment right through to graduation. You can use our team as often or as little as you like either via email or the telephone at no extra cost.
Upon successful completion of this course you will receive a nationally recognised, accredited Certificate IV in Dental Assisting (Radiography and Oral Health Promotion Specialisation) (HLT45015) from Foundation Education.
By completing two key specialisations in dental assisting services, you will have greater career flexibility and the skills to pursue employment in oral health.
Wondering where a career as a dental assistant could take you? Potential job outcomes may include; Dental Assistant, Dental Nurse.
Foundation Education offers affordable payment options to help you start your studies sooner.
Payment Option 1: Pay in weekly instalments
Payment Option 2: Pay in full when you enrol and save up to $800 on your course fees
Tuition fees include course delivery and assessment, study materials and dedicated student support services. Please note this course does not qualify for assisted government funding.
Fill out this form and a Study Consultant from Foundation Education will get back to you.


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User Reviews
Displaying all 2 1958 Chevrolet Corvette reviews.
CarGurus https://www.cargurus.com
1958 Chevrolet Corvette Convertible RWDReview
A Dream Com True — She was a short track racer that had been hit in the front right corner. the old man that owed it lost his eye sight and could't race any more. he advertised it as a race car only, but i reconized it right away from my boy hood dreams. She currently has 425 hp and tires from a dodge caravan. she is the greatest car ever made, with the HD suspension there are only 144 of these to grace the earth
Pros: 6 Miles to the gallon
Cons: Tires don't last long ( that may be my fault)
CarGurus https://www.cargurus.com
1958 Chevrolet Corvette Convertible RWDReview
One Vette In The Works — This pic of the Corvette is not one of mine. This is what mine is bound to look like though. My dad and I are fixing it up and plan to put a 383 stroker in it and possibly nos later on. The car i know is going to be a light blue, maybe a little darker than this blue. We plan to add some type of flames in the white coves. The flames will most likely be blue. The frame is all restored and the body has been re-fiberglassed, so all we need to do is join the two halves and will have one killer Vette.
Pros: Great Styling and Lots of Chrome
Cons: Not many that are important.
Displaying all 2 1958 Chevrolet Corvette reviews.
Reviews From Other Years
2015 Chevrolet Corvette Reviews
Impressive Value
Sexy car, I do not stop hearing people voice their enthused opinions at me. Every red light is a chance to hear what the next guy has to say about this impressive car. The fuel economy mode is impecca... Read More
2014 Chevrolet Corvette Reviews
This is the best car I've driven, and I've driven a lot of sports and luxury cars. This is hands down better than my $94,000 08 Viper. Drove the west coast from Washington down thru California and it ... Read More
All Muscle And So Much Fun To Drive!
It's hard to say anything bad about this car! My first sports car could not have been a better fit! I have a need for speed and this was the perfect car for me! Things I love - heads up display, hi... Read More
2013 Chevrolet Corvette Reviews
I love everthing about my vette,stylethree color black the yellow brack pads and i put the car in a show an win a best in show for the frist, time and i`m 65 yours young and love it when i take it out... Read More
Best Car!
I love the ride, looks and handling! With the top down in the evenings, there is nothing like it! This is the perfect car for the Carolina's, drive year round! This is my 7th Corvette, but the first c... Read More
2012 Chevrolet Corvette Reviews
I Love It, I Really Do, The Siblings Love It, So Im A Keeping It!
This car is a very fun car, it may be expensive, but she is my other women! she has always been there, she has always let me process my anger threw her pedals, and her gears. Her name is Treasure, for... Read More
I Am Extremely Impressed With The Car.
If looking for a performance car that is also a great daily driver, you can't beat this car. For a car with 430HP the mileage of 19-24MPG is excellent. Outstanding cargo capacity with the hatch rea... Read More
2011 Chevrolet Corvette Reviews
Best Non Aspirated Corvette
What can I say, I really cannot say enough. This Z06/Z07 is equipped the same as the Carbon Edition with subtle differences in paint scheme and interior trim, however from a performance perspective th... Read More
Only Drivin To Car Shows
This is an outstanding example of a Grand Sport. The color which is Supersonic Blue draws your attention. In fact, I've been told it's one of the most attractive colors of that model year. I get a lot... Read More


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I sit down with Joel De Jong—a life long entrepreneur, a scholar, an adventurer, a mystic, and a genuinely beautiful human being. You may know him from being one of the innovators behind Cheasty Greenspace, the largest contiguous forest in the Rainier Valley set in the urban corridor there in Seattle near his families home. He's also a musician and a singer, coordinating an exciting new project called Crowd Sourced Choir, which is dedicated to restoring collective singing as a core cultural compentency.
Our conversation centers on "The Dark Night of the Soul" in his own life, and the way of decent, in which Joel lost many of the things he believed were crucial to his identity, and ultimately discovered his most essential self, and highest purpose. This is a powerful interview relating to a man who is a turn-around artist and warrior of the soul. Check it out!


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It took me a while to realize how many comics about love, dating, and romance I was reading and enjoying… even though I read romance novels! I mentally classified most of these graphic novels as "slice of life" comics, I guess? Once I realized I had a theme going, clearly I had to to round them up on my blog. So here's the list of romance graphic novels I love and recommend. Hope you find something new to read!
Before we jump in:
- All comics here can be bought as graphic novels/collections, not only as single issues. Your library may own many of these!
- Amazon links are affiliate links.
- If you find this post helpful, please SHARE it, thank you!
- Any questions, corrections, recommendations? Let me know via the comments or my contact form.
- If you need to know whether a book has certain content that would make it a bad fit for you, I'm happy to check!
Bonnie N Collide (Lipstick Kiss Press / Amazon / Comixology / Read as a webcomic / Goodreads) By Monica Gallagher.
Gallagher's own words describe this better than I ever could: "The story of Bonnie N. Collide, a roller-girl, and the adventures she has at a humdrum day job. Bonnie's inability to separate her vibrant roller derby life from her normal working life means she gets to gleefully crash from one aspect of her life into another, seamlessly, and using the same amount of gusto. Oh, and one of her coworkers is a werewolf named Herb."
I absolutely can't resist the combo of sweet (and awkward) romances, girl-power derby bonding, office hijinks, and the occasional werewolf humor in this long-running webcomic, now available in print. The people are all drawn so cute! Bonnie herself is exuberant, a steadfast friend, and bi (yay!). There are many queer characters, a significant woman of color character, and the hot derby girls aren't all tiny. It's a soap opera, and I mean that in a good way, so if you need some suds in your life, this is a great one to pick.
Love Not Found (Strawberry Comics / Read as a webcomic / Goodreads) By Gina Biggs.
Charming, adorable, diverse, and queer sci-fi comic set in a future when people have almost completely given up directly touching each other. Sex is a recreational activity that happens in virtual reality, and anyone who expresses interest in romantic love or touch is regarded with extreme suspicion. Our two main characters are Abeille and Miel. Yes, Abeille means bee, and Miel means honey, so even if you knew nothing else, you can see where this is going. They meet by literally running into each other, and Abeille can't stop thinking about how that human contact felt. What follows is her and Miel's journey towards reclaiming touch as part of their sexuality, and the impacts their relationship has on various people around them.
I'm enjoying this series, especially how the writing and the character relationships and cast grow over time. I will definitely be supporting the Kickstarter for the second book whenever it begins!
Band Vs Band Comix (Band Vs Band Shop / Read as a webcomic / Goodreads) By Kathleen Jacques.
Jacques describes this as a "retro-cartoon-inspired, queer, handlettering-obsessed comic series about rival girl-fronted rock bands." One of the bands is daisies and kittens and all things wholesome. One of the bands is punk AF, never saw a sign they didn't want to deface, and snarls a lot. So of course, the gay lead singers have crushes on each other. We supported this on Kickstarter mostly on a whim, not having a clue if we'd like it, and I was immediately hooked. The characters are so funny and the art is so dang cute. It's a super slow burn, with plenty of band hijinx, song lyrics, and cute cats. TRY IT!
Diversity note: Jacques is gay.
Breaks (Amazon / Read as a webcomic / Goodreads) By Emma Vieceli and Malin Rydén.
I really loved this queer YA comic about two young men falling in a very tangled attraction. Neither of our two heroes is initially that easy to like, but as the story developed, I couldn't help but hope they'd figure themselves out and find a way to de-complicate everything so they have a chance together. Which will apparently be happening only in a future volume, because teenagers, what can you do? The art style is distinctive, with super clean penciling and a huge variety of paneling styles. I'm definitely looking forward to a next book.
Content note: violence and homophobia. Also, one of the characters has a girfriend so he's cheating – again with the whole "teenager" thing, they don't always make the best choices. (Granted, neither do adults.)
Diversity note: Malin Rydén is queer.
Taproot (Amazon/Comixology / Goodreads) By Keezy Young.
Quiet little queer romance between a ghost and a gardener who can see ghosts, with a bit of supernatural drama arising from the gardener's powers. It's structured a bit strangely IMHO, like one complete story and then a very long epilogue. Fine with me, though, since I found the art, the atmosphere, and the emotional connection between the two characters compelling. The color scheme feels very different and distinctive, and I would love to shop at Hamal's plant nursery. (If I could keep a plant alive ever.) If you like the cover and quiet love stories about adorable people, definitely give it a try.
Diversity note: Keezy Young is queer and nonbinary.
Hopeless Savages (Amazon / Comixology / Goodreads) By Jen Van Meter, with various artists.
Jen Van Meter has my undying gratitude for writing these stories about the happy, loving Hopeless-Savage family, a clan comprised of two parents who met during their careers as punk rock musicians, then settled down to have four children in a suburban setting which isn't always the best match. The first arc is centered on the youngest daughter, Zero Hopeless-Savage. Whether foiling her parents' kidnapping, de-brainwashing her oldest brother who is working for a corporate coffee chain, or living through a documentary film crew following them around, Zero is one of the most genuine and wonderful teenagers I've had the pleasure to read about… and did I mention she meets a great guy? Oh yes! And he is awesome! We also get good arcs about each older sibling, including Zero's gay older brother Twitch. His lost romance and his little sister's fledgling one collide beautifully at one point, and it's so sweet it practically brought me to happy tears. Sometimes love is hard, and sometimes it's awesome, and often you just have to rock out.
I also enjoyed the followup graphic novel Hopeless Savages: Break, with art by Meredith McClaren, for more adventures with Zero.
What Did You Eat Yesterday? (Amazon / Goodreads) By Fumi Yoshinaga.
The covers don't do this manga justice, but trust me on this one.
Shiro is a middle-aged lawyer who spends his nights cooking gourmet multi-course meals at home. He shares the meals, and his home, with his hairdresser boyfriend Kenji – but Shiro isn't out at work. The series bounces around between different happenings in Shiro and Kenji's lives, past and present. Foodies will love the in-depth step-by-step recipes and Shiro's explanations of why he prepares the food as he does. The rest of us can skim those parts and get to the next "adventure" as Shiro stays closeted at work, Kenji doesn't, ex-partners pop up, legal cases arise, and grocery prices are tracked very closely. (If there was extreme couponing in Japan, Shiro would do it.) There are super-touching moments, some absolutely hilarious parts, and it's such an interesting look into a facet of gay life in Japan.
I've read up through the latest volume out right now, which is volume 12. It's very slow-moving for character and relationship development, and I know some folks have had trouble with that, but it's worth trying the first several volumes to get a real feel for the series. Shiro's character, especially, has a lot going on with his family and his identity as a gay man in a culture that isn't welcoming, and sometimes he does things that seem to under-value his relationship with Kenji. I never worry, though, because by now I know the author is going to circle back around and allow him to keep growing.
The Less Than Epic Adventures of TJ and Amal (Amazon / Comixology / Goodreads) By E.K. Weaver.
Oh, dude, Amal! Don't get blind drunk two weeks before med school finals and right after you come out to your conservative parents and break off your engagement. That's just not good. And if you wake up hungover with a stoner in your apartment and he says you agreed to drive him cross country and he's going to pay for everything with cash? RUN AWAY. I don't care if he's cute and you really need to change your headspace and he's good in bed and actually listens to what you say and accepts who you are as a person. Seriously, I can see bad things happening! Where did all TJ's money come from?! Aw hell never mind. Y'all are too cute. I'm just going to hope for the best.
More seriously, this is a stunningly well-crafted love story / road trip comic! It's one of those books where if I talk about it in person, I just end up flailing my hands. It's that good. I'm trying very hard not to pick a top favorite from my favorites, but if I did…
Check Please! (Amazon / Goodreads) By Ngozi Ukazu.
An adorable, funny graphic novel about a collegiate men's hockey team. Our POV character is Eric Bittle, nicknamed Bitty, a gay former junior figure skating champion who's afraid of being body-slammed ("checked"). He loves to bake, he has a vlog, and oh yeah, he's probably falling in love with team Captain Jack Zimmerman. This volume mostly follows the ups and downs of college sports life, with the romance aspect ramping up towards the end. I don't know much about hockey aside from vague info gleaned from reading hockey romance novels, but that was fine, because this is more about the people than the game. The characters are engaging, and the art is cute. This the first of two volumes collecting the webcomic, and I am so there for the next book.
My Love Story (Amazon/Comixology / Goodreads) By Kazune Kawahara, with art by Aruko.
C-Man brought this high school romantic comedy manga series home from the library and almost fell off the couch laughing. I started reading it and also almost fell off the couch laughing. So, protective gear is advisable?
It starts by subverting your expectations. Takeo, the giant bruiser who would normally be the sidekick, is the romantic lead. The glamorous looking cool guy is a secondary character, Takeo's infinitely patient best friend Sunakawa. When Takeo falls in love with Yamato, this tiny, cute girl in his grade, it becomes clear that both parties are 100% clueless, in the most mutually devoted and entertaining way. If it weren't for the periodic intervention of Sunakawa, it's anybody's guess what would become of this poor couple.
It's refreshing, sweet, and well worth the 12 not-overly-long volumes. This was our most anticipated series for a couple of years until it concluded.
Sunstone (Amazon/Comixology / Goodreads) By Stjepan Šejić.
Lisa and Ally thought they could get together for some BDSM entertainment with no emotional strings attached. Hahaha, you silly girls, that would be too easy. Five volumes worth of feeeelings later, the series concluded, and Sunstone fans everywhere almost freaked out until we were told there would be spinoffs. It's too hard to let go of such a geeky, adorable, complex-because-people-are-dorks romance vibe like this! If you have triggers for BDSM elements, clearly this won't work for you, but if you're even neutral and you want more stories in your life about two ladies falling in love? Check this out.
ALSO: This comic proves that superhero comics don't *have* to objectify women. Because if you can create a five volume series about two women who do BDSM and are often in lingerie, and they're not objectified? Then there's no excuse for so many women who punch supervillains to be drawn the way they sometimes are.
Archie (Amazon/Comixology / Goodreads) By Mark Waid. Across the first four volumes: Illustrated by Fiona Staples, Annie Wu, Veronica Fish, Joe Eisma, and Pete Woods. Art assistance by Thomas Pitilli and Ryan Jampole. Colors by Andre Szymanowicz and Jen Vaughn. Letters by Jack Morelli.
I had no exposure to the Archie universe before starting this high school soap opera, so I didn't know what to expect, but I've found Mark Waid's take on Archie and his pals great fun. If you're not familiar with the Archie-verse, the setup is that our hapless main character, Archie, is caught between two girls: wholesome blonde Betty and scheming rich brunette Veronica. Love triangles rarely interest me, but I've been impressed with how not-triangle-y Waid had made this, with all three characters being humans rather than stereotypes. The series had antics and hijinks galore, a refreshingly diverse cast of secondary characters, and it's something to read when you're in the mood for fun teen drama.
So far, there are five collected volumes published, and it's an ongoing series.
Nana (Amazon/Comixology / Goodreads) By Ai Yazawa.
One of my blog readers suggested this series to me, so if you're still reading, thank you! I've been enjoying this series that follows two young women with the same name, Nana, who meet by chance and end up as roommates. Enough material was published for twenty-one volumes before the creator became ill and had to put the series on hiatus, and I'm only up through the seventh.
It feels very slice of life and new adult soap opera. Rock star hopeful Nana and no-career-yet Nana (who ends up nicknamed Hachi) interact with a continuously shifting group of lovers and friends clustered around members of Nana's band. I had to steel myself early on not to get attached to any of Hachi's boyfriends, or really any other pairing that seems like a good idea, because it seems inevitable that the author will break them up almost as quickly as they get together. (And am I misinterpreting that the over-arching narration, with Hachi telling us the story from some future point, starts to confirm a very slow arc of Hachi falling in love with Nana?)
So yeah, I have 14 more volumes before it potentially cliffhangs… but I know that's an occupational hazard in episodic storytelling like comics and television, and I enjoy each volume's (melo)drama so much that I won't be freaking out about "what happens next?!" Hopefully.
I'm kind of stunned that according to a source linked from Wikipedia, the series overall had sold over 43,600,000 collected volumes as of 2008. That's a lot of manga.
Crossplay (Amazon / Goodreads) By Niki Smith.
I couldn't summarize this better than the publisher did, so I won't try: "Close friends and new acquaintances at an anime convention confront their crushes, challenge their hang-ups, and question their once-comfortable identities in this erotic graphic novel about discovering who you're meant to truly be and who you're meant to love."
It's NSFW, sexy, fun, heartbreaking at times, and also incredibly compassionate and kind to all its very diverse and queer cast. The interconnected short stories are each very distinct, and I liked every single one. if it sounds at all interesting to you, give it a try.
That concludes today's roundup of comics about love, romance, and dating that I love and recommend! If you have any suggestions for me, please leave them in the comments – and thanks for sharing this post on social media or with friends.


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Do you know what's most important in life? Do you feel too busy to make time for those things?
This study gleans wisdom from the Book of Proverbs and from the example of Jesus to show you how to center your life on God's priorities. You will examine the things that matter most to God and to you as a follower of Christ. Then you will learn to apply godly wisdom that will help you incorporate these activities into your busy life. In some cases, you will find that you are doing the wrong things. Or you might find that you are doing the right things for the wrong reason or in the wrong way, so they are not life-giving or fulfilling.
Each week's session will function as a stand-alone message, providing great opportunities to invite friends and engage in conversations about a given topic through the week topically (for example, a neighborhood dinner to discuss parenting together or a focused time to build a budget).
Wisdom is learning how to do the right things at the right time in the right way. Allow God to show you what is most important in life and when and how to engage in those activities.
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Company receives CE marking; customer evaluations have begun for first multiplex panel, delivering healthcare providers in U.S. and Europe a new resource for antimicrobial stewardship
Salt Lake City, September 20, 2016 – Great Basin Scientific, Inc. (Nasdaq:GBSN), a molecular diagnostics company, announced today the U.S. and European commercial launch of its Staph ID/R Blood Culture Panel, the Company's first multiplex panel. This sample-to-result, automated panel benefits hospitals and laboratories by detecting, in about two hours, bloodstream infections caused by MRSA and other Staphylococcus species, enabling accurate, timely, and cost-effective diagnosis and treatment of patients. The panel detects the mecA gene, a major drug resistance marker that confers resistance to methicillin and other beta-lactams and creates the superbug MRSA. The assay is unique in its ability to distinguish Coagulase-negative Staphylococci (CoNS), which may not require treatment with antibiotics, from pathogenic Staphylococci, and can therefore assist healthcare organizations with antimicrobial stewardship programs and campaigns.
"The commercial launch of our Staph ID/R Blood Culture Panel marks a major milestone in our growth as a company as we deliver on our unique capability of providing low-plex, direct-from- specimen and multiplex testing on a single platform," said Ryan Ashton, co-founder and chief executive officer of Great Basin Scientific. "Additionally, we're pleased to be able to offer a significant tool for hospitals and labs as they implement antimicrobial stewardship programs into their broader healthcare strategy. The need for a solution to help clinicians battle antibiotic resistance is so great, nearly half of our installed customer base has already expressed an interest in our Staph ID/R Blood Culture Panel. Further, this panel is opening doors for our sales team at new larger sites not previously open to the Great Basin platform. Our Staph ID/R Blood Culture Panel will allow the clinical laboratory to play an important role in achieving the antimicrobial stewardship goals of improving patient care, reducing costs, and combating emerging antimicrobial resistance."
The Centers for Disease Control (CDC) reports that between 20 and 50 percent of all positive blood cultures are likely false positives due to contamination caused by CoNS, which are not usually pathogenic. However, some species have been associated with a growing number of hospital-acquired infections as well as antibiotic resistance caused by overuse or incorrect dosing or duration of these drugs. By clearly identifying CoNS, the Great Basin Staph ID/R Blood Culture Panel equips healthcare providers with actionable data they can leverage to quickly determine appropriate treatment paths, while helping eliminate false positives and inaccurate information that can lead to antibiotic resistance and outbreaks at a broader level. Informing disease-appropriate treatment for patients and curbing the unnecessary use of broad- spectrum antibiotics directly supports the larger fight against antibiotic-resistant infections, while lowering patient treatment costs, reducing potential readmissions and extending those savings to a hospital's bottom line.
Staph ID/R Blood Culture Panel has received CE marking designation under the European Directive on In Vitro Diagnostic Medical Devices, extending the availability to facilities across 32 countries in the EU and Great Britain. Great Basin Scientific currently has four commercially available assays, the Staph ID/R Blood Culture Panel, Shiga Toxin Direct Test, Group B Streptococcus (GBS) Test and Toxigenic Clostridium difficile (C. diff) Test. The Company has five additional tests and panels in development phase; Nasal S. aureus Pre-Screen, Candida Blood Infections Panel, Stool Bacterial Pathogens Panel, CT/NG Test and Bordetella Direct Test. All commercially available assays, as well as those in development run on the Great Basin analyzer.
The Company expects the Staph ID/R Blood Culture Panel to contribute meaningfully to revenue beginning in the second half of 2017, due to the panel's longer customer evaluation period.
About Great Basin Scientific
Great Basin Scientific is a molecular diagnostics company that commercializes breakthrough chip-based technologies. The Company is dedicated to the development of simple, yet powerful, sample-to-result technology and products that provide fast, multiple-pathogen diagnoses of infectious diseases. The Company's vision is to make molecular diagnostic testing so simple and cost-effective that every patient will be tested for every serious infection, reducing misdiagnoses and significantly limiting the spread of infectious disease. More information can be found on the company's website at www.gbscience.flywheelsites.com.
Forward-Looking Statements
This press release includes forward-looking statement regarding events, trends and business prospects, which may affect our future operating results and financial position, including but not limited to statements regarding anticipated current customer uptake of the Staph ID/R Blood Culture Panel, the effectiveness and role of the Staph ID/R Blood Culture Panel in patient care decisions, the expected contribution of the Staph ID/R Blood Panel to Company revenue, the Company's general development plans of sample-to-result technology and products. Forward- looking statements involve risks and uncertainties, which could cause actual results to differ materially, and reported results should not be considered as an indication of future performance. These risk and uncertainties include, but are not limited to: (i) our limited operating history and history of losses; (ii) our ability to develop and commercialize new products and the timing of commercialization; (iii) our ability to obtain capital when needed; and (iv) other risks set forth in the Company's filings with the Securities and Exchange Commission, including the risks set forth in the company's Annual Report on Form 10-K for the year ended December 31, 2015 and Quarterly Report on Form 10-Q for the quarter ended June 30, 2016. These forward-looking statements speak only as of the date hereof, and Great Basin Scientific specifically disclaims any obligation to update these forward-looking statements, except as required by law.


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Egyptian police closed a commercial passage with Israel on Sunday in support of colleagues who shut down a crossing with Gaza to protest the abduction of policemen, according to the AFP news agency, which based its report on reports in Egyptian state media.
The protesters say the commercial Al-Ouga crossing between Egypt and Israel will remain closed until three policemen and four soldiers abducted by gunmen in the Sinai peninsula are released, the official MENA news agency reported.
Police had shut down the Rafah crossing with Gaza on Friday, a day after gunmen abducted the policemen, who worked in the crossing, and soldiers.
State media has said security officials were in talks with the abductors via local Bedouin leaders who hold sway in the increasingly lawless peninsula.
A spate of hostage-taking has rocked the Sinai, which borders Israel as well as Gaza, but they usually last for no longer than 48 hours and are often carried out by Bedouin seeking the release of jailed relatives. Gas pipelines to Jordan and Israel have been bombed repeatedly.
Such abductions have been on the rise since the 2011 uprising that toppled veteran president Hosni Mubarak.


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Abhinav S wrote:Ashwin -
Abhinav S wrote:He should be out!
Abhinav S wrote:If we need a batsman, we will use find else.
Sandeep Mewara wrote:Yeah.. give chance to some 16yr old new comer and see
Abhinav S wrote:- however, there are some in the range 20-23.
Abhinav S wrote:, we will win in the future (even if we loose now).
_AK_ wrote:Every one bowled bad topday.
_AK_ wrote:Guy who use to be our trupcard sometime back become toothless.
Sandeep Mewara wrote:Except Yuvraj!
_AK_ wrote:He took his exclusion from the test team rather too seriously
_AK_ wrote:Anyway he was at his best in bowling in both the matches.
General News Suggestion Question Bug Answer Joke Praise Rant Admin
Use Ctrl+Left/Right to switch messages, Ctrl+Up/Down to switch threads, Ctrl+Shift+Left/Right to switch pages.


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Monday, June 23, 2008
F#ck Real Estate Agents!!!
4 comments:
- gingercat said...
My guess of someone laundering money is 1 in 10,000. Collect more personal information from law abiding people to catch the very odd person. Cons are not tough on law and order just content on taking people's personal information and rights away.
- Jun 23, 2008, 7:40:00 PM
- real estate said...
Fewer "Eyes" on Your Listing - Mispricing your home can prevent it from ever being seen by a certain percentage of potential buyers who might otherwise be interested in your home. Savvy buyers today research the local market even before acquiring an agent.
- Jul 8, 2010, 11:54:00 PM
- agence immobiliere auch said...
Check around first before you chose which real estate agency you go to. List down at least 3 best agencies and do a little research of their background.
- Apr 19, 2011, 8:41:00 PM
- Tiago said...
When we compare real estate agents we find people much more professional than others. So I think that if someone wants to spend dirty money buying a house just have to go to an unprofessional agent, so what is this policy´s point?
- Apr 15, 2013, 10:58:00 AM


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ASBMB President Steven McKnight writes in his second column: "I submit that the demise of the review process can be attributed to two changes. First, the quality of scientists participating in CSR study sections two to three decades ago was, on average, superior to the quality of study section participants today. Second, study sections have become highly specialized such that they narrowly define a differentiated club of biomedical research."
Find out what's happening in the lives of ASBMB members. Have news you'd like to share? Email us at firstname.lastname@example.com.
Considering the demonstrated importance of cilia for human health, many research groups have sought to characterize the molecular mechanisms of cilia biogenesis and function.
ASBMB Public Affairs Director Benjamin Corb writes, in part: "Doing advocacy is like doing regular maintenance on your car. It's not cheap and at times it's frustrating, but not taking the time to do it can result in catastrophic damage."
Find out about the latest round of winners of the Journal of Biological Chemistry/Herbert Tabor Young Investigator Award. And read about members who passed away recently.
Updates from the ASBMB journals: three thematic series (on prion disease, phospholipase D and cancer, and ketogenic diets) and more!
Mary L. Kraft and Raehyun Kim write about a new chemical imaging technique to visualize the distributions of sphingolipids and cholesterol in the plasma membranes of fibroblasts.
The 14 themes of the meeting symposia cover the broad range of biomolecules and systems that interest the ASBMB membership. We are certain that you will find new twists in the recent discoveries as well as new areas in which the science is advancing rapidly.
More from the current issue


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The pastoral nature of the campus is perhaps its most remarkable quality. It should be permitted to grow outwards, or inwards in this case, covering the campus center with trees, flowers, and grass. The park is re-drawn. An idylic garden is the result, creating a special place for gathering, conducting informal classes, and simply relaxing. The notion of weaving together the campus is again evident, blurring the distinction between old and new, building and landscape.
An interior seam slices through the topography creating an active void, a hybrid place, in which a variety of activities can unfold. It is an avenue, gently descending, controlling the flow of traffic, leading to a monumental stair carrying visitors upwards like the Champs Elysees or the Campidoglio in Rome.
The central seam serves many purposes;
- An entry court, from which accessto the various departments exist
- A node, or point of trajectory to another destination
- A forum for the exchange of ideas as students gather after class to discuss their views
- A pizza, with the cafeteria spilling out creating a real destination to stop and relax
- An outdoor theatre, as the stair can be used in an amphitheater like fashion
- A sculpture garden where indoor gallery events can push outwards


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After setting up your account online at https://fsc.cincweb.com, you can manage your own automatic draft payment information as needed.
To access your account online, visit https://fsc.cincweb.com Click the register button. Enter your information. Your name, complete address and email are required. Click submit. You will receive an email with your password once your registration has been approved. You may want to check your junk/spam folder. If your information does not match our database, it may take up to 24 hours to process your registration.


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Tag Archives: International Travel
Apr 29 2019
Traveling with a group can be extremely rewarding. Friends Club handles the details, keeping everything simple for our guests so they can relax and enjoy the sights, sounds, and experiences of group travel to its fullest!
Mar 08 2017
Another quarter full of new Friends Club events is quickly approaching, but we still have some great activities planned for the next two months. Take a look at what's still available and keep an eye out for our second quarter …
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Posted in Friends Club, News and Events Tagged International Travel
Feb 08 2017
We are looking forward to more wonderful Friends Club events and activities and invite you to take a look at what we have planned for the coming months. Simply click the title of each event to find more information and learn …
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Posted in Friends Club, News and Events Tagged International Travel
It's 5 p.m., our first evening in the African bush, and the three open Jeeps of 15 Iowans take a slight turn off the beaten path, leading us to a pair of male elephants shaking a tree and pulling at the …
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Did you know Portugal accounts for approximately half of cork produced annually worldwide, and that it takes 9 years for the bark on the oak tree to grow before it can be harvested? I bought a cork purse while visiting …
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Mar 02 2016
Cuba, Our Neighbor Cuba is more than cigars, rum, and coffee—it is an underdeveloped country filled with friendly, welcoming, and optimistic locals who embrace all travelers—especially Americans. Recently, the Hills Bank Friends Club took two groups to Cuba for an educational …
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Posted in News and Events Tagged Friends Club, International Travel
Nov 03 2015
Watch Karissa's video on tips for packing your suitcase for your next trip on YouTube: https://www.youtube.com/watch?v=hYAFyS2ZCbw
Posted in Organization Tagged International Travel, Time Management
Jul 09 2014
If you are like most people, you look forward to summer vacation with family and friends. Don't let what should be a fun time of memories in the making turn into an opportunity for thieves. If you are planning a …
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Mar 19 2014
Don't let fraud, pickpockets, identity theft, or a lost purse or wallet ruin your next vacation. The following tips will help protect your wallet, valuables, identity, and financial information while traveling.
Posted in Personal Banking Tagged International Travel, Money Management, Organization, Travel Tips


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In the latest issue of Cancer Research Update, AICR's biweekly email newsletter on the science of cancer prevention, treatment and survival, we asked cancer researchers and educators to answer one, simple question:
What do we know today that we didn't know just 10 years ago?
Their answers might surprise you – they surprised us. Although epidemiologists, clinicians, basic researchers and health professionals differ on what they believe to be the most important achievement in the past ten years, they agreed on one thing: It's never been clearer that diet, physical activity and a healthy weight all play an important protective role.
Check out the article — and subscribe to Cancer Research Update today.


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As we draw ever closer to Christmas and being constantly reminded by my daughter every waking minute, its made me reflect on how much more aware of it all she is this year. The excitement is bursting out of her like a can your friend at school would shake in a bid to soak you in humiliation – Unfortunately in this situation I can't 'tap her three times' to minimise the pent up hysteria.
This time last year she could barely speak, had an offensive barnet, tiny gappy teeth and as the title would suggest, if you asked her to say Christmas her response was "Piss".
It seems we've gone in full circle this past year, from telling everyone "Merry Piss" … to her Tangled obsession where she proceeded to tell adults I'd "locked her in the garden", her inability to pronounce her 'R's has had her repeating "rank" in a less than appropriate manner, dropping something in a supermarket and getting very shouty "oh for fzzks Sake!" to telling the entire nursery staff that "Daddy has a tail" – I will never live that down! to very matter-of-factly tell random adults at parties "I don't like you" and to just now, when Casey's milk bottle leaked all over him (damn these anti colic contraptions) when she said "Look, piss! its pissed all over him!" –
Note to self – We really must get that swear jar fixed.
Merry almost Piss-mas!


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Tory Johnson recommended our company as one of the direct sell company – "Passion Into Cash" on July 6th.
Join this growing creative storybooking company on my team & start making extra income this month. Check out http://www.KimsBookshelf.com! I LOVE what I DO!
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Financial Director, FCCA
Darren is a Certified Accountant and has been working in finance for over 20 years. He is the Financial Director of Sahara Care Limited and responsible for ensuring that the company is economically sound, viable, and cost efficient.
Darren spent over 13 years as an Audit manager in a firm of Chartered Accountants so has vast experience and knowledge in safeguarding not only the company's assets but also, most importantly, those of the service users.
Darren also has experience working directly with people who have learning disabilities. He takes great pride in his work which goes a huge way towards positioning Sahara Care Limited as an economically and socially responsible company.