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Natural killer (NK) cells hold promise for improving the therapeutic potential of allogeneic hematopoietic transplantation, but their effectiveness is limited by inhibitory HLA types. We sought to overcome this intrinsic resistance by transducing CD56+CD3- NK cells with chimeric receptors directed against CD19, a molecule widely expressed by malignant B cells. An abundance of NK cells for transduction was secured by culturing peripheral blood mononuclear cells with K562 cells expressing the NK-stimulatory molecules 4-1BB ligand and interleukin 15, which yielded a median greater than 1000-fold expansion of CD56+CD3- cells at 3 weeks of culture, without T-lymphocyte expansion. Expression of anti-CD19 receptors linked to CD3ζ overcame NK resistance and markedly enhanced NK-cell-mediated killing of leukemic cells. This result was significantly improved by adding the 4-1BB costimulatory molecule to the chimeric anti-CD19-CD3ζ receptor; the cytotoxicity produced by NK cells expressing this construct uniformly exceeded that of NK cells whose signaling receptors lacked 4-1BB, even when natural cytotoxicity was apparent. Addition of 4-1BB was also associated with increased cell activation and production of interferon γ and granulocyte-macrophage colony-stimulating factor. Our findings indicate that enforced expression of signaling receptors by NK cells might circumvent inhibitory signals, providing a novel means to enhance the effectiveness of allogeneic stem cell transplantation.
B-cell malignancies of children and adults, such as acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), and non-Hodgkin lymphoma (NHL), are often incurable even with intensive chemotherapy. For many patients, bone marrow ablation followed by allogeneic hematopoietic stem cell transplantation is the only potentially curative option, but the disease may return after transplantation.1 The well-documented association between T-cell-mediated graft-versus-host disease (GvHD) and a delay or suppression of leukemic relapse after allogeneic stem cell transplantation2-4 has led some investigators to manipulate GvHD by infusion of donor T lymphocytes. Although this procedure can induce a measurable antineoplastic response,5-8 it carries the risk of severe GvHD, particularly in those patients (> 70%) who lack an HLA-identical donor. Moreover, in some B-cell malignancies, such as ALL, the effect of lymphocyte infusions is often inadequate.6,9,10
Besides T lymphocytes, natural killer (NK) cells also exert cytotoxicity against cancer cells.11 Recent studies have emphasized the potential of NK-cell therapy in recipients of allogeneic hematopoietic stem cell transplants. In animal models of transplantation, donor NK cells could lyse leukemic cells and host lymphohematopoietic cells without affecting nonhematopoietic tissues,12 suggesting that NK-mediated graft-versus-leukemia responses may occur in the absence of systemic disease. Because NK cells are inhibited by self-HLA molecules, which bind to killer immunoglobulin-like receptors (KIRs), these findings have led to the clinical practice of selecting hematopoietic stem cell transplantation donors with an HLA and KIR type that favors NK-cell activation and thus could be expected to promote an antileukemic effect.13-15 However, selection of the “best” donor is limited to patients who have more than one potential donor and the capacity of NK cells to lyse lymphoid cells is generally low and difficult to predict.13,15-17
Emerging evidence indicates that T lymphocytes genetically modified with chimeric receptors able to recognize a surface molecule of target cells and transduce activation signals can specifically enhance T-cell cytotoxicity against cancer cells both in vitro and in vivo.18-21 The studies presented here are based on the concept that expression of chimeric receptors on NK cells could overcome HLA-mediated inhibitory signals, thus endowing the cells with cytotoxicity against otherwise NK-resistant cells. To test this hypothesis, we first developed a novel method that allows specific and vigorous expansion of NK cells lacking T-cell receptors (CD56+CD3- cells) and their highly efficient transduction with chimeric receptors. Then, we tested the relative antileukemic effects of genetically modified NK cells bearing chimeric receptors (directed against CD19, a molecule widely expressed by malignant B cells) that deliver different primary and costimulatory signals.
Materials and methods
The CD19+ human B-lineage ALL cell lines RS4;11, OP-1, 380, 697, and KOPN57bi; the T-cell line CEM-C7; and the myeloid cell lines K562 and U-937 were available in our laboratory.21 Cells were maintained in RPMI 1640 (Gibco, Grand Island, NY) supplemented with 10% fetal calf serum (FCS; BioWhittaker, Walkersville, MD) and antibiotics.
Primary leukemia cells were obtained from 9 patients with B-lineage ALL, with appropriate informed consent and Institutional Review Board (IRB) approval; from 4 of these patients, we also studied (with Saint Jude IRB approval) cryopreserved peripheral blood samples obtained during clinical remission. An unequivocal diagnosis of B-lineage ALL was established by morphologic, cytochemical, and immunophenotypic criteria; in each case, more than 95% of the cells were positive for CD19. Peripheral blood was obtained from 8 healthy adult donors. Mononuclear cells collected from the samples by centrifugation on a Lymphoprep density step (Nycomed, Oslo, Norway) were washed twice in phosphate-buffered saline (PBS) and once in AIM-V medium (Gibco).
Plasmids and retrovirus production
The anti-CD19-ζ, anti-CD19-BB-ζ, and anti-CD19-truncated (control) plasmids are described elsewhere.21 The pMSCV-IRES-GFP, pEQ-PAM3(-E), and pRDF constructs were obtained from the Saint Jude Vector Development and Production Shared Resource. The cDNA encoding the intracellular domains of human DAP10 and 4-1BB ligand (4-1BBL), and interleukin-15 (IL-15) with long signal peptide were subcloned by polymerase chain reaction (PCR) with a human spleen cDNA library (from Dr G. Neale, St Jude Children's Research Hospital) used as a template. An anti-CD19-DAP10 plasmid was constructed by replacing the sequence encoding CD3ζ with that encoding DAP10, using the splicing by overlapping extension by PCR (SOE-PCR) method. The cDNA encoding the signal peptide of CD8α, the mature peptide of IL-15 and the transmembrane domain of CD8α were assembled by SOE-PCR to encode a “membrane-bound” form of IL-15. The resulting expression cassettes were subcloned into EcoRI and XhoI sites of murine stem-cell virus-internal ribosome entry site-green fluorescent protein (MSCV-IRES-GFP).
The RD114-pseudotyped retrovirus was generated as previously described.21 We used calcium phosphate DNA precipitation to transfect 293T cells with anti-CD19-ζ, anti-CD19-DAP10, anti-CD19-BB-ζ, or anti-CD19-truncated; pEQ-PAM3(-E); and pRDF. Conditioned medium containing retrovirus was harvested at 48 hours and 72 hours after transfection, immediately frozen in dry ice, and stored at -80°C until use.
Development of K562 derivatives, expansion of NK cells, and gene transduction
K562 cells were transduced with the construct encoding the “membrane-bound” form of IL-15. Cells were cloned by limiting dilution, and a single-cell clone with high expression of GFP and of surface IL-15 (K562-mb15) was expanded. This clone was subsequently transduced with human 4-1BBL and designated as K562-mb15-41BBL. K562 cells expressing wild-type IL-15 (K562-wt15) or 4-1BBL (K562-41BBL) were produced by a similar procedure. Peripheral blood mononuclear cells (1.5 × 106) were incubated in a 24-well tissue-culture plate with or without 106 K562-derivative stimulator cells in the presence of 10 IU/mL human IL-2 (National Cancer Institute BRB Preclinical Repository, Rockville, MD) in RPMI 1640 and 10% FCS.
Mononuclear cells stimulated with K562-mb15-41BBL were transduced with retroviruses, as previously described for T cells.21 Briefly, 14 mL polypropylene centrifuge tubes (Falcon, Lincoln Park, NJ) were coated with human fibronectin (100 μg/mL; Sigma, St Louis, MO) or RetroNectin (50 μg/mL; TaKaRa, Otsu, Japan). Prestimulated cells (2 × 105) were resuspended in the tubes in 2 to 3 mL virus-conditioned medium with Polybrene (4 μg/mL; Sigma) and centrifuged at 2400g for 2 hours (centrifugation was omitted when RetroNectin was used). The multiplicity of infection (4-6) was identical in each experiment comparing the activity of different chimeric receptors. After centrifugation, cells were left undisturbed for 24 hours in a humidified incubator at 37°C, 5% CO2. The transduction procedure was repeated on 2 successive days. After a second transduction, the cells were restimulated with K562-mb15-41BBL in the presence of 10 IU/mL IL-2. Cells were maintained in RPMI 1640, 10% FCS, and 10 IU/mL IL-2.
Detection of chimeric receptor expression and immunophenotyping
Transduced NK cells were stained with goat anti-mouse (Fab)2 polyclonal antibody conjugated with biotin (Jackson ImmunoResearch Labs, West Grove, PA) followed by streptavidin conjugated to peridinin chlorophyll protein (PerCP; Becton Dickinson, San Jose, CA). For Western blotting, cells were lysed in RIPA buffer (PBS, 1% Triton-X100, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate [SDS]) containing 3 μg/mL pepstatin, 3 μg/mL leupeptin, 1 mM phenylmethylsulfonyl fluoride (PMSF), 2 mM ethylenediaminetetraacetic acid (EDTA), and 5 μg/mL aprotinin. Centrifuged lysate supernatants were boiled with an equal volume of loading buffer with or without 0.1 M dithiothreitol (DTT), and then separated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on a precast 10% to 20% gradient acrylamide gel (Bio-Rad, Hercules, CA). The proteins were transferred to a polyvinylidene fluoride (PVDF) membrane, which was incubated with primary mouse anti-human CD3ζ monoclonal antibody (clone 8D3; PharMingen, San Diego, CA). Membranes were then washed, incubated with a goat anti-mouse IgG horseradish peroxidase-conjugated second antibody, and developed by using the enhanced chemiluminescence system (Amersham, Little Chalfont, United Kingdom).
The following antibodies were used for immunophenotypic characterization of expanded and transduced cells: anti-CD3 conjugated to fluorescein isothiocyanate (FITC), to PerCP, or to energy-coupled dye (ECD); anti-CD10 conjugated to phycoerythrin (PE); anti-CD19 PE; anti-CD22 PE; anti-CD56 FITC, PE, or allophycocyanin (APC); anti-CD16 CyChrome (antibodies from Becton Dickinson, PharMingen, or Beckman-Coulter, Miami, FL); and anti-CD25 PE (Dako, Carpinteria, CA). Surface expression of KIR and NK activation molecules was determined with specific antibodies conjugated to FITC or PE (from Beckman-Coulter or Becton Dickinson), as previously described.15 Antibody staining was detected with a FACScan or an LSR II flow cytometer (Becton Dickinson).
Cytotoxicity assays and cytokine production
Target cells (1.5 × 105) were placed in 96-well U-bottomed tissue-culture plates (Costar, Cambridge, MA) and incubated with primary NK cells transduced with chimeric receptors at various effector-target (E/T) ratios in RPMI 1640 supplemented with 10% FCS; NK cells were cultured with 1000 U/mL IL-2 for 48 hours before the assay. Cultures were performed in the absence of exogenous IL-2. After 4 hours and 24 hours, cells were harvested, labeled with CD10 PE or CD22 PE and CD56 FITC, and assayed by flow cytometry as previously described.21-23 The numbers of target cells recovered from cultures without NK cells were used as a reference.
For cytokine production, primary NK cells (2 × 105 in 200 μL) expressing chimeric receptors were stimulated with various target cells at a 1:1 ratio for 24 hours. The levels of interferon γ (IFN-γ) and granulocyte-macrophage colony-stimulating factor (GM-CSF) in cell-free culture supernatants were determined with a Bio-Plex assay (Bio-Rad).
A test of equality of mean NK expansion with various stimuli was performed using analysis of variance for a randomized complete block design with each donor considered a random block. The Tukey honest significant difference procedure was used to compute simultaneous confidence intervals for each pairwise comparison of the differences of treatment means. Differences in cytotoxicities and cytokine production among NK cells bearing different chimeric receptors were analyzed by the paired Student t test.
Culture conditions that favor the expansion of primary NK cells
To transduce chimeric receptors into primary NK cells, we searched for stimuli that would induce specific NK-cell proliferation. In preliminary experiments, we depleted peripheral blood mononuclear cells of CD3+ T lymphocytes and stimulated the remaining cells with IL-2 (1000 U/mL) or IL-15 (10 ng/mL). Under these culture conditions there was no expansion of NK cells, which, in fact, progressively declined in numbers. With phytohe-magglutinin (PHA; 7 μg/mL) and IL-2 (1000 U/mL) as stimuli, we observed a 2- to 5-fold expansion of CD56+ CD3- NK cells after 1 week of culture. However, despite the low proportion of contaminating CD3+ cells (< 2% in 2 experiments) at the beginning of the cultures, these cells expanded more than NK cells (> 30-fold expansion), and after 1 week of culture represented approximately 35% of the cell population.
NK cells can be stimulated by contact with the human leukemia cell line K562, which lacks HLA-antigen expression,24 and genetically modified K562 cells have been used to stimulate cytotoxic T lymphocytes.25 We therefore tested whether the NK-stimulatory capacity of K562 cells could be increased through enforced expression of additional NK-stimulatory molecules, using 2 molecules that are not expressed by K562 cells and are known to stimulate NK cells. One molecule, the ligand for 4-1BB, triggers activation signals after binding to 4-1BB (CD137), a signaling molecule expressed on the surface of NK cells.26 The other molecule, IL-15, is a cytokine known to promote NK-cell development and the survival of mature NK cells.27-30 Because IL-15 has greater biologic activity when presented to NK cells bound to IL-15 receptor α (IL-15Rα) on the cell membrane of stimulatory cells, rather than in its soluble form,31-35 we made a construct containing the human IL15 gene fused to the gene encoding the human CD8α transmembrane domain and used it to transduce K562 cells. Expression of IL-15 on the surface of K562 cells was more than 5 times higher with the IL-15-CD8α construct than with wild-type IL-15 (not shown).
To test whether the modified K562 cells expressing both 4-1BBL and IL-15 (K562-mb15-41BBL cells) promote NK-cell expansion, we cultured peripheral blood mononuclear cells from 7 donors in the presence of low-dose (10 U/mL) IL-2 as well as irradiated K562 cells transduced with 4-1BBL or IL-15 (or both), or with an empty control vector. As shown in Figure 1, expression of either 4-1BBL or IL-15 by K562 cells improved the stimulation of NK-stimulatory capacity of K562 in some cases but not overall, whereas simultaneous expression of both molecules led to a consistent and striking amplification of NK cells (median recovery of CD56+CD3- cells at 1 week of culture, 2030% of input cells [range, 1020%-2520%] compared with a median recovery of 250% [range, 150%-640%] for K562 cells lacking 4-1BBL and IL-15; P < .001). In 24 experiments with cells from 8 donors, NK-cell expansion after 3 weeks of culture with K562 cells expressing both stimulatory molecules ranged from 309-fold to 12 409-fold (median, 1089-fold). Importantly, neither the modified nor unmodified K562 cells caused an expansion of T lymphocytes (Figure 1).
Among expanded CD56+CD3- NK cells, expression of CD56 was higher than that of unstimulated cells (Figure 2); expression of CD16 was similar to that seen on unstimulated NK cells (median CD16+ NK cells in 7 donors: 89% before expansion and 84% after expansion). We also compared the expression of KIR molecules on the expanded NK cells with that on NK cells before culture, using the monoclonal antibodies CD158a (against KIR 2DL1), CD158b (2DL2), NKB1 (3DL1), and NKAT2 (2DL3). The prevalence of NK subsets expressing these molecules after expansion resembled that of their counterparts before culture, although the level of expression of KIR molecules was higher after culture (Figure 2). Similar results were obtained for the inhibitory receptor CD94, whereas expression of the activating receptors NKp30 and NKp44 became detectable on most cells after culture (not shown). In sum, the immunophenotype of expanded NK cells reiterated that of activated NK cells, indicating that contact with K562-mb15-41BBL cells had stimulated expansion of all subsets of NK cells.
Transduction of NK cells with chimeric receptors
Before transducing peripheral blood mononuclear cells with retroviral vectors containing chimeric receptor constructs and GFP (Figure 3), we stimulated them with K562-mb15-41BBL cells. In 27 experiments, the median percentage of NK cells that were GFP+ at 7 to 11 days after transduction was 69% (43%-93%). Chimeric receptors were expressed at high levels on the surface of NK cells and, by Western blotting, were in both monomeric and dimeric configurations (Figure 4).
To identify the specific signals required to stimulate NK cells with chimeric receptors, and overcome inhibitory signals mediated by KIR molecules and other NK inhibitory receptors that bind to HLA class I molecules, we first compared 2 types of chimeric receptors containing different signaling domains: CD3ζ, a signal-transducing molecule containing 3 immunoreceptor tyrosine-based activation motifs (ITAMs) and linked to several activating receptors expressed on the surface of NK cells,11,36 and DAP10, a signal transducing molecule with no ITAMs linked to the activating receptor NKG2D and previously shown to trigger NK cytotoxicity.11,36,37 As a control, we used NK cells transduced with a vector containing an anti-CD19 receptor but no signaling molecules or containing GFP alone. NK cells were challenged with the CD19+ leukemic cell lines 380, 697, and RS4;11, all of which express high levels of HLA class I molecules by antibody staining. By genotyping, RS4;11 is Cw4/Cw3, Bw4 and A3; 380 is Cw4/Cw4, Bw4; and 697 is Cw3/Cw3. Hence, these cell lines were fully capable of inhibiting NK-cell cytotoxicity via binding to NK-inhibitory receptors.
Expression of receptors without signaling molecules did not increase NK-mediated cytotoxicity over that exerted by NK cells transduced with the vector containing only GFP (Figure 5). By contrast, expression of anti-CD19-ζ receptors markedly enhanced NK cytotoxicity in all experiments (Figure 5), regardless of the intrinsic ability of donor NK cells to kill leukemic targets. For example, 380 cells were highly resistant to NK cells from donors 2 and 3, but were killed when these donor cells expressed anti-CD19-ζ receptors. Similar observations were made for RS4;11 cells and the NK cells of donor 1 and for 697 cells and NK cells of donor 2. Moreover, the anti-CD19-ζ receptors led to improved killing of target cells even when natural cytotoxicity was present. In all experiments, the cytotoxicity triggered by the anti-CD19-ζ receptor was enhanced over that achieved by replacing CD3ζ with DAP10 (P < .001; Figure 5).
4-1BB-mediated costimulatory signals enhance NK cytotoxicity
Previous studies have shown that the addition of costimulatory molecules to chimeric receptors enhances the proliferation and cytotoxicity of T lymphocytes.21,38-43 Of the 2 best known costimulatory molecules in T lymphocytes, CD28 and 4-1BB, only 4-1BB is expressed by NK cells.26,44,45 We therefore determined whether the addition of 4-1BB to the anti-CD19-ζ receptor would enhance NK cytotoxicity. In a 4-hour cytotoxicity assay, cells expressing the 4-1BB-augmented receptor showed a markedly better ability to kill CD19+ cells than did cells lacking this modification (Figure 6A-B). The superiority of NK cells bearing the anti-CD19-BB-ζ receptor was also evident in 24-hour assays with NK cells from different donors cultured at a 1:1 ratio with the leukemia cell lines 697, KOPN57bi, and OP-1 (not shown).
Next, we determined whether the antileukemic activity of NK cells expressing anti-CD19-BB-ζ receptors extended to primary leukemic samples. In 5 samples from children with different molecular species of ALL, NK cells expressing the 4-1BB receptors exerted strong cytotoxicity that was evident even at low E/T ratios (eg, < 1:1; Figure 7) and uniformly exceeded the activity of NK cells expressing signaling receptors that lacked 4-1BB. Even when donor NK cells had natural cytotoxicity against ALL cells and CD3ζ receptor did not improve it (patient no. 3 in Figure 7), addition of 4-1BB to the receptor significantly enhanced cytotoxicity. Consistent with their increased cytotoxicity, NK cells expressing anti-CD19-BB-ζ mediated more vigorous activation signals. As shown in Figure 8A, 46% of NK cells bearing this receptor expressed the IL-2Rα chain CD25 after 24 hours of coculture with CD19+ ALL cells, compared with only 17% of cells expressing the anti-CD19-ζ receptor and less than 1% for cells expressing receptors that lacked stimulatory capacity. Moreover, anti-CD19-BB-ζ receptors induced a much higher production of IFN-γ and GM-CSF on contact with CD19+ cells than did receptors without 4-1BB (Figure 8B).
We asked whether the expression of signaling chimeric receptors would affect spontaneous NK activity against NK-sensitive cell lines not expressing CD19. Spontaneous cytotoxicity of NK cells from 3 donors against the CD19- leukemia cell lines K562, U937, and CEM-C7 was not diminished by expression of chimeric receptors, with or without 4-1BB (not shown).
Anti-CD19 chimeric receptors induce NK cytotoxicity against autologous leukemic cells
To determine whether the NK-cell expansion and transduction system that we developed would be applicable to clinical samples, we studied peripheral blood samples that had been obtained (and cryopreserved) from 4 patients with childhood B-lineage ALL in clinical remission, 25 to 56 weeks from diagnosis. NK-cell expansion occurred in all 4 samples; after 1 week of culture with K562-mb15-41BBL cells, recovery of CD56+CD3- NK cells ranged from 1350% to 3680% of the input.
After transduction with chimeric receptors, we tested the cytotoxicity of the NK cells against autologous leukemic lymphoblasts obtained at diagnosis. As shown in Figure 9, expression of anti-CD19-BB-ζ receptors overcame NK-cell resistance of autologous cells; NK cells expressing the receptors exerted cytotoxicity, which was as powerful as that observed with allogeneic targets.
In this study, we demonstrated that the resistance of cancer cells to NK-cell activity can be overcome by chimeric receptors expressed on primary NK cells. The stimulatory signals triggered by the receptors on contact with target cells predominated over inhibitory signals and induced powerful cytotoxicity against NK-resistant leukemic cell lines and primary leukemic cells. We found that the type of stimulatory signal delivered by the chimeric receptor was a key factor in inducing cytotoxicity. Although DAP10 signaling can elicit NK cytotoxicity,37 chimeric receptors containing this molecule in our study induced weaker NK-cell activity than that generated by CD3ζ-containing receptors, despite identical levels of surface expression. We also found that addition of the costimulatory molecule 4-1BB to the chimeric receptors markedly augmented cytotoxicity and that receptors containing both CD3ζ and 4-1BB triggered a much more robust NK-cell activation and cytokine production than did those containing only CD3ζ. The important contribution of 4-1BB signals agrees with findings that anti-4-1BB antibodies activate murine NK cells,46 and enhance their antitumor activity.26 Leukemic lymphoid cells usually do not express 4-1BBL21 ; only 2 of 284 diagnostic B-lineage ALL samples studied by gene arrays at our institution expressed 4-1BBL transcripts.47 Hence, 4-1BB signals can be delivered to NK cells only if the molecule is incorporated into the receptor.
Efficient and stable transduction of primary NK cells is notoriously difficult, prompting us to devise a new gene transduction method for the present study. Most investigators have demonstrated efficient gene transfer only in continuously growing NK-cell lines48-54 or reported methods yielding only transient gene expression.37,55,56 We achieved stable expression of chimeric receptors in primary CD56+CD3- NK cells by using an RD114-pseudotyped retroviral vector and specifically expanding primary CD56+CD3- NK cells before they were exposed to the retrovirus, a step that allowed highly efficient gene expression. Although several cytokines such as IL-2, IL-12, and IL-15 stimulate NK cells,27,57,58 their capacity to induce proliferation of resting CD56+CD3- cells has been poor, unless accessory cells are present in the cultures.24 Perussia et al59 found that contact with irradiated B-lymphoblastoid cells induced as high as a 25-fold expansion of NK cells after 2 weeks of stimulation, and Miller et al60 reported an approximate 30-fold expansion of NK cells after 18 days of culture with 1000 U/mL IL-2 and monocytes. However, these culture conditions are likely to promote the growth of CD3+ T lymphocytes as well as NK cells. Because our ultimate aim is to generate pure preparations of donor NK cells devoid of CD3+ T lymphocytes that can be infused into recipients of allogeneic hematopoietic stem cell transplants, we searched for methods that would maximize NK-cell expansion without producing T-cell mitogenicity.
Contact with K562 cells (which lack major histocompatibility complex [MHC] class I molecule expression and hence do not trigger KIR-mediated inhibitory signals in NK cells) is known to augment NK-cell proliferation in response to IL-15.24 We found that membrane-bound IL-15 and 4-1BBL, coexpressed by K562 cells, acted synergistically to augment K562-specific NK-stimulatory capacity, resulting in vigorous expansion of peripheral blood CD56+CD3- NK cells without concomitant growth of T lymphocytes. After 2 to 3 weeks of culture, we observed NK-cell expansions of up to 10 000-fold, and virtually pure populations of NK cells could be obtained, even without the need for T-cell depletion in some cases. NK cells expanded in this system retained the immunophenotypic diversity seen among peripheral blood subsets of NK cells, as well as their natural cytotoxicity against sensitive target cells, even after transduction with different chimeric receptors. Hence, this system should help studies of NK-cell biology that require specific cell expansion or gene transduction or both, but it should also be adaptable to clinical applications after generating K562-mb15-41BBL cells that comply with current good manufacturing practices for clinical trials. Recently, Harada et al reported that expansions of CD56+CD3- cells (up to 400-fold after 2 weeks) were apparently superior after contact with another HLA class I-negative cell line, the Wilms tumor cell line HFWT.61 Future studies should determine whether HFWT cells express 4-1BBL or whether enforced expression of 4-1BBL together with IL-15 results in a greater specific expansion of NK cells than seen with modified K562 cells.
In the context of allogeneic hematopoietic stem cell transplantation, infusions of activated donor T cells would carry an unacceptably high risk of severe GvHD, particularly in recipients of haploidentical or mismatched transplants. By contrast, infusions of pure CD56+CD3- NK cells should not impose that risk.12 Most clinical studies of the therapeutic effects of NK cells have been performed in an autologous setting and have yielded only moderately promising results.11,62 This is not surprising because NK-cell activity is inhibited by surface receptors that recognize autologous HLA molecules expressed by both normal and neoplastic cells. Allogeneic NK cells may be more effective, but even in an allogeneic setting the capacity of NK cells to kill malignant lymphoid cells is generally modest and often negligible.17 Leung et al15 detected NK cytotoxicity against an ALL cell line expressing particularly low levels of inhibitory HLA molecules, but cytotoxicity was much lower than that observed against the NK-cell target K562; only about 50% of the ALL cells were killed at an effector-target ratio of 40:1. In that study, RS4;11 cells, which express HLA-C alleles that bind the most commonly expressed KIRs, were NK resistant, whereas these cells, as well as autologous leukemic cells, were highly sensitive to NK cells expressing anti-CD19 signaling receptors in our study. Thus, NK cells expressing signaling chimeric receptors have much more powerful antileukemic activity than unmodified NK cells and can kill target cells irrespective of their HLA profile.
An increased understanding of the signals leading to immune cell activation, together with progress in gene cloning and transfer, have made the treatment of cancer with “adoptively acquired immunity”63 a realistic goal. Clinical precedents, such as administration of T-cell clones that target cytomegalovirus epitopes64 or Epstein-Barr virus-specific antigens,65 attest to the clinical feasibility of adoptive immune cell therapy. Nonetheless, potential limitations may affect the effectiveness of cell therapy guided by chimeric receptors. One is that the murine scFv portion of the chimeric receptor or the fusion sites of the human regions that compose it may trigger a host immune response leading to elimination of the modified cells.18 Although the impact of such an event in a clinical setting remains to be determined, we anticipate that immune responses against modified NK cells will be limited in immunosuppressed patients after hematopoietic stem cell transplantation. Another potential limitation is that adoptively transferred cells may have inadequate persistence in vivo, although a recent study showed that NK cells obtained from haploidentical donors and activated ex vivo could expand in patients when infused after administration of high-dose cyclophosphamide and fludarabine, which caused an increase in endogenous IL-15.66 We speculate that such expansions would also occur with genetically modified NK cells and suggest that further studies to identify signaling molecules that promote NK-cell proliferation when incorporated into chimeric receptors are warranted. In patients at a high risk of leukemia or lymphoma relapse, the expected benefits of genetically modified NK cells will outweigh the risk of insertional oncogenesis posed by the use of retroviruses for chimeric receptor transduction.67 We also predict that the coexpression of suicide genes will become a useful safety measure in clinical studies68 ; this strategy would also ensure that the elimination of normal CD19+ B-lineage cells is only temporary.
Novel therapies that bypass cellular mechanisms of drug resistance are urgently needed for patients with refractory leukemia and lymphoma. NK-cell alloreactivity is a powerful new tool for improving the therapeutic potential of allogeneic hematopoietic stem cell transplantation. The results of this study indicate that signaling receptors can enhance the efficacy of NK-cell alloreactivity and widen its applicability. We envisage initial clinical trials in which donor NK cells, collected by apheresis, are expanded ex vivo as described here, transduced with chimeric receptors, and then infused after transplantation in patients with B-lineage ALL. The target molecule for the chimeric receptors, CD19, was selected because it is one of the most widely expressed surface antigens among B-cell malignancies, including ALL, CLL, and NHL. In these malignancies, CD19 is highly expressed on the surface of virtually all cells but has limited or no expression in normal tissues.69 However, the NK-cell strategy of immunotherapy we describe would not have to be directed to the CD19 antigen but could be applied to any of the numerous molecules identified as potential targets for chimeric receptor-based cell therapy in patients with cancer.18
Prepublished online as Blood First Edition Paper, March 8, 2005; DOI 10.1182/blood-2004-12-4797.
Supported by grants CA58297 and CA21765 from the National Cancer Institute and by the American Lebanese Syrian Associated Charities (ALSAC).
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 U.S.C. section 1734.
We thank Martha Holladay and Jim Houston for KIR expression analysis, Victoria Turner for cell line genotyping, Stanley Pounds for assistance with statistical analysis, the St Jude Vector Development and Production Shared Resource for retroviral vectors and the St. Jude Tissue Resource laboratory for providing patient samples.


text:
Opposing the Bedroom Tax
- 09:52 am, Wed 13th Nov 2013
The full transcript of my speech to the House of Commons during the debate on the abolition of the Bedroom Tax
You can watch a video of the full speech here: http://youtu.be/bPDAtyuj0cA
12th November 2013 - 2.41 pm
Mr David Crausby (Bolton North East) (Lab): I intend to use the term “bedroom tax” today and not “under-occupancy penalty” or “single room subsidy”. If that offends anyone, I can assure them they will not be anywhere near as offended as thousands of my constituents have been by this repulsive Government attack on disadvantaged and disabled people. The conflict surrounding the description of this despicable act reminds me of Margaret Thatcher’s attempt to force the term “community charge” down the throats of the British people. Not surprisingly, she failed, and history damned it as the poll tax. The same fate awaits the single room subsidy.
The policy itself will also fail because, like the poll tax, it is based on mean-mindedness and political dogma. It will be also rejected by Conservative and Liberal leaderships yet to come, and once it has gone—as it will do after the next election—it will be disowned. Even as Margaret Thatcher was being driven away from Downing street in tears, John Major was planning to ditch the poll tax. He had remembered what Mrs Thatcher had clearly forgotten: that, regardless of how big someone gets for their boots, if they want to win elections and stay in power they should keep in touch with public opinion. They should also bear in mind that our electorate are, I am proud to say, for the most part decent and fair-minded people who know a pig in a poke when they see one. John Major understood that, which is why he went on to win the next election in 1992.
Prime Ministers have their albatrosses, however: John Major’s was the exchange rate mechanism; Margaret Thatcher’s was the poll tax; Tony Blair’s was Iraq; and Jim Callaghan’s was the winter of discontent. The bedroom tax will belong to the present incumbent. As with the ancient mariner, it will hang round his neck in shame.
Dr Coffey (con): The hon. Gentleman might wish to check the recent opinion polls. We would appreciate it if he were more consistent about changing the rules for people on local housing allowance. If they were so bad for private sector rented flats, why is the situation so different for the public sector?
Mr Crausby: I do not think that my political principles have changed, to be perfectly honest. I would have put forward these same arguments prior to the election as well.
On a more serious point, nearly 2,500 people back home in Bolton have been affected, and more than 75% of them have fallen into arrears since April—so much for this being a money-saving measure.
Sir Tony Cunningham (Workington) (Lab): On the difficulty of moving, I have a constituent who has got into arrears because of the bedroom tax. The only way in which she can get out of arrears is to move to a smaller property but, guess what, she cannot move because she is in arrears. Does not this demonstrate the madness of this policy?
Mr Crausby: Absolutely. I will come to that point in a moment.
The fact is that millions of pounds will be lost. That represents much-needed cash that needs to be spent on making living conditions better, not worse. The demand for debt advice and financial service advice is bound to soar, and housing staff will concentrate most of their efforts on chasing rent arrears and helping people to move—when they can, that is. Legal expenses will escalate, and the potential cost of evicting decent families is enormous. This additional expense might not come directly from the Government’s coffers, but it will come from British people’s pockets and, frankly, we have better things to spend it on.
One example of the measure’s inflexibility involves constituents of mine who have two children, a boy and a girl. The girl was nine when they moved in, just before the bedroom tax was implemented. They were not entitled to live in a three-bedroom house until she was 10, when she could no longer be expected to share a room with her brother. As a result, the family were penalised for months until she was 10. The problem with this cruel measure is that it is focused on punishing people, and not on dealing with the problems of under-occupancy.
Under-occupancy is plainly a problem, but the bedroom tax is definitely not the solution. Where is the justice in denying tenants the opportunity to move to smaller, more energy-efficient properties when there are hardly any available—that is certainly the case in my constituency —and at the same time penalising them financially? The sensible solution involves helping people and building affordable homes for rent. It also involves giving tenants an incentive to downsize, not making them suffer because they are poor and in receipt of welfare benefits. Back in Bolton, the Conservative and Liberal councillors actually get it, and they have voted for a council motion to abolish the bedroom tax—then again, maybe they understand what John Major understood when he abolished the poll tax.
The Payday Loans Charter
- 04:43 pm, Fri 1st Nov 2013
Local residents have made their voices heard this year about the problems caused by the payday loan industry.
Action by local people helped to convince Bolton Wanderers to drop QuickQuid as their new sponsor and I was proud to support the Fabulous Finance Fayre in town in September.
I have said many times that I think it is time for this industry to be regulated, and we need to do more to back positive alternatives like credit unions. It is important to keep promoting that idea in order to bring about change.
I was very pleased to see that some of the leading anti-poverty and consumer advice organisations share this opinion and have developed the Charter to Stop the Payday Loan Rip-off. Among others, the Charter is supported by Which?, Citizens Advice, StepChange Debt Charity, Church Action on Poverty, and the Centre for Responsible Credit.
I signed the Charter in support at the launch event in Parliament and you can show your support at change.org/paydayloancharter
The full text of the Charter to Stop the Payday Loan Rip-off reads:
We believe irresponsible payday lending and other high cost credit is damaging the health and wealth of our country. Payday lenders are exploiting millions of people across the UK, trapping them in spirals of debt, and the problem is getting worse.
Payday lenders are breaking promises they made in their own customer charter. Self-regulation has failed. We call for effective regulation of payday lenders and high cost credit, which is properly enforced, to:
Stop them giving loans to people who can’t realistically afford to pay them back
Stop them repeatedly rolling over loans and creating spiralling debt
Stop hidden or excessive charges
Stop them raiding borrowers’ bank accounts without their knowledge and leaving them in hardship
Stop irresponsible advertising and instead provide clear and transparent information
Require lenders to promote free and independent debt advice, and ensure they co-operate with other services to help people get out of debt.
We also want action to support the growth of credit unions and other forms of more responsible lending; we want banks to increase the availability of credit to people on low and middle incomes: and we want new research on capping the total cost of credit undertaken now.

domain: economics


text: I got my start in a garage in Raleigh NC as a member of a high school band called Quest. In college I was the singer for a band called The Stegmonds, this is where i really learned a lot about music and my place in it. Way led to way and I met a young woman who would become my wife. I climbed into a UPS truck to help make a living for us and our two children. Now our children are older and I have begun to re-explore music and start writing. What this will produce only time will tell.

domain: artistic


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- Friendly Staff
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Share Your Experience
Have you seen Dr. Alan Burgess?
August 22nd, 2016
March 20th, 2016A true Care-Giver
November 19th, 2015
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December 15th, 2014
I seen Maureen, Dr Burgess's NP but I didnt see her name in the system. The office asked if I can give a review and I told them absoulity because this location is awesome! They are friendly and have great energy there. I would recommend anyone to go there.
December 9th, 2014Great location
I only been to this location a couple of times but the practice asked if I would write a review. I was more than happy to write a review because everyone there is so professional and Dr Burgess is great. He takes time to listen to my issue verse rushing me out the door. This is a great practice.
May 7th, 2014
April 22nd, 2014Nurse Practicianer
April 13th, 2014
Dr Burgess is such a great guy. I love this location. Everyone there is so nice.
April 2nd, 2014Awesome doctor
He is very friendly and willing to listen. He addressed all my issues and gave me suggestions on my health to improve myself. He is awesome!
Dr. Alan Burgess is similar to the following 3 Doctors near Denver, CO.
Dr. Eric Exelbert
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- Birth Control Facts about birth control to help decide which type is right for you.
- Type 2 Diabetes Basic facts about type 2 diabetes & risk factors to be aware of.
- COPD The facts about chronic obstructive pulmonary disease (COPD).
- View All Care Guides Prepare for your next visit with our extensive library of Care Guides

domain: medical


– Kofi Kingston says he doesn’t care what people think about his in-ring performances.
The latest issue of WWE Magazine (cover-dated January 2014) features the 2013 WWE Magazine Awards and the seven-time mid-level champion was voted by fans as “Most Underrated.” Kingston, however, doesn’t care for the moniker.
“I don’t really care what people think about me,” Kingston remarked to WWE Magazine about being called underrated. “I think if someone’s underrated, that they’re under appreciated. I’m fully confident in my abilities, I know every night, in my mind, that I’m going to have a good match. People feel underrated when they think they’re not getting the credit they deserve. That’s why I don’t care that much about what people think.”
– Chris Jericho recently spoke with WWE.com about a potential return and more. Here is what he had to say…
“I don’t really have the itch. I’m super busy with all the stuff I’m doing, between Fozzy tours, we’re getting ready to record a new record, the podcast, “But I’m Chris Jericho,” which is another big hit. I’m also finishing my third book. There’s a lot of stuff going on. Having said that, I’d be open to it, but I haven’t had any conversations or anything like that. It’s WrestleMania. Anybody that’s been in WWE wants to be involved. If I’m there, it’ll be awesome and I’ll do everything I can to have the best match on the show. If I’m not, I’ll have a great time watching it, I’m sure.”


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I was excited to see the Steens, a fault-block mountain of the basin and range geology, not like the volcanoes of the Cascade Mountains by my home. I had driven from the Hart Mountain Antelope Reserve and when I reached the valley rim above Frenchglen the Steens looked like a huge gentle giant of a hill that just kept going skyward. There was no clue that the backside abruptly drops to the Alvord valley.
I was headed to the BLM Page Springs campground; a campground that my boondocking brother had recommended. It must be nice if he recommended it. It was centrally located for exploring the vast area with a nice amount of space between each campsites. I found the campground a happy place with a comfy camp site that I settled into for a three night stay.

domain: geography


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Funny Valentine's Card, Valentines Day Card, Tom Hardy, Funny Love Card, Boyfriend Card, Funny Him Card
Let someone know how you feel with this funny greeting card with the message ‘Roses are Red, Violets are Blue, Tom Hardy's taken, so you'll do’. A great way to show you care to a loved one or Friend, for Valentines, Anniversary or just because! This cheeky card is guaranteed to make your friend, boyfriend or girlfriend smile!
There are a variety of celebrity crushes you can choose from (Tom Hardy, Ryan Gosling, David Beckham, Danny Dyer) or add your own! Just select 'Add your own name' and include the required name in the notes section at checkout.
Finished by hand in a beautiful gold foil, this lovely card glistens! Due to the hand-made nature of this item, please note that no two items are the same, so please do allow for individual finishes.
- Handmade item
- Materials: ink, recycled board, recycled envelopes, card stock, print, giclee print, cello bags, hardbacked envelope, illustration, design, professional printing, environmentally friendly.
- Gift message available


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As the leaves change and temperatures get cooler, what better way to celebrate than with a weekend fall hike. Here are the best areas for fall hikes to suit any experience level. Give yourself a boost of autumn spirit and enjoy the great outdoors this autumn.
New England has such diverse landscape, starting in Maine (one of my “bucket list” vacations) and running the length of Connecticut. The four states in between are Massachusetts, Vermont, New Hampshire, and Rhode Island. Every state offers up moderate to strenuous hikes near water or wilderness depending on your endurance.
An easy to moderate hike in Massachusetts loved by artists and nature lovers is Bish Bash Falls. Two miles to enjoy scenery and the biggest waterfall in the state! For a more challenging hike, Mount Mansfield in Vermont will give you breathtaking views near Lake Chaplain. The 6-mile roundtrip hike has been known to have the very best views in the state. With the elevation change be prepared with warmer clothes as you reach the summit and colder temperatures.
Great Smoky Mountains National Park
The Great Smoky Mountains have an abundance of wilderness trails mixed with amenities to make your fall excursion perfect for any age. Trails range from level walking paths to bike and board trails for all ages. There are also nature walks, to help you and your family identify plants and the surrounding wildlife easily. Finally, due to the elevation of the park, your hike can show you some of the best fall foliage near the East Cost of the United States.
If you live in the southwestern region of the United States, you have many choices to experience the changing leaves. Enjoying the crisp fall weather is only a short drive away to northern Arizona. Sedona, Flagstaff and along the Mogollon Rim all have the advantage of being close the urban center of Phoenix. These towns offer some of the best nature walks with only a two-hour drive north.
Humphrey’s Peak Trail is considered an easy 4.5-mile hike reaching an elevation gain of about 3500 feet. The highest point of the peak is 12,000 feet, so be aware of the elevation change and plan accordingly. Temperatures will also be much cooler when you reach the summit so for all hikes pack warm clothing and the appropriate amount of water and food.

domain: travel


text:
How do you get a 20% smaller cerebellum than normal? I am interested in this topic because of Temple Grandin’s struggle with autism. This relates to Temple Grandin because when we as a class were reading her life story right in the beginning she states that she has a 20% smaller cerebellum than normal. My question to find out the answer to is how do you get a 20% smaller cerebellum than normal? Families and teachers can help children with a smaller cerebellum by taking it slow and parents need to look at what they are good at and the strategies that doctors have to offer. What we should do to support the children and families that need help to overcome a this deficit children need to be helped slowly. This is a very common birth disease and defect.
If you were to have a smaller cerebellum than normal, then you might have autism or something similar to that. Your cerebellum is the main part of your body that controls almost everything that you do. “The cerebellum controls walking, balance, and your eyes”(Chiari Malformation). So this would mean you would have incoordination. “If your cerebellum is smaller than normal your movement is more unsteady and clumsy”(What is the cerebellum and what does it do). “When you have a smaller cerebellum this also means that you can get easily distracted and that you have shaky hands”(Living with Cerebellar Hypoplasia). I actually have someone in my family that can relate to this because my cousin’s daughter has a brain disorder just like the one i’m talking about now she does have autism and I seen her recently and I can tell how her mind works. Having a smaller cerebellum is a disability that makes it hard to keep, balance, focus, and clumsy.
The effect of having a smaller visual cortex is that movements and seeing is more difficult than normal. This all connects the the hemispheric part of the mind which is the biggest part of the mind.“Some things that come from having a small cerebellum is that you can get dyslexia or your motor movements aren’t able to function”(The Cerebellum. Does Size Matter). “Your cerebellum also controls memory, cognition, and you would have a chance of learning disabilities”(The Cerebellum. Does Size Matter). This is where people go in with the Cine Mri to study the cerebrospinal fluid.“This happens when you’re born and a syndrome that I found is Chiari Malformation”(Cerebellar Hypoplasia). The thing that they also do to help these children with this deficit is they use decompression to help, decompression is when they release pressure from your cranium and spinal cord.Even if kids ever get this they will usually have some struggles ahead of them because this controls most of their bodies and how their bodies work. In some ways you can overcome this but in other ways you can’t but if you have faith the sky’s the limit. However, children with a smaller cerebellum usually have a smaller visual cortex and this controls the motor movements in the body.
For example is you have a smaller cerebellum means that we can overcome this by running tests/scanning the brain. “The things the doctors use to look at the brain are mri, ssep scans, cat scans”(Cerebellar Hypoplasia). What these things mean are magnetic resonance imaging, somatosensory evoked potential, computed tomography. “Some other type of brain scans are baer and etv”. Baer mean brainstem auditory evoked response and etv means endoscopic third ventriculostomy. “There is another disease called truncal ataxia and this disease is when you body falls backwards and is caused by a midline cerebellar lesion”(Chiari malformation). The reason why I chose these words is because these words deal with talking about the mind and scanning it which it very helpful if you want to find out if something is wrong. In addition, specific strategies like using technology can be used to study the brain.
Families, teacher, and supporters can help children with a smaller cerebellum by taking it slow you need to look at they are good at you have to be strategic. The research that I have done for my project has changed my perspective so much because I shouldn’t be that person who does not support children with disabilities. I have to be mindful because they deal with everyday and I do not. So when I go to school I try so hard to support them because they have been threw much more than me. People in the world society and in life are or can be disrespectful to people with this disability. Knowing what I now, things I want to do is be mindful about what I say.
Work cited section
1.What is the cerebellum and what does it do?
3.The cerebellum does size matter
5.Living with cerebellar hypoplasia (video)


text:
Showing 2 Holiday Packages
02 Nights Nainital / 01 Night Ranikhet / 02 Nights Corbett
Day 01 : Arrival at Delhi transfer to Nainital, Day 02 : In Nainital, Day 03 : Nainital to Ranikhet, Day 04 : Ranikhet to Corbett, Day 05 : In Corbett and Day 06 : Corbett to Delhi Departure.
Jim Corbett National Park, which is a part of the larger Corbett Tiger Reserve, a Project Tiger Reserve lies in the Nainital district of Uttarakhand. The ever popular wildlife Tourist destination Jim Corbett National Park was established by noted hunter and naturalist Edward James Jim Corbett. If you love watching tigers in their natural habitats then leave the town behind look forward to Corbett in search of majestic tigers and wealth of wildlife species.


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The Hong Kong based Asian Human Rights Commission (AHRC) has urged the Judicial Commission of Pakistan to stop the blame game between different institutions of the state.
“Full confidence must be restored in the judiciary and the matter concerning the son of the Chief Justice must be dealt with in a transparent and unbiased manner.
Published in The Express Tribune, August 11th, 2012.
Comments are moderated and generally will be posted if they are on-topic and not abusive.
For more information, please see our Comments FAQ

domain: law


2014’s Hottest Hardware, Are You Ready For Some Epic Upgrades?
Peter Donnell / 9 years ago
2014 is already proving to be a great year for tech lovers, and with the PC market currently offering a mixture of incredible value and high performance components, there has never been a better time to be a PC enthusiast. Yet for all the high-end hardware on the market that we have available to us today, we can’t help but look forward at what is coming next. There’s always something just around the corner that makes you want to delay your next upgrade, something bigger and better that promises to be a real game changer. Today we thought it would be great to take a quick look at some of the hottest things that could hit the market this year.
Improved Performance And Power Consumption From DDR4 Memory
First up we have DDR4 memory, this one has been a long time coming and while it may not offer huge performance increases over current offerings, it’s improved reliability and lower power usage is certainly a tempting option. With new hardware that supports DDR4 slowly working its way towards the consumer market it’s nice to see that there may be some changes in the memory market later this year, and that’ll hopefully pave the way to big performance increases further down the line.
Cheaper 4K Capable Monitors And Devices
4K gaming is quite a popular topic these days, but 4K isn’t just about gaming, it can be used for movies, content creation or the most popular thing in the internet world,
porn doge memes. The only issue with 4K is that it’s insanely expensive compared to its 1080p and 1440p counterparts. The screens are very expensive, and even recent price cuts to sub $1000 still leave it out of the reach of many. We expect this year will see not only the screens coming down in price, but to also be followed by price cuts in 4K capable graphics cards.
Faster And More Powerful Graphics Hardware
Speaking of new graphics cards, we love the current line up of graphics cards that are on the market. AMD are pushing some gorgeous hardware and Nvidia have some serious powerhouses of their own, but the grass is always greener on the other side and we would be crazy to not be excited about what is coming next from the red and green camps. With Maxwell and Volta from Nvidia and further developments from AMD with their GCN (Graphics Core Next) and APU (Accelerated Processing Unit) hardware, PC gaming is set for some big changes. Getting ultra-high graphics on 4K hardware is far from easy, so more powerful and more efficient hardware, as well as huge changes in memory management will help a lot.
High Performance Next-Generation CPU’s
Going hand in hand with the new graphics cards and memory products we’re looking forward to, we’re even more excited about the next generation of CPU hardware from both Intel and AMD. With Broadwell and Skylake, as well as an even more advanced range of APU’s on the horizon, we can expect support for DDR4, unified memory, faster data transfer speeds and advancements in power management and a whole lot more. Personally I’m looking forward to Skylake more than anything, but for now my Ivy Bridge will do me just fine. If AMD keep boosting the power in their APU’s enough, we may also have a great solution for compact gaming rigs and SteamBox style builds.
Ultra-High Performance And Capacity SSD’s
It’s not often that I can get too excited about storage, SSD’s are great for boosting system performance and they’re endlessly more powerful than their mechanical counterparts. However, their performance seems to be hitting its limits and short of RAID configurations, there isn’t much else that can be done. New memory controllers, as well as other advances should see some nice improvements later this year, but we’re hoping there is another big change ahead that will see massive gains in terms of read and write times, not to mention power consumption and price reductions.
That about wraps it up for our most wanted tech and when combined you can see that we’re hoping for a pretty broad improvement in performance across the whole spectrum of PC components and devices. With 4K working it’s way to becoming common place and prices of UHD displays dropping we’re going to need lightning fast, and large capacity storage devices to handle all the rendering, massive textures and more that come with it, just look at the 48GB needed to install Titanfall and you can see where we are heading! Then you’ll need more powerful graphics cards to enjoy them to their full potential, and the added performance and power management of new CPU’s and memory products certainly couldn’t hurt either.
What tech are you looking forward to later this year? Will you be upgrading now, or are you planning on holding out for the next generation of hardware? Let us know in the comments section below.


HORNADY 308 STEEL MATCH 155 GRAIN
Featuring Hornady Match rifle bullets or HAP (Hornady Action Pistol) handgun bullets, Steel Match ammunition is loaded at the Hornady factory with optimized propellant for each load that provides consistency and the highest levels of accuracy. Utilizing coated steel cartridge cases, and non-corrosive berdan primers, Steel Match ammunition delivers Hornady quality and performance, but with an economical price (saves as much as 40% when compared to brass case match ammo).
Steel Match 40 S&W, 45 Auto and 308 Win loads all meet IDPA/ IPSC/USPSA “Major” power factor performance criteria, but are kept to moderate velocities to deliver only mild recoil for reduced muzzle jump and fast recovery on target. Additionally, Steel Match 9mm and 223 Rem both meet “Minor” power factor criteria.
Whether plinking, practicing, or competing, the economical solution for accurate, consistent performance, is STEEL MATCH ammunition from Hornady.


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Report says Army's operational command failed to protect Polish airspace
A report into a mysterious military object, believed to be a missile, found in a forest near the northern city of Bydgoszcz has concluded that the Polish Armed Forces operational command was negligent in its duties to protect Polish airspace, a Defence Ministry statement has said.
The unidentified object was found in late April and its origins remain a mystery although there are suspicions it could be an unarmed Russian cruise missile that went off course when it was fired to confuse Ukrainian air defences.
According to the Polish Defence Ministry, the missile fell to earth on December 16, 2022. But an investigation was only launched after the missile was found.
"The conclusions are convergent with the results of a review, which has been carried out by the Ministry of National Defence and handed over to the Supreme Audit Office (NIK)," the ministry wrote on Friday evening.
The ministry also said that the report, which had already been handed over to the president and the prime minister, "is a detailed analysis of reports and statements by commanders of the security services who are connected to the discovery of a mystery military object in Zamosc near the northern city of Bydgoszcz."
The ministry wrote that the conclusions are proof of negligence on the part of the operational command of the Polish Armed Forces, who, in accordance with appropriate laws, is responsible for the protection of Poland's airspace.
Mariusz Blaszczak, the Polish defence minister, said on Thursday that the operational command had failed to inform him about an unidentified object entering Polish airspace.

domain: politics


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From Beer to Energy: Researchers Turn Brewery Waste To Fuel Cell Technology
Researchers from Boulder, Colorado just came up with a unique way for turning brewery wastes into low-cost lithium-ion battery electrodes. Beer manufacturers are not allowed to dump these waste products into sewers as they require extra filtration. Now, however, this waste product just got a lot more useful.
These run-offs are quite abundant, too. "Breweries use about seven barrels of water for every barrel of beer produced," said Tyler Huggins, a graduate student in CU Boulder's Department of Civil, Environmental and Architectural Engineering and lead author of the new study.
Beer Wastewater - An Excellent Source Of Battery Production
Currently, converting biological mass such as timber into carbon-based battery electrodes has been used by some energy industry sectors, according to Utility Dive. However, biomass that's naturally-occurring is innately in short supply, making it costly and difficult to effectively take advantage of.
But as the researchers recently found, this abundant byproduct makes for an excellent and freely available material for battery production. Their process is akin to beer-making in that they use a fast-growing fungus called Neurospora crasaa that is found thriving in the wastewater.
By cultivating their feedstock in the brewery waste, it allowed them to better manipulate the fungus' chemical and physical process from the start, explained Science Daily. The end result is one of today's most efficient, naturally-obtained lithium-ion battery, with the researchers simultaneously cleaning the wastewater in the process.
Win-Win Situation For Researchers And Beer Makers
"The wastewater is ideal for our fungus to flourish in, so we are happy to take it," Huggins said. Large scale application of the process may result in breweries significantly decreasing the cost of treating wastewater, while the researchers would gain an abundant amount of cost-effective materials needed in developing battery tech components.
The findings are so promising that Huggins and Justin Whiteley have already filed a patent for their research and started Emergy, a company that aims to monetize this remarkable discovery. They have also partnered with Avery Brewing in Boulder to explore possible pilot programs that would take advantage of the technology.
"We see large potential for scaling because there's nothing required in this process that isn't already available," said Huggins. Considering the creativity and resourcefulness that was exercised, it certainly is a rare and beautiful moment when beer and science work hand in hand.
Who Is Going To Win Artificial Intelligence Race?
While major tech companies are in a race to develop real-life artificial intelligence applications, self-aware AI might come sooner than you think.
You Can Choose To Replace Your Note 7 With Upcoming Huawei Mate 9
Huawei will launch next month its Mate 9 line of flagship smartphones composed of two models.
Can Crowdsourcing Lead To Better Smartphone Concepts Design?
Initiatives such as ZTE's crowdsourcing smartphone design may lead to better products.
A Pint Of Beer A Day Can Increase The Risk For Your Heart
With the number of beer lovers all around the world, could your love for beer pave the way for your health problem in the future? What’s the truth behind claims that consuming a pint of beer a day can actually increase your risk of developing heart problems? Find out what health professionals have to say
Nuclear Fusion Core Foundation of Clean Energy
With the earth's natural resources in the brink of running out, scientists found a new hope that can provide a new energy source for the coming generations.
MORE IN ITECHPOST
4 Reasons To Secure Your PDF Files
The PDF format is one of the most commonly used document types for sharing files. It can be opened by a variety of programs and maintains formatting between devices and programs. Additionally, PDFs are less likely to be edited by any unwanted parties.
How Do Personal Emergency Response Systems Work?
Personal emergency response systems, known as PERS for short, are systems that help people to raise the alarm and get immediate help when a medical or personal emergency occurs. They are ideal for older people and anyone with a mobility issue or an injury or illness that can cause falls.
The 9 Most Popular (And Fun) Internet Games of 2019
Are you bored of board games? What are the most popular internet games from this year? These nine are popular for a reason. Check 'em out!

domain: news


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domain: food


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May We Recommend: Parchment Paper
Rachel Ringler bakes her Shabbat challah on parchment paper
Parchment paper is a relatively recent addition to my kitchen supplies. Six years ago, I learned about it from a friend who is a professional cook. But now that I have found it, I can’t imagine my kitchen without it.
Why, I wondered, was I so late to the parchment game?
According to Susan Reid, food editor of King Arthur Flour’s magazine, Sift, “People who worked in restaurants knew about parchment paper forever. Once the amateur cook began looking behind the swinging [restaurant kitchen] doors, they wanted what the professionals had." And the professionals all used parchment paper.
King Arthur Flour, said Reid, began selling parchment paper to the consumer in 1991. It is now, she said, one of the top ten most popular items that this Vermont based purveyor of all things baking-related sells.
What’s to love? First of all, parchment paper is coated in silicone making it both non-stick and heat resistant. Cookies slide off the paper-lined pan, breads do, too. And you can turn roasting vegetables with ease when the sheet pan is lined with the stuff.
You don’t have to throw the paper away after one use: good, heavy parchment paper can be re-used for up to a dozen times and, once you have finished your roasting and baking, clean up is as easy as crumpling the paper and tossing it in the trash.
The first parchment paper I purchased came in rolls and I grew frustrated with the way the paper would spool up on itself and with the fact that the paper never fit exactly within the parameters of my pans.
I now use pre-cut sheets of the paper because they are designed to nestle perfectly within the confines of a flat, metal sheet pan. You can buy flat parchment paper in a wide selection of shapes and sizes – half-sheet, quarter-sheet, round papers to fit into round baking pans.
In addition, you have the choice of the white parchment paper or the unbleached variety. Unbleached parchment paper is free of chlorine, the chemical that turns the paper white. Both types of paper offer the same properties of being non-stick and heat resistant.
Rachel Ringler bakes cookies on parchment paper
While most uses for parchment paper involve lining surfaces, to keep baked and roasted foods from sticking to the cooking surface and to make clean-up easier, don’t stop there. Parchment can also be used as a cooking tool. Fish, roasted “en papillote” is cooked inside a parchment paper packet, sealing in flavor and juices. Cookbook writer Julia Turshen has a to-die-for brisket recipe that calls for scrunching up moistened parchment paper, covering the brisket with it, and cooking away. You can even cut it into triangles and fold it up to use for piping chocolate on cakes. And when presenting home-baked breads as gifts, you can wrap them in parchment paper.
You can find the paper in your local supermarket or on-line. When you try it, you will feel like a professional chef while making your cooking life easier, too.


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Sen. Joe Manchin (D-W.Va.) lied about the Tax Cuts and Jobs Act while speaking in Davos, Switzerland on Monday. Manchin tried to claim the Trump tax cuts were “regressive.”
Manchin is wrong, and in pushing tax increases he is violating his tax pledge to his constituents.
Here are the facts, according to the latest official IRS data.
26% tax cut for West Virginians making between $25k – $50k. Thanks to the Tax Cuts & Jobs Act — also known as the Trump tax cuts — West Virginia households with adjusted gross income between $25,000 and $50,000 saw their average federal income tax liability drop from $2,190 in 2017 to $1,783 in 2019, a 26% reduction in federal income tax liability.
24.8% tax cut for West Virginians making between $50k – $75k. West Virginia households with adjusted gross income between $50,000 and $75,000 saw their average federal income tax liability drop from $5,348 in 2017 to $4,268 in 2019, a 24.8% reduction in federal income tax liability.
23% tax cut for West Virginians making between $75k – $100k. West Virginia households with adjusted gross income between $75,000 and $100,000 saw their average federal income tax liability drop from $8,720 in 2017 to $7,064 in 2019, a 23% reduction in federal income tax liability.
Just a 2.9% tax cut for West Virginians making over $1 million. Contrary to Manchin’s dishonest assertions, middle income West Virginians saw a significantly greater tax cut than those earning over $1 million. West Virginia households earning over $1 million saw their federal income tax liability drop just 2.9%.
Official data from the Congressional Budget Office also shows that high-earning Americans pay a greater share of taxes than before enactment of the Trump tax cuts.
In other words, the Tax Cuts and Jobs Act actually made the tax code more progressive, though you won’t hear Manchin admit it.
18,320 WV households are no longer stuck paying the Obamacare mandate tax. The TCJA zeroed out the Obamacare individual mandate tax penalty effective 2019. In 2017, 22,960 West Virginia households paid the Obamacare individual mandate tax penalty. 20,940 (91%) households earned less than $75,000. 18,320 households paid the Obamacare individual mandate tax penalty in 2018. 16,180 households (88%) earned less than $75,000.
186,230 WV households are benefiting from TCJA’s expanded child tax credit. The TCJA doubled the child tax credit from $1,000 to $2,000. 187,660 WV households took the child tax credit in 2019 including 180,710 households earning less than $200,000. 186,230 households took the child tax credit in 2018 including 179,600 households earning less than $200,000.
739,000 WV households are benefiting from TCJA’s doubled standard deduction. The TCJA doubled the standard deduction from $12,000 to $24,000 for taxpayers filing jointly and $6,000 to $12,000 for single filers. 723,830 WV households took the standard deduction in 2018 including 712,380 households earning less than $200,000. 739,300 households took the standard deduction in 2019 including 722,600 households earning less than $200,000.
20% tax deduction for WV small businesses. The TCJA created a new, 20% deduction for small businesses organized as passthrough entities (LLCs, sole proprietors, S-corporations, partnerships). 71,030 WV taxpayers claimed the small business deduction in 2019 including 61,630 taxpayers earning less than $200,000. 62,510 taxpayers claimed the small business deduction in 2018 including 54,390 taxpayers earning less than $200,000.
Utility Bill Savings: If not for TCJA, utility bills would be even higher today. Utility companies pay the corporate income tax and must build the tax burden into the billing rates. Higher tax rate = higher bills. Lower tax rate = lower bills. See WV citations in the list below. A national compilation can be found here and local media coverage here.
Manchin wants to raise the combined federal-state corporate income tax to a rate much higher than the Europe average of 19%. Such rate increases will make the U.S. less competitive and will further increase utility bills for WV households.
West Virginia businesses cite Trump tax cuts as a driver of new job creation, business expansion and pay increases:
Eagle Manufacturing (Wellsburg, West Virginia) – Creating new jobs and purchasing new equipment:
The company traditionally has sought to buy used equipment at low prices when replacement of machinery was necessary, according to Trimmer.
“Because of that tax break, we can invest and buy our next machines,” he said. “My department is now looking to buy a $2 million machine, rather than a 1990 classic. It will pay for itself in a short manner of time.”
Trimmer said the company is about to sign some government contracts that will put Eagle cabinets and metal products on the next generation of U.S. naval destroyers.
This could result in as many as 25 jobs being created, he said. – May 3, 2018, The Intelligencer article excerpt
Doss Enterprises (Jane Lew, West Virginia) – Job creation, pay raises and equipment purchases:
The benefits of the Tax Cuts and Jobs Act are evident just from looking around the Doss facility, Capito said.
“When I saw their board of (new) hires, there must have been 20 or 30 names on there, just April and May,” she said.
The company has chosen to reinvest its tax savings in its employees and into new equipment, Capito said.
“All of the employees are seeing (it) in their paychecks,” she said. “(Doss) has also bought some new equipment with his money, which is a great investment. He’s going to be hiring at least another 30 people.” – May 2, 2018, WVNews.com article excerpt
Warwood Tool (Wheeling, West Virginia) – New product line:
The tax cuts have given the company and its customers enough optimism for the future that Warwood Tool is developing a new line of products. – May 3, 2018 The Intelligencer article excerpt
Panhandle Cleaning and Restoration (Wheeling, West Virginia) – New trucks and equipment:
While Panhandle is often called upon for clean-up services after national disasters, Contraguerro told Jenkins the company relies on its day-to-day operations. Their employees, trucks and equipment are called to local homes and businesses each day following water-line breaks or other incidents, and Panhandle doesn’t wait for hurricanes to hit, he said.
The company plans to invest the money it receives from the tax cuts into more trucks and equipment.
“We will be reinvesting in the business,” he told Jenkins. “There is a huge benefit to reinvesting.” — May 3, 2018 The Intelligencer article excerpt
Davis Trust Company (Elkins, West Virginia) – 3% across the board pay raises (on top of existing compensation structure.)
Potomac Edison (Martinsburg, West Virginia) – The utility is passing on tax savings to customers:
‘More than 85,000 Potomac Edison customers in the Eastern Panhandle should see lower bills in the coming weeks thanks to federal tax reforms adopted in December.
The West Virginia Public Service Commission announced Friday that it approved rate reduction settlements for utility companies totaling almost $85 million annually, starting next month. – August 24, 2018 Herald Mail Media
West Virginia American Water (Charleston, West Virginia) – The utility is passing along tax savings on to customers:
West Virginia American Water Company announced a settlement plan last week which — if approved by the PSC — would result in an average savings of $3.77 a month for water and sewer customers in the state.
“The recent federal tax reform will save our customers an estimated $4.6 million annually, so we are passing these savings on to our customers beginning next month,” Brian Bruce, president of West Virginia American Water. – August 21, 2018 Bluefield Daily Telegraph excerpt
Appalachian Power Co. (Milton, West Virginia) – the utility is passing along tax reform savings to customers.
Appalachian Power Company saved $235 million dollars from the federal tax cuts and the company is proposing passing the money back to its customers in a variety of ways.
The multi-pronged proposal is in a filing with the state Public Service Commission due Wednesday. The PSC is requiring all utilities to tell it their tax cut savings and what they plan to do with it.
West Virginia Consumer Advocate Jackie Roberts told MetroNews the money clearly belongs to the customers.
“They (the utilities) had taxes in their rates and now the taxes in their rates have significantly decreased—so they shouldn’t be able to keep collecting and keeping those higher taxes in their rates,” Roberts said.
Appalachian Power Company Communications Director Jeri Matheney agrees–the $235 million Appalachian Power will save belongs to its customers.
“It is customer money. What we propose to do is provide a method to keep rates as stable as possible over the long term and as much as possible eliminate the need for rate increases,” Matheney said.
The Appalachian Power distribution proposal for West Virginia customers includes:
–$131 million to completely offset the company’s fuel and vegetation control program funding request that was part of an April filing with the PSC
–$19 million reduction in the company’s base rate case filed earlier this month (taking the $115 million request down to $96 million)
–$51 million to reduce next year’s fuel recovery cost rate case
–$1 million for a pilot economic development grant program
–$30.1 million to return to customers over the next three years – May 30, 2018, MetroNews article excerpt


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HipHopDX – This week in Hip Hop, Mase dropped a diss record titled “The Oracle” aimed at Cam’ron. Also, 21 Savage penned an Instagram note berating the “OG rappers” who don’t appreciate the artistry of the younger generation and Eminem Revival’s gets another rumored release date.
Mase Drops “The Oracle” Dissing Cam’ron
Mase fired back at Cam’ron with “The Oracle” over JAY-Z’s “The Blueprint 2” instrumental as a response to Cam name-dropping him on his new mixtape The Program Friday (November 24).
Mason Betha accuses Cam of things like snitching, holding Juelz Santana in a bad deal and incest. Just to make things ultra-clear, Mase annotated the lyrics with Genius.
Rap Twitter went nuts and Cam’ron’s online response was short and sweet: “Good morning to u to betha…lol, Let’s play nigga..,” complete with a kissy face emoji. He previewed his response to Mase’s diss on Friday (November 24).
Read more about Mase and Cam’ron’s beef and how it all began here.
21 Savage Calls Out OG Rappers
There seems to be a strange cold war of sorts between the older generation of Hip Hop artists and the younger generation. 21 Savage has something to say to that and shared his thoughts with an Instagram post, questioning if rapping about drug dealing was any better than rapping about doing drugs.
“Why do so many of these ‘Og Rappers’ judge the new generation?,” Savage wrote. “They say we make drug user music like making drug selling music is better what’s the difference? What about the fact that rap is the number one genre of music right now none of y’all acknowledge that. Artists been snorting cocaine and smoking crack since the 70s and 80s did y’all forget that? Instead of judging how about y’all make a change.”
As the old adage goes, “People who live in glass houses shouldn’t throw stones” and T.I. seems to agree with 21’s note to the OGs.
When Will Eminem Release Revival?
Eminem has his fans anxiously waiting for his ninth studio album Revival. They’ve been let down a few times and now there seems to be more possible dates for Em’s upcoming release.
According to HITS Daily Double, Revival will be released on December 8. According to one savvy Reddit user who apparently works at Target, the physical copies for Revival will be available on December 15.
Read more about the release of Eminem’s Revival here.
#DXCLUSIVES: 2017 Turkey Awards, CyHi The Prynce, Flavor Flav, Krayzie Bone & Teddy Walton
The 2017 HipHopDX Turkey Awards
HipHopDX’s annual Turkey Awards roasted a few of this year’s craziest moments, including XXXTENTACION’s drama-fueled social media accounts , the hyped-up fight that never happened between Chris Brown and Soulja Boy, R. Kelly’s alleged sex cult and so much more.
CyHi The Prynce Explains Split With G.O.O.D. Music
CyHi The Prynce explained to DX why he split with G.O.O.D. Music on #DXLive as well as details on his musical relationship with Kanye West. He says its nothing but love between the two.
“Different songs like ‘I Am A God’ and ‘New Slaves’ and ‘Father Stretch [My Hands]’ — those were songs I was really a part of and it kind of like molded the community to understand when I come out it wouldn’t be so off of what’s the norm of music,” he says. “I think that’s something I got to do that’s dope is instill a few of my ideologies through other people’s music, so now when I come out with mine, it’s a place where it can live versus me just being out there stranded by myself.”
Flavor Flav On Chuck D: “We Can Iron Out These Wrinkles”
Flavor Flav also spoke with DX on his relationship with Public Enemy comrade Chuck D. Flav says like most family members, they have fights, but always come back together one way or another.
“But as years go on, all families have conflict, but usually they end up working it out, and moving on,” he told DX’s Kyle Eustice. “So that’s the situation with me and my partner Chuck D, you know? It’s just a few wrinkles in the situation, but we can iron out these wrinkles and we can keep it moving. That’s all.”
Krayzie Bone Gives JAY-Z “Real MC” Honor For Rocking Over “Thuggish Ruggish Bone”
JAY-Z spit a freestyle over Bone Thug-n-Harmony’s 1994 hit “Thuggish Ruggish Bone” during his set on his current 4:44 Tour and Krayzie Bone appreciated the love. Speaking with DX about the moment, Krayzie crowned Jigga with a “Real MC” honor for the move.
“I looked at it as him doing what real MCs do, [which is] pay homage,” Krayzie said to DX. “At the same time, he knows that we’re one of the biggest acts to ever come out of Cleveland and that resonates with a lot of artists that come thru Cleveland. They always mention, shout out, or do something in reference to Bone. It’s definitely flattering and cool to see somebody of his stature acknowledge that Bone Thugs-n-Harmony did in fact exist and was a great contribution to Hip Hop since the industry seems to forget that a lot.”
#DXHitList: The Teddy Walton Session
Fresh hitmaker Teddy Walton curated a special Spotify playlist for HipHopDX with some of his favorite tunes and detailed his come-up from Memphis to Hollywood as well as his upcoming EP, Mental Health.
“The reason I named it Mental Health is because when I go home some people I know have it,” Walton shared. “I don’t feel like I have it, but I’ve talked to people without knowing that they have a mental illness. My mom works in that field, that’s what she wakes up every day to do. I got mad one day my mom gave away some of my Forces.
“But one day, I was just chilling and Lil Yachty hit me like, ‘Hey, I got your verse just send me some beats,'” he continued. “Yachty is a huge influence on the youth. If I get Yachty on my project, I’m definitely naming it something that means something. It’s going to sound crazy but it’s the fact that you’re going to learn something from it. Dom [Kennedy], Lil Yachty, Juicy J. It’s going to be crazy.”

domain: celebrity


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High-profile self-defense trials elsewhere in the country, notably Rittenhouse's, have prompted some introspection in the legal community, with some advocating abolishing, or at least modifying, the state's self-defense regime.
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The Denver school board member is suing Black Lives Matter 5280 and others for $1 million for their roles in publishing the allegations.
Deborah Ramirez’s associates are standing with the intensely private Boulder woman who launched herself into the national spotlight Sunday with allegations of sexual misconduct by U.S. Supreme Court nominee Brett Kavanaugh.
Just as the nation’s attention to underserved communities and sexual assault victims has increased, the U.S. House of Representatives has — acting purely along partisan lines — taken steps that have the potential to significantly harm these groups. The House is currently considering a bill, inaptly named the “PROSPER Act,” that would drastically limit the ability of students with financial need to pay for higher education and weaken protections for sexual assault victims.
U.S. Rep. Diana DeGette, D-Colo., is one of nearly 145 members of Congress calling for an investigation into allegations of sexual misconduct against President Trump.
As Alabama voters prepared to head to the polls Tuesday, U.S. Rep. Mike Coffman, a Colorado Republican, denounced Roy Moore, the state’s Republican U.S. Senate nominee, as “morally bankrupt” and called a Moore win “the absolute last thing Washington needs.”
“His words are weightless to me. You cannot bargain or barter the truth.” It has only been a few months since I last wrote about a sexual assault case from Boulder, but yet another tragedy is under the spotlight. The quote above is taken from a letter the victim wrote and read aloud in court, but not in a trial. Instead, he agreed to a plea bargain offered by the Boulder District Attorney’s office in exchange for a more lenient sentencing recommendation.
The Sixth Amendment to the U.S. Constitution guarantees “an impartial jury” to all defendants. But what seems to be a clear and concise rule becomes much less so in practice. Most dramatically, whatever is said among deliberating jurors cannot be used to prove bias. No matter how far from impartial such statements may remove a jury. That is the case of Miguel Angel Pena-Rodriguez recently heard by the U.S. Supreme Court. Charged with sexual assault against two teenage girls, the jury convicted him 12-0. The unanimous opinion of his peers found him guilty, just as the system is supposed to work.


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It is therefore inevitable that some people will be annoying; indeed, to expect otherwise, Marcus says, is like expecting a fig tree not to yield its juice. Thus, if we find ourselves shocked or surprised that a boor behaves boorishly, we have only ourselves to blame: We should have known better. Page 137
I just recently finished reading A Guide to the Good Life: The Ancient Art of Stoic Joy and no recent book has resonated quite as strongly with me. I enjoyed so many parts of this book that I didn't want to write a traditional review but instead wanted to share direct quotes that will explain much better than I what Stoicism is and is not.

domain: literature


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The opinion of the court was delivered by: Gregory G. Hollows United States Magistrate Judge
LAW OFFICES OF SCOTT L. TEDMON SCOTT L. TEDMON, CA BAR NO. 96171 A Professional Corporation 1050 Fulton Avenue, Suite 218 Sacramento, California 95825 Telephone: (916) 482-4545 Facsimile: (916) 482-4550 Email: firstname.lastname@example.org Attorney for Defendant CHARLES HEAD
APPICATION FOR TRANSPORTATION
ORDER FOR TRANSPORTATION
PURSUANT TO 18 U.S.C. 4285
In this matter, defendant Charles Head has a status conference hearing on December 8, 2011 at 10:00 a.m. concerning the disposition of Mr. Head's motions to dismiss based on a violation of the Speedy Trial Act. Mr. Head is indigent and his financial statement is included herewith as Attachment 1. Application
Application is hereby made for an Order for Transportation and subsistence for defendant Charles Head to enable him to attend his hearing in the above-referenced matters scheduled for December 8, 2011 at 10:00 a.m. before the Honorable Kimberly J. Mueller. While this Application for Transportation Order is only being filed in case number Cr. S-08-093-KJM, and only one Order from the Court is needed to provide the necessary transportation for Mr. Head, this Application for Transportation is equally applicable to case number Cr. S-08-116-KJM as the status conference hearing regarding Head's motion to dismiss for violation of the Speedy Trial Act in that case will also be heard on December 8, 2011.
Mr. Head has advised counsel that he is unable to afford travel and is requesting that transportation be arranged for his one-way travel from Pittsburgh, Pennsylvania to Sacramento, California on December 7, 2011 so that he may confer with counsel after his arrival and before the status conference hearing. Mr. Head requests that transportation be arranged for his one-way travel after the hearing from Sacramento, California to Pittsburgh, Pennsylvania on December 8, 2011.
Wherefore, in the interests of justice, the Court is requested to arrange for Charles Head's means of non-custodial transportation or furnish the fare for such transportation to the place where his appearance is required; to direct the United States Marshal to furnish Charles Head with an amount of money for subsistence expenses, not to exceed the amount authorized as a per diem allowance for travel under Section 5702(a) of Title 5, United States Code, and pursuant to 18 U.S.C. Section 4285.
Defendant seeks round trip air fare and subsistence to travel from Pittsburgh, PA to Sacramento, CA. so that he can attend a status conference or possible change of plea. He makes the request pursuant to 18 U.S.C. § 4285. No matter how incongruous the words of the statute may seem to defendants in criminal actions, the statute permits disbursement of only one-way funds for appearances in the district of prosecution. "[The court may]...direct the United States Marshal to arrange for that person's non-custodial transportation or furnish the fare for such transportation to the place where his appearance is required..." The undersigned is unaware of any case interpretation of § 4285 which would permit defendant's round trip request. See cases only permitting payment for one-way travel: United States v. Centeno, 2009 WL 3334144 (S.D. Cal. 2009); United States v. Miracle, 2009 WL 3211104 (E.D. Ky. 2009); United States v. Headden, 2009 WL 2960382 (D. Ariz. 2009); United States v. Birdhorse, 2007 WL 2358634 *2 (D.N.D. 2007); United States v. Vaughn, 2002 WL 1067456 *1 (D. Kan. 2002); United States v. Sandoval, 812 F.Supp. 1156, 1157 (D. Kan. 1993); United States v. James, 762 F.Supp. 1, 2 (D.D.C. 1991; United States v. Gonzales, 684 F.Supp. 838, 841 (D. Vt. 1988).
While defendant may be seeking a judicial wink at the statute for logical reasons, the court is not authorized to expend funds except as directed by statute.
Therefore, the Marshal is authorized and directed to furnish the above named defendant, Charles Head, with transportation and subsistence expenses for travel from Pittsburgh, PA to Sacramento, California on December 7, 2011 to timely make his appearance on December 8, 2011 at 10:00 a.m. The United States Marshal's Office must furnish Charles Head with money for subsistence expenses to his destination, not to exceed the amount authorized as a per diem allowance for travel under section 5702(a) of Title 5, United States Code.
The Clerk shall serve a copy of this order on the United States Marshal.


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NBA Commish: 'I will talk to Congress and push for legislation' for sports betting
Thu, Dec 4, 2014
NBA Commissioner Adam Silver is saying promising things about legalizing and regulating sports betting again, this time while on a trip to India to promote the NBA there with Sacramento Kings owner Vivek Ranadive.
“I will talk to Congress, and will push for a legislation that will allow our states to decide whether it is something they want to participate in. In the US, all the states have lotteries. I’m not trying to encourage sports betting, I’m just saying there is a lot of sports betting, and if it’s going to happen, it must be done in a regulated environment,” Silver said, according to a report in the Indian Express.
It’s just the latest vote of confidence from Silver in the fight to legalize sports betting and we’re now hearing him say he’ll be actively lobbying Congress on the issue. It was a tad surprising considering Silver and Ranadive are there to celebrate the Jr. NBA program in India, which has reached more than 750,000 youth in the country.
But we’ll take it wherever we can get it and it’s nice to hear Silver’s support sounding stronger than ever. We can also add another owner to the mix who is speaking out in support of legalization.
“A part of what gets fans involved in games is to let them bet on matches. So this is a logical evolution of the sport,” said Ranadive.
Mark Cuban has also spoken out in supporting the effort to legalize sports betting, saying "I think we're the world's biggest hypocrites” when the leagues benefit from sports betting while at the same time denigrating it.
The latest comments from Silver go even a step further and now suggest the NBA not only fully supports legalization, but it intends to lead the charge.
This is the kind of break sports bettors have been waiting decades for. With New Jersey’s efforts stalled at the court level, it’s looking more and more like changing the federal law will be the only avenue to regulating and legalizing sports betting in America. Now that a major pro sports league is stepping up, that actually seems possible.


Click here to list ALL Makes
Maserati Shamal, 1994. For Sale (1991)The Maserati Shamal was another iconic design out of the hands of Marcello Gandini, the famous designer who was also responsible for the Lamborghini Miura, the Lancia Stratos and the Bugatti EB110.
Maserati only made a total production of 369 cars. The cars were fitted with a 3.2 litre twin-turbocharged V8 DOHC-32 Valve engine with 326 BHP. That made the Shamal capable of doing 270 Km/h.
This black Shamal was first road-registered in 1994 and today it has a total mileage on the clock of 33.000 KM's from new. It has always been serviced according to the instructions and kept in excellent order. Comes with EEC title and MOT.


Rescue Effort to Get 5 People Out of Gold Canyon 'too dangerous'
5 people who did not heed mandatory evacuation orders to leave their homes in the Gold Canyon area in the path of the Station Fire are now "trapped in a canyon and it's too dangerous to rescue them," the Los Angeles County Sheriff's Department has announced, according to cbs2.com. The four men and one woman called for help today, after having elected against orders to remain in the area. Plans to send in a large sheriff's rescue helicopter have been abandoned, as "the flames are too intense and authorities must wait for the fire to pass by before going in."
During his press conference yesterday morning, Gov. Arnold Schwarzenegger took an opportunity to remind those who find themselves in the path of this fast-moving blaze to do as they are told and leave their homes upon orders.