1Department of Infectious Diseases, St. Jude Children's Research Hospital, Memphis, Tennessee, USA. 2Department of Microbiology and Immunology, University of Texas Health Science Center at San Antonio (UTHSCSA), San Antonio, Texas, USA. 3Department of Hematology, St. Jude Children's Research Hospital, Memphis, Tennessee, USA.
Address correspondence to: Elaine I. Tuomanen, Department of Infectious Diseases, St. Jude Children's Research Hospital, 262 Danny Thomas Place, Memphis, Tennessee 38105-3678, USA. Phone: (901) 595-3114; Fax: (901) 595-3099; E-mail: elaine.tuomanen@stjude.org.
Authorship note: Carlos J. Orihuela and Elaine I. Tuomanen are co–senior authors.
J Clin Invest. 2010;120(2):627–635. https://doi.org/10.1172/JCI39843.
Sickle cell disease (SCD) is the most common genetic disorder worldwide, with an estimated 300,000 affected individuals born each year (1). SCD causes a chronic hemolytic anemia characterized by distorted erythrocytes that aggregate and occlude blood flow in the microvasculature. Individuals with SCD experience a wide spectrum of vascular complications, including vaso-occlusive crisis, stroke, and hyposplenism. In addition, children with SCD have a 400-fold greater risk of fulminant, lethal pneumococcal sepsis than their healthy peers or patients with other hemolytic anemias (2). This invasive disease rate greatly exceeds the 2- to 3-fold increased risk of sepsis from other encapsulated bacteria in SCD, suggesting a unique vulnerability to the pathogenic mechanisms of pneumococci in particular.
The pneumococcus is the single most frequent cause of lethal pneumonia in children worldwide (3). The bacteria is carried in the nasopharynx by approximately 20% of children at any time, a rate that is unchanged in children with SCD (4). Invasive pneumococcal disease develops during pneumonia when inflammation promotes receptor-mediated translocation of bacteria from alveoli into the bloodstream. This invasive process involves 2 steps. First, pneumococci adhere to host cells by interactions mediated by adhesins such as bacterial CbpA (5). Subsequent to adherence, bacteria bind by surface phosphorylcholine to platelet-activating factor receptor (PAFr) and cross the host cell by receptor-mediated endocytosis (6). Preexisting inflammation accelerates both adherence and invasion as host receptor expression is positively regulated by inflammation (7), a setting recapitulated in the mouse model of SCD (8–10). To mitigate the very high risk of sepsis in SCD, young children are treated prophylactically with penicillin (11). However, the emergence of resistant strains underscores the importance of identifying additional preventive options (12). We hypothesized that the antiinflammatory activity of statins could be used prophylactically to decrease baseline cellular activation in the SCD lung and vasculature and potentially mitigate bacterial invasion.
Statins, 3-hydroxy-3-methylglutaryl CoA reductase inhibitors, are among the most widely prescribed drugs in the world and are used to treat elevated levels of cholesterol and heart disease. Statins inhibit the synthesis of cellular cholesterol, resulting in a compensatory increase in cholesterol uptake by cells and concomitant decrease in plasma cholesterol (13). Statins also have potent antiinflammatory properties that are independent of their lipid-lowering ability and are suggested to be of benefit in the setting of sepsis (14–18). Currently, it is believed that statins inhibit lipid raft formation and prenylation of signaling molecules, thereby disrupting cellular signaling networks (17). Furthermore, statins have been shown to reduce inflammation in response to lipopolysaccharide (19). The potential protective effect of statins during bacterial respiratory infections has been suggested (20, 21), a setting in which the inflammatory status of the host is an important aspect of pathogenesis and disease progression.
Using a mouse model of SCD, we tested whether the antiinflammatory properties of statins could be applied to SCD to confer protection against pneumococcal challenge. Herein, we demonstrate that statin therapy reduced bacterial adherence and invasion of host cells in association with decreased activation-induced expression of host receptors. Additionally, statins protected host cells from the cytotoxic effects of the cholesterol-dependent pneumococcal toxin pneumolysin.
Simvastatin improves survival in SCD mice. The development of transgenic knockout mice that exclusively express human sickle hemoglobin has greatly enhanced the understanding of SCD (22). Transplantation of bone marrow from sickle transgenic mice effectively recapitulates the manifestations of SCD, including erythrocytic sickling, multiorgan infarcts, anemia, vascular inflammation, and heightened white blood cell counts. Similar to children with SCD, the transplant mouse model of SCD showed heightened mortality following pneumococcal challenge (9). Consistent with these previous results, all SCD mice died within 48 hours of pneumococcal challenge of the respiratory tract, whereas there were no deaths in the WT group; 40% of WT animals showed long-term survival extending beyond the 6-day duration of the study (Figure 1A). To determine the effect of statins on disease severity, both WT and SCD mice were treated with simvastatin daily for 5 days prior to bacterial challenge. Treatment with simvastatin had no significant effect on the survival of infected WT mice (Figure 1A). In contrast, the SCD mice receiving simvastatin showed a significantly delayed time to death (P = 0.023) compared with SCD mice receiving carrier only (Figure 1A). These data indicated that prophylactic statin treatment prolonged survival in the face of invasive pneumococcal disease in SCD mice.
Impact of statin therapy on survival of pneumococcal infection in SCD mice. (A) Survival of WT versus SCD mice receiving mock treatment (n = 26 and n = 24, respectively) or simvastatin (n = 27 and n = 32, respectively) following intranasal challenge with pneumococci; *P = 0.023 versus SCD, **P = 0.0001 versus SCD. (B) Blood and lung bacterial titers at 24 hours after challenge of SCD mice receiving mock or simvastatin treatment. (C) Blood and lung bacterial titers at 24 hours after challenge of WT mice receiving mock or simvastatin treatment. Each symbol represents an individual mouse; horizontal lines indicate the mean; dashed lines indicate limit of detection. *P < 0.01, mock versus simvastatin; Mann-Whitney U test.
The intranasal challenge model of infection mimics the course of human disease, including colonization of the nasal passages, aspiration of bacteria into the lungs, development of pneumonia, and translocation of bacteria from the lungs into the bloodstream, resulting in bacteremia, sepsis, and ultimately death. To further understand at what stage in the disease statin intervention was active, we determined bacterial burden in the lungs and blood following intranasal challenge. Statin treatment significantly decreased the amount of bacteria in the lungs and blood at 24 hours after infection (P = 0.0024 and P = 0.0001 respectively, Figure 1B). Moreover, in more than half of the animals, statin treatment completely blocked the appearance of bacteria in the bloodstream at 24 hours (Figure 1B). This protection was limited to SCD mice, as no significant difference in recoverable bacteria in the lungs or blood was observed in WT mice receiving simvastatin (Figure 1C). One possible explanation for the difference between WT and SCD mice is that the statins may have a more pronounced protective effect in models with significant preexisting airway or vascular inflammation such as SCD (23). This is in agreement with recent clinical data indicating that prior statin use leads to improved outcome in community-acquired pneumonia particularly in the setting of underlying chronic pulmonary inflammation (20).
Effect of statins on severity of pneumonia. The effect of statin treatment on lung pathology was examined in greater detail at 18 hours after infection, the time point at which bacteria began to enter the bloodstream in mock-treated animals. Examination of lung sections revealed a marked reduction in lung consolidation in statin-treated SCD animals when compared with the mock-treated SCD group (Figure 2). Lungs of mock-treated animals exhibited a spectrum of moderate to severe pneumonia. For example, moderate cases showed widespread areas of leukocyte infiltration into alveoli and interstitial spaces accompanied by extensive thickening or obliteration of alveolar walls (Figure 2A), while severe cases showed dense consolidation with intense leukocyte infiltration and intra-alveolar hemorrhage resulting in red and gray hepatization (Figure 2B). In contrast, lungs of statin-treated mice demonstrated only mild pneumonitis, with minimal perivascular leukocyte infiltration and either normal alveolar architecture or patchy, mild thickening of the alveolar walls (Figure 2, D and E). To confirm these observations, lung pathology was blindly scored from 1 (mild) to 5 (severe), with at least 6 representative images examined from 6 mice per group. The statin-treated group had a mean score of 2.4 ± 0.2, whereas the untreated group had a significantly higher pathology index of 4.0 ± 0.5 (P = 0.0034). Lung sections were also examined for apoptosis using TUNEL staining. A high degree of apoptosis was observed in the mock-treated group (Figure 2C), whereas the statin-treated lungs showed only an occasional, more localized apoptotic response (Figure 2F). Thus, the degree of apoptosis paralleled the severity of histopathology, with the mice having the most severe pneumonia showing the highest degree of apoptosis.
Impact of statin therapy on histopathology of pneumonia in SCD mice. SCD mice were pretreated daily for 5 days with vehicle control (A–C) or simvastatin (D–F) and then challenged intranasally with pneumococci. Lungs were harvested at 18 hours for histopathology (H&E staining: A, B, D, and E) or apoptosis (TUNEL: C and F). See text for full description; sections are representative of at least 6 mice per group. Scale bars: 100 μM.
Individuals with SCD have been shown to have increased levels of proinflammatory cytokines in the blood (10, 24, 25), and levels of proinflammatory cytokines are reduced in statin-treated patients with bacterial infections (26). In uninfected SCD mice, lung levels of IL-6 but not TNF-α, IL-1β, IL-10, IFN-γ, or IL-12 were elevated, and statin therapy decreased IL-6 from 34 ± 9.1 pg/ml to 7.0 ± 5.4 pg/ml (P = 0.046). Six hours after intratracheal challenge with nonreplicating bacteria, TNF-α increased in the lungs to 321 ± 215 pg/ml in untreated SCD mice, a change that was attenuated in statin-treated SCD mice to 154 ± 97 pg/ml (P = 0.03). No other cytokines (IL-1β, IL-10, IFN-γ, IL-12, IL-6, or IL-17) showed significant decrease with statin treatment in the infected SCD mice (data not shown). Collectively, these data suggest that significant amelioration of lung pathology in statin-treated SCD occurs with relatively modest changes in cytokines, suggesting that the statins may afford benefit through other non-antiinflammatory mechanisms.
Effect of statins on bacterial-host cell interactions. The progression of disease from pneumonia to sepsis requires that Streptococcus pneumoniae interact with host cells, particularly those of the pulmonary epithelium and the vascular endothelium. Invasion into host cells occurs when bacteria bind to PAFr and are engulfed into vacuoles; upregulation of PAFr increases bacterial invasion (6, 27). Statin treatment is reported to lower the amount of Ptafr transcript in experimental aortic aneurysms, as well as improving various aspects of endothelial function (28–31). We first documented statin effects on PAFr on endothelial cells in vitro and then examined lung and vasculature in vivo.
Using vascular endothelial cells activated with TNF-α to model the inflammation seen in SCD mice, we observed decreased expression of PAFr with simvastatin treatment as determined by immunohistochemistry and Western blot analysis (Figure 3, A and C). The decreased expression of PAFr was biologically relevant, as treatment with simvastatin also significantly lowered the number of bacteria associated with activated endothelial cells (Figure 3B). Importantly, the inhibitory effect of simvastatin on bacterial binding was reversed by the addition of mevalonate, the immediate downstream metabolic product of the acetyl-CoA/3-hydroxy-3-methylglutaryl CoA reductase reaction (Figure 3, B and C).
Effect of statin treatment on TNF-α–induced upregulation of PAFr on endothelial cells in vitro. (A) Immunohistochemical detection (purple) of PAFr expression on HBMECs activated with TNF-α with or without simvastatin. Original magnification, ×200. (B) Relative bacterial adhesion to activated HBMECs with or without pretreatment with simvastatin, and reversal by mevalonate. *P < 0.01 versus no treatment; †P < 0.01 versus statin; ‡P < 0.01 versus TNF plus statin. Error bars represent SD. (C) Detection of PAFr by Western blot analysis in activated HBMEC lysates and infected lung homogenates of SCD mice with or without simvastatin treatment. β-Actin served as the loading control. (D) Uninfected lung sections of SCD (with or without simvastatin), WT, and SCD Ptafr–/– mice showing bronchial epithelium stained for PAFr (purple). Arrowheads indicate staining in sections showing patchy expression. Original magnification, ×100.
To correlate the in vitro findings with in vivo effects in SCD, we examined lungs from uninfected mice for PAFr expression with and without simvastatin treatment. WT mice showed patchy PAFr expression on airway epithelia (Figure 3D). SCD mice (mock-treated) exhibited strong, confluent staining for PAFr, a pattern that returned to WT levels upon treatment with statin; SCD Ptafr–/– mouse lung showed minimal staining (Figure 3D). Western blot analysis of cellular PAFr levels in the infected lung homogenates of SCD mice indicated that statin treatment reduced PAFr expression (Figure 3C). These results suggested that statins downmodulate PAFr expression in vivo in the lungs of a SCD model.
To investigate effects of statin against bacterial-vascular interactions in vivo, we fitted SCD mice with cranial windows and quantitated adherence of bacterial components to the vasculature. Fluorescent latex beads coated with pneumococcal CbpA only bind to activated cerebral endothelium (5). While WT mice showed minimal adherent beads, SCD mice showed significantly increased bead adherence (Figure 4, A and B), consistent with an activated endothelium. Treatment of the SCD mice with statins resulted in a significant decrease in the number of CbpA-coated beads adherent to the SCD vasculature (Figure 4A). Negative control beads coated with BSA showed virtually no binding (Figure 4A). These data indicated that statins attenuated the bacterial-host cell adherence interaction to lung and vasculature in vivo.
Effect of statin treatment on bacterial interactions with vasculature and survival of SCD mice. (A) WT and SCD mice fitted with cranial windows were injected intravenously with fluorescent beads coated with the pneumococcal adhesin CbpA to assess activation-dependent adhesion to cerebral vessels (representative images at ×4 magnification). Negative control beads coated with BSA were also utilized in both WT and SCD mice as indicated. (B) Quantitation of bead adherence is presented as average ± SD of beads/×4 field for 10 fields per animal from at least 3 independent experiments. *P < 0.01 versus SCD mock; **P < 0.01 versus WT mock. (C) Ptafr–/– mice were transplanted with WT (squares; n = 11) or SCD (circles; n = 11) bone marrow. Survival following intranasal pneumococcal challenge was monitored after mice received mock (open symbols; n = 11) or simvastatin treatment (filled symbols; n = 11). Survival of Ptafr+/+ SCD mice (triangles; from Figure 1) is shown for comparison.
Statin activity in Ptafr–/– mice. Pneumococci utilize PAFr for invasion into host cells, and mice lacking PAFr show a dramatic attenuation of invasive pneumococcal disease (6, 32). Since PAFr is upregulated in SCD mice (9) and statins downregulate PAFr in the lungs of SCD mice (Figure 3), we hypothesized that protection against mortality by statins occurred by downregulation of PAFr expression. If that were the only mechanism, statins should not add to the protection observed in SCD mice lacking PAFr. SCD mice WT and null for Ptafr were treated with and without statin. As expected from previous studies (9), the absence of PAFr improved the survival of SCD mice upon pneumococcal challenge (Figure 4C). Surprisingly, however, statin treatment of Ptafr–/– SCD mice further improved survival (Figure 4C, P = 0.02) compared with Ptafr–/– SCD mock controls. The protective effect of the statins resulted in the mean time to death being statistically indistinguishable from that of the Ptafr–/– WT mock group (P = 0.35) but still distinct from that of the WT Ptafr–/– statin group (P = 0.04). No significant differences were observed in time to death in the WT Ptafr–/– between the mock- and statin-treated groups (P = 0.31). Bacterial titers in the blood were consistent with these findings, in that statin lowered the level of bacteremia in Ptafr–/– SCD mice but not Ptafr–/– non-SCD mice (mean log titers: SCD, 4.2 ± 1.8 vs. SCD/statin, 3.0 ± 2.2; non-SCD, 3.2 ± 1.9 vs. non-SCD/statin, 3.2 ± 2.2; n = 11 per group). These data indicate that, although downregulation of PAFr by statin was operative in the SCD setting, it was not the only mechanism of protection.
Statins protect cells against cholesterol-dependent cytotoxins. Pneumolysin is a pore-forming toxin that binds to cholesterol in lipid rafts on the host cell membrane (33). Since statins inhibit cholesterol synthesis and the formation of cholesterol-rich lipid rafts, we sought to determine whether statins modulated the ability of pneumolysin to cause cell death in vitro. Cytotoxicity of recombinant pneumolysin on endothelial cells was evidenced by rounding of spindle-shaped endothelial cells within 30 minutes, followed by detachment from the well. In contrast, simvastatin-treated cells exhibited some rounding at 30 minutes but regained spindle shape without detachment by 60 minutes. This protection was further measured by determination of the percentage of dead cells after pneumolysin treatment (Figure 5A) and by cytolysis-induced release of cytoplasmic lactate dehydrogenase (LDH) (Figure 5C). Western blot analysis of whole-cell lysates from endothelial cells exposed to pneumolysin demonstrated that statin had no effect on the amount of pneumolysin present in the cell membrane (Figure 5B), suggesting the mechanism of protection related to effects on activation of the toxin after binding to cholesterol in host cell membranes. Furthermore, toxin-induced red blood cell hemolysis was not inhibited by statins; addition of simvastatin (101%) and mevalonate (98%) alone or in combination (98%) did not alter the hemolytic capacity of pneumolysin, indicating that the drugs did not directly interfere with pneumolysin activity. Statin-mediated protection extended to challenge with live bacteria, and the addition of mevalonate counteracted the protective effect of the statin (Figure 5C). Further experiments indicated that the protective effects of the statins extended to other cholesterol-dependent cytolysins, including tetanolysin and streptolysin O (Figure 5D).
Effect of simvastatin on cytolysin-mediated endothelial cell death. (A) PI staining of HBMECs following exposure to pneumolysin. Cells were treated with increasing concentrations of simvastatin overnight (0.1, 1.0, and 10 μM, respectively) or with simvastatin (1 μM) supplemented with mevalonate. For each condition, 20,000 cells were analyzed by FACS for PI-positive staining. Percentages of PI-positive cells in controls were: mock, 1.5%; simvastatin, 0.1%; mevalonate, 0.2%; simvastatin plus mevalonate, 0.1%. Data represent the mean ± SD from 2 independent experiments. *P < 0.01 versus mock. (B) Western blot of pneumolysin (arrowhead) in HBMEC membranes with or without simvastatin and/or mevalonate pretreatment (molecular weight markers indicated). (C and D) Release of LDH as a measure of HBMEC lysis after 1 hour of exposure to living bacteria (C) or cholesterol-dependent toxins pneumolysin (C), tetanolysin (D), or streptolysin O (D). Pretreatment with simvastatin and reversal by mevalonate as indicated. LDH released from HBMEC monolayer completely lysed by 1% Triton X-100 was assigned a value of 100%. *P < 0.01 versus mock. Error bars represent SD. (E) SCD and WT mice with or without simvastatin treatment were challenged intratracheally with 500 ng recombinant pneumolysin in 0.5% glycerol. Lung histology was assessed at 5 hours. Effects of pneumolysin toxoid and 0.5% glycerol alone are shown as negative controls. Data are representative images from at least 3 mice per group. Scale bar: 100 μM.
The protective effect observed in vitro was investigated in vivo in mice pretreated with statins and subsequently inoculated with purified pneumolysin into the lungs. Histological examination of lungs 5 hours after pneumolysin exposure showed a marked reduction in lung injury in the statin-treated SCD animals compared with the mock-treated control group (Figure 5E). Administration of either PBS or a nonhemolytic mutant pneumolysin protein showed no marked lung damage (Figure 5E), indicating that statin-mediated protection was directed to active toxin. Such a mechanism should be observable also in WT animals. Despite the fact that statins had no effect on survival of WT animals (Figure 1), a significant reduction in lung injury was observed upon pulmonary challenge of statin-treated WT mice with pneumolysin (Figure 5E). In summary, statins alter cellular susceptibility to cholesterol-dependent cytotoxins (CDCs) in WT and SCD settings, but attenuation of mortality is seen only in SCD.
As a result of vascular occlusion and subsequent hypoxia, SCD is characterized by chronic activation of vascular endothelial cells, with marked increases in the expression of many inflammation-responsive cell surface receptors (10, 22). In the setting of pneumococcal challenge, chronic upregulation of PAFr promotes bacterial invasion and predisposes SCD individuals to invasive disease (7, 9, 27). Statins are potent inhibitors of systemic inflammation (16, 26, 29) and thus might serve as a prophylactic measure for SCD individuals, potentially reducing the incidence and severity of pneumococcal infection. Our data indicate that statin therapy prolonged survival in experimental pneumonia and sepsis. This protective effect arose by several mechanisms. Statins reduced PAFr expression on the chronically activated SCD vascular endothelium as well as in the lungs. This decrease was coupled to reduced bacterial adherence and invasion in vitro and in vivo. It is reasonable to hypothesize that the effects of statins in the context of SCD arise from a combination of effects on cholesterol-containing membrane rafts. These membrane microdomains organize and compartmentalize G protein–coupled receptors, such as PAFr and their signaling components. Our results indicate that statins also disrupt microdomain functions important for β-arrestin–mediated, PAFr-directed endocytosis of pneumococci. This activity would potentially also apply to infection by the many invasive bacteria that use activation of host receptors in membrane microdomains as a mechanism for endocytosis.
While the SCD mouse model clearly indicated that statins interfered with PAFr-dependent pneumococcal invasion, an additional protective mechanism was suggested by the beneficial activity of statins in infected SCD mice lacking Ptafr. As expected in this setting, bacteria failed to invade the bloodstream, indicating that the additional benefit to survival must have arisen within the alveolar compartment. Pneumococci and several other Gram-positive invasive bacteria produce a class of CDCs that bind to host cell lipid rafts and oligomerize to form pores (34). Direct testing of the effect of statins on cells treated with purified pneumolysin toxin or toxin-producing bacteria indicated that statins prevented cytolysis. The effect was not mediated by direct statin-toxin binding or decreased toxin binding to cells, suggesting that statin-treated lipid rafts contained cholesterol sufficient to ligate pneumolysin, but the interaction failed to support the pore formation required for cytotoxicity. Recent evidence indicating a role for cholesterol in toxin activation rather than cell binding (35) supports this contention. Furthermore, attenuation of CDC-induced lung damage by statins was also apparent in WT mice. This protection from injury, however, did not translate to increased survival, as only SCD mice experienced a survival benefit. The ability of statins to protect against cytolysis by tetanus toxin and streptolysin O indicate that statins might be of broader benefit in arresting tissue damage from fulminant Gram-positive toxemias.
Collectively, the multiple points of inhibition of pathogenesis afforded by statins in the context of pneumococcal challenge suggest that statin prophylaxis might not only decrease the heightened susceptibility of children with SCD to lethal pneumococcal disease but also attenuate progression of bacterial invasion in other settings of chronic inflammation, such as in individuals with chronic pulmonary or vascular inflammation that elevates the risk of pneumonia. These findings support clinical investigation of the protective efficacy of statins in children with SCD.
Preparation of bacteria and statins. S. pneumoniae strains D39, R6, TIGR4, or T4R were grown on tryptic soy agar (EMD Chemicals) supplemented with 3% sheep blood or in Todd Hewitt broth (TH) or defined semisynthetic casein liquid medium (36, 37) supplemented with 0.5% yeast extract. Simvastatin or lovastatin (10 μM; Calbiochem) and mevalonate (100 μM; Sigma-Aldrich) did not affect bacterial growth. No major changes were observed in gene expression by array despite dosage of bacteria with simvastatin at levels 10-fold greater than that used in our infection models (Supplemental Table 1; supplemental material available online with this article; doi: 10.1172/JCI39843DS1).
Simvastatin was resuspended in ethanol and sodium hydroxide as an inactivated stock solution stored at –20°C for up to 3 weeks. Immediately prior to use, the stock solution was activated by addition of hydrogen chloride and subsequently diluted in sterile PBS (38). For animal experiments, simvastatin was administered at 1 μg/g body weight via intraperitoneal injection based on previous studies (38). Mock-treated controls were injected with PBS supplemented with ethanol as for the simvastatin group. Injections were performed daily for 5 days prior to pneumococcal challenge and each day subsequent to challenge. Serum cholesterol was measured 5 days after statin intervention and was found to decrease from 107 ± 8 mg/dl to 96 ± 7 mg/dl (P = 0.012, Mann-Whitney U test), indicating that the administration route was successful. Simvastatin had no effect on bacterial growth rate or pneumolysin secretion at the concentration used in these studies (Supplemental Figure 1).
Adhesion assay. Assays were performed using human brain microvascular endothelial cells (HBMECs; ScienCell Research Laboratories). Cells were grown to 80%–90% confluence in 24-well plates (Costar Corning Inc.). Prior to the experiment, cells were treated overnight with fresh growth medium containing 1.0 μM simvastatin with or without 100 μM mevalonate (Sigma-Aldrich). This dose of simvastatin was selected based on previous investigations on the effect of simvastatin on endothelial cells (31, 39). The following day, cells were washed and incubated for 2 hours with serum-free F12 medium (Invitrogen) or F12 containing recombinant TNF-α (10 ng/ml; Sigma-Aldrich). Cells were washed and incubated with F12 containing 1 × 107 CFU/ml T4R for 1 hour at 37°C in 5% CO2. Cells were then washed 3 times with F12 to remove nonadherent bacteria, and the cells were lysed by 100 μl 0.1% Triton X-100. The number of bacteria in the cell lysate, representing adherent and intracellular bacteria, was determined by serial dilution, plating, and colony counting.
Western blot analysis. HBMECs were pretreated with 1 μM simvastatin overnight and activated with TNF-α for 2 hours, followed by lysis in 0.1% Triton X-100. To ensure equal loading, protein concentration was determined for each lysate and loaded accordingly. Lysates were run on 4%–12% NuPAGE Bis-Tris gels (Invitrogen). Proteins were subsequently transferred to PVDF membranes by Western blot and probed with antibody to PAFr (1:500; Santa Cruz Biotechnology Inc.) and against β-actin (1:2,000; Santa Cruz Biotechnology Inc.). Various concentrations and durations of simvastatin treatment were analyzed (Supplemental Figure 2), with the overnight exposure at the low dose providing the most consistent results.
Cytokine analysis. To model early events in inflammation, intratracheal challenge with 106 ethanol-fixed bacteria was utilized so as to eliminate differential bacterial outgrowth in the lung tissues. Mice were challenged after 5 days of statin treatment, and lungs were harvested at 6 hours and resuspended in 500 μl PBS supplemented with protease inhibitors. Lungs were then homogenized and debris was removed by centrifugation. Supernatants were frozen at –80°C, and cytokine levels were measured in duplicate for each sample using a Milliplex cytokine assay (Millipore). A standard curve was generated for each cytokine to ensure that values were within the linear range of detection for the assay. Two independent quality controls of cytokines of known concentration were also used to ensure validity of the data.
Immunohistochemistry. Mice were perfused with 3% paraformaldehyde, after which lungs were excised and kept in 3% paraformaldehyde overnight. Lungs were then transferred to 30% sucrose for 3–5 days, frozen in OCT freezing medium (Tissue Tek, Electron Microscopy Sciences) in dry ice, and sectioned (20 μm). For PAFr staining, sections were submerged in 0.6% hydrogen peroxide for 30 minutes, washed, permeabilized with 0.4% Triton X-100 for 20 minutes, and washed. Sections were blocked with 1% BSA and 5% goat serum for 30 minutes and incubated with primary anti-PAFr antibody (Santa Cruz Biotechnology Inc.) at 1:100 overnight at 4°C. After washing, sections were incubated with goat anti-rabbit IgG-HRP (Bio-Rad) at 1:500 for 1 hour at room temperature. Sections were washed and visualized with VIP substrate kit and counterstained with methyl green (Vector Laboratories).
Generation of SCD mice. Lethally irradiated 8-week-old female C57BL/J6 mice (The Jackson Laboratory) were transplanted as described previously with 2 × 106 bone marrow cells from either BERK SCD mice or WT mice (40). Baytril (enrofloxacin; 2.27% solution, diluted 1:100 in drinking water; Bayer) was administered as an antimicrobial prophylaxis for 3 weeks after transplantation. One hundred days after transplantation, the sickle phenotype was confirmed by hemoglobin cellulose acetate electrophoresis of red cell lysates (41). Complete blood counts were determined to ensure that the white blood cell number, hematocrit, hemoglobin, and red blood cell distribution were within expected values for both WT and SCD mice.
Intravital fluorescence microscopy. All experiments using animals were performed with the prior approval of and in accordance with guidelines of the St. Jude Institutional Animal Care and Use Committee. Lack of availability of intravital microscopy in BL2 animal facilities precluded the use of fluorescently labeled bacteria; hence, one of the major proteins required for pneumococcal adherence to the brain endothelium, CbpA, was selected for these studies. Cranial windows were installed overlying the cerebral cortex as described previously (5, 42). WT and SCD mice were immediately imaged after surgery due to challenges in the long-term survival of the SCD mice after the procedure. Mice with cranial windows were anesthetized (2.5% isoflurane; Baxter Healthcare), immobilized on a stereotaxic frame, and placed under an industrial-scale microscope (model MM-11, Nikon) with a camera assembly and bright-field and fluorescent light sources. Yellow-green fluorescent 2-μm microspheres (Polysciences) were coated with recombinant CbpA according to the supplier's instructions and injected intravenously (2.5 μl/g body weight of a 108 sphere/ml suspension). Video images and still pictures were captured during injection and at 3 and 6 minutes after injection at an excitation wavelength of 490 nm using MetaMorph software (Molecular Devices Analytical Technologies). To ensure that beads were adherent and not merely passing through the vasculature, imaging was done at multiple time points, and only beads remaining stationary from the previous frames were included in the enumeration of adherence.
Mouse challenge. Bacterial challenge studies were performed as previously described (43). All mice were maintained in BSL2 facilities, and all experiments were done under inhaled isoflurane (2.5%). Bacteria (strain D39) were introduced by intranasal administration of 106 CFU in 25 μl PBS. Mice were monitored daily for signs of infection, and differences in time to death were compared via Kaplan-Meier survival estimates. Bacterial density in blood was quantified at 24 hours after infection and compared by Mann-Whitney U test. Bacterial density in lungs was measured by harvesting lungs at 24 hours after infection followed by homogenization in 500 μl PBS and subsequent serial dilution and enumeration of CFU. For histopathology, lungs were collected at 18 hours after infection and immediately perfused with 10% formaldehyde and embedded in paraffin. Sections were stained with H&E for tissue morphology and with TUNEL for indications of cell death (ApopTag Plus Peroxidase in Situ Apoptosis Detection Kit, Chemicon International).
Effect of simvastatin on cytotoxin activity. The ability of pneumolysin to kill statin-treated cells in vitro was determined using fluorescence microscopy to detect acridine orange (AO) and ethidium bromide (EtBr) staining (44). HBMEC monolayers in 24-well plates at 20%–40% confluence were treated overnight with 1 μg/ml simvastatin, washed, and incubated with serum-free medium containing 0.5 μg/ml pneumolysin, AO (100 mg/ml), and EtBr (100 mg/ml). At the designated times, images of the cells were taken using a Leica DMIM fluorescence microscope. The percentage of dying cells was determined by dividing the number of necrotic and apoptotic cells (orange) by the total number of cells present per field of vision (orange plus green). At least 8 fields were counted from 3 independent experiments.
The ability of pneumolysin, streptolysin O, or tetanolysin (Sigma-Aldrich) to form lytic pores was determined by the release of intracellular LDH into the cell culture medium (45). His-tagged pneumolysin, nonhemolytic pneumolysin toxoid (34), and streptolysin O (46) were expressed in E. coli BL21(DE3) induced by 1 mM IPTG for 4 hours and purified by metal affinity chromatography. Streptolysin O expression plasmid was provided by R. Tweten (University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA). Endotoxin was removed by gentle mixing with Endotoxin Decontamination Beads (Thermo Scientific) for 2 hours, and its removal was confirmed using Lonza's colorimetric LAL assay. Endotoxin levels were similar for all constructs, approximately 5 EU/ml of purified protein. This corresponds to approximately 0.001 EU for the in vivo lung administration. HBMEC monolayers at 80% confluence were treated with simvastatin overnight, washed, and incubated at 37°C in 5% CO2 with serum-free medium containing 1 μg/ml recombinant cytolysin for 90 minutes. The supernatant was removed and centrifuged to remove cellular debris, and LDH in the supernatant was measured using the LDH-Cytotoxicity Assay Kit II (BioVision). Pneumolysin cytotoxicity was quantified as a percentage of the total LDH in the entire monolayer determined by complete lysis of control monolayers with 1.0% Triton X-100. Both cell viability and LDH experiments were performed in triplicate, with a minimum of 8 wells quantified for each lysin per experiment. Values for each experiment were based on the average of a minimum of 3 separate wells per experimental condition tested. Binding of toxin to cells was determined by Western blot analysis using rabbit anti-pneumolysin antibody (1:20,000).
Hemolysis assay. The hemolytic activity of pneumolysin and streptolysin O was determined using washed, defibrinated sheep red blood cells (Hemostat Laboratories) suspended in PBS to a final concentration of 3%. In a 96-well plate, 100 μl of the 3% blood solution was added to 100 μl of serially diluted toxin and incubated at 37°C. The unlysed blood cells were centrifuged at 200 g for 5 minutes. The supernatant (180 μl) was then transferred to a new plate, and the absorbance at 540 nm was measured. Controls included PBS only (0% lysis), simvastatin alone at 1–10 μM (0% lysis), mevalonate alone (0% lysis), and distilled water (100% lysis). Hemolytic activity was determined by the amount of protein required to lyse 50% of the cells within the allotted time.
Live/dead staining. HBMECs were seeded in 6-well plates and treated overnight with 0, 0.1, 1, or 10 μM simvastatin. The next day, cells were treated with serum-free medium containing hemolytic units lysing 50% of cells (HU50) of 3 μg/ml of pneumolysin for 4 hours at 37°C in 5% CO2. After 4 hours, the cells and cell culture supernatants were collected. Washed cells were suspended in 500 μl HEPES buffer and 5 μl propidium iodide (PI) and assessed for the percentage of live/dead cells on a BD FACSCalibur. Control staining of cells treated with simvastatin, mevalonate, or simvastatin and mevalonate in combination did not exhibit levels of PI-positive staining above background of the untreated controls (data not shown).
Statistics. Kaplan-Meier analysis and generalized Wilcoxon tests were performed to determine statistical significance of mortality data. Mann-Whitney U tests were performed to determine statistical significance of bacterial lung and blood titers, and adherent bead counts for cranial windows. Student's 2-tailed t test was used to determine significance of cytokine levels. A P value less than 0.05 was considered statistically significant.
We thank the Animal Imaging Facility at St. Jude Children's Hospital for excellent technical assistance with the cranial windows, Heather Forrest for histological scoring of lung pathology, and Karla Gorena of the UTHSCSA Flow Cytometry Core Facility for data processing and analysis. This work was supported by National Institute of Allergy and Infectious Diseases grant 27913 and National Heart, Lung, and Blood Institute grant U54HL070590 to E.I. Tuomanen, National Institute on Aging grant AG29313 to C.J. Orihuela, and the American Lebanese Syrian Associated Charities. Funding agencies had no role in conduct of the study or interpretation of the data.
Citation for this article: J Clin Invest. 2010;120(2):627–635. doi:10.1172/JCI39843.
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OUR YOUNGEST AGE DIVISIONS
2017 & 2018 BIRTHS
PEE-WEES DIVISION
2018 Birth Boys and Girls
Players meet on SUNDAYS at Gladwyne Playground
Boys sessions are at 9:15, 11:45 and 2:15
Girls sessions are at 10:30 and 1:00
ATOMS DIVISION
2017 Birth Boys
Players meet on SATURDAYS at Gladwyne Playground
Boys sessions are at 11:45 and 2:15
NEUTRONS DIVISION
2017 Birth Girls
Girls sessions are at 10:30 and 1:00 PM
The emphasis of our Youngest Age Divisions is on enjoyment of the game and basic skill development.
Players meet once a week. They will be led through a 25-30 minute skills clinic. This will be followed by games of 4 vs. 4 on scaled down fields to maximize player participation. Games will last approximately 30 minutes.
A size 3 ball is used.
These age groups use a "pickup soccer" format where players are randomly placed on different teams each week. This is done in order to minimize the pressures of winning and losing, thereby allowing each child to enjoy the game in a stress-free environment.
Heading a ball and slide tackles are illegal in these age groups.
Because of the small numbers of players on each team, we need to have a large number of coaches in each division. Over 90 percent of our coaches are parents of the players. Parents are asked to give consideration to volunteering to coach. You do not need to have playing experience to be a coach. At LMSC, we set aside a large percentage of our budget for coaching materials, coaching clinics, etc. We believe that the best way to have a successful program is to have coaches who are enthusiastic and understand their roles and responsibilities.
Atoms, Neutrons and Pee-Wees Divisions
At Gladwyne Playground
Our Gladwyne Playground Directors
Franky Carrozza and Henry Farr
CLICK HERE TO SEE A VERY SHORT VIDEO OF OUR PEE-WEES
STAFF COACHES FOR OUR YOUNGEST AGE GROUPS
Franky Carrozza - Franky was a full time preschool and kindergarten teacher for 10 years. She played youth soccer for the New-Hope Solebury Soccer Club. She later played field hockey and lacrosse at Lehigh University and has coached soccer, field hockey, lacrosse and floor hockey for a wide range of age groups. Franky also coached in the US Field Hockey Futures Olympic Development Program.
Henry Farr - Henry is co-head coach for the LMSC Wolves, our Under 9 boys travel teams. He is also the assistant coach for the Harriton High School Boys Varsity. Henry played high level travel team soccer in Lower Merion Soccer Club for many years, helping lead his team, the LMSC Magic, to two state championships. He was a standout soccer player at The Shipley School and later played soccer at Trinity College. Henry started out at LMSC as one of our youngest players and is now helping lead the Pee-Wees, Atoms and Neutrons Divisions.
Art Hernandez - Art joined LMSC as the Director of the club's Developmental Academy. He is also head coach for the LMSC Mayhem, our Under 16 boys team and runs our "Little All-Stars" program for 2 and 3 year olds. Coach Art was raised in Los Angeles, California and has been living in Chester County for over 20 years. He has coached in this area for the past 12 years and has developed under terrific coaches and mentors. He has coached and trained teams that have won State Cups, along with National and Regional competitions and tournaments. Coach Art is very vested in his coaching education and, in addition to being a current A-License candidate, he has all his grassroots licenses through US Soccer. Additionally, he holds his Advanced National, Advanced National Goalkeeper and FUTSAL diplomas from United Soccer Coaches. He has attended courses through ANEFS and TOVO, based out of Spain. In addition to coaching, he currently volunteers his time to collaborate and network with various soccer coaching groups (locally and nationally) to produce and host different online content.
Anthony Haines - Anthony is the current head coach for our Under 9 A Rovers and Under 10 B Irm�os Red boys travel teams. He is the head coach for the Harriton High School Boys Junior Varsity team and he also helps coach LMSC summer camps, Developmental Academy, indoor training programs, and our youngest age groups in the fall and spring season. Anthony was named the Boys Travel Team coach of the year in Fall 2019 and Intramural Program coach of the year in 2017. He currently holds his US Soccer D coaching license. Anthony grew up playing for LMSC Magic, helping the team win two outdoor and three indoor State Championships. Anthony played varsity soccer at Lower Merion High School for three years where he was captain his senior year, receiving All Main Line second team and All Central League honors. He played on the Club soccer team at Temple University where he graduated with a BBA in both Economics and Finance at Temple's Fox School of Business. Anthony is the LMSC Director of Operations for LMSC. He primarily acts as travel team registrar, compliance officer, and database manager utilizing Amazon Web Services RDS for PostgreSQL.
Web Page Designed And Maintained By: Biff Sturla

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Chicago Sun-Times works in the Printing & Publishing industry, specialising in Chicago news, Custom Content, Digital Solutions, Marketing Services, Politics, Sports. The company was incorporated in 1948 and is located in Chicago, United States. Chicago Sun-Times has an estimated revenue and employee in the range of $10 to $50 million and 201-500 respectively.
The purpose of this report is to provide vital facts about Chicago Sun-Times and help in key decision making process, via market and competitive benchmarks and key metrics. The report will surely add value to your business development process, deal sourcing and should save countless hours of research and analytical work for the buyer.


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The University of West Florida, also known as West Florida and UWF, is a mid-sized public university located in Pensacola, Florida, United States. Established in 1963 as a member institution of the State University System of Florida, the University of West Florida is a comprehensive research university without faculties of law or medicine, a designated space-grant institution, and sits on the third largest campus in the State University System
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Celebrating the life and work of Luise Rainer (1910 – 2014)
Art & Exhibitions
Candid Shots & Press Photos
Studio Portraits & Publicity Photos
Gallery: Escapade (1935)
Gallery: The Great Ziegfeld (1936)
Gallery: The Good Earth (1937)
Gallery: The Emperor's Candlesticks (1937)
Gallery: Big City (1937)
Gallery: The Toy Wife (1938)
Gallery: The Great Waltz (1938)
Gallery: Dramatic School (1938)
Gallery: Hostages (1943)
Gallery: Television
Gallery: Theatre
Gallery: Magazine Covers
Luise (2010)
by Hans Ulrich Obrist
This article is taken from AnOther magazine (Spring 2010). Cover portrait: Marion Cotillard by Craig McDean.
The celebrated 1930s actress Luise Rainer was the first to win an Oscar in two consecutive years. But Rainer fell out with "brainless" Hollywood, famously giving away
her second golden statuette to a removal man. Here, on her 100th birthday, Rainer talks exclusively to co-director of the Serpentine Hans Ulrich Obrist about studying theatre with Reinhardt, standing up to Brecht, and turning down Fellini.
My interview project is a kind of transdisciplinary school, incorporating lots of different sub groups. The beginning of this Rainer adventure started five years ago, when the legendary German artist Rosemarie Trockel told me she thought it was important for me to focus on pioneers, and particularly the idea of someone's eyes having seen a whole century. Very often we don't know where these great pioneers are because they live discreetly. Although I was familiar with some of her work from the 30s, I had no clue that Rainer was still active and about to celebrate her 100th birthday. And I certainly had no clue that she was my neighbour in London
We started what has become a series of three interviews, which took place at her astonishing home in Kensington, London, once also home to another Hollywood Legend, Vivien Leigh. Wolfgang Tillmans told me it had been one of his unrealised projects to photograph Rainer, so it was wonderful that that this story produced this interview but also this new portrait. I found that Rainer had met almost everyone of renown in theatre and cinema in the 20th century, and not only that, but she remembered everything with a photographic memory. It's amazing that someone who met all the greats is still alive to tell us about it first hand.
Luise Rainer, photo by Wolfgang Tillmans, taken for this article to mark her 100th birthday
After her career in theatre, Rainer was discovered by Hollywood, but she always said that nothing was as exciting as Berlin in the 1920s and 1930s. Rainer's career has been far from linear, with many unrealised projects. If she didn't agree with a role, she felt it was okay to say no, and I think this is what makes her such a fascinating actress. The things you don't do are as important as the things you do, which I think is true for all of us. How can you tell a life in an interview? Rainer has said it would take her a century to tell it all. This interview was about a testimony of a century, but it's not about nostalgia. The future is always made out of fragments of the past.
Hans Ulrich Obrist: I read a wonderful interview with you in the German newspaper, Die Zeit.
Luise Rainer: You know, they just write, write and write whatever they want, it's crazy. I am not an actress, I am just a person.
HUO: I was wondering, what were your beginnings, how were you introduced to theatre?
LR: I am asking about you and you want me, immediately to tell you about my life?
HUO: Well, I can tell you about me more, but I thought it might bore you?
LR: What are you writing?
HUO: I write about contemporary art and architecture, but it's a transdisciplinary project. If we want to understand the forces in art it's important to understand what's happening in other fields.
LR: I'm not contemporary art, I am last-century art.
Luise at her Strathmore Apartment home, designed by architect Richard Neutra.
HUO: You have a lot of artworks on the wall.
LR: Well, it is a normal thing, artists love beauty. I knew a lot of marvellous architects, Richard Neutra; I knew the big one in California, Walter Gropius, and I knew Frank Lloyd Wright.
HUO: You knew him personally?
LR: Yes. I lived in a house by his son, and Frank lloyd Wright said to me, 'how can you live in a house by that untalented man!'
HUO: Amazing!
LR: So, what do you want to know?
HUO: I wanted to know how you became introduced to theatre. Did you have an epiphany, a revelation?
LR: It was a lot more human, I had an enormous urge to give out, but I didn't know in what form. I grew up in Hamburg, Germany. My father was wonderful, but very strict, my mother was a fantastic pianist. My father and I went to the music rehearsal in the evening every Monday but I was never permitted to go to the theatre, because I was too young. In the 20s we didn't have anything, television didn't exist. Finally, when I was 14, a play called Der Kreidekreis, in English Circle of Chalk, and I thought, 'I can do that too'. Of course, I could not let on, my father thought a young girl should go to finishing school. You must understand, these were the olden times. When my girl's college finished, I was 16 at the time, parents were invited and various pupils were brought to the front. The head of the college congratulated me for having the ability to be a leader. My father was quite impressed with what this woman said about me, and they permitted me to have a desire, What wish could I have? I was very clever, and I knew that Berlin was a fortress for theatre.
HUO:With Max Reinhardt
LR: With Reinhardt. We had relatives in Berlin, and I asked if I could go and visit. I didn't go as a young girl was supposed to, to museums and what not, I made my way to the Reinhardt Theatre. I found out that Reinhardt was on holiday, but I was told there was an audition in the Kammerspiele [a theatre in Berlin that Reinhardt built in 1906]. I made my way there, and about ten young people were waiting in a room. I was horrified because I was very unsophisticated and very thin, and dressed like a girl of 12. One by one the people were called out for the audition, and the last one was me. I sat down and I forgot that I had a straw hat on, with a rim. And the straw hat went down further and further down over my face, I forgot all of my lines, and all I could say was 'yes, yes, yes'. There was a big laugh from the auditorium. There were three men sitting there, just like shadows in the dark, and they said, 'girly, you had better go home and learn your lines!' That was my first experience. Let's get a drink right away. We have sherry.
HUO: Wonderful. That's a great idea, I love sherry.
LR: Me Too.
HUO: Is it your favourite drink?
LR: No, I don't drink very much, but actually, I really like champagne.
HUO: It would be wonderful to talk about Brecht because I think you knew him very well?
Luise (centre) on stage in an unknown Max Reinhardt production, early 1930s.
LR:Not very, but I knew him well. Brecht asked me to help him to come to America. At the time I had been successful in Hollywood and became, I hate the word famous, but I was very known. So he came, and one day we went for a walk along the beach and he said, 'I would like very much to write a play for you. What do you think?' I said, 'of course.' In the first play I saw in my life Gustav Gründgens, who is a famous German actor, was playing the main part of Joan and I was fascinated. I mentioned this play to Brecht, and Brecht, I can still see him doing it, spread his arms and said, 'it isn't possible! I was going to write a play about that same story, but I will write my version.' Which was The Caucasian Chalk Circle. And that is what happened.
HUO: But you never played in it?
LR: I went into the Second World War. I had a producer and I said 'look, the man has no money, I would like you to pay him while I'm away because he is going to write that play. I had had Malaria in Africa, and then I went to Italy after the troops, I came back to New York and then, low and behold, I got Jaundis. I was very sick, and that poor producer told me, 'Miss Rainer, Miss Rainer, I don't know what to do. I paid this man Brecht every week and I never saw a single page of what he wrote.' So I called Brecht and said, 'I must see what you have written.' He sent me just two double-spaced pages, He called me a day or two after and he said, 'what do you think?' and I said, 'what do you mean what do I think? I think that it's nothing, because what you have sent me is nothing!' And he was, of course, very taken aback and said, 'why Miss Rainer, maybe we should talk about it.' A friend of mine, a young Danish ambassador who had to go on an assignment, had an apartment, so I arranged for Brecht to stay there. Anyway, I came to that apartment and it was like a French Comedy, I don't know how many doors there were, but it seemed to me when I arrived, out of every door, a lady fled! I thought he was completely nuts. He said 'Elisabeth Bergner' – who was at the time a famous actress – 'would be on her knees to me for this.' And I said, 'Brecht, I will not be on my knees to you, take your play and do whatever you want with it. I don't want to be in it'. And that is The Circle of Chalk. [laughter].
HUO: One other theatre great with whom you worked was Pirandello?
LR: That was before I came to Hollywood. He was a marvellous man, like a dancer, he was, like his plays, quite pixelated. He was full of beautiful stories, he and Reinhardt were of course two great people, that's how I met him. He travelled with us until we reached Holland, then he went to Milan, and I stayed in Holland doing the play Six Characters In Search of an Author.
HUO: So you acted in Pirandello's plays?
LR: In Six Characters.
HUO: I wanted to ask you how you went from you career in theatre to the cinema? What was your first movie?
LR: My first film in America was called Escapade [directed by Robert Z Leonard, 1935], with William Powell, and it was a very lovely film. Unfortunately it was never shown. Many of my films, even The Good Earth [1937] were just so timely, so they were not shown.
William Powell and Luise Rainer in Escapade (1935)
HUO: But the BFI London showed some of your films to celebrate your 100th birthday this year.
LR: Yes it was very nice, wonderful.
HUO: How was it for you to come from the theatre into the cinema?
LR: No difference, except I would turn around and stand with my back to the camera. Dear William Powell, he just gently turned me. That was the only difference to me between theatre and films: position. You see, I worked from inside out, it was not putting on an act, I must feel what I did, and when you feel what you do, you can connect with people, they listen to you. I wanted to give, that was my whole life, always.
HUO: Before you made movies, and before you were an actress, what was the first movie you saw?
LR: One of the first was A Farewell to Arms [directed by Frank Borage, 1932] with Helen Hayes, a beautiful film.
HUO: Did you find racism in Hollywood?
LR: No, I was never a political person. I knew good from bad, but it didn't go into politics, it went into human behaviour and human feeling. I was also married to a man [playwright Clifford Odets, 1937-1940], who was very determined in certain political directions, but I never accepted it. I remember once he gave me Marx to read, I read a few pages and I gave it back to him.
HUO: I read somewhere that you were involved in some political activist committees in the 1930s?
LR: I was very Left, as I felt very deeply about the difference between good and bad, which is an idiotic definition. I felt very strongly about the difference between poor people and rich people. But it didn't go into politics, I didn't transfer it.
HUO: So you didn't adhere to a party?
LR: It was purely an emotional concept.
HUO: In the war you did a lot of travelling to gather information on troop moral, in Italy and Africa, What was your position in the war?
LR: I travelled alone with a minister, he did all the religious side, and I the social welfare. Wherever I was stationed, and I went from place to place; from the islands over to Africa, I found out what the men were asking for, and I was able to report back what they were in need of, but much more, in the evening I would do my part as an actress and I did scenes from plays I had done, to entertain.
HUO: Do you have any unrealised projects?
LR: Yes, many. I broke my contract with Louis B Mayer, who did not permit me to do
One that got away: Luise coveted the lead role in MGM's biopic, Madame Curie.
certain projects. What I wanted to do would have been the role of Marie Curie [Madame Curie, directed by Melvyn LeRoy, 1943], or For Whom the Bell Tolls [1943], but I couldn't do either of them. Much, much later came Love is a Many-Splendored Thing [1955] by Han Suyin, which was a marvellous story. I'm even mentioned in her novel before she knew me, but I never was permitted to do any of them. I had all kinds of offers, Fellini also, that I didn't do.
HUO: You met Fellini?
LR: Yes, that's big story.
HUO: Which film did he propose to you, Fellini?
LR: He wanted me to be in La Dolce Vita [1960].
HUO: Wow.
LR: I didn't want to be in it.
HUO: Why?
LR: I met Fellini in Rome at a lunch party and he took a great shine, not to me as a woman but to me as a person, or an actress. We became great friends, and on that same day, after lunch, he showed me around Rome. A few months later he wrote to me saying: 'you will receive some papers from me.' Like a fortune teller. It was Christmas time, and a great piece, which he later made into La Dolce Vita was sent. I couldn't make head nor tail of it, which is typical for Fellini. He works sporadically and takes practically whatever the actor or actress does out of their mouths and puts it into his story. I wrote back saying, 'forget it.' But he didn't forget it! I was bombarded with telegrams.
HUO: He sent you telegrams?
LR: Yes I have a whole stack of them. He said, you know Italians, 'I need your poetic face' and whatever nonsense. He was absolutely definite that I be in the play. When I was back in London he sent a wire saying, 'I invite you for lunch in Rome, I'll pay your round trip'. I went, and Fellini said, 'Luise, I want you to write your own scene, and I did. In La Dolce Vita, every woman he meets, he goes to bed with, but I said there should be one woman that says, 'look, if you want to be a writer, why don't you write, instead of all the time whoring about?' That was the essence of the scene I wrote. It was a very poetic and beautiful scene. So I made a contract for La Dolce Vita, I was in Rome to do the scene, I think it was in July, and it was terribly hot. It was delayed an delayed, and I was melting
Marcello Mastroianni, Anouk Aimée, Luise Rainer, Federico Fellini, Anita Ekberg and Yvonne Furneaux – the would-be cast of 'La dolce vita'
away. Every time I contacted Fellini, I said, 'Federico, when are you going to make my scene?' He said, 'very soon, but first I have to make that scene and that scene…' One day he said to me, 'Luise, I am going to look for a location for my next scene, come with me.' We were driving out to the country, he always has these funny locations somewhere, I don't know how many cars we had, we were like a funeral procession, I aid to Fellini, 'so when are we doing my scene?' He said, 'we are doing your scene, but he has to fuck her like the other women', and I said, 'no, I am telling you that I feel that one woman should say to him go to hell!'
HUO: In both stories of Brecht and Fellini you gave resistance.
LR: Because it was against my scene.
HUO: Tell me about your experiences with The Good Earth.
LR: I knew that Louis B Mayer was very much against it because he wanted me to be very much one of his… let's call it 'beautiful horses' instead of a little Chinese woman. Irving Thalberg, who was a very important part of Metro [Metro-Goldwyn-Mayer film studios] who unfortunately died in the making of that film, insisted on me doing the part. When I read he script I thought, 'my god, the woman doesn't talk at all, I will be a hilarious bore! What am I going to do?' So I went to Thalberg's office and said, 'there is another actress dying to play the part of O-Lan, and I think she would be far better then I am', and Thalberg took me by the scruff of my neck and pushed me out of the door. I did it, as I have said before, it is all I do. Today I could do it exactly the same, no difference. When I did my last film The Gambler, the director said, 'you can still do it,' and I replied, 'I could still do it for the rest of my life!'
HUO: You could do it right now?
LR: People can identify themselves with you, and that is what I wanted. Like when you read a book and you say, 'oh, that character, I know her, I wonder what the character is going to do next'. If I can give those ideas to any body, so that they can identify that is what I hoped for. I can't tell you more.
HUO: And that is why you never stopped, you came back to the cinema in 2003, with the wonderful appearance in The Gambler. You never stopped.
Luise with Dominic West in The Gambler (1997).
LR: It was 60 years after I had done my last film, but also I did a stage play in New York, and I did an Anton Chekhov.
HUO: I wanted to ask you about your drawings and collages.
LR: Well there was a period when I painted, and it came very easily, I had a one-woman show here in London. Mainly my collages were of the mountains, we used to be climbers, my husband an I [Luise's second husband, publisher Robert Knittel, 1945-1989]. Everything was so fabulous. We would pick flowers… two or three o'clock in the morning, we got up and went walking into the mountains.
HUO: What is your favourite movie you did?
LR: That's like asking if I had eight children, 'which one do you love best?' I have no favourite, but I love most what gave the most to people, which is probably The Good Earth, The Great Waltz or The Great Ziegfeld. The film I really liked but was not appreciated is The Toy Wife [1938].
HUO: Yes, it was dismissed by most people, but I think it was a good film. You said that the Academy Awards you won were a bad thing for you.
LR: Yes, it is very different to stand on a pedestal, because it goes back and forth. It's very dangerous and I didn't like it.
HUO: I have only a couple more questions to ask.
LR: We haven't talked about 100,000 things.
HUO: No, because your life is so rich, it would take 100 years.
LR: It is only my life, but I can't remember the details. There has been so much.
HUO: There is a lovely little book by Rainer Maria Rilke called Letters to a Young Poet. What would your advice be to a young actor or actress?
LR: To observe, and to really watch life, because only out of life can you create. Columbia University in New York asked if I would teach acting and I said, 'I would wring their neck!' Why? Because you can't teach acting. You can teach speech, movement even, but acting, it comes out of your experience.
HUO: We also haven't spoken about your other artistic endeavours; your dancing, for example.
LR: I had an enormous search inside, I had to express myself and I wanted to, it didn't matter what it was… I didn't think of acting. I danced. I modelled, I made little things, I had started drawing and things like that already as a youngster, I had to get it out of myself. It sounds idiotic, because right now I cannot tell you what I felt.
HUO: No, it makes perfect sense.
LR: I had written a lot and it all went to Boston University.
HUO: And we contacted the university, as you recommended, and they agreed that we could publish some documents but they said that we should ask you which documents you would like us to publish?
LR: I can't tell you. I have a heap of nonsense that was put together in an enormous box, and sent to the Boston University archives, and I don't know which is what anymore.
HUO: Are there any letters, correspondence from the people you knew or met in the business?
LR: No, I didn't have an organised archive like that. They are little notes that I wrote here and there, or every night I would remember certain things and I would get up and start writing, but I didn't make the extraordinary effort of a writer. I just had ideas and I put them together, well that is what writers do obviously. But I have so many personal things, i don't even know what is in that archive. They are all my love stories and I had so many, I was always in love, all my life. So, I can't tell you what to take from the collection.
HUO: But that's so great.
LR: I have of course many particular things, I wasn't joking when I said all my life was love. It was, and there was also a great deal of sorrow. It was living, like everybody. Life is not even, it depends how sensitive you are, and how observant and how many human beings can enter your life or touch you, or whatever happens. You, know, life is full, it is enormous, and I sniffed in many directions.


WORSHIP LEADER, Paul Wilbur, emphatically states, "I was born for this project!" Paul, who attended synagogue before his conversion, had the opportunity to learn about live ministry as a member of the contemporary Christian music group, "Harvest."
Paul later spent ten years leading worship in a messianic congregation, and founded the successful messianic music group, "Israel's Hope" along with Marc Chopinsky and Rene Bloch.


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FP | National Govt: UNP submits resolution to parliament Sec. Gen.
National Govt: UNP submits resolution to parliament Sec. Gen.
Minister and Leader of the House, Lakshman Kiriella today submitted a resolution to the Parliament Secretary General on the formation of a national government thus enabling it to increase the Cabinet of ministers to 48 and the deputy, state and non-Cabinet ministers to 45.


Jesse S. By: Jesse S.
I poured the coke from the original coke bottle from the top right and it must have gave it this effect. I was lucky enough to capture it!
Strobist: Nikon Sb-24 Fired behind subject w/ diy gridspots. 85mm zoom setting at 1/2 power.


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Places to Visit around Georgetown, Ontario
Great War Flying Museum
13691 McLaughlin Road, Brampton, ON, L7C 3L7, Canada
Come visit us at the Great War Flying Museum, Canada's only Great War flying aircraft museum. We have airplanes, uniforms, medals and much more.
Lorne Scots Regimental Museum
2 Chapel St, Brampton, ON, L6W 2H1, Canada
The Regimental Museum reflects the strong history of The Lorne Scots through the display of unique artefacts and telling images. Uniforms, weapons, musical instruments, maps, medals, and documents trace the heroic adventures of the thousands of soldiers who have...
Peel Heritage Complex
9 Wellington St. E., Brampton, ON, L6W 1Y1, Canada
he Peel Heritage Complex is generously funded by the Region of Peel with additional support from the Ontario Arts Council and the Ontario Ministry of Culture.
Crawford Lake Conservation Area
3115 Conservation Road , Campbellville, ON, L9T 2X3, Canada
The pristine waters of Crawford Lake have drawn people to its shores for hundreds of years. The rare lake, with surrounding boardwalk, is nestled in lush forests atop the stunning Niagara Escarpment where visitors can watch soaring turkey vultures glide through the...
Halton Region Museum
5181 Kelso Rd. RR3, Milton, ON, L9T 2X7, Canada
The Halton Region Museum is located in the Kelso Conservation Area in Milton, Ontario. The museum is focused on the area's cultural history, and also includes a library and archives of local historical documents and genealogical materials.
Country Heritage Park
8560 Tremaine Road , Milton, ON, L9T 2Y3, Canada
Country Heritage Park, an interactive heritage park depicting agriculture and rural life over the last 150 years, is located on an 80 acre site with over 30 exhibit buildings and 20,000 artifacts. Nestled in the shadow of the Niagara Escarpment south of Highway 401,...
Halton County Radial Railway
13629 Guelph Line, Milton, ON, L9T 5A2, Canada
Since the beginning, the vision of the HCRR was to inform, educate and inspire the public about the electric railway history of Ontario and Canada. Today, the museum displays and operates a variety of historic streetcars, radial cars and work cars, and maintains a...
Old Brittania Schoolhouse
5576 Hurontario Street, Mississauga, ON, L5R 1B3, Canada
"The Britannia Schoolhouse is recommended for designation on the architectural grounds that, in design, it represents a typical nineteenth-century Ontario schoolhouse and, in its attention to details such as the blind rose window, the imbrication and the decorative...
Blackwood Gallery
Kaneff Center, University of Toronto Mississauga, 3359 Missisauga Road North, Mississauga, ON,...
The Blackwood Gallery seeks to be a catalyst for current debates, and a laboratory for contemporary artistic and curatorial experiments. Through its activities, it aims to disturb preconceptions, foster discussion, and engage the intellect as well as the senses....
Art Gallery of Mississauga
300 City Centre Drive , Mississauga, ON, L5B 3C1, Canada
The Art Gallery of Mississauga (AGM) is a public, not-for-profit, art gallery generously sponsored by the City of Mississauga, the Ontario Arts Council, the Canada Council for the Arts, the Ontario Trillium Foundation, private citizens and its membership. The AGM is...
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Fun in New Hamburg


to Edith a home worthy of her charms, her virtues, and her love.
almost immediately after his brother had quitted London.
persuaded Coningsby, whom he saw depressed, to dine with him alone.
must be a harmony, a keeping, in disbursement, which very few men have.
is let alone. But the instant you have a large fortune, duties commence.
after reflection, they evinced some intention of plundering the house.
black, adorned with the escutcheon of the deceased peer.
the superior members of the household, among them M. Villebecque.
Coningsby, then unknown to his grandfather; the same sum to Mr. Rigby.
annuities to faithful servants, decayed actors, and obscure foreigners.
of the will, all would have consequently centred, was Mr. Rigby.
Coningsby was left sole residuary legatee.
therefore in his own disposition, was left to Coningsby. The legacy to Mr.
The solicitor paused and begged permission to send for a glass of water.
he as yet exchanged any recognition with Coningsby.
Castle, from the amiable motive that after Lord Monmouth's decease Mr.
Rigby might wish, perhaps, to present it to some other friend.
Lord Eskdale and Mr. Ormsby took care not to catch the eye of Mr. Rigby.
the sight of this instrument that sustained Rigby at this great emergency.
Rigby; he read the latent malice of that nevertheless anxious countenance.
foe to their union was departed.
however, any expression of condolence.
'This time next year you will not think so,' said Sidonia.
'I have been thinking all this time of your position,' said Sidonia.
'A sorry one, I fear,' said Coningsby.
'What have I?' said Coningsby, despondingly.
knowledge, a fine courage, a lofty spirit, and no contemptible experience.
'You console me,' said Coningsby, with a faint blush and a fainter smile.
scrapes. But you can do nothing if you be in debt. You must be free.
excited. I am hardly surprised. In your position it would not satisfy me.
had no trust. Why should he succeed? Success was the most rare of results.
of course been duly digested by all who knew him. What a contrast to St.
with absolute ruin, that could alone have inspired such a hallucination!
noble-hearted Coningsby delivered himself up to despair.
roused himself, dressed, and wrapping his cloak around him, sallied forth.
success and his fame would sound and be re-echoed.
is prepared to rise on the morrow collected for the great human struggle.
brought him a note. The handwriting was feminine. The note was from Flora.
convenience, at the hotel in Brook Street where she now resided.
her lot? He replied, therefore, that in an hour he would wait upon her.
destruction of all his prospects, and the balk of all his aspirations.
'You are very good to come to me,' she said, faintly smiling.
own, looking down much embarrassed.
the first awkwardness of their meeting.
tears stole down her pale cheek.
nothing; and I shall never see you again. Let me tell you what I mean.
will soon see you again.' Mournfully he bade her farewell.
the hall, and Henry Sydney and Buckhurst were ushered in.
it was a providential dispensation to make him a Lord Chancellor.
dreams, and their dazzling adventures in the world.
which it ill became the destitute to prefer.
on a subject that was consecrated in his inmost soul.
the catastrophe of the government would be financial.
administration and fluctuating opinions, energy and a creed.
Conservative party beat the shattered and exhausted Whigs.
him, but he was out of sight, and did not wish to be remembered. Mr.
lawyer. Mr. Rigby cut Coningsby.
had made Lady Wallinger acquainted with the whole history.
friends and old companions were candidates, and with sanguine prospects.
unfortunately been too long absent from the public life of this country.
by calling out Buckhurst for his personalities.
was headed, 'Extraordinary Affair! Withdrawal of the Liberal Candidate!
of their confidence than HARRY CONINGSBY, ESQ.
local than political circumstances, liked the name of Coningsby. Mr.
accomplished youth, who had been wronged, and who deserved to be rewarded.
to that Providence that had so signally protected him.
which gradually had brought about so singular a revolution in his favour.
to others the convictions which he acknowledged in unison with herself.
Sir Joseph spoke to Mr. Millbank, who stared; but Sir Joseph spoke feebly.
circumstances were sufficiently ripe to urge his father on the subject.
with that irresistible frankness which a strong faith can alone inspire.
Coningsby. A Coningsby working for his bread was a novel incident for him.
could have dared to desire.
intelligence that Oswald conveyed to Coningsby during their rapid travel.
morning. At Birmingham, therefore, they remained.
British Parliament; public characters, responsible agents, with a career.
styling his opponent a Venetian, and all their companions of early days.
a sudden become elements of power, springs of the great machine.
bearing, and his intelligent brow, at once won every female heart.
the head of his procession.
The car stopped before the principal hotel in the High Street. It was Mr.
Edith, who was leaning on her father's arm.
like this heard in Darlford before.
even a formal declaration of his sentiments.
believed in Coningsby: and a very good creed.
warmth, and offered his hearty congratulations.
'It is to you, dear sir, that I am indebted for all this,' said Coningsby.
hands, and blessed them with words of tenderness.
which place was in future to be the residence of the member for Darlford.
with the sanction, but at the express instance, of his son.
believing in their own energies, and daring to be great?


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Principal » Análisis » blog de ebastida
UN Role in Reforming International Finance for Development
Mié, 08/30/2017 - 13:57 -- ebastida
Jomo Kwame
Growing global interdependence poses greater challenges to policy makers on a wide range of issues and for countries at all levels of development. Yet, the new mechanisms and arrangements put in place over the past four decades have not been adequate to the growing challenges of coherence and coordination of global economic policy making. Recent financial crises have exposed some such gaps and weaknesses.
Multilateral UN inclusive
Although sometimes seemingly slow, the United Nations (UN) has long had a clear advantage in driving legitimate discussion on reform because of its more inclusive and open governance. Lop-sided influence in the current international financial system is a principal reason why many countries lack confidence in existing arrangements. Rebuilding confidence in such arrangements will require that all parties feel they have a stake in the reform agenda.
But the UN is also suited to drive the discussion because of its long tradition of reliable work on international economic issues. The UN secretariat has developed and maintained a coherent and integrated approach to trade, finance and sustainable development, with due attention to equity and social justice issues.
The ongoing 'secular stagnation' has again highlighted the interdependence of global economic relations, exposing a series of myths and half-truths about the global economy. These include the idea that the developing world has become "decoupled" from the developed world; that unregulated financial markets and the new financial instruments had ushered in a new era of "great moderation" and "stability"; and that macroeconomic imbalances — due to decisions made in the household, corporate and financial sectors — were less dangerous than those involving the public sector.
UN Secretariat different, but competent
The UN secretariat has long doubted such arguments, and warned that any unravelling of global macroeconomic imbalances would be unruly. Also, persistent asymmetries and biases in global economic relations have particularly hit developing countries, both emerging markets and the least developed countries.
Not surprisingly, the UN Secretariat also drew attention to the close links between the financial crisis and the food and energy crises of recent years. A more integrated approach to handling these threats is needed, particularly to alleviate the downside risks for the poorest and most vulnerable communities.
The UN Secretariat has a strong track record of identifying systemic threats from unregulated finance, warning against a misplaced faith in self-regulating markets and offering viable solutions to gaps and weaknesses in the international financial system. Special drawing rights (SDRs), the 0.7 per cent aid target and debt relief, for example, were all conceived within the UN system during the 1960s and 1970s.
From the 1980s, the UN secretariat – both in New York and Geneva — has consistently warned against the excessive conditionalities attached to multilateral lending, promoted the idea of rules for sovereign debt restructuring, and cautioned that the international financial institutions were moving away from their traditional mandates of guaranteeing financial stability and providing long-term development finance.
UN has more than earned leadership role
During the 1990s, UN agencies warned against the dangers to economic stability, particularly in developing countries, from volatile private capital flows and the speculative behaviour associated with unregulated financial markets. The UN was among the very few warning Mexico in 1994, the East Asian countries in 1997 and the world in 2008 that excessive liberalization threatened crisis. The UN system was also almost alone among international institutions to identify growing inequality as a threat to economic, political and social stability, and insisted early on measures for a fairer globalisation.
Many of these concerns culminated in the 2002 Financing for Development Conference in Monterrey, Mexico. More recently, the UN has insisted on the importance of policy space for effective development strategies and particularly on the need for macroeconomic policies to support long-term growth, technological upgrading and diversification.
The combination of a strong track record and a core secretariat steeped in its tradition of an integrated multilateral approach to policy-oriented research places the UN in the best position to advance discussions to reform the international financial architecture if given the chance to do so.
arquitectura financiera internacional
Tema de investigación:
Arquitectura financiera
http://www.ipsnews.net/2017/08/un-role-reforming-international-finance-developme...
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blog de ebastida
Reserva de derecho al uso exclusivo del título: 04-2012-110916392700-203.


Student Ambassadors are students who assist EPCC with recruitment, retention and public relations. Student Ambassadors reach out to both prospective and current students and make them aware of the programs and services we have available. Student Ambassadors promote campus life activities and involvement. By being a Student Ambassador, you will have a unique opportunity to work with students, staff, and the community and be a positive role model to other students.
Various hours representing EPCC at all campuses and community events. Some off campus activities required.


Today you will learn how to make the German Hamburger, Frikadelle or Bulette. This German meat dish has different names depending on the location in Germany. Common names are "Frikadelle" or "Bulette". The German Hamburger looks almost like an Hamburger but is thicker and contains aside meat, some bread and egg. In the North of Germany they call them "Bulette", in the South "Fleischpflanzerl" or "Fleischkuechle" as they call them in the Swabian region. The classic side dish for the Bulette is potato salad.
Enjoy them warm or cold. They make a perfect snack as well. Just place them into a roll (Broetchen), and add some mustard. It's a delicious snack! You also can use bison meat instead of beef, or a mix of half beef and half pork. Happy Cooking!
- Place the meat into a bowl.
- Soak the bread or bun in cold water, when soaked squeeze out the water.
- Add soaked bread and egg to meat.
- Peel onion, chop it into very small pieces, add to meat.
- Spice with salt, pepper and paprika powder to taste.
- Knead the meat with the hands, it's the best, until all ingredients are mixed.
- Form round to oval hamburgers out of the meat (diameter about 6-8cm). The German burgers are not super flat and 50 percent smaller than the American burgers. They also shrink during the frying process a little bit.
- Turn the meat in natural bread crumbs (no spices added).
- In a pan heat the clarified butter or oil.
- Fry the hamburgers on medium heat for 5-10 minutes each side (depends how thick they are).
Serve them warm or cold potato salad, some like to serve it with French Fries or fried potatoes, and a mixed green salad.
Im from Malaysia and currently working on traditional way to eat frikadelle.so is it just serve with salad and mustard only?
warm: with salad or any side dish like french fries, fried potatoes.
The way my German mother served them was in a beef broth or brown gravy. We always ate them alongside mashed potatoes and peas or corn.


Samsung One fonts for Sailfish OS. You must have UI Themer installed to be able to use it.
This is just fan made, unofficial stuff, I am not linked anyhow with Samsung. All the trademarks, trade names and fonts belong to their respective owners.
Add Chinese Please.Thank you very much.


text:
Streaming Station
Sky Radio 00's Hits
Sky Radio Love Songs
Sky Radio Summer Hits
Sky Radio Hits
685 All Stations
Load Station
There are two artists with this name. 1. Aubrey "Drake" Graham (born October 24, 1986 in Toronto, Ontario) is a Grammy award nominated Canadian actor, rapper and singer signed to Young Money/Universal Motown. Drake shot to global fame upon the release of his critically acclaimed 2009 mixtape So Far Gone and Best I Ever Had, a single from the mixtape which peaked at #2 on the Billboard Hot 100. Drake's music career began in 2006 with the release of his debut mixtape room for improvement. The tape featured collaborations with Trey Songz, Lupe Fiasco and Nikelus F and marked the beginning of a long-time partnership with Canadian producer boi-1da. The mixtape sold 6,000 physical copies in the year of its release. comeback season, Drake's second official mixtape, was released in 2007 and featured the single replacement girl. The track featured vocals from Trey Songz and received heavy rotation on MTV and BET television networks. robin thicke, clipse and rich boy featured on the mixtape, however it was Man of the Year - a track recorded with future mentor lil wayne - which signified the start of a close affiliation between Drake and Lil' Wayne's recording label Young Money Entertainment. In 2009, Drake released his highly anticipated third mixtape so far gone to critical acclaim and commercial success. Its lead single, Best I Ever Had, peaked at #2 on the Billboard Hot 100 and has since been certified double platinum by the RIAA. The success of the track lead to a highly pu...
Joe Budden
Royce da 5'9"
Best I Ever Had
Hold On, We're Going Home
Houstatlantavegas
The Calm
Started from the Bottom
Marvins Room
The Motto
Shot for Me
Too Good
In My Feelings
Over My Dead Body
Make Me Proud
Fake Love
Worst Behavior
10 Bands
0 To 100 / The Catch Up
Doing It Wrong
Furthest Thing
Wu-Tang Forever
I'm Goin In
HYFR (Hell Ya Fucking Right)
Tuscan Leather
Under Ground Kings
© 2016 iMusicPlayer.com
iMusic Player


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Trains from Stafford to Garston (Hertfordshire)
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Garston (Hertfordshire)
Home Destinations Garston (Hertfordshire) Trains from Stafford to Garston (Hertfordshire)
From Stafford, our trains run up to 1 times per hour to Garston (Hertfordshire).
Travel during the off-peak and you'll have more choice of trains and a cheaper ticket as you avoid the commuter rush hour. Off-Peak or Super Off-Peak tickets are our cheapest 'pay on the day' tickets. Don't worry though, when you search for your journey, we'll show you the cheapest options in the results.
If you need ultimate flexibility an Anytime ticket is your best option. You can travel on any train at any time. If your ticket is marked 'London Northwestern Railway or West Midlands Railway' just make sure you get on our trains!
Book with us and the ticket price is all you pay – no hidden booking fees!
Frequent train travellers can save up to 1/3 off every journey with a National Railcard, head over to our National Railcards page and check out the options available to you.
Free Wi-Fi is available on our long distance services and some trains around the West Midlands.
Download our app for quick journey planning and mobile tickets.
Journey information from Stafford to Garston (Hertfordshire)
How much is a train ticket from Stafford to Garston (Hertfordshire) by train?
Catch the train from Stafford to Garston (Hertfordshire) from just £8 one-way when you book in advance.
How long does it take to travel from Stafford to Garston (Hertfordshire) by train?
Our fastest journey to Garston (Hertfordshire) takes 2 hours 25 minutes. Journey times may be slightly longer on weekends or public holidays. Use our Journey Planner to access downloadable timetables, live departures and arrivals. Stay connected while on the move with our free on-board WiFi.
How many trains a day are there from Stafford to Garston (Hertfordshire)?
There are up to 17 trains per day from Stafford to Garston (Hertfordshire).
Are there any direct trains from Stafford to Garston (Hertfordshire)?
Yes, you can travel from Stafford to Garston (Hertfordshire) without having to change trains. Find direct trains to Garston (Hertfordshire) using our Journey Planner.
What time is the first train from Stafford to Garston (Hertfordshire)?
The first train departs from Stafford to Garston (Hertfordshire) at 05:24. Book your tickets in advance and enjoy a few more minutes of hitting snooze.
What time is the last train from Stafford to Garston (Hertfordshire)?
The last train from Stafford to Garston (Hertfordshire) departs at 23:06.
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Stafford to Milton Keynes Central
More trains from Garston (Hertfordshire)
Garston (Hertfordshire) to Malvern Link
Garston (Hertfordshire) to Shrewsbury
Garston (Hertfordshire) to St Albans Abbey
Garston (Hertfordshire) to Cradley Heath
Garston (Hertfordshire) to Perry Barr
Adderley Park to Wood End
Flint to Ridgmont
Fenny Stratford to Oxenholme Lake District
Stoke-on-Trent to Kidderminster
Llandudno Junction to Leamington Spa
More tickets types between Stafford to Garston (Hertfordshire)