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The ligand for the receptor tyrosine kinase fms-like tyrosine kinase 3 (flt3), also referred to as fetal liver kinase-2 (flk-2), has an important role in hematopoiesis. The flt3 ligand (flt3L) is a growth factor for hematopoietic progenitors and induces hematopoietic progenitor and stem cell mobilization in vivo. In addition, when mice are treated with flt3L immature B cells, natural killer (NK) cells and dendritic cells (DC) are expanded in vivo. To further elucidate the role of flt3L in hematopoiesis, mice lacking flt3L (flt3L−/−) were generated by targeted gene disruption. Leukocyte cellularity was reduced in the bone marrow, peripheral blood, lymph nodes (LN), and spleen. Thymic cellularity, blood hematocrit, and platelet numbers were not affected. Significantly reduced numbers of myeloid and B-lymphoid progenitors were noted in the BM of flt3L−/− mice. In addition a marked deficiency of NK cells in the spleen was noted. DC numbers were also reduced in the spleen, LN, and thymus. Both myeloid-related (CD11c++ CD8−) and lymphoid-related (CD11c++ CD8+) DC numbers were affected. We conclude that flt3L has an important role in the expansion of early hematopoietic progenitors and in the generation of mature peripheral leukocytes.
The continual production of blood cells during an individual's lifetime is mediated by the coordinated effects of a number of cytokines acting on hematopoietic stem and progenitor cells in the bone marrow (BM).1,2 The fms-like tyrosine kinase 3 ligand (flt3L) is 1 of the cytokines that affects the development of multiple hematopoietic lineages.3 These effects are mediated through the binding of flt3L to the flt3 receptor, also referred to as fetal liver kinase-2 (flk-2)4 and STK-1,5 a receptor tyrosine kinase that is expressed on hematopoietic stem and progenitor cells.4 Flt3L is expressed as a membrane-bound protein on the surface of cells and can be proteolytically cleaved to generate a soluble protein. Both the membrane-bound and soluble forms are biologically active.6Although a weak growth stimulator as a single factor in vitro, flt3L synergizes with numerous hematopoietic growth factors to stimulate myeloid progenitors to proliferate and differentiate.7-11Flt3L synergizes with interleukin (IL)-7, c-kit ligand (KL), IL-3, and IL-11 to stimulate B-cell lymphopoiesis in vitro.12-15 Flt3L augments T-cell development from BM-derived precursors cultured in the presence of thymic stroma plus IL-1216 or BM stroma,17 and increases primitive thymus-derived precursor expansion induced in vitro with IL-3 plus IL-6 plus IL-7.18 In this respect, flt3L shares many functional properties with KL, augmenting the response of multipotent and lineage-committed progenitor cells to a variety of cytokines. In addition to the results obtained from in vitro experiments, studies of hematopoietic disorders in humans support the concept that flt3L has an important role in hematopoiesis. Healthy individuals have low levels of circulating flt3L in their plasma.19 However, in the case of hematopoietic disorders that affect the stem cell compartment (pancytopenias), specifically Fanconi anemia and acquired aplastic anemia, soluble flt3L levels are highly elevated. Flt3L may be produced in an effort to compensate for the deficiency in stem cells in these anemias.19,20
When soluble flt3L is administered to mice, hematopoietic progenitors in the BM and spleen are expanded, and potent mobilization of stem and progenitor cells into the peripheral blood (PB) occurs.21The mobilization includes cells with long-term, multilineage reconstitution potential, demonstrating that pluripotent stem cells are mobilized.22 In addition to the effect on stem and progenitor cells of the myeloid lineage, expansion of immature B cells is noted in the BM and spleen.21 Splenomegaly and lymphadenopathy result, and examination of these tissues indicates that administration of flt3L induces a large increase in the number of dendritic cells (DC) in the spleen and lymph nodes (LN) as well as in the PB, lungs, liver, Peyer patches, and thymus.23-25 The flt3L-expanded DC can process and present antigen to naive T lymphocytes in vitro and in vivo.23 Administration of flt3L to tumor-bearing mice results in the generation of specific antitumor T-cell responses, revealing a potent antitumor activity of the protein.26-28 The role of the expanded DC in the generation of this antitumor response is currently under investigation.
Mice lacking the receptor for flt3L (flk-2−/−) exhibit hematologic defects.29 Flk-2−/− mice have reduced numbers of B-cell precursors (pro-B cells) in the BM, though normal numbers of functional B cells are present in the periphery. Transplantation experiments have revealed a defect at the multipotent stem cell level. In competitive repopulation experiments, stem cells from flk-2−/− mice transplanted to irradiated recipients do not effectively reconstitute the hematopoietic system, especially the T-lymphoid lineage.29 The interaction of KL with its receptor c-kit is important for regulation of early events in hematopoiesis. The complete lack of KL or c-kit expression in mice is lethal, but some nonlethal mutations in this cytokine/receptor combination have been described.30 These mice (eg, W/Wv) have severe defects affecting hematopoietic progenitors and erythropoiesis, in addition to nonhematopoietic defects. Mice homozygous for mutations in both c-kit and flk-2 (W/Wv flk-2−/− mice) display reduced hematopoiesis and a life expectancy of 6 weeks.29 It was concluded that flt3L and KL are both required for normal hematopoiesis.
In an effort to further elucidate the role of flt3L in hematopoiesis, we have generated mice lacking flt3L and have found that flt3L−/− mice have a more profound hematologic phenotype than that noted for flk-2−/− mice. Flt3L−/− mice have significantly reduced cellularity in a number of hematopoietic tissues, reduced numbers of myeloid and lymphoid progenitors in the BM, and reduced numbers of DC and NK cells in the lymphoid organs.
Materials and methods
Targeting of the flt3L gene by homologous recombination and generation of flt3L-deficient mice
Genomic clones encoding murine flt3L were isolated from a C57BL/6 genomic library (Stratagene, La Jolla, CA) and mapped by a combination of polymerase chain reaction (PCR) and restriction analyses. A targeting vector was constructed by replacing a 2.8-kb NotI-SpeI fragment encoding exons 1 to 5 (amino acids 1-164) with a PGK-neo cassette. A thymidine kinase cassette (MC-TK) was inserted into the 3′ end of the vector. C57BL/6-derived embryonic stem cells were electroporated with the flt3L targeting construct and selected as described previously.31 Approximately 1/80 G418 and ganciclovir-resistant clones carried an flt3L allele disrupted by homologous recombination as determined by both PCR and genomic Southern blot analyses.
Flt3L-targeted embryonic stem cell clones were injected into day 3.5 BALB/c blastocysts and transferred to day 2.5 pseudopregnant Swiss Webster recipients. Resulting male chimeras were bred to C57BL/6 females and offspring were analyzed for germline transmission of the mutant alleles by PCR and genomic Southern blot analyses. C57BL/6 mice heterozygous for the flt3L mutation (flt3L +/−) were intercrossed to yield mice homozygous for the mutation (flt3L−/−). The lack of flt3L expression in flt3L−/− mice was confirmed by PCR analysis of first strand complementary DNA (cDNA) synthesized according to manufacturer specifications (Amersham Pharmacia Biotech, Piscataway NJ) from spleen and BM total RNA and amplified using the primers 5′-CACCTGACTGTTACTTCAGCC and 5′-CCTGGGCCGAGGCTCTGG, spanning the complete extracellular domain (481 nucleotides) of mature flt3L.6 Amplified products were analyzed by Southern blot analysis using a radiolabeled flt3L cDNA probe. Mouse β-actin reverse transcriptase PCR (RT-PCR) analysis was performed as a control for uniform RNA integrity and first strand synthesis using the primers 5′-AGGTAGTCCGTCAGGTCC and 5′-CGGAGTCCATCACAATGC.
Flt3L−/− mice, maintained on a C57BL/6 background, were bred at Immunex Corporation (Seattle, WA). C57BL/6 mice were purchased from Taconic Farms Inc (Germantown, NY), housed for a minimum of 1 week, and used as age- and sex-matched controls. DBA/2 mice were purchased from Jackson Laboratory (Bar Harbor, ME) and Swiss Webster mice were purchased from Taconic Farms. Data are presented from mice aged between 5 and 14 weeks. Mice were housed under specific pathogen-free conditions.
Peripheral blood was harvested by cardiac puncture and collected into heparinized tubes (Venoject, Fischer Scientific, Pittsburgh, PA). Hematocrits were performed as previously described.21 White blood cell (WBC) counts, blood differentials, and platelet counts were analyzed using a Hemavet Multispecies Hematology Analyzer (CDC Technologies, Oxford, CT). BM cell suspensions were prepared by flushing cold phosphate-buffered saline (PBS) containing 5% fetal bovine serum (FBS) (Intergen, Purchase, NY) through isolated femurs. Cell suspensions were prepared from spleens, LN (2 inguinal plus 2 brachial plus 2 axillary), and thymi by gently breaking up the tissues between frosted glass slides.
In vitro hematopoietic progenitor assays
Colony-forming units–granulocyte-macrophage (CFU-GM), burst-forming units–erythroid (BFU-E), and B-cell colony-forming units (B-CFU) lymphoid assays were performed by plating BM cell suspensions in methylcellulose (Stem Cell Technologies, Vancouver, BC, Canada) supplemented with 30% FBS (Intergen) and 50 U/mL penicillin, 50 μg/mL streptomycin, and 2 mmol/L glutamine (JRH Biosciences, Lenexa, KS). Methylcellulose for CFU-GM and BFU-E assays was supplemented with 100 ng/mL recombinant murine (rmu) IL-3, 200 ng/mL rmu KL, plus 2 U/mL recombinant human (rhu) erythropoietin. CFU-GM and BFU-E were scored in the same assay. Methylcellulose for CFU-B lymphoid assays was supplemented with combinations of 50 ng/mL rhu IL-7, 100 ng/mL rhu flt3L, and 200 ng/mL rmu KL. All cytokines were produced and purified at Immunex Corporation except erythropoietin (R & D Systems, Minneapolis, MN). Cells were plated in quadruplicate wells in 24-well plates (Costar, Cambridge, MA) at 0.5 mL/well. Plates were incubated at 37°C, 6.5% CO2. B-lymphoid colonies and myeloid colonies containing more than 50 cells were scored after 7 and 9 days, respectively.
Colony-forming units-spleen13 (CFU-S13) assays
The BM cells were isolated from the femurs of 4 to 5 mice per experiment and pooled. Adult female C57BL/6 mice (9-20 weeks old) served as recipients and were lethally irradiated (1000 rads) using a137Cs source (Mark 1 irradiator, J. J. Shephard and Associates, Glendale, CA) at a rate of 89 to 115 rads/min. BM cells (2.5 × 104) were injected intravenously (IV) in a volume of 0.2 mL PBS, 3 to 4 hours after irradiation (n = 10/group). After 13 days, the mice were killed, the spleens isolated and fixed in Tellyesniczky solution, and the macroscopic colonies were scored.
Monoclonal antibodies and flow cytometry
The following antibodies were used for flow cytometry: anti-CD3 (145-2C11), anti-NK1.1 (PK136), anti-B220 (RA3-6B2), anti-Gr-1 (RB6-8C5), anti-CD11b (M1/70), anti-CD4 (RM4-5), anti-CD8α (53—6.7), anti-CD11c (HL3), anti-IAb (AF6-120.1), (Pharmingen, San Diego, CA), polyclonal goat-anti-IgM, and anti-IgD (SBA 1) (Southern Biotech, Birmingham, AL). Isotype-matched controls used included mouse IgG2a, rat IgG2a, rat IgG2b, and hamster IgG labeled with the appropriate fluorochromes (Pharmingen). Up to 1 × 106 cells per sample were first incubated in FACS buffer (PBS containing 2% FBS and 0.02% sodium azide), containing 2% mouse serum (Biocell, Rancho Dominguez, CA), 2% hamster serum (Jackson Immunoresearch Laboratories, West Grove, PA) and 10 μg/mL anti-CD16/32 (2.4G2) (Immunex) to minimize nonspecific binding. Cells were then labeled with the appropriate antibodies for 30 minutes at 4°C in FACS buffer. Finally propidium iodide (2.5 μg/mL) (Boehringer Mannheim, Indianapolis, IN) was added to exclude dead cells from the analysis. Samples were analyzed on a FACScan (Becton Dickinson, San Jose, CA), or FACSCalibur (Becton Dickinson). Between 20 000 and 100 000 events were collected for analysis. Data were analyzed using Cellquest software (Becton Dickinson).
Isolation of DC
The DC were enriched as previously described.32 Briefly, spleens, thymi, and LN (2 inguinal plus 2 brachial plus 2 axillary nodes) were harvested. Spleens and thymi were infused with 100 U/mL of type III collagenase (Worthington Biochemicals, Lakewood, NJ) and incubated in 400 U/mL of type III collagenase for 30 minutes at 37°C. Intact LN were incubated in 400 U/mL of type III collagenase for 30 minutes at 37°C. Tissues were further dissociated in Ca++-free media (Hanks balanced salt solution [HBSS]) (Life Technologies, Grand Island, NY) containing 10 mmol/L EDTA (Life Technologies), and cells were separated into low- and high-density fractions on a Nycodenz gradient (density 1.080 gm/mL) (Nycomed Pharma, Distributor in the United States, Life Technologies). The low-density cells, enriched for DC, were harvested from the interface. The cells were incubated with anti-CD11c and anti-CD8α and the DC purified on a FACSVantage (Becton Dickinson).
Mixed lymphocyte reaction
The CD4+ T lymphocytes from the LN and spleens of DBA/2 adult female mice were purified as previously described.23One hundred thousand purified CD4+ T cells were incubated in 10% CO2 in air at 37°C with serially diluted purified DC in 96-well round-bottomed plates (ICN Biomedicals, Aurora, OH) in complete Dulbecco's modified Eagle's medium (DMEM) supplemented with 5% FBS (Life Technologies), 50 U/mL penicillin, 50 μg/mL streptomycin and 2 mmol/L l-glutamine (JRH Biosciences), 5 × 10−5 mol/L 2-ME, 0.1 mmol/L nonessential amino acids, and 1 mmol/L sodium pyruvate (Life Technologies). On day 4, 0.5 μCi of tritiated thymidine was added to the wells and after 5 hours the cells were harvested onto glass fiber sheets for counting on a gas-phase β counter.
Cytotoxicity assays
Mice were injected intraperitoneally with PBS or 100 μg of polyinosinic-polycytidylic acid (poly-I:C) (Sigma, St Louis, MO) in PBS and their spleens were isolated the following day. Cell suspensions were prepared and cytotoxicity was measured in a standard 4-hour51Cr-release assay. Yac-1 cells were labeled with Na251CrO4 (100 μCi/3 × 106 cells) for 1 hour at 37°C. Serial dilutions of effector cells were mixed with labeled targets (5 × 104/well) in 96-well round-bottomed plates and centrifuged at 400g for 3 minutes. Assays were performed in RPMI media (Life Technologies) containing 10% FBS (Intergen), 50 U/mL penicillin, 50 μg/mL streptomycin and 2 mmol/Ll-glutamine (JRH Biosciences), 5 × 10−5 mol/L 2-ME, 0.1 mmol/L nonessential amino acids, and 1 mmol/L sodium pyruvate (Life Technologies). In some experiments, labeled P815 target cells were included as a control for NK-specific lysis. Cell-free supernatants were harvested after 4 hours of incubation at 37°C, 5% CO2 using a cell harvester (Skatron, Sterling, VA). Percent specific lysis was calculated as ([experimental release-spontaneous release]/[maximum release-spontaneous release]) × 100.
Statistical analyses
Data were analyzed using a 2-tailed Student t test.
Generation of flt3L−/− mice
Mice genetically deficient in flt3L (flt3L−/−) were generated by gene targeting using a vector that deletes the majority of the flt3L extracellular domain (Figure 1A). Southern blot analyses using diagnostic restriction digests in conjunction with probes lying beyond the regions of homology contained within the targeting vector confirmed that the flt3L gene was disrupted by homologous recombination (Figure 1B), and a sensitive RT-PCR analysis confirmed the lack of flt3L expression in flt3L−/− mice (Figure 1C). Mice homozygous for the mutation were generated from crosses of heterozygotes at the expected mendelian frequency, displayed no overt phenotype, and bred normally. The flt3L−/− mice used throughout these studies were maintained on a C57BL/6 inbred genetic background.
Flt3L−/− mice have reduced numbers of leukocytes in their hematopoietic tissues and PB
We examined leukocyte cellularity in the PB and a number of hematopoietic tissues in the flt3L−/− mice. Total WBC numbers were reduced by 45% in the PB from a mean of 11.0 × 106 cells/mL in C57BL/6 mice (flt3L+/+) to 6.1 × 106 cells/mL in flt3L−/− mice (Figure 2A). The relative proportion of lymphocytes, neutrophils, and monocytes was examined in the PB of flt3L−/− mice using a Hemavet blood analyzer (n = 8). The proportion of monocytes was unchanged, but the proportion of lymphocytes was significantly reduced from a mean of 84.4% in the flt3L+/+ mice to 75.8% in the flt3L−/− mice. Conversely, the proportion of neutrophils was significantly increased from 12.1% in the flt3L+/+ mice to 20.3% in the flt3L−/− mice (Figure2B). This translates to an absolute lymphocyte count of 9.2 × 106/mL in flt3L+/+ PB compared to 4.6 × 106/mL in flt3L−/− PB, and an absolute neutrophil count of 1.4 × 106/mL in flt3L+/+ PB compared to 1.2 × 106/mL in flt3L−/− PB. The hematocrit (percent packed cell volume) was examined and appeared normal in the flt3L−/− mice (Figure 2C). Platelets were also quantified using the Hemavet blood analyzer. There was no difference in the mean number of platelets in the PB of flt3L−/− mice compared to flt3L+/+ mice (Figure2D). Eosinophils in the PB were quantified and no difference was noted between flt3L−/− and flt3L+/+ mice (flt3L−/−, 4.1 ± 2.6 × 104/mL; flt3L+/+, 2.8 ± 2.5 × 104/ml; n = 8).
Cellularity of the BM was reduced by 27% in flt3L−/− mice (Figure 3). In flt3L+/+ mice, mean total leukocyte cellularity from 2 femurs was 47.9 × 106cells, compared to a mean of 34.8 × 106 cells from the femurs of flt3L−/− mice. Analysis by flow cytometry revealed that the reduction resulted from a decrease in the number of immature B lymphocytes (B220+, IgM−) and immature myeloid cells (CD11b+, Gr-1+) (data not shown). Spleen cellularity was reduced by 30% from a mean of 140.0 × 106 cells/spleen to 98.0 × 106 cells/spleen in flt3L−/− mice (Figure 3). LN cellularity was reduced by 35% in flt3L−/− mice, from a mean of 27.7 × 106 cells/6 nodes to 18.0 × 106 cells/6 nodes (Figure 3). Analysis of spleen and LN cells by flow cytometry revealed that both T cells and B cells were reduced in number, but that the ratio of T cells (both CD4+ and CD8+) to B cells was unchanged in the flt3L−/− mice when compared to flt3L+/+ mice (data not shown). In contrast flt3L−/− thymus cellularity was not significantly affected (Figure 3). Histochemical analyses of spleen, thymi, and LN revealed no disruption to normal architecture in the tissues of flt3L−/− mice (n = 8, data not shown).
We conclude that targeted disruption of the flt3L gene results in reduced leukocyte cellularity in primary hematopoietic tissue, secondary lymphoid organs, and the PB. Thymic cellularity, blood hematocrit, and platelet numbers were not affected. Although leukocyte cellularity was significantly reduced in the flt3L−/− mice, these animals do not appear to suffer an increased frequency of infection, and their lifespan in a specific pathogen-free animal facility appears to be equivalent to flt3L+/+ mice (data not shown).
Reduced numbers of hematopoietic myeloid and lymphoid progenitors in the BM of flt3L−/− mice
We hypothesized that the reduced numbers of leukocytes observed in the periphery of flt3L−/− mice resulted from reduced numbers of lymphoid and myeloid hematopoietic progenitors in the BM. The frequency and numbers of myeloid precursors were examined in vitro and in vivo. Although the frequency of precursors for the myeloid lineage (CFU-GM) in the BM was not significantly different (Figure 4A), the absolute number of CFU-GM was reduced from a mean of 80.8 × 103/2 femurs in the flt3L+/+ mice to a mean of 47.2 × 103/2 femurs in the flt3L−/− mice (Figure 4B).
In addition to CFU-GM, the frequency and total number of multipotent CFU-S13 progenitors in the femurs were determined. BM cells from flt3L+/+ and flt3L−/− mice were injected IV into lethally irradiated C57BL/6 recipients, and after 13 days, the spleens were isolated, fixed in Tellyesniczky solution, and the macroscopic spleen colonies were scored (Table 1). The frequency of CFU-S13 progenitors in the BM was not significantly different in the flt3L+/+ and the flt3L−/− mice; however, the absolute number of CFU-S13 progenitors in the BM of flt3L−/− mice was reduced 39% from a mean of 9777 CFU-S13 progenitors per 2 femurs in the flt3L+/+ mice to a mean of 5951 CFU-S13 progenitors per 2 femurs in the flt3L−/− mice.
|Experiment number
|BM cellularity (×106/2 femurs)
|CFU-S13 frequency (/105)
|Total CFU-S13 (per 2 femurs)
|10 579
|Mean ± SD
|45.5 ± 2.4
|28.4 ± 1.9
|21.7 ± 2.0
|21.1 ± 2.2
|9777 ± 723
|5951 ± 324
|Experiment number
|BM cellularity (×106/2 femurs)
|CFU-S13 frequency (/105)
|Total CFU-S13 (per 2 femurs)
|10 579
|Mean ± SD
|45.5 ± 2.4
|28.4 ± 1.9
|21.7 ± 2.0
|21.1 ± 2.2
|9777 ± 723
|5951 ± 324
BM cells were isolated from the femurs of flt3L+/+ and flt3L−/− adult mice (n = 4-5/experiment), and pooled for each group. Groups of 10 lethally irradiated C57BL/6 mice were injected intravenously with 2.5 × 104 BM cells. After 13 days, the mice were sacrificed, the spleens isolated and fixed in Tellyesniczky solution. In each experiment, macroscopic colonies (≥1.0 mm in diameter) on the spleens were scored by 2 individuals, and the results averaged. Data are presented as the mean ± SD from the 3 experiments. BM cellularity: P = .0007; CFU-S13 frequency: not significantly different; Total CFU-S13 per 2 femurs:P = .0011.
The numbers of BFU-E were scored as a measure of the number of erythroid progenitors in the BM. The number of BFU-E per 2 femurs was slightly reduced in the BM of flt3L−/− mice but did not reach significance (5.8 ± 3.0 × 103 in flt3L+/+ BM compared to 4.3 ± 2.8 × 103 in flt3L−/− BM; n = 12).
The B-lymphocyte precursors (B-CFU) present in the BM were also examined in vitro. B-cell precursors responsive to IL-7 alone were reduced 90% from 13.5 × 103/2 femurs in flt3L+/+ mice to 1.4 × 103/2 femurs in flt3L−/− mice (Figure 5). The frequency of IL-7-responsive precursors was reduced from 28/1 × 105 BM cells in flt3L+/+ mice to 4/1 × 105 BM cells in flt3L−/− mice. B-lymphocyte precursors responsive to the cytokine combinations IL-7 plus KL or IL-7 plus flt3L were reduced 81% and 75%, respectively (Figure 5). Unlike the myeloid precursors, which were reduced in absolute number in the BM, but not in frequency, the absolute numbers and the frequency of B-cell precursors were significantly reduced in the BM of flt3L−/− mice.
Because the number of B-cell precursors in the BM was severely reduced, we examined the peripheral B-lymphocyte populations to determine whether these were affected qualitatively or quantitatively. B cells present in the spleen and LN expressed normal levels of surface IgM and IgD (data not shown), and normal levels of circulating IgA, IgG1, IgG2a, IgG2b, IgG3, and IgE were present in the serum (n = 7 mice, data not shown). However, significantly increased levels of IgM were noted in the serum of flt3L−/− mice. The mean concentration of serum IgM in flt3L+/+ mice was 63 ±12 μg/mL, as compared to 178 ± 66 μg/mL in flt3L−/− mice. Flt3L−/− mice were immunized with trinitrophenol-conjugated keyhole limpet hemocyanin (TNP-KLH) in alum, and 3 weeks later rechallenged with TNP-KLH. Concentrations of various immunoglobulin isotypes were measured in the serum after the primary and secondary challenge. No differences in the concentration of TNP-specific IgA, IgG1, IgG2a, IgG2b, IgG3, IgE, or IgM were noted between flt3L−/− and flt3L+/+ mice (data not shown). In addition, the spleens of immunized animals were examined for the presence of peanut agglutinin-positive germinal centers and no differences were noted in the number formed in the spleens of flt3L−/− mice compared to flt3L+/+ mice (data not shown). These results indicate that although the numbers of B-cell precursors present in the BM of flt3L−/− mice are severely reduced, peripheral B cells are present and appear to function normally in response to immunization with a T-cell-dependent immunization protocol.
Flt3L−/− mice have reduced numbers of DC
Administration of flt3L to mice for 9 to 10 days induces a dramatic expansion of DC in the PB as well as in multiple tissues23 suggesting a role for flt3L in the development of DC. We examined the spleens, thymi, and LN of flt3L−/− mice for the presence of DC. To enrich for DC, the tissues were treated with collagenase and the resulting cell suspension was centrifuged over Nycodenz (1.080 g/mL) to enrich for low-density cells. Cells were incubated with antibodies to CD11c and CD8α. Myeloid-related DC were identified as low-density cells expressing high levels of CD11c and lacking expression of CD8α, whereas lymphoid-related DC were identified as cells expressing high levels of CD11c and CD8α. All CD11cbright cells expressed high levels of major histocompatibility complex class II (IAb) (data not shown). Representative examples from the spleen, thymus, and LN are presented in Figure 6. The absolute numbers of CD8α− and CD8α+ DC in each tissue are presented in Table 2. The numbers of myeloid-related and lymphoid-related DC were reduced in the spleen, thymus, and LN of flt3L−/− mice. In some experiments the number of DC present in the LN of flt3L−/− mice was below the level of detection (0.1 × 104/6 LN) (Table 2).
|flt3L+/+ CD11c++ CD8α−
|flt3L−/− CD11c++ CD8α−
|Fold reduction
|flt3L+/+ CD11c++ CD8α+
|Flt3L−/− CD11c++ CD8α+
|Fold reduction
|Experiment 1 (n = 3)
|Experiment 2 (n = 4)
|Experiment 3 (n = 3)
|Experiment 1 (n = 5)
|Experiment 2 (n = 8)
|Experiment 3 (n = 4)
|Experiment 4 (n = 3)
|Experiment 1 (n = 5)
|Experiment 2 (n = 8)
|Experiment 3 (n = 4)
|Experiment 4 (n = 5)
|Experiment 5 (n = 7)
|flt3L+/+ CD11c++ CD8α−
|flt3L−/− CD11c++ CD8α−
|Fold reduction
|flt3L+/+ CD11c++ CD8α+
|Flt3L−/− CD11c++ CD8α+
|Fold reduction
|Experiment 1 (n = 3)
|Experiment 2 (n = 4)
|Experiment 3 (n = 3)
|Experiment 1 (n = 5)
|Experiment 2 (n = 8)
|Experiment 3 (n = 4)
|Experiment 4 (n = 3)
|Experiment 1 (n = 5)
|Experiment 2 (n = 8)
|Experiment 3 (n = 4)
|Experiment 4 (n = 5)
|Experiment 5 (n = 7)
DC were enriched from the spleen, the thymus, and 6 LN (2 inguinal plus 2 brachial plus 2 axillary) as described in "Materials and methods." Tissues were digested with collagenase, and the low-density cells were isolated by centrifugation on a Nycodenz gradient. The proportion of myeloid-related DC (CD11c++, CD8α−) and lymphoid-related DC (CD11c++, CD8α+) were analyzed by flow cytometry. The data are presented as the absolute number of DC per spleen, per thymus, or per 6 LN, from 3 to 5 experiments in which tissues from 3 to 8 mice were pooled for analysis.
The CD8α+ DC and the CD8α− DC from the spleens of flt3L−/− and flt3L+/+ mice were purified by cell sorting, and their ability to stimulate proliferation of allogeneic naive T cells was examined in a mixed lymphocyte reaction (MLR) (Figure 7). The CD8α− DC from the flt3L−/− mice were equivalent to those isolated from flt3L+/+ mice at stimulating allogeneic T-cell proliferation (Figure 7). The CD8α+ DC were less efficient than the CD8α− DC at stimulating allogeneic T-cell proliferation; however, no difference between the CD8α+ DC from the flt3L+/+ and the flt3L−/− mice was noted. We conclude that the number of splenic DC is markedly reduced in flt3L−/− mice, but the DC that are present are able to efficiently stimulate naive allogeneic T-cell proliferation in vitro.
Flt3L−/− mice are deficient in NK cells
The presence of NK cells (NK1.1high, CD3−) in the spleen was examined by flow cytometry and yielded an unexpected finding. The number of NK cells in the spleens of flt3L−/− mice was dramatically reduced from a mean of 4.14 × 106 cells in the spleens of flt3L+/+ mice (range 2.31-6.37 × 106) to a mean of 0.78 × 106 (range 0.39-1.64 × 106) in the spleens of flt3L−/− mice (Figure 8A). The number of natural T (NT) cells (NK1.1low, CD3low) present in the spleens of flt3L−/− mice did not appear to be affected (data not shown). Administration of poly-I:C to mice induces quiescent NK cells to become activated, which can be quantified by their ability to lyse Yac-1 lymphoma cells. Flt3L+/+ and flt3L−/− mice were injected intraperitoneally with poly-I:C and the following day their spleens were isolated. NK cytotoxicity was measured by culturing splenocytes with51Cr-labeled Yac-1 cells. Poly-I:C treatment stimulated NK cell activity in flt3L+/+ mice (Figure 8B). Poly-I:C treatment of flt3L−/− mice stimulated a small increase in the percent specific lysis of Yac-1 cells, compared to the activity observed from flt3L−/− splenocytes from mice treated with PBS. However the level of killing was below the level of background killing observed with splenocytes from flt3L+/+ mice treated with PBS. An example of the levels of specific lysis observed is presented in Figure 8B. We conclude that targeted disruption of the flt3L gene affects development of NK cells.
We have generated mice deficient for the gene encoding the hematopoietic growth factor flt3L. Although these mice are viable, breed normally, and appear healthy, at least when maintained in a specific pathogen-free facility, we have observed a number of deficiencies in hematopoietic lineages. Specifically, leukocyte cellularity was significantly reduced in the PB, BM, spleen, and LN and the numbers of DC and NK cells were dramatically reduced.
Examination of the PB revealed a decreased proportion of lymphocytes and a corresponding increase in the proportion of neutrophils. The proportion of monocytes was not changed (Figure 2B). Conversely, erythrocyte and platelet production was not affected in the flt3L−/− mice (Figure 2C and D). In addition to reduced leukocyte cellularity, decreased numbers of hematopoietic progenitors were present in the BM (Figures 4 and 5, Table 1). The absolute numbers of myeloid progenitors were reduced, but the frequency of these cells was not different from that observed in flt3L+/+ mice (Figure 4, Table1). Similar observations have been made in W/Wvflk-2−/− mice.29 These mice, which lack flk-2 and have a mutated c-kit receptor, have severely reduced hematopoiesis, including reduced absolute numbers of progenitors in the BM, but the frequency of the myeloid progenitors is normal. This implies that a mechanism to maintain normal myeloid progenitor frequencies in vivo exists, which does not involve flt3L or KL. In contrast to the myeloid lineage, both the absolute number and the frequency of B lymphoid progenitors in the BM were reduced in the BM of flt3L−/− mice, implicating a critical role for flt3L in B lymphopoiesis (Figure 5). The reduced numbers of lymphoid and myeloid progenitors in the BM of flt3L−/− mice are consistent with the numerous observations that flt3L synergizes with myeloid and lymphoid cytokines to augment expansion of these progenitors.3 Interestingly, mice with mutations in KL (Sl/Sld) have reduced numbers of hematopoietic progenitors in their BM, along with reduced BM cellularity, as described here for the flt3L−/− mice, but, unlike flt3L−/− mice, peripheral leukocyte numbers are close to normal.30
The reduced numbers of DC noted in the spleens, LN, and thymus of flt3L−/− mice (Figure 6, Table 2) support our hypothesis that flt3L is an important cytokine in the generation of DC. This hypothesis was based on our previous observations that flt3L administration to mice for 9 to 10 days induces a profound expansion of DC in multiple tissues.23 Analysis of DC from the spleens, LN, and thymus of flt3L−/− mice revealed a marked reduction in both myeloid-related and lymphoid-related DC. However, there were detectable numbers of DC in the flt3L−/− mice, and on purification the splenic DC were as efficient at stimulating naive allogeneic T-cell proliferation in an MLR as DC isolated from the flt3L+/+ mice (Figure 7). As has previously been described,33 the lymphoid-related DC were less effective than the myeloid-related DC at stimulating CD4+ T-cell proliferation. It remains to be determined whether the reduced numbers of DC will have an effect on infectious agent or tumor cell challenge in vivo. In vitro studies show that the addition of flt3L to a combination of cytokines can augment the generation and expansion of myeloid-related34,35 and lymphoid-related DC.36A number of other cytokines have been implicated in DC development including GM-CSF, tumor necrosis factor-α (TNFα), and transforming growth factor-β1 (TGFβ1). DC can be generated in vitro with GM-CSF alone37 or TNFα plus GM-CSF,38-40 but mice lacking GM-CSF do not have reduced numbers of lymphoid tissue DC.41 Mice deficient in TGFβ1 lack Langerhans DC, which reside in the epithelia, implying a critical role for TGFβ1 in the development of this subset of DC.42 One or more of these cytokines may substitute for the lack of flt3L to generate the residual DC present in flt3L−/− mice.
The flt3L−/− mice have a marked reduction in NK cells in the spleen (Figure 8A) demonstrating that flt3L is critical for NK cell generation. NK numbers were reduced 5.3-fold and this resulted in an almost complete loss in the ability of poly-I:C-stimulated flt3L−/− splenocytes to lyse Yac-1 lymphoma targets (Figure8B). Administration of flt3L to C57BL/6 mice results in increased numbers of NK cells in numerous tissues,43 and administration of flt3L to severe combined immunodeficient (SCID) mice (which lack T and B cells) also expands splenic NK cells (D.H.L., R.E.M., and K.B. unpublished observation). These flt3L-expanded NK cells appear to be functional because SCID mice inoculated with fibrosarcoma cells and treated with flt3L exhibit slower tumor growth26 and this effect can be abrogated if the mice are treated in vivo with an anti-NK cell antibody (PK136) (D.H.L. and R.E.M., unpublished observation).
A number of reports have demonstrated that flt3L has a role in the expansion of immature B cells that develop in the BM.13-15Flt3 receptor expression has been detected in the earliest B-cell progenitors, the pre-progenitor B cells, and as development progresses, flt3 receptor expression is down-regulated.44 The significant reduction in BM-derived B-cell progenitors we noted in the flt3L−/− mice is similar to that described for the flk-2−/− mice.29 However, preliminary immunization experiments with TNP-KLH plus alum (T-cell-dependent antibody response) indicated that flt3L−/− mice generated TNP-specific antibodies at levels similar to flt3L+/+ mice. Normal B-cell function was also indicated by the presence of normal levels of circulating polyclonal immunoglobulins in the serum of unchallenged flt3L−/− mice, with the exception of IgM, which was elevated (2.8-fold increase). The significance of this is unclear. Peripheral B cells in the spleen and LN expressed normal levels of cell-surface IgM (data not shown), and after immunization no differences in serum IgM concentrations were noted.
Thymic cellularity in the flt3L−/− mice was unaffected. Flt3L is expressed in the thymus45 and primitive thymocytes express the flt3 receptor4 implying a role for flt3L in thymocyte development. However, like the flt3L−/− mice described here, the flk-2−/− mice have normal thymic cellularity, though reduced numbers of primitive thymocytes are noted in neonatal mice.29 Although thymic cellularity was normal, the absolute number of T cells present in the lymphoid tissues was reduced. Factors produced by other circulating cells, which are reduced in the flt3L−/−mice, may be required to maintain normal numbers of circulating T cells.
There is no evidence that there are other ligands for the flt3 receptor, nor is there any evidence that flt3L binds to any other protein besides the flt3 receptor. Thus it would be expected that mice carrying targeted disruptions in either the ligand or the receptor gene would have an identical phenotype. It is therefore not clear why such differences are seen between the flt3L−/− mice described here and the flk-2−/− mice previously described.29 We have observed that 10 days of flt3L administration to flt3L−/− mice partially restores DC and NK numbers (data not shown), confirming that these defects are due specifically to the flt3L mutation. The DC and NK lineages were not discussed in the manuscript describing the flk-2−/− mice, so it remains possible that there are defects in these lineages in the flk-2−/− mice. However, the decreased leukocyte cellularities in the PB and hematopoietic tissues, as well as decreased numbers of myeloid precursors, were not observed in the flk-2−/− mice, though the more primitive stem cells, as assayed by competitive repopulation, are deficient. The possibility exists that the differences are strain dependent, because the flk-2−/− mice were created on a 129 background, and the flt3L−/− mice were created on a C57BL/6 background. Strain-dependent phenotypes have been described for gene knock-out mice. Epidermal growth factor receptor knock-out mice have varying phenotypes and life expectancies depending on the strain in which the mutation is introduced.46-48
We conclude that flt3L is a multipotent cytokine with effects on the expansion of hematopoietic progenitors of both the myeloid and lymphoid lineage. Flt3L appears to be particularly important for the generation of B-cell precursors, splenic NK cells, and lymphoid tissue DC because these cell types were the most severely reduced in flt3L−/− mice.
The authors acknowledge the expert editorial assistance of Anne Aumell, the contributions of the animal facility technical staff and Joann Schuh for reading the histology slides. In addition we thank Doug Williams for helpful discussions.
Reprints:Hilary McKenna, Immunobiology Department, Immunex Corporation, 51 University St, Seattle, WA 98101; e-mail:firstname.lastname@example.com.
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.


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When I drive and start accelerating, the transmission begins slipping out of gear. The engine has difficulty picking up speed. The shifting is also rough. What might be the problem?
Hello, thanks for writing in about your 1990 Toyota Cressida. The transmission is most likely the issue in this case. The fluid level and condition need to be checked. If the fluid level is extremely dark or exhibits a burnt smell this may indicate an extremely worn clutch packs in a worst case scenario. A transmission service where the fluid and filter are replaced may also correct the issue especially if the transmission has not been recently serviced. You may want to enlist the help of a mechanic to check the fluid level and condition in order to diagnose the slipping problem and rough shifting.
Our certified mechanics come to you ・Backed by 12-month, 12,000-mile guarantee・Fair and transparent pricing

subset: commoncrawl


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Wild Mushroom and Spinach Lasagne Recipe
- Vegetarian
Prep:40 min
Cooking:20 min
Nutrition per portion
|422 kCal
|- Saturates
- 25g (1oz) dried porcini mushrooms
- 25g (1oz) butter
- 3 garlic cloves, crushed
- 1kg (2lb 4oz) mixed mushrooms
- 3tbsp dry white wine
- 300g (10oz) spinach, wilted and seasoned
- 150g (5oz) fresh lasagne sheets
- for the béchamel sauce
- 75g (3oz) butter
- 75g (3oz) plain flour
- 700ml (1¼pt) whole milk
- 100g (4oz) Parmesan, plus extra for grating
Preheat oven to 200 C, 180 C fan, 400 F, gas 6. For the sauce, melt butter in a pan; stir in flour. Cook for 1 min; turn heat down, slowly pour in milk while whisking. Cook, stirring, until you have a thick, smooth sauce. Remove from heat, stir in Parmesan and season. Put clingfilm on surface of sauce.
Soak porcinis in hot water for 15 mins; drain. Melt butter in a frying pan; cook garlic for 2 mins. Increase heat; throw in all mushrooms, mix, season, sauté for 5 mins. Add wine; allow the excess liquid to evaporate. When mushrooms have softened slightly, remove from heat, place in a sieve and drain off liquid.
In an ovenproof dish, make 2 layers of: mushrooms, then spinach, lasagne and sauce. Sprinkle top with Parmesan; bake in top third of oven for 20 mins. Remove from oven; rest for 10 mins before serving. You can make the sauce the day before and refrigerate, or freeze the made-up lasagne before cooking.


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Harvest Celebration at the Farm!!!!
On behalf of New Mercies Farm I cordially invite you to the New Mercies Farm 2nd annual Farmer's Ball!!! It's a harvest celebration. A festival to celebrate the bounty of our efforts and toils throughout the year.
It will be a potluck style banquet. Bringing crafted seasonal drinks and dishes that support the seasonal theme of the Harvest season are deeply encouraged. Please prepare whatever your contribution may be to nourish 10-12 people.
CSA Members will receive a complimentary jar of our very own honey!!!
There will be all sorts of activities for children and adults:
-Archery contest with prizes from the farm like sauerkraut, honey, canned tomatoes, maple syrup, winter squash and garlic!!
-Crown Making Table
-Dream Catcher Making
-Apple Bobbing
-Circus/Acroyoga Workshops
-Harvest Altar
-Pagan ritual/storytelling (The idea here is to re-acquaint
our senses to the Moon phases and seasonal agrarian
rhythms through story, prose and song)
We will also be featuring local musician and dear friend Jen Taylor's release of her latest CD Holy Cow! As well as other live performances from Cosmos Sunshine, Someone You Can Xray and LionBird. It will be a dance party extravaganza ya'll!!!!
I graciously accept all of your contributions and talents and ideas/suggestions to make this a festival to remember. Please contact me personally if you have specific talents you'd like to contribute. Are you an artist and want to help decorate? Are you a storyteller? What gift of yourself can you share with us? Any contribution is greatly accepted!!! Please contribute your uniqueness to this once a year celebration.
Specific contributions I am looking for:
-Bow and Arrow for children and adults
-Decorations/ Altar design
-Film recording
-Story telling
-Harvest Altar preparation
-Welcoming Portal - a festive entry way for guest to enter
our sacred space
-Crafts/craft making tables
There will be places throughout the farm to pitch a tent to spend the night and join us for brunch sunday.
Rain Date will be the following day Sunday Oct 19 2pm
Please Please RSVP ASAP!!! so I can know how many chairs, dishes etc. to have on hand. Hope to see each and everyone of you!
Michael Melillo
Land Steward
New Mercies Farm


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Green Goddess Juice/
I LOVE this recipe! The flavours go so well together and the subtle hint of mint makes it delisiously unique!
- 1 Cucumber
- 4 Celery Stalks
- 1 Pear
- 4 Kale Leaves
- 2 Cups Spinach
- 1/2 Cup Fresh Mint Leaves
Run all ingredients through your juicer and drink!
***Juicy Tip - The celery and cucumber are very hydrating and you get loads of deep green veggies at the same time. YUM! Live Juicy!


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Kit Kats are produced by Hershey in the US, but they are made by Nestlé everywhere else, often in unusual flavors. more...
November 20 , 1945 : The Nuremberg Trials began. more...
Learn something new every day
More Info... by email
Holographic tape is tape that has highly reflective properties or some variety of holographic image printed along its non-adhesive side. Like regular tape, it comes on spools and has a side that is adhesive, allowing you to affix it to various surfaces. Unlike regular tape, the non-adhesive side is printed using holography technology to give it the appearance of having 3D or three-dimensional images on a two-dimensional surface. These holographic images give the tape a unique quality that is very useful for certain applications.
The images on holographic tape might depict something recognizable, like an animal or corporate logo, or they might contain language. Sometimes they show only a complex three-dimensional pattern. The tape is printed using special technology that includes high-resolution dot matrix printers and holography devices. Images or patterns can be customized by the buyer and made to be very unique and almost impossible to copy. In some cases a laser reader or magnifier is needed to read micro text contained within the image.
When holographic tape is made specifically to be unique, it is often used for security purposes such as ID badges, currency, official documents, laboratory samples, or tamper seals. Sometimes it is imprinted with words like "genuine" or "safety sealed." If someone were to attempt to remove the tape, it might be made to leave behind words indicating it had been tampered with, such as "void" or "tainted." Probably the most popular use for holographic tape is for security.
Holographic tape is highly reflective and its surface can be rendered with any background color. This includes extremely bright colors outside of the normal spectrum, or which show up well at night. Even lower resolution varieties will show all the colors of the rainbow and brightly stand out from moderate distances. The tape is therefore frequently used in any number of eye-catching applications calling for reflective surfaces, including signs marking hazards and warnings, road signs, motor vehicles, bicycles, and pedestrian clothing and accessories.
The many adaptations of holographic tape are not limited to security, brand protection, anti-counterfeiting, transportation, and aviation. In addition, holographic tape can used to scare off birds and animals, and it can be used to catch fish. Various types of holographic tape make an excellent lure that resembles minnow skin and other bait fish. Due to the presence of certain harmful materials used to produce the tape, one of the goals of certain manufacturers is to produce tape that is environmentally friendly.
One of our editors will review your suggestion and make changes if warranted. Note that depending on the number of suggestions we receive, this can take anywhere from a few hours to a few days. Thank you for helping to improve wiseGEEK!


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Top sellers!
|www.Aviapress.com > Books > Frontline Illustrations series >
FRI-005 T-35, SMK and T-100 Red Army heavy multi-turret tanks (1930 - 1941) book
Special order
FRI-005 T-35, SMK and T-100 Red Army heavy multi-turret tanks (1930 - 1941) book
The book is dedicated to the development and employment of various versions of the T-35 heavy multi-turret tanks. It dwells on the scientific efforts made by Soviet tank designers in developing the TG, SMK and T-100 experimental tanks. This issue offers a detailed account of the Soviet heavy tank designs, their drawings and data on their tactical use, as well as 105 photographs and eight colour drawings.
80 pages, 105 photo, 19 schemes, 8 paintjob variants, with captions in English
1. T-35-1 during the May Day pparade. 1 may 1933
2. SMK heavy tank. Leningrad, August 1939
3. T-100 heavy tank. Vic. Leningrad, August 1939
4. T-35s are rolling downb the Red Square. 1 May 1937
5. tanks of the 5th heavy armoured brigade during an exercise. Vic. Kharkov, summer 1936


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In order to benefit from the new UEFI features (including faster system startup), you will have to prepare your USB device slightly different.
Main take-away here: UEFI accepts boot loaders in files placed a the FAT formatted boot volume
To prepare your USB stick (4GB minimum) manually follow these steps:
- Open an elevated command
- Start diskpart
- Enter list disk
Locate your USB stick, no room for mistakes here or you might destroy the wrong thing.
- Enter select disk # (where # is the number of your USB drive, in the example above it's number 3)
- Enter the following commands, keep in mind that all data on the USB device will be deleted:
- clean
- create partition primary
- select partition 1
- active
- format quick fs=fat32
- assign
- exit
- xcopy S:\* D:\ /s /e (S = the ISO drive or contents, D = the USB device)
Do you want to check if Windows is installed in such a way that its actually using UEFI, please consult my other blog post.


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If you take a look at the outdoor GPS navigation units produced by Garmin to date, you'll have a hard time picking out the best. More so, majority of the outdoor Garmin GPS units have been in the market for quite some time, and selection of new versus old may be yet another tough task. So, let us take a look at some of the best outdoor Garmin GPS units that are worth your money.
Important Parameters for Choosing the Right Garmin Outdoor GPS
Choosing the right Garmin outdoor GPS isn't just about finding a Garmin GPS that fits your budget, and works well. It is more about finding the best Garmin outdoor GPS navigator that offers the desired set of features that suit your requirements in the best possible manner.
So, here are the top 3 parameters for choosing the right Garmin outdoor GPS unit –
1. Preferred Mode of Usage: If you're planning to go on a long biking trip, a good car GPS isn't something that is going to help the cause. As such, it is necessary that you pick an appropriate Garmin outdoor GPS that really serves your purpose seamlessly.
2. Pricing: Pricing always plays a key role in the final decision. If you can't afford to shred off say $400-500 on a fully featured GPS navigator, you may have very little options at your disposal. More so, if you're running on a low budget, you don't find too many options either.
3. Robustness: If you're planning to use your Garmin outdoor GPS unit inside the car, you may not be really worried about its water-resistant capabilities. However, if you're required to step out, and navigate when there's heavy rainfall, it may make a world of difference. Hence, it is advisable to pick an outdoor Garmin GPS unit, which is robust enough to survive few bad falls, water/liquid spills, and rough handling.
Apart from the abovementioned factors, style, and class, are also two more significant aspects to be considered as well.
Top Garmin Outdoor GPS Units
Garmin Oregon Series – If you want an absolutely rugged next generation outdoor Garmin GPS that offers cool looks as well as plethora of features, the Garmin Oregon series is definitely for you.
Garmin Oregon 400T has already been reviewed in detail separately, and you can take a look at this review here.
All Garmin Oregon units come preloaded with the U.S. topographies, and top notch features like barometric altimeter, fascinating color display, cool-looking touch screen, a high-sensitivity GPS receiver, wireless data sharing, and the likes of them.
However, the Garmin Oregon GPS units are a costly affair, as they come with a starting price tag of $400+.
Garmin Dakota Series
The Garmin Dakota models are extremely lightweight GPS devices, which combine rich set of features, and rugged waterproof body in a sleek compact design.
They also have a high sensitivity GPS receiver like that of the Garmin Oregon series GPS navigators.
What's more, you get 20 hours of battery life, and much more; so they're the way to go, if you need a waterproof GPS in lightweight and trendy design.
They're also priced around the same range as that of the Garmin Oregon series navigators.
GPSMAP 60 CSx
Coming to the high performance GPSMAP 60CSx navigator, it is one of the best handheld GPS outdoor navigators that has the features of the latest generation GPS devices, along with added goodies that are missing in the other 60–series units.
As far as the pricing goes, Garmin Oregon, Dakota, as well as GPSMAP 60 CSx are quite close; the 60 CSx may cost you about $50 greater than the other 60-series products, and the Dakota series units.
However, the GPSMap 60 CSx scores over the Oregon and Dakota series with its portable nature, and capability to be used as a handheld GPS under all circumstances. They're also ideal for trekking purposes.
Garmin eTrex Series
For the adventure-lovers, the Garmin eTrex GPS units are simply a superb option, which are ideal for adventurous trips like hunting, and trekking.
The versatile Garmin eTrex series of outdoor handhelds features rugged shell, and they come loaded with high-sensitivity GPS receivers to help you in holding on to your position even in the deepest cover.
Apart from these outdoor Garmin GPS units, you may also be interested to take a look at the Best Future GPS Devices, and the Garmin Nuvi 885; a worthy GPS device.


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Virgin Cleansing Myth/
Ayanna Nahmias, Editor-in-ChiefLast Modified: 13:16 PM EDT, 9 February 2012
NEW DELHI, India – Yesterday we wrote about the scandal of three Indian politicians watching pornography during a parliament session. Today, Indian is once again in the news but in a slightly more positive light.
South African peace activist, Archbishop Desmond Tutu, the 1984 Noble Peace Prize winner, is now chairman of 'The Elders." This group is comprised of prominent people of diverse backgrounds and heritage who are dedicated to addressing humanitarian issues from around the world.
Tutu, 80, is spearheading a global movement called "Girls Not Brides" which is aimed at ending child marriages. We have focused a lot of attention on this issue because this practice has such a deleterious impact on its victims. Child brides are subjected to rape, fistulas, physical and psychological abuse, and murder often condoned by the community as the right of the husband because of a lack of a dowry or as an honor killing.
Tutu told Reuters late Wednesday that "India is doing fantastically." But intimated that the country's growth and role as a significant world player could increase exponentially if it "enlisted the participation of 50 percent of the population,' which means Women. The problem of marginalization, discrimination, abuse and murder of women is not unique to India.
Child marriages are most prevalent in Africa, the Middle East and South Asia, but also occurs in the United States. Though many countries have laws on the books prohibiting this practice, most of the families that engage in this type of behavior live in remote regions of the country where the police have, in their opinion, more pressing concerns than what they consider to be a 'family matter.'
According to the International Center for Research on Women (ICRW), "100 million girls will be married before the age of 18 in the coming decade. Most will be in sub-Saharan Africa countries, some of which are (Mali, DRC, Mozambique, Eritrea, Ethiopia) and the Asian Subcontinent countries, some of which are (Nepal, India, Pakistan, and Bangladesh). In Niger, for example, 74.5% of women in their early 20s were married as children. In Bangladesh, 66.2% were. Child marriage also occurs in parts of the world including the United States and the Middle East. (Source: ICRW)
According to UNICEF, an estimated 14 million girls between the ages of 15 and 19 give birth each year. Because their bodies have not fully developed they are twice as likely to die during pregnancy or childbirth as women in their 20s. Girls who marry between the ages of 10 and 14 are five times as likely to die during pregnancy or childbirth, and their infants are 60 percent more likely to die. (Source: UNICEF)
In India, 47 percent of women between the ages of 20 and 24 are married before the legal age of 18 according to the government's latest National Family Health Survey. Tutu, who is traveling in India with other Elders, including former Irish President Mary Robinson and Gro Harlem Brundtland, who was Norway's first prime minister, believes that this type of inequality is a definite impediment to increased socioeconomic development.
Tutu has been a vociferous campaigner on the issues of fighting HIV/AIDS, an epidemic that has plagued his own country, South Africa. In numerous interviews he asserts his belief that girls married off to older men, have little control over their sex lives and thus are more likely to be infected by HIV/AIDS as a consequence.
This is especially true in South Africa, where older men who lack access to proper healthcare resort to raping female babies and infant girls. This abhorrent practice is known as the Virgin Cleansing Myth "that if a man infected with HIV, AIDS, or other sexually transmitted diseases has sex with a virgin girl, he will be cured of his disease." (Source: Wikipedia)
There are many issues that must be addressed worldwide in an effort to achieve gender equality. We don't believe that 'gender equality' equates with 'gender sameness.' Women and men are uniquely created to complement each other and we believe this is healthy. It is only when one or the other, but in the case of this post, when a man chooses to exert control over a woman and to rob her of her natural right to self-determination, that we must stand up in one voice and denounce the perpetrators.
Martin Luther King, Jr. said it best, "Injustice anywhere is a threat to justice everywhere."
Tutu, The Elders, and NGOs are doing their part to increase global awareness of the practice of child marriages. We can support these campaigns at a grassroots level through donations, writing and blogging about this issue, or just reaching out to a woman in need in your community. To achieve gender equality at all levels of society we must do all that we can in support of the development of 50 percent of humanity. Women.Follow Nahmias Cipher Report on Twitter Twitter: @nahmias_report Editor-in-Chief: @ayannanahmias
Related articles
- What Difference Does Goodness Make? Desmond Tutu and his Daughter Mpho Tell Us How it Changes Everything in 'MADE FOR GOODNESS' (prweb.com)
- Grassroot Diplomat: The 26-Year Old Entrepreneur with Desmond Tutu, Kofi Annan & Nick Clegg In Her Network (thenextwomen.com)
- HarperOne to Publish 'Tutu: Authorized' in Celebration of Desmond Tutu's 80th Birthday (prweb.com)
- Ending child marriage can boost India's rise: Tutu (thehimalayantimes.com)


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In an actual survival situation, what blade would you prefer to have on your person in order to help you survive? Think about that perfect blade, now forget about it. Survival situations do not happen on your schedule, and unless you EDC that perfect survival blade on you every day of your life, it is not going to help when the situation calls for it. Natural disasters or the inopportune flat tire in the middle of no where arejust a couple examples of when an emergency survival blade could be employed.
An emergency survival blade is any blade you happen to have on you when chaos strikes, for most individuals this would usually be a folding knife of some sort. There are pocket survival kits that are available which are normally housed in an Altoids tin and have just enough room for a small knife or razor blade, along with a way to start a fire, etc. However, considering one of your most important tools in a survival situation is a cutting device, a tiny knife or a razor blade can only be used to a certain point. A much more beneficial option is the folding knife you carry every day.
On a regular basis, your EDC may only be used for light tasks such as opening letters, cutting boxes, etc. Although this may be the image you think of, your EDC is capable of so much more. Nearly any folding knife can be put into the role of an emergency survival knife, because honestly, at the time you have no choice. You would have to use what you have available.
The edge itself is the most important aspect of the knife, which is why we use knives in the first place. But once the user knows the physical limitations of the knife, the strengths of the model's design, and efficient techniques in using the knife for which ever type of task, the capability of the knife is increased one hundred fold. The basis for this knowledge and ability comes down to personal experience with the particular knife, skills, and knowledge of edged weapons.
Manual folders, spring assisted, old design styles, automatics, and well made OTF knives are all capable of filling the role as an emergency survival blade. This is not to say that the user should simply buy which ever knife is the cheapest, though it can be compared to the old firearm saying, "a .22 in the hand is better than a .45 at home."
Which ever knife you choose to carry will fill multiple roles in its lifetime, from every day utility tasks to self defense. Look at your EDC and ask yourself, "If I had to, could I use this in an survival situation?" Do you have the experience to use the blade to it's full potential? If not, what do you have to learn to make it so? A knife is limited only by the skill of the user.


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802 3D photos uploaded
Amaze Your Buddies Together With Your Photos with Such Photography Tips
Are you aware you will find the talent if this involves taking intriguing and creative pictures? Would you like enhance your natural photography abilities? You've found the best place if you are ready to get this done. This short article consists of advice to get began taking better still photos.
Snap your shots as rapidly and intuitively. Should you wait a moment to accept perfect shot, the topic may move or something like that may relocate to bar your view.
Framework is really a photo is essential. Focus in your subject or even the key to the photo and eliminate any surrounding distractions. This could eliminate any unnecessary points of interest and clutter inside your photographs away.
Choose just the best photography to focus on and display. Do not show a lot of photos and vary the identical subject. It may end up with boring for other people to determine exactly the same factor over and again. Keep that which you show others fresh by revealing a number of photos.
This tip can help you may use in photography. Become knowledgeable about shutter speed is and just how to control it. The digital camera has configurations labeled A, M, P, and P modes. The label "P" may be the setting means program mode. This mode will completely automate the shutter speed ad aperture already set. If you do not understand what you will take images of, you need to choose P.
Start shooting pictures immediately whenever you depart! You need to look out for interesting photos to consider throughout your vacation, the trip itself should be regarded as an opportunity to try taking some original shots. Document each phase of the vacation — your way you might have the ability to have some interesting things in an airport terminal.
Take photos of the travel souvenirs you bought whenever you travel. This creative project helps produce a more memorable reference to the objects and sights out of your souvenirs that you could enjoy whenever you go back home.
Composition is a vital component that every beginning digital photographer should think about and master when diving into photography. Like other artistic representations, when the composition inside your photo isn't right, then your outcome won't be the very best.
Do you want images of objects which are drenched or misted with rain? You may make this look if you take a sprig bottle and "mister" some rain in your subject just before taking photos of.
Take time to read and comprehend the manual that is included with the digital camera manual. Manuals are frequently a lengthy read and thick. They often get shoved towards the bottom drawer or simply disposed of. Rather than allowing it to gather dust, use time for you to read its contents. The manual can train you in taking greater quality pictures and stop dumb mistakes.
The greater the ISO is placed to, the greater grain is going to be in your pictures. This could completely ruin the picture mandates that kind of effect.
After you have reviewed this short article, you ought to be ready and excited to test something totally new and experiment. These pointers will help you enhance 802 3D photos uploaded your photography abilities. Try another thing if these pointers fail to work.


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The management of HR plays an important role in the management of a successful business. They administer all the work related to the HR department. But you can not manage everything at once. It will be difficult for them to manage everything accordingly. The HR Software in Pakistan is designed to make the process for all contracting parties more manageable. From candidates to hiring managers, it is important that Recruitment Software in Pakistan meets the needs of your organization.
So, what characteristics do recruiters need for their recruitment software?
Job application:
It goes without saying that one of the main job functions for recruiters is to raise and publicize jobs. Often, this will be in conjunction with HR manager to ensure that the work goes through the approval process.
Recruiters want to be able to do this quickly and efficiently, publishing on as many channels as possible. PeopleQlik also provides Perfect integration with Performance Management Software in Pakistan is vital to ensure that the details of the work are accurately distributed.
In times of shortage of candidates, talent groups become valuable recruiting tools. Recruiters should create talent pools of candidates segmented by skills, location, experience or availability.
By communicating with candidates directly through the Recruitment Software in Pakistan, recruiters can nurture talent groups and keep existing candidates updated with recruitment software, news and cultural information from the company.
Candidate evaluation:
Recruiters should be able to assess the suitability of the candidate and progress through the Recruitment Software in Pakistan workflow easily.
During communication of workflow with candidates, recruiters should be able to schedule and record details of phone calls, emails and SMS communications with candidates for a variety of reasons.
PeopleQlik Solutions:
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- Core HR Software – HRMS
- Cloud Payroll Management Software
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|Rhys Coren, Biancazzurri, 2012
Gloria Maria Gallery
22 Marzo - 30 Giugno
private view: 21 Marzo 2013, 7 – 10 pm
8.30 pm, Lorenzo Senni presents AAT, live performance
Guide to the Galaxy
Rhys Coren, Paul Flannery, Aude Pariset, Yuri Pattison,
Lorenzo Senni, Travess Smalley, Priscilla Tea, Amalia Ulman
Curated by Attilia Fattori Franchini
Curated by Attilia Fattori Franchini
A guide, is a collection of information leading through unknown topics or places often utilized with the aim of exploration.
The exhibition takes its title from the 1977's science-fiction comedy series - The Hitchhiker's Guide To The Galaxy - created by Douglas Adams and used as inspiration for the first web open encyclopaedias.
The concept of classification and presentation of worldly knowledge is taken as a point of departure to reflect about perception, reality and the internet in a post-digital age.
The show revolves around the constitutive textual and visual elements of digital interfaces, as layout, audio and content accessibility, overlooking at their legacy and influence within contemporary artistic practice.
Computer, i-phone and i-pad rapidly evolving formats and mass-producing images and (mis)-information contribute to a new configuration of the world often biased, constantly shifting between physical and digital, fictional and personal, influencing the way we absorb and understand reality.
This duality and its contextualisation is explored through the work of eight international artists, looking at the convergence between embodied media and mixed reality in social and physical communication, contemporary artistic production and digital vision.


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The long-awaited trailer for Pitch Perfect 2 has finally arrived and it's aca-mazing. The Bellas are preparing to say good-bye to College as well as their time in the Acapella group, but not before one final blow-out competition.
All the ladies are back, as well as Adam DeVine, Skylar Astin, and the hilarious Acapella judging duo of John Michael Higgins and first-time director Elizabeth Banks. Side note: the world hates Americans.
Pitch Perfect 2 hits theatres on May 15, 2015, which feels like an eternity. What can we do to pass the time? Throw on some pjs, pop some popcorn, and enjoy these fierce female friendship movies we love as much as Pitch Perfect.
The Sisterhood of the Travelling Pants
This two-part movie series based on the books by Ann Brashares is a one-size fits all kind of deal. It's easy to find yourself represented by the four main characters, who share a bond that reminds us what it really means to be a good friend and support one another in the darkest times. And it's cool to think one pair of jeans can look good on anyone.
Now And Then
Roberta, Samantha, Teeny, and Chrissy show us that growing up in the 1970s wasn't so different from growing up in 2014. Parents get divorced, movies are awesome, boys are confusing, and having a good group of friends can mean a much-needed second family.
All I Wanna Do/Strike!
Depending on whether you watched the film in Canada (Strike!) or the US (All I Wanna Do), the title may be different, buy the storyline is always awesome. A group girls at a Preparatory school fight back when the Head Master announces the school is going coed. Sex, bras, spiking drinks, and periods are all on the table. Side note: We just realized Momo is Merritt Weaver from Nurse Jackie and New Girl. Very cool.
Divine Secrets of the Ya-Ya Sisterhood
Sidda (Sandra Bullock) has never had a good relationship with her mother and when she gives an unflattering interview on her mother in the New York Times, things go from bad to worse. Sidda's mother's friends, who make up the 70 year-old friendship group the Ya Ya Sisterhood, bring the two women together. In addition to lessons about family and mother-daughter relationships, a big lesson from the film (and the Rebecca Wells books they're based on) is that true friends have seen you in your worst piece-of-crap-state and stick it out with you.
A League of Their Own
Where do we even start with the amazingness of a League of Their Own? Women taking on men's roles during World War II extended to the baseball field, where they got to find the silver lining in a terrible period in history. Madonna gives one of her greatest performances, Jon Lovitz and Tom Hanks deliver some of the greatest lines in sports movie history, and Geena Davis is too cool for words. Batter up, hear that call, it's time again for one and all to plaaaaay ball!
Mona Lisa Smile
Julia Roberts plays Katherine, a 30 year-old Art History teacher at Wellesley College in 1953. With a strong cast rounded out by Kirsten Dunst, Ginnifer Goodwin, Julia Stiles, and Maggie Gyllenhaal, issues of marriage, family expectations, education for females in the 1950s, love, and art are examined at length within the polarizing group.
The Babysitters Club
Okay, we know it's a movie about young girls who LOVE babysitting way more than anyone in the history of time has ever loved babysitting, but it's more than that. It also had long-lost dads, teenagers who ate junk food non-stop and never gained weight or got pimples, and a 17 year-old boy who was shocked to find out he was dating a 13 year-old girl (how was this storyline not a red flag for movie execs?). But The Babysitters Club is also about friendship, family and first loves, something everyone can relate to.
Pitch Perfect
Let's not forget why we're here today – the aca-mazing, aca-wesome, aca-riffic Pitch Perfect. Beca is ready to be the modoy goth girl at college until she's discovered singing in the shower by Acapella member Chloe. After joining the Bellas, Beca must endure gaining a group of weird and wonderful new friends, a super cute crush, and a reason to be a college other than studying. College would have been way better if we could sing.


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Real estate
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Our client base includes many UK and overseas based investors, developers and occupiers.
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The 12 Most Feminist '90s Shows Prove The Past Was More Progressive Than You Remember
Way back when, there was a decade when women started rocking out en masse, when a teen battled vampires and when women in hip hop started getting more attention than their male counterparts (Missy, I'm looking at you). So, obviously, it's your go-to decade for those The Future is Female vibes. Reference the most feminist shows from the '90s, and you'll be surprised at how curiously modern so many shows were in terms of their gender politics.
Obviously, there are exceptions to this. We're still talking about two and a half decades ago, and the '90s — despite our preoccupation with it — wasn't perfect. And, again, the below isn't a definitive guide as to how to be a feminist, and I've tried to highlight the weaknesses of each show as well as their considerable strengths. But it's hard to imagine the shows that currently exist and that deal, in a nuanced way, with what it means to be a woman (Insecure, Transparent, Broad City) coming into being without the TV shows of the '90s doing some heavy lifting first.
So, if you haven't done a pop culture rewind, take a browse. You might end up finding a new favorite to marathon.
1. 'Sex and the City'
Sure, this show's feminism had profound flaws. The SATC model of empowerment is mostly about buying yourself a fabulous new pair of Manolos, and there's no way the show's feminism even tried to be intersectional (the less we talk about the episode where Samantha dates a black man and her behavior, the better). But it did something revolutionary. It discussed the limitations of searching for a fairytale ending, and it focused on a group of women who regularly enjoyed sex outside of relationships without shaming them.
2. 'Full House'
OK, so the show technically premiered in the '90s, but it ran through the '90s so I'm keeping it. Aunt Becky made it seem like it might be possible to have it all; she was a wife and mother, who always put her career first. When she got pregnant with twins, she forced her husband to wear a fake pregnancy belly so he could experience the stresses of pregnancy with her. Plus, D.J. and Kimmy's friendship was one of the show's main focus after Kimmy became an unexpected fan favorite.
3. 'Married... with Children'
Though it, like Full House, first premiered in the late '80s, Married... with Children still needs to be discussed. Despite the show's retro sounding name, its gender politics were intriguing — mainly thanks to the Bundys' neighbor, the inimitable Marcy D'Arcy. If Marcy D'Arcy was a real person, she'd definitely be retweeting those "male tears" memes on Twitter. She was a feminist who started her own group FANG (Feminists Against Neanderthal Guys), she made enough money to support her family, and she watched porn (though, admittedly, was so ashamed of this that she disguised herself while browsing the adult section of the video store).
4. 'The Powerpuff Girls'
They may have just been three tiny girls, but, without their smarts and strength, the city of Townsville would have been destroyed by villains and giant monsters long ago.
5. 'Recess'
Whether you think of Miss Grotke's progressive takes on history, Vince deciding he didn't really care about the construct of gender and rocking some high heels because he felt like it, or TJ becoming an Ashley because "it's the 90s," this show had a more left-wing take on feminism and gender than you remembered.
6. 'Judge Judy'
OK, this one's a little controversial, since Judge Judy herself has publicly declared that she does not consider herself to be a feminist. But I beg to differ. The TV show focuses on a strong woman who gives zero f*cks, dispensing her own brand of unedited wisdom. She's immune to the idea that women should be more pleasant or diplomatic or sweeter than their male counterparts, and she's clearly never thought of her gender as being something that would get in the way of her professional ambition (which is why, arguably, she's still a feminist in action, if not in how she identifies).
7. 'Daria'
She's more preoccupied with books than her appearance. Despite her cynicism, she tries to support the women around her, even those that lead wildly different lives to her — like her fashionista sister Quinn. And her friendship with Jane Lane is total #goals. Daria Morgendorffer, everybody.
8. 'My So-Called Life'
This show's feminism was light years ahead of its time — not just because of its focus on female friendship and healthy perspective on sexuality, but because mopey teen protagonist Angela Chase was forced to check her privilege on the regular. Her best friend Rayanne had substance abuse issues, while Rayanne's other best friend Rickie was physically and emotionally abused by his uncle and, as a queer Latinx boy, highlighted issues of prejudice that Angela would have never noticed without him being around.
9. 'The Fresh Prince Of Bel-Air'
While Will's endless pick up lines might not make this an obvious source for feminist wisdom, Vivian Banks was a goddess who took no sh*t, while fashion-obsessed Hilary, despite being extremely different from her mother in most ways, boasted a similar strength and DGAF attitude.
10. 'The X-Files'
Scully was a woman working in a male-dominated field, who was so competent that she was recruited to the FBI while still in medical school. We got to see both the power of feminism (Scully's incredible smarts and professionalism) and what toxic aspects of the patriarchy it needs to tackle (Mulder's mansplaining, and the fact that he often makes decisions for her).
11. 'Clarissa Explains It All'
This was Nickelodeon's first female-led comedy, and it's hard not to love. Sure, Clarissa can veer into "Cool Girl" territory (her best friend's a guy, she owns a pet crocodile, she's so incredibly chill), but she's also independent, has her own style, is so competent at tech that she can design her own computer games, and, refreshingly for a TV show about a teenage girl, isn't obsessed with finding a boyfriend.
12. 'Moesha'
She set her own boundaries (like in that one episode with Usher where he suggests they "get to know each other" in a quieter spot at the party), spoke up for other women, and she earned her own money. Remember when her dad bought her a Saturn, and she wasn't into it, so she worked hard to buy a car that was more her style?
Feminism isn't a contemporary invention — and these shows prove it. The '90s might feel like a long time ago, but so much of its television pre-empted what we'd come to see in our era in terms of females (and males) sticking up for women's rights, setting boundaries, and supporting each other and a more fluid idea of gender.


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Sunday, November 30, 2008
MESSAGE IN THE SAND Online
Read and listen to a reading of our latest children's book here. The flash file is a little heavy, but the wait is worth it.
Thursday, November 27, 2008
The Business of Art
Transforming Art into a More Lucrative Career Choice
Monday, November 24, 2008
Juanito Torres is It!
CANVAS is pleased to announce that Juanito Torres has been selected as our Fellow, and is the designated artist for CANVAS Romeo Forbes Children's Literature Initiative for 2009-2010.
An artwork (or our contest piece) has now been commissioned, and once completed, its image will be used as the inspiration for the next CANVAS' annual Romeo Forbes Children's Storywriting Competition. The image of the contest piece, together will the rules of the competition and other details, will be released to the public in the first week of January 2009.
* Show for Hope by Juanito Torres. 48"x48" Oil on canvas (2008). Image courtesy of Britania Art Projects. Note: This is not yet the contest piece. :-)
What Will the World Look Like in 2025?
Some of their predictions:
- ...the U.S. dollar, while remaining important, will decline to "first among equals" among other national currencies. Similarly, the US will continue to be the most powerful country in the world, but its days as "the world's only superpower" will be over. China and India, following a "state capitalism" economic model, were likely to join the United States atop a multipolar world and compete for influence.
- Russia's potential was less certain, depending on its energy wealth and internal investment. But Iran, Turkey and Indonesia were also seen gaining power.
- A world with multiple power centers has been less stable than one with a single or two rival superpowers, and there was a growing potential for conflict.
- Global warming will be felt, and water, food and energy constraints may fuel conflict over resources.
- Global wealth was seen shifting from the developed West to the energy-rich Gulf States and Russia, and to Asia, the rising center of manufacturing and some service industries.
- A shift away from an oil-based energy system will be underway or complete by 2025. Better renewable technologies such as solar and wind power offer the best opportunity for a quick and low-cost transition.
- The risk that militant groups would use biological weapons was greater than the risk of nuclear terrorism.
- India, China and Brazil will rise, the Korean peninsula will be unified in some form, and new powers are likely to emerge from the Muslim non-Arab world.And where does the Philippines fit in all these possible scenarios? Well, the Philippines merits only a line - mainly as an example of a country with potential conflicts between a large Christian population and a minority Muslim constituency. Which really is a reflection of our relative influence in the world.
But reading the report itself, taking it as a jump-off point for stepping back and trying to see the big picture, and thinking about where we are and where we are headed - as a country and as a people and even as individuals - is a very useful exercise.
We highly recommend it.
You can download the report for free here.
Thursday, November 20, 2008
Michael Cacnio : Eclectic
In "Eclectic," Michael continues to take the noted social realist turn seen in his works of late and exploits the malleability of his material to explore the material needs and desires of his fellow man. In this collection, Michael also shows, for the first time, his paintings on canvas. Their debut, whether as backdrop to his relief or scene against which a small sculpture rests, is evidence of his versatility as an artist.
His works have earned critical acclaim as well as commercial success in the Philippines, Singapore, Europe and USA. Testaments to his strong following in the art world are his successful exhibitions (more than fifty both locally and abroad) and the consistently positive reviews from critics and collectors alike. Among his recent successful outings were his solo exhibits in New York and Brussels in 2007 and his marked participation in the anchor show of the Philippine Art Trek in Singapore in June, 2008.
"Eclectic" runs from November 24 to December 7, 2008 at 1/of Gallery, 2/L Shops at Serendra, Bonifacio Global City. Cocktails on Monday, November 24, 2008 at 7pm. For more information call 901-3152 or email firstname.lastname@example.com.
"Subok" by Michael Cacnio (2008).
Wednesday, November 19, 2008
Press Freedom = Less Corruption?
Two words that are can be said to apply to the Philippines today are "press freedom" and "corruption."
The World Bank says, and many people agree, that there is a correlation between the two: greater press freedom equals less corruption. Or as Severino says, sunlight is the best disinfectant.
But, as you will probably surmise, the Philippines is an outlier - we undoubtedly have freedom of the press, and yet corruption continues not only to exist, but actually to worsen.
Severino continues:
I've been looking at trends captured in the annual Corruption Perception Index (CPI) of Transparency International. The following have been the Philippines' rankings among 170-180 countries in the CPI through the years, with 1 the least corrupt.
1996: 44
1997: 40
1998: 55
1999: 54
2000: 69
2001: 65
2002: 77
2003: 92
2004: 102
2005: 117
2006: 121
2007: 131
2008: 141 (out of 180 countries)
1996-98 were the latter years of Fidel Ramos, 1998-2001 was the Estrada era. It slightly improved the year Gloria took over, then went progressively downwards. The Philippines hit three digits in the election year 2004 (not coincidentally, the year of "Hello Garci"), and its ranking has been worsening by leaps and bounds since then.
So, what accounts for us bucking the trend? What do you think?
Sunday, November 16, 2008
Is the Price Too High?
Do they have any choice? As a people and as a country, can we really have it any other way?
Check out this perceptive report from Time. Really something to think about...
Saturday, November 15, 2008
The Boy Who Touched Heaven, by Iris Gem Li (with artworks by Sergio Bumatay III) is our first National Book Award Winner as the best children's book of the year!
Inspired by the Banawe Rice Terraces, "The Boy Who Touched Heaven" won CANVAS' Elias Dakila Children's Storywriting Competition on Environment and Culture, and was published in partnership with Ang Ilustrador ng Kabataan and Adarna House.
To read the story, click here.
Signed first edition hardbound copies are still available at 1/of Gallery. To inquire, please call 901-3152, or email email@example.com.
Tuesday, November 11, 2008
Shoot to Move the Nation
An exquisite art book for children, MESSAGE IN THE SAND introduces the young eco-champion, Miguel, who loves the sea and sets out to save it. He is armed with nothing but faith, imagination and his mother's love and encouragement; his only weapons are those that the sea has provided for its own preservation – seashells, seaweed and sand. With these simple tools, Miguel must battle two giants: the powerful mining company, whose toxic waste is ruining the beach and killing the beautiful fish and seabirds; and the powerful ocean that keeps washing away his words. The book teaches gentle lessons in perseverance, and in finding unusual and peaceful means to effect change. It inspires and empowers children to act against seemingly insurmountable odds, showing us how even one child can make a difference.
Proceeds from the sale of the paintings and the book will benefit and be used to further CANVAS efforts to promote Philippine art, culture and the environment. The book, and a complete range of giftware featuring the paintings from MESSAGE IN THE SAND will also be available at its online store CANVAS Downstream.
First edition hardbound versions of MESSAGE IN THE SAND are available through CANVAS, the Ayala Museum and 1/of Gallery in Serendra. A special e-book version of MESSAGE IN THE SAND, together with an accompanying teaching module, is being prepared by CANVAS, and will be made available for free downloading on its website.
Charmaine Aserappa is the author of IN A JAPANESE GARDEN (with woodcuts by Akiko Naomura; Council Oak Books, San Francisco, U.S.A 1999). A Publishers Weekly Top Ten Gift Book, an Amazon.com Five Star, Most Wished For and Most Gifted Book, IN A JAPANESE GARDEN appears on Bestselling Lists all over the world in categories as diverse as philosophy, architecture, and poetry, and is used in many schools. Charmaine Aserappa said, "It was an honor to be invited to write MESSAGE IN THE SAND for CANVAS. I am delighted with the book and hope its message will spread across the world. We all share this wonderful but fragile planet, and children must be taught that caring for it is the sacred duty of every human being."
Roel Obemio's glowing acrylic paintings for the book have been inspired by the art of Fernando Botero (Colombia), and Mauro Malang Santos (Philippines), but are entirely original, and the unique and endearing characters in MESSAGE IN THE SAND are tenderly and amusingly rendered.
For more information, please email firstname.lastname@example.com, or text/call +63 917.890.6160.
Thursday, November 6, 2008
Who Will be Our Obama?
Perhaps the better question is, do we really need our own version of Obama - someone Filipinos can see as a transformative leader, a savior... burdened with expectations so high it would make most others dizzy?
We had one such leader before, of course, in Corazon Aquino. Talk to those who were there, especially in the giddy months of February, March and April 1986, capped by her triumphant speech in the US Congress - and the chills, tears, hopes, inspiration and dreams - not just here in the Philippines but on a global scale (as country after country will invoke her name and use people power to oust leader after leader in the late 1980s) will sound familiar.
Fast forward to today, when the hopes and dreams of EDSA have long been dashed for many (but not us - a preternaturally optimistic bunch - at CANVAS), and still we continue to look for inspiration or salvation from singular personalities.
The danger of Obama's victory for the Philippines is that many will now look to see if any in our current crop of political leaders offers similar inspiration... and we will be doomed to look in the least likely place to find one. Scan the list of likely and even probable presidentiables for 2010, and no one - absolutely no one - offers a vision of change that we can truly call, well, visionary.
And this now, is the challenge then for Juan... if we cannot find inspiration in our leaders, where will we now look?
We in CANVAS know and say that we need not look far. We only have to look within ourselves, to the people close to us and to the people we run into everyday in our everyday lives.
Obama, speaking about his recently deceased grandmother, said "She was one of those quiet heroes that we have all across America. They're not famous. Their names are not in the newspapers, but each and every day, they work hard." Anyone could just as well have been speaking about the countless Filipinos across the country, and indeed, all over the world.
Everyday Filipino heroes are all out there... and it is in this light that CANVAS, through its Looking for Juan Program, have long been conceptualizing, and will soon launch a new major event, centered on this very theme.
Details to follow...


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A report released this week by the Urban Institute at UNC Charlotte offers 12 strategies for expanding the supply of affordable housing in the City of Charlotte and Mecklenburg County. Prepared for the Charlotte-Mecklenburg Housing Advisory Board, the 66-page report outlines policies that 'can be used to finance and facilitate the development of quality affordable housing, with a focus on long-term affordability.'
You can download the full report HERE.
Filed under: Home Builder Issues, MultiFamily Issues, Realtor® Issues, Uncategorized | Tagged: Affordable Housing | Leave a comment »


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Do you have uncluttering or organizing projects on your mind? Consider one or more of these three easy projects:
- Pull a weed. You can do this either literally if you have a garden, or figuratively if there is a small task on your to-do list that will take you just a few seconds to complete. Do it and be done with it. There is no need for that pesky item to bother you any longer.
- Plan ahead. Many people in the U.S. have next Monday off from work in observance of Memorial Day. If you want to spend the three days relaxing and not tackling a giant list of to-do items, create a list now of the things you need to do before Saturday morning arrives. Then, make a plan for your week for how you're going to accomplish these tasks. Three days without a giant list of responsibilities hanging over your head will be good for you.
- Pack a suitcase. There isn't a reason to really pack a suitcase, but now is a great time to put together a packing list for the next time you head out on a summer trip. Having a checklist is a terrific way to pack wisely and not forget anything when you travel, and making the list now gives you time to get your list in shape. I have 10 packing lists saved on my computer: Romantic weekend with husband, 4-day conference for work, 3-day consulting with client in business casual environment, 3-day consulting with client in corporate business environment, 3-day trip with extended family, 7-day trip with extended family, 3-day relaxing trip with friends, 7-day beach/mountain trip with friends, 3-day sight-seeing trip, and 7-days as a tourist in a foreign city. The lists are all built on the same foundation (toothbrush, toothpaste, etc.), but each is tailored to meet the experience.


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Kvichak Delivers Debris Removal Vessel
Kvichak Marine Industries recently constructed and delivered the M/V John A.B. Dillard, Jr., a Debris Removal Vessel for the U.S. Army Corps of Engineers (USACE) San Francisco Bay District. The USACE San Francisco Bay District has a unique mix of duties, including keeping the navigation and shipping lanes of the greater San Francisco Bay region and its lower tributaries free of floating debris deemed as hazardous to marine vessels. This mission is especially demanding based on the diverse topography and ever changing weather conditions. This vessel will allow shorter response times, increased capacity and greater range. Key to this enhanced capability is the large aft deck, pedestal mounted grapple crane and robust 15 ton deck load capacity. This allows operators to recover large heavy objects in a single "grab" rather than the hazardous and time consuming process of towing objects to base, or reducing their size while working over the side of smaller vessels.
With an 86.5-ft overall length and overall beam of 26 ft, the all welded aluminum catamaran is powered by twin Caterpillar C-32 ACERT engines, coupled to ZF 3050 marine gears. Additional vessel control is provided by two Wesmar V2-12E double propeller bow thrusters. The hull design is from Incat Crowther Ltd of Australia.
This vessel is capable of a multitude of missions including bottom profile surveying, submerged object identification and recovery, floating command center for emergency operations and vessel assist and emergency response.
Specifications/Main particulars:
Length, o.a. 86.5 ft
Beam, o.a. 26 ft
Draft 6 ft
Fuel capacity 2,500 gal
Top speed ~27.5 kn
Two Alaska Diesel Electric Generators, 92 kW
Kinshofer Grapple & Rotator
Effer 4400 3S-L Crane