Glucocorticoids are a mainstay of anti-inflammatory therapy, but significant adverse effects ultimately limit their utility. Previous efforts to design glucocorticoid structures with an increased therapeutic window have focused on dissociating anti-inflammatory transcriptional repression from adverse effects primarily driven by transcriptional activation. An alternative to this medicinal chemistry approach is a systems biology based strategy that seeks to amplify selectively the anti-inflammatory activity of very low dose glucocorticoid in immune cells without modulating alternative cellular networks that mediate glucocorticoid toxicity.
The combination of prednisolone and the antithrombotic drug dipyridamole was profiled using in vitro and in vivo models of anti-inflammatory activity and glucocorticoid-induced adverse effects to demonstrate a dissociated activity profile.
The combination synergistically suppresses release of proinflammatory mediators, including tumour necrosis factor-α, IL-6, chemokine (C-C motif) ligand 5 (RANTES), matrix metalloproteinase-9, and others, from human peripheral blood mononuclear cells and mouse macrophages. In rat models of acute lipopolysaccharide-induced endotoxemia and delayed-type hypersensitivity, and in chronic models of collagen-induced and adjuvant-induced arthritis, the combination produced anti-inflammatory activity that required only a subtherapeutic dose of prednisolone. The immune-specific amplification of prednisolone anti-inflammatory activity by dipyridamole did not extend to glucocorticoid-mediated adverse effects, including corticosterone suppression or increased expression of tyrosine aminotransferase, in vivo after repeat dosing in rats. After 8 weeks of oral dosing in mice, treatment with the combination did not alter prednisolone-induced reduction in osteocalcin and mid-femur bone density, which are markers of steroid-induced osteoporosis. Additionally, amplification was not observed in the cellular network of corticotroph AtT-20/D16v-F2 cells in vitro, as measured by pro-opiomelanocortin expression and adrenocorticotropic hormone secretion.
These data suggest that the multi-target mechanism of low-dose prednisolone and dipyridamole creates a dissociated activity profile with an increased therapeutic window through cellular network selective amplification of glucocorticoid-mediated anti-inflammatory signaling.
The robust anti-inflammatory effects of glucocorticoids are applied broadly in the clinical setting to treat diverse conditions, including rheumatic diseases, allergy, skin disorders, pulmonary conditions, cancer, transplant rejection, and even spinal cord injury. Unfortunately, the long-term clinical utility of glucocorticoids is limited by undesirable adverse effects, including suppression of the hypothalamus-pituitary-adrenal (HPA) axis, increased serum glucose, induction of osteoporosis and glaucoma, altered electrolyte balance, insomnia, and other behavioral alterations. Chronic treatment with even relatively low doses (for instance, 7.5 mg/day prednisolone) can lead to a subset of glucocorticoid-induced adverse effects [1, 2]. Development of glucocorticoids with an improved therapeutic window has therefore been an area of focus for multiple groups.
The diverse effects of glucocorticoids are mediated by the glucocorticoid receptor (GR). Unliganded GR is retained in the cytosol by heat shock proteins that are released upon binding and activation by glucocorticoid . Once activated the GR translocates to the nucleus and can bind directly to glucocorticoid response elements (GREs) as a homodimer, resulting in both activation and repression of transcription, depending on promoter structure and interaction with various co-activators and co-repressors. Additionally, activated GR can affect transcription through mechanisms independent of DNA binding that modulate the activity of other transcription factors, including nuclear factor-κB, activator protein-1, and STAT (signal transducer and activator of transcription) . Finally, activated GR elicits a variety of transcription-independent effects through modulation of mRNA stability via 3'-untranslated region binding . It is the integration of these diverse mechanisms, which affect multiple molecular targets, cells, and tissues, that results in the desirable anti-inflammatory activity and undesirable adverse effects of glucocorticoid treatment.
Efforts to dissociate the anti-inflammatory activity from the adverse effects of glucocorticoids have focused primarily on separating the DNA-binding-dependent (transcriptional activating) and DNA-binding-independent (transcriptional repressing) activities of activated GR. Dimerization-defective mutants of GR that lack DNA-binding activity can repress activator protein-1 mediated transcription, but they cannot activate transcription of GRE-regulated genes . Glucocorticoid treatment can suppress local and systemic inflammation in homozygotic mice that express this GRdim mutation, underscoring the importance of DNA-binding-independent mechanisms to the anti-inflammatory effect observed in vivo . In contrast, many adverse effects of glucocorticoid treatment are due to DNA-binding-dependent activation (hyperglycemia, hypertension) or repression (suppression of HPA axis, osteoporosis) of transcription through activated GR homodimer binding to GREs [8, 9]. A number of selective GR modulators or selective GR agonists (SEGRAs) have been developed that can dissociate anti-inflammatory activity from some of the classical glucocorticoid adverse effects [10–15].
Early attempts at steroid dissociation using medicinal chemistry have yielded mixed degrees of success because the anti-inflammatory activity and adverse effects of glucocorticoids do not break cleanly along the mechanistic lines of transcriptional repression and transcriptional activation. For example, adverse glucocorticoid effects including suppression of HPA axis, osteoporosis, and skin atrophy are probably induced, at least in part, by DNA-binding-independent repressive effects . Similarly, the anti-inflammatory targets annexin-1 (lipocortin-1) , glucocorticoid-induced leucine zipper [17, 18], and tristetraprolin  are positively regulated by DNA-binding-dependent transcriptional activating effects of glucocorticoids. Finally, macrophages from GRdim mice exhibit a decreased potency of glucocorticoid suppression of IL-1β, monocyte chemotactic protein-1 (CCL2), macrophage inflammatory protein-2 (CXCL2), and interferon-gamma-inducible protein-10 (CXCL10) . It is likely that effective dissociation of glucocorticoid action to enhance therapeutic index will require a careful tuning of both DNA-binding-dependent (transcriptional activating) and DNA-binding-independent (transcriptional repressing) effects to achieve an improved balance of desirable anti-inflammatory activity over induction of adverse effects . This type of multi-parametric optimization presents a significant challenge to the medicinal chemistry approach to glucocorticoid dissociation.
An alternative approach to dissociation makes use of synergistic multi-target biology to amplify selectively the anti-inflammatory activity of glucocorticoids in immune cells without affecting glucocorticoid-induced adverse effects in alternative cellular networks [22, 23]. The combined molecular effects of the antithrombotic agent dipyridamole and a very low dose of the glucocorticoid prednisolone create such an activity profile. Dipyridamole inhibits the activity of equilibrative nucleoside transporters and phosphodiesterases to increase cAMP and cGMP that block platelet activation, and it is used therapeutically in combination with low-dose aspirin for secondary stroke prevention . Dipyridamole has also demonstrated anti-inflammatory activity using cell-based in vitro models . The synergistic combination of prednisolone and dipyridamole suppresses tumor necrosis factor (TNF)-α secretion by lipopolysaccharide (LPS)-stimulated human peripheral blood mononuclear cells, as well as secretion of a unique set of cytokines, chemokines, and proteases by mouse bone-derived macrophages  (Fraser CC, unpublished data). In addition to suppressing the rheumatoid arthritis (RA)-modifying target TNF-α, the combination inhibits additional targets, including chemokine (C-C motif) ligand 5 (RANTES) and matrix metalloproteinase-9 (gelatinase-B), which are upregulated in RA synovium [27–30], and IL-6, which has been validated as a new target for the treatment of RA . In RA, low-dose prednisolone treatment is generally considered to be a daily dose of 7.5 mg [32, 33]. The combination of very low dose prednisolone (3 mg/day) and dipyridamole (400 mg/day) has exhibited statistically significant effects in human clinical trials of hand osteoarthritis  and RA (Kirwan JR, unpublished data). The selectivity of this synergistic combination was demonstrated by first measuring the activity of the combination in both acute and chronic models of inflammation in rats. The combination was then tested in various in vivo models of glucocorticoid-induced adverse effects, including suppression of the HPA axis marker corticosterone, induction of the gluconeogenesis gene tyrosine aminotransferase (TAT), and effects on markers of bone homeostasis. These data support a selective activity profile for this combination, in which dipyridamole amplifies the desired anti-inflammatory activity of prednisolone in immune cells without enhancing glucocorticoid action in alternative cellular networks that mediate adverse effects to generate an improved therapeutic index.
Compounds were obtained from Sigma-Aldrich (St. Louis, MO, USA), and stock solutions of appropriate concentration (in dimethyl sulfoxide) were serially diluted on master plates using liquid-handling automation and transferred to assay plates. Human buffy coat was obtained fresh daily from healthy donors and diluted in media supplemented with 10% fetal bovine serum (FBS; HyClone (Logan, UT, USA)) prior to stimulation with LPS (catalog number L-4130; Sigma-Aldrich) at 2 μg/ml and addition to assay plates. Plates were incubated for 18 hours at 37°C and 5% carbon dioxide. Supernatants were transferred to an ELISA plate coated with anti-TNF-α antibody (catalog number 551220; BD Pharmingen (San Diego, CA, USA)). Plates were then washed before probing with a second antibody (catalog number 554511; BD Pharmingen) and europium-labeled detection reagent (catalog number 1244-360; PerkinElmer (Waltham, MA, USA)). Raw data values of time-resolved fluorescence were converted to relative fractional inhibition (I = 1 – T/U) by comparing compound or combination treated values (T) with the median vehicle-alone level (U). Synergy is determined by comparing the combination's response to the Loewe additivity standard , and comparisons were made numerically using the combination index (CI) . For example, CI70 = (CX/IC70X) + (CY/IC70Y), where (CX/IC70X) for a mixture is the ratio of compound X concentration in a 70% effective mixture (CX) over its 70% inhibitory concentration when applied alone (IC70X).
Lewis (LEW/SsNHsd) rats (n = 8/group) were administered the appropriate test or control agent via oral gavage. Two hours after test or control substance administration (time = 120 minutes), animals were injected intraperitoneally with Escherichia coli serotype 0111:B4 LPS (Sigma-Aldrich). Control animals received a saline injection. Animals were euthanized by carbon dioxide asphyxiation 90 minutes after LPS administration. Serum samples were assayed for TNF-α levels using an ELISA (BioSource, Camarillo, CA, USA). All study procedures were approved by the CombinatoRx, Inc. Institutional Animal Care and Use Committee.
CD-1 mice (n = 5/group) were sensitized with 2,4-dinitrofluorobenzene (DNFB) solution by application to the abdomen. Five days after application of DNFB, mice were administered test agents by oral gavage at the indicated doses (mg/kg). Two hours after the administration of the test agents, animals were challenged by painting the outer and inner surface of the left ear with DNFB. The right ear was painted with diluent (4:1 acetone/olive oil) as a control. Twenty-four hours after challenge, mice were anesthetized and the thickness of the DNFB-treated ear and the control ear were measured using electronic precision calipers to determine the change in thickness (mm).
Lewis (LEW/SsNHsd) rats (n = 12/group) were immunized with type-II collagen from newborn calf joints (Elastin Products Company, Inc., Owensville, MO, USA) emulsified in incomplete Freund's adjuvant (product number F5506; Sigma-Aldrich). Approximately 2 mg/kg collagen was given to all animals via intradermal injection on day 1 of the study. Two injections of 100 μl of collagen/adjuvant were made, one into the base of the tail and the other further up the back, separated by approximately 1.5 cm. A boost injection of the same material was given intradermally on day 6 of the study. Vehicle and test agents were administered via oral gavage. Dosing volume was 5 ml/kg and was adjusted weekly based on body weight measurements. Treatment period was from day 10 through day 27. Tibiotarsal joint thickness was measured using an electronic caliper on days 3, 6, 8, 13, 15, 17, 20, 22, 24 and 27. Change in joint thickness was calculated relative to the day 3 measurement. All study procedures were approved by the MDS Pharma Services (Bothell, WA, USA) Institutional Animal Care and Use Committee.
Louvain rats (n = 12/group) were immunized with solubilized type II collagen (1 mg/ml) in incomplete Freund's adjuvant injected intradermally into 15 sites on the back. Collagen-induced arthritis (CIA) developed over the next 10 days and test agents were administered every day by intragastric gavage from days 10 to 28 at the doses indicated (mg/kg). Arthritis severity was recorded daily for each hind paw using an integer scale from 0 to 4 to quantify the level of erythema and swelling (0 = normal; 4 = maximum). The sum of both hind paws (maximum score of 8) represented the severity of arthritis. Hind limbs were harvested at sacrifice (day 28) and scored by radiographic joint index on a scale from 0 to 3, based on soft tissue swelling, joint space narrowing, periosteal new bone formation, and presence of erosions and/or ankylosis (0 = normal; 3 = maximum joint destruction). The radiographic joint index represented the sum of both hind paws with a maximum score of 6. The experimental protocol conformed to the approved protocols of the UCLA Animal Care and Use Committee.
Lewis (LEW/SsNHsd) rats (n = 5/group) were weighed and placed into one of the six study groups. Body weights were recorded every other day throughout the study. Animals were dosed daily via oral gavage with test agents at volumes based on body weight progression throughout the study. On day 10, 2 hours after oral dosing, animals were euthanized via carbon dioxide asphyxiation. All study procedures were approved by the CombinatoRx, Inc. Institutional Animal Care and Use Committee.
Blood was collected and separated for corticosterone determination from serum (Diagnostic Systems Laboratories, Inc., Webster, TX, USA). Thymus and spleens were collected and weighed. Liver samples were removed and stored in RNAlater (Ambion, Austin, TX, USA) at 4°C. Liver samples were homogenized using TissueRuptor (Qiagen) and total RNA was isolated using the RNeasy-Plus Mini kit (Qiagen, Valencia, CA, USA). Equal amounts of total RNA were used for one step RT-PCR (QuantiTect Probe; Qiagen). Commercially available assay reagents (Taqman Gene Expression Assays; Applied Biosystems, Foster City, CA, USA) were used for detection of TAT and β-actin (endogenous control) mRNA, using the Applied Biosystems 7300 Real-Time PCR System.
Mice (C57Bl/6) were randomized (n = 10/group) to treatment groups based on body weight before the start of dosing. Test agents were administered via oral gavage, with the exception of dexamethasone, which was administered via subcutaneous injection. All agents were dosed twice daily and new formulations prepared weekly for a total treatment period of 8 weeks. Animals were given two doses of calcein 10 mg/kg intraperitoneally 6 and 2 days before necropsy for fluorochrome labeling. Animals were anesthetized with isoflurane before necropsy, a terminal blood sample was collected, and serum was separated and stored frozen until analysis for bone markers. Femurs and lumbar vertebrae were also collected for dual energy X-ray absorptiometry, peripheral quantitative computed tomography and histomorphometry. All study procedures were approved by the MDS Pharma Services Institutional Animal Care and Use Committee.
AtT-20 cells were seeded at a density of 60,000 cells per well in a 96-well plate for determination of changes in cAMP levels in response to various treatment conditions. Cells were allowed to recover for 18 hours and then treated with dipyridamole (10 μmol/l), rolipram (10 μmol/l), or dimethyl sulfoxide control for 30 minutes at room temperature. Cells were then stimulated with corticotrophin-releasing factor (37.5 nmol/l), or control (vehicle), for 30 minutes at room temperature. cAMP levels were quantitated using the LANCE cAMP Detection Kit (PerkinElmer).
The murine anterior pituitary corticotroph cell line AtT-20/D16v-F2 was obtained from the American Type Culture Collection (Manassas, VA, USA) and maintained in Dulbecco's minimal essential medium (American Type Culture Collection) with 10% FBS, at 37°C with 5% carbon dioxide. To determine relative pro-opiomelanocortin (POMC) expression in AtT20 cells treated with prednisolone and/or dipyridamole, real-time RT-PCR was performed on cell lysates using the FastLane Cell RT-PCR kit (Qiagen). Commercially available assay reagents (Taqman Gene Expression Assays; Applied Biosystems) were used for detection of POMC β-actin (endogenous control) mRNA. Real time RT-PCR was done using the Applied Biosystems 7300 Real-Time PCR System. For ACTH secretion experiments, AtT20 cells were seeded in 24-well plates at a density of 125,000 cells/well in Dulbecco's minimal essential medium supplemented with 10% charcoal/dextran FBS (HyClone), and treated with prednisolone and/or dipyridamole. After 24 hours, the medium was refreshed with the same compound treatment in the absence or presence of corticotropin-releasing factor (CRF; 100 nmol/l). Three hours later (27 hours), culture medium was collected for evaluation of ACTH by ELISA (MD Biosciences, St. Paul, MN, USA).
The combination of prednisolone and dipyridamole synergistically suppresses production of proinflammatory markers in vitro. The combination was discovered based on the observation of synergistic suppression of TNF-α from phorbol myristate acetate and calcium ionophore stimulated human peripheral blood mononuclear cells (PBMCs [see Figure S1 in Additional data file 1]). In a secondary assay the combination was found to synergistically suppress TNF-α secretion from LPS-stimulated PBMCs with a CI of 0.31 ± 0.02 (Figure 1a, left panel). Combinations with CI about 1 interact additively, such as would be expected when combining a drug with itself, and CI values below 1 indicate a synergistic interaction between the components . Isobolographic analysis indicates that the synergistic effect of the combination allows reduction of the drug concentrations required to achieve 70% inhibition of TNF-α secretion by ten-fold for prednisolone and five-fold for dipyridamole (Figure 1a, right panel).
Synergistic anti-inflammatory activity of prednisolone and dipyridamole in vitro. (a) Dipyridamole and prednisolone were diluted orthogonally using a twofold serial dilution, and then combined to produce a drug combination dose-response matrix. The dose responses for prednisolone and dipyridamole as individual agents are located in the bottom row and left column, respectively. Combination doses fill out the matrix and component concentrations can be read from the row and column labels. The combination dose-response matrix was applied to lipopolysaccharide (LPS)-stimulated human peripheral blood mononuclear cells (PBMCs), and tumor necrosis factor (TNF)-α in the supernatant was measured by ELISA after 18 hours. Percentage inhibition of TNF-α secretion relative to vehicle-treated controls is indicated in each cell of the matrix and represented by a color scale, where warm colors indicate more inhibition (left). Isobolographic analysis of the inhibition matrix (blue line) compares the activity of the combination with a theoretical additive interaction (red line) at the 70% inhibition level (right). Synergistic interactions fall below the additivity threshold and approach the origin, and an antagonistic interaction would lie above the red additivity line. (b) Combination matrices were measured including a fixed dose of RU486 at the indicated concentration at each point in the corresponding dose response matrix. Percentage inhibition of LPS-induced TNF-α secretion relative to vehicle-treated controls is indicated in each cell of the matrix.
The activity of prednisolone and the combination effect is abolished by treatment with the GR antagonist RU486 at a concentration of 50 nmol/l, but dipyridamole activity is unaffected (Figure 1b). Antagonism by such a low dose of RU486 suggests that the effect may be mediated primarily by the GRE-dependent transcriptional-activating activity of dimerized GR.
The combination of prednisolone and dipyridamole suppresses TNF-α in models of acute inflammation, and disease activity in CIA in rats. High-dose prednisolone (10 mg/kg) administered orally 2 hours before LPS challenge was able to significantly reduce serum TNF-α in an acute model of rat endotoxemia . Prednisolone at 1 mg/kg, and dipyridamole at 150 mg/kg, yielded nonsignificant reductions in TNF-α release compared to the LPS control. The two agents combined yielded reduction in TNF-α release that was intermediate between the effects of low-dose and high-dose prednisolone (Figure 2a). These trends were also observed in repeats of the LPS challenge model.
Prednisolone and dipyridamole combine to suppress acute inflammation in vivo. (a) Lewis rats were treated orally with compounds as indicated (mg/kg) for 2 hours before challenge with lipopolysaccharide (LPS). Serum was collected 90 minutes later and tumor necrosis factor (TNF)-α quantitated by ELISA. *P < 0.01 versus the LPS control; aP = 0.06, bP = 0.98, cP = 0.81, dP = 0.59 versus the combination. (b) Mice were sensitized with the chemical irritant 2,4-dinitrofluorobenzene (DNFB). Five days later animals were administered test agents by oral gavage at the indicated doses (mg/kg) and challenged on the ear with DNFB solution. Change in ear thickness was measured 24 hours after challenge (Δ ear thickness [mm]). *P < 0.05 versus the vehicle control; aP = 0.02, bP = 1.0, cP = 0.02, dP = 0.05 versus the combination. Dipyridamole was dosed at 150 mg/kg. Error bars are + standard error of the mean. Statistical comparison is by analysis of variance with Tukey. Dp, dipyridamole; Pd, prednisolone.
High-dose prednisolone dosed orally at 30 mg/kg was able to suppress chemical hypersensitivity induced ear swelling (Figure 2b). A tenfold lower dose of prednisolone (3 mg/kg) and dipyridamole (150 mg/kg) as individual agents had no effect relative to vehicle-treated controls. The combination demonstrated efficacy equal to high-dose prednisolone, suggesting ten-fold amplification by dipyridamole of the anti-inflammatory activity of low-dose prednisolone in this acute model.
Disease activity was also suppressed by the combination of prednisolone and dipyridamole in CIA models in Lewis and Louvain rats. To test the activity of the combination in this model of chronic inflammation, rats were sensitized with collagen and hind paw arthritis developed over the next 10 days [38, 39]. Lewis rats treated with prednisolone (3 mg/kg orally) exhibited negligible tibiotarsal joint swelling compared with animals that were not subjected to collagen induction, and rats treated with 0.3 mg/kg prednisolone or 150 mg/kg dipyridamole showed similar levels of tibiotarsal inflammation to CIA-induced, vehicle-treated controls (Figure 3a,b). Tibiotarsal joint swelling trends were maintained throughout the study. The combination of prednisolone and dipyridamole (0.3/150 mg/kg) yielded a significant reduction in swelling compared with dipyridamole alone, and tibiotarsal swelling in the combination group was intermediate between the low-dose and high-dose prednisolone groups at all points in the study.
Prednisolone and dipyridamole combine to suppress collagen-induced arthritis in vivo. Collagen-induced arthritis (CIA) was developed in Lewis rats for 10 days before oral daily dosing with compounds as indicated (mg/kg) for the next 17 days. Change in hind limb tibiotarsal joint diameter relative to the day 3 measurement is reported (a) over the course of the study and (b) at study completion. *P < 0.001 versus the CIA control; aP = 0.29, bP = 0.10, cP = 0.14, dP = 0.004 versus the combination. (c) CIA was induced in Louvain rats for 10 days and test agents were administered orally once daily from days 10 to 28, as indicated (mg/kg). Arthritis severity was scored daily based on erythema and swelling. *P < 0.001 versus the CIA control; aP = 0.0003, bP = 0.001, cP = 0.15, dP = 0.12 versus the combination at day 28. (d) Hind limbs were scored by radiographic joint index after the completion of the study. **P < 0.0001, *P < 0.01 versus the CIA control; aP = 0.005, bP = 0.17, cP = 0.84, dP = 0.01 versus the combination at day 28. Dipyridamole was dosed at 150 mg/kg. Error bars are ± standard error of the mean, and statistical comparison is by analysis of variance with Tukey. Dp, dipyridamole; Pd, prednisolone.
The CIA model was also repeated in Louvain rats , and arthritis score was measured daily from days 10 to 28 (Figure 3c,d). At the conclusion of the study (day 28) animals treated with vehicle or dipyridamole had arthritis scores of about 6.5, which were significantly different from the scores in the combination and high-dose prednisolone groups. Combination treated animals (0.3/150 mg/kg) had an average arthritis score of 2.8, which was intermediate between the effect of low-dose prednisolone (4.9) and high-dose prednisolone (0.7), suggesting that dipyridamole can amplify the activity of low-dose prednisolone in suppressing erythema and joint swelling. Radiographic analysis of the hind limbs at the conclusion of the study indicated that the combination significantly reduced tissue damage relative to the vehicle control, and was similar to low-dose steroid alone on measures of joint space narrowing and the presence of erosions and/or ankylosis (Figure 3d).
An additional test of the combination was conducted in an adjuvant-induced arthritis model , and similar anti-inflammatory activity of the combination was observed. A higher dose of dipyridamole (300 mg/kg) was required to observe the effect in this particular model. Tissue was collected and prepared for histologic evaluation of tarsal and phalangeal joints based on inflammatory infiltrate, pannus formation, and cartilage and bone degeneration. The combination of prednisolone and dipyridamole achieved reduction in cartilage damage in the tibiotarsal joint similar to that observed for the high-dose prednisolone (5 mg/kg) positive control group. The combination strongly suppressed cartilage damage in the phalangeal joints as well as inflammation, pannus formation, and bone damage, as indicated by histologic analysis [see Figure S2 in Additional data file 1].
The observed amplification by dipyridamole of prednisolone anti-inflammatory activity did not extend to classical glucocorticoid adverse effects. Lewis rats were treated once daily for 10 days with oral dose groups of prednisolone identical to those used in the CIA model. The amplifying dose of dipyridamole was increased two-fold to 300 mg/kg for the safety studies. This increased dose of dipyridamole had demonstrated increased anti-inflammatory effect in some models. At the conclusion of dosing, appropriate tissues were harvested to measure safety parameters. Liver mRNA levels of TAT, a marker of glucocorticoid-activated glucose metabolism, were evaluated by RT-PCR after repeated treatment with prednisolone and dipyridamole alone, and in combination. Animals treated daily with 3 or 5 mg/kg of prednisolone for 10 days experienced a 2.6-fold increase in the expression of TAT mRNA in the liver (Figure 4a). In contrast, animals treated with 0.3 mg/kg prednisolone exhibited a 1.7-fold increase in TAT mRNA with oral daily dosing. Treatment with the combination of prednisolone and dipyridamole (0.3/300 mg/kg) resulted in significantly lower TAT mRNA levels than in the high-dose prednisolone (3 mg/kg) treatment group, but was no different from the effect of the component prednisolone dose (0.3 mg/kg) alone (Figure 4a).
Dipyridamole does not alter the low-dose prednisolone safety profile. (a) Tyrosine aminotransferase (TAT) mRNA levels from liver were evaluated by RT-PCR after 10 days of repeat dosing in Lewis rats as indicated. β-Actin was used as the endogenous control, and results are shown as fold increase in TAT mRNA over vehicle. *P < 0.01 versus the vehicle control, aP = 1.0, bP = 0.01, cP = 0.90, dP = 0.30 versus the combination. (b) Corticosterone levels in serum were evaluated by ELISA after 10 days of chronic dosing. **P < 0.0001, *P < 0.05 versus the vehicle control; aP = 0.03, bP = 0.01, cP = 0.44, dP = 0.98 versus the combination. (c) Thymus weight was measured at the conclusion of the study. **P < 0.0001, *P < 0.01 versus the vehicle control; aP = 0.83, bP = 0.14, cP = 0.06, dP = 0.03 versus the combination. (d) Adrenal weights were also evaluated upon study completion. *P < 0.05 versus the vehicle control; aP = 0.21, bP = 0.98, cP = 1.0, dP = 0.50 versus the combination. Dipyridamole was dosed at 300 mg/kg. Error bars are + standard error of the mean, and statistical comparison is by analysis of variance with Tukey. Dp, dipyridamole; Pd, prednisolone.
Glucocorticoids can suppress products of the HPA axis including, serum corticosterone. After chronic treatment of Lewis rats with the combination or individual components for 10 days, serum was collected to quantitate levels of corticosterone. The 3 and 5 mg/kg prednisolone groups exhibited significantly suppressed serum corticosterone (Figure 4b). Prednisolone at 0.3 mg/kg had less of an effect on corticosterone, and this effect was not amplified when applied in combination with dipyridamole (300 mg/kg). Thymus and adrenal gland weights were also measured following chronic dosing. Prednisolone at 3 and 5 mg/kg suppressed thymus weight, but 0.3 mg/kg prednisolone alone or in combination had no significant effect on thymus weight compared with vehicle control (Figure 4c). The effect of the combination (0.3/300 mg/kg) on adrenal weight was identical to the effect of 0.3 mg/kg prednisolone alone, and neither was significantly different from the vehicle control group (Figure 4d).
Chronic treatment with glucocorticoids can alter expression of various markers of osteoporosis, including osteocalcin and structural measures of bone density and quality. Female mice were treated orally twice daily with various doses of prednisolone alone, dipyridamole, or the combination of varying doses of prednisolone with dipyridamole, for 8 weeks before quantitation of these surrogate markers of osteoporosis. Dexamethasone (5 mg/kg once daily) significantly reduced osteocalcin and mid-shaft femur bone density compared with vehicle-treated controls. Prednisolone was associated with a dose-dependent reduction in osteocalcin and mid-femur bone density that was not altered by the addition of dipyridamole (37.5 mg/kg twice daily; Figure 5).
Dipyridamole does not amplify prednisolone effects on surrogate markers of osteoporosis. BL/6 mice were dosed twice daily with test agents for a total of 8 weeks to measure effects on markers of bone homeostasis. (a) Serum was collected at the end of the study and osteocalcin was measured by ELISA. (b) End of study mid-shaft femur bone density was measured by flurochrome labeling, sectioning, and peripheral quantitative computed tomography. Prednisolone alone (grey curve); prednisolone in combination with dipyridamole twice daily (black curve); dipyridamole alone and vehicle control are indicated with open triangle and open circle, respectively; sub-cutaneous dexamethasone (5 mg/kg once daily) positive control is indicated with a black square. *P < 0.05 versus the vehicle control. Dipyridamole was dosed at 37.5 mg/kg twice daily in this study (allometrically scaled from a rat total daily dose of 150 mg/kg). Error bars are ± standard deviation, and statistical comparison is by analysis of variance with Tukey.
The observed amplification by dipyridamole of the anti-inflammatory activity of prednisolone did not extend to suppression in vitro of the POMC gene and suppression of secreted ACTH from corticotroph cells. The effects of prednisolone, dipyridamole, and the combination were measured in the murine anterior pituitary cell line AtT-20/D16v-F2 (AtT-20), a well studied corticotroph model system . The relative amount of cAMP was increased in the AtT-20 cells by 1.5-fold after treatment with dipyridamole and CRF stimulation (Figure 6a). The prototypic phosphodiesterase (PDE) 4 inhibitor rolipram increased cAMP by three-fold under these conditions. CRF stimulation increased ACTH secretion in untreated control cells after 3 hours, and pretreatment with dipyridamole (10 μmol/l) significantly increased ACTH release compared with CRF stimulation alone (Figure 6b). AtT-20 cells were pretreated with prednisolone, dipyridamole, or the combination for 24 hours, and then stimulated with CRF (100 nmol/l) to induce ACTH secretion. Pretreatment with prednisolone reduced ACTH secretion compared with the CRF-stimulated control, and the stimulatory effects of dipyridamole (10 μmol/l) on ACTH secretion were not observed in combination with any dose of prednisolone (Figure 6c). Prednisolone decreased POMC mRNA expression, with a maximum decrease of about 50% observed at the 24-hour time point (Figure 6d). The addition of dipyridamole (10 μmol/l) did not amplify the effect of prednisolone on POMC mRNA levels, and was able to compensate for the suppressive effect of very-low-dose prednisolone.
Dipyridamole does not amplify suppression of markers of the HPA axis in vitro. (a) AtT-20 corticotroph cells were pretreated with dipyridamole (Dp; 10 μmol/l) or rolipram (Rol; 10 μmol/l) as indicated. Cells were stimulated with corticotropin-releasing factor (CRF) or vehicle control before quantitating cAMP levels. Error bars are + standard deviation (SD). *P < 0.01 versus CRF alone. (b) AtT-20 cells were pretreated with Dp or control for 24 hours. Medium was refreshed with compounds, and then stimulated with CRF or vehicle control for an additional 3 hours. Medium was collected for determination of ACTH levels by ELISA. Error bars are + SD. *P < 0.001 versus CRF + Dp. (c) AtT-20 cells were pretreated with increasing doses of prednisolone in the absence (grey curve) or presence (black curve) of Dp (10 μmol/l) for 24 hours. Medium was refreshed with compounds plus CRF. After 3 hours, ACTH levels were determined by ELISA. *P < 0.001 versus vehicle alone. (d) AtT-20 cells were incubated with increasing doses of prednisolone in the absence (grey curve) or presence (black curve) of Dp for 24 hours. POMC mRNA levels were determined by RT-PCR analysis, using β-actin as the endogenous control. Dp alone (10 μmol/l) and vehicle control responses are indicated with an open triangle and circle, respectively. Error bars are ± SD, and statistical analysis is by analysis of variance with Tukey. HPA, hypothalamus-pituitary-adrenal.
The potent anti-inflammatory activity and disease-modifying effects of glucocorticoids are well documented , but safety concerns observed with chronic dosing  have created a desire for safer glucocorticoids with an expanded therapeutic window. Many groups are pursuing this goal with a medicinal chemistry approach. Significant progress has been made by identifying novel GR ligands that retain substantial anti-inflammatory activity while reducing key glucocorticoid-induced adverse effects [10–15]. The success of the dissociated ligands developed to date is impressive, given the extreme complexity of the GR system and the challenge of developing low-molecular-weight compounds that retain desirable activities of the native ligand while selectively eliminating undesirable effects. Unfortunately, these ligands generally retain unacceptable activity on one or more adverse effect measures, suggesting that an alternative approach to glucocorticoid dissociation may be required.
A multi-component therapeutic can be envisaged that leverages systems biology to amplify glucocorticoid activity selectively in the network context of inflammatory cells over alternative cellular networks that mediate adverse effects. In this multi-target approach, an enhancing agent is used to sensitize the immune cell network to the effects of very-low-dose prednisolone by modulation of intersecting signaling pathways that selectively amplify the anti-inflammatory activity of the glucocorticoid. The divergent molecular context of cellular networks that mediate glucocorticoid adverse effects (for example, corticotrophs or hepatocytes) do not support amplification, and therefore the safety profile of the very-low-dose glucocorticoid is maintained. At the organism level, the effects of the combination produce the desired anti-inflammatory activity of a glucocorticoid with an enhanced therapeutic window.
The combination of prednisolone and the antithrombotic agent dipyridamole synergistically suppresses the secretion of TNF-α and other proinflammatory mediators from human PBMCs stimulated with LPS (Figure 1a), but also phorbol myristate acetate/Ionomycin-stimulated and α CD3/α CD28-stimulated cultures [see Figure S1 in Additional data file 1]. Because of the multi-target action of the components, the anti-inflammatory activity is achieved at doses where prednisolone and dipyridamole have marginal activity as individual agents. The components of the combination, including dipyridamole, are known to suppress TNF-α individually , but the synergistic combination effect was unexpected . In secondary assays the combination was found to inhibit production of a unique profile of cytokines, chemokines, and proteases from LPS-stimulated mouse bone-derived macrophages, including synergistic suppression of TNF-α, IL-6, and chemokine (C-C motif) ligand 5, which are validated targets in rheumatic disease (Fraser CC, unpublished data).
The activity of prednisolone and the synergy with dipyridamole was blocked in vitro by the GR antagonist RU486 (Figure 1b), demonstrating that the anti-inflammatory effect of the combination is, at least in part, GR dependent and may require dimerization and transcriptional activation. GRE-independent repression of nuclear factor-κB mediated transcription by glucocorticoid is not significantly antagonized by 10–100 nmol/l RU486, but GRE-dependent transcription is strongly inhibited . This result potentially differentiates the activity of the combination from the dissociated GR ligands that are more effective at transrepression than transactivation [12, 13, 15]. The combination of prednisolone and dipyridamole, but not the components alone, has been shown to upregulate expression of glucocorticoid-induced leucine zipper and dual-specificity phosphatase-1 (DUSP1/MKP1) mRNA in LPS-stimulated mouse macrophages (Fraser CC, unpublished data). DUSP1 induction has been shown in some systems to require GR dimerization , but this was not observed in GRdim mouse macrophages . Together, these data suggest that transcriptional activation may make an important contribution to the anti-inflammatory activity of the combination.
The combination of prednisolone and dipyridamole was found to have anti-inflammatory activity in both acute and chronic models of inflammation in vivo. Low-dose prednisolone and dipyridamole had minimal effect as single agents, but the combination was able to suppress serum TNF-α in an LPS-challenge model, and ear swelling in the delayed-type hypersensitivity model (Figure 2). These data suggest that the molecular effect of dipyridamole may amplify the anti-inflammatory activity of low-dose prednisolone between three- and ten-fold in vivo. The ability of low-dose prednisolone to suppress disease activity in chronic models of arthritis was also amplified by dipyridamole. The arthritis score reduction for the combination was 2.1 units (4.9 for low-dose prednisolone and 2.8 for the combination), or one half the reduction observed between low-dose (0.3 mg/kg) and high-dose (3 mg/kg) prednisolone (4.9 – 0.7 = 4.2). These results suggest that amplification may be in the range of five-fold for the arthritis models, assuming a linear dose response (Figure 3). Prednisolone and dipyridamole also combine to suppress histologic markers of inflammation in an adjuvant-induced model of arthritis. Interestingly, the effect of combination was most pronounced in the phalangeal joints, where clinically relevant suppression of histologic markers of inflammation, and bone and cartilage degeneration were observed (see Figure S2 in Additional data file 1). However, this model required a higher dose of dipyridamole (300 mg/kg) that also demonstrated significant activity. The tissue-sparing effect of the combination may stem from the potent suppression of proinflammatory mediators from macrophages (Fraser CC, unpublished data). The activity of the combination observed in these animal models suggests that the molecular effects of dipyridamole can amplify the subtherapeutic anti-inflammatory activity of very-low-dose prednisolone to produce a disease-modifying effect.
Dipyridamole was not found to amplify the effect of prednisolone on markers of glucocorticoid-induced toxicity under the stringent conditions of repeat daily dosing in vivo. Increased gluconeogenesis leading to hyperglycemia is a common side effect of chronic glucocorticoid treatment . The gluconeogenic potential of prednisolone and dipyridamole was evaluated by measuring the effect on the classical transcriptional activation target TAT by RT-PCR analysis of liver tissue from rats treated with the combination or its components. The effect of low-dose prednisolone was not altered by simultaneous dosing with 300 mg/kg dipyridamole (Figure 4a), suggesting no enhancement of the effect of prednisolone in hepatocytes. This result suggests that positive GRE-mediated glucocorticoid adverse effects, including hyperglycemia, may not be amplified by simultaneous dipyridamole treatment. Adrenal insufficiency is another common adverse effect of chronic glucocorticoid treatment , caused by the complex negative feedback control of the system. Prednisolone dose-dependently reduced serum corticosterone, and dipyridamole did not further suppress the effect of low-dose prednisolone (Figure 4b). This result suggests that the anti-inflammatory synergy observed between prednisolone and dipyridamole does not extend to the undesirable suppression of the HPA axis. Thymus and adrenal weights were also monitored in this study, and high-dose steroid was found to reduce these organ weights after once-daily dosing for 10 days. No significant effects relative to the vehicle control were observed for the component doses alone or the combination.
Dipyridamole did not amplify prednisolone-induced effects on markers of bone degradation in vivo. Osteoporosis is a common adverse effect of chronic treatment with glucocorticoids, and loss of bone from spine and hip is estimated at 1.5% per year with a daily dose of 9 mg prednisolone or equivalent . Dipyridamole demonstrated no effect as a single agent and did not alter the prednisolone dose response for osteocalcin or mid-shaft femur bone density, suggesting a lack of combination effect on these markers (Figure 5) in mice treated for 8 weeks. The dipyridamole dose of 37.5 mg/kg twice daily in this study was allometrically scaled from the total daily dose of 150 mg/kg used in the rat CIA studies, in which dipyridamole was found to amplify the anti-inflammatory activity of low-dose prednisolone. Osteocalcin is a marker of bone formation that is known to be suppressed by glucocorticoids, caused by a negative GRE in the promoter . These data suggest that dipyridamole does not amplify DNA-binding-dependent (GRE-mediated) transrepressive effects of low-dose glucocorticoid treatment. The results from this mouse study indicate that the immune-selective action of dipyridamole may not amplify the osteoporotic effect and fracture risk associated with very-low-dose prednisolone  in the clinical setting with chronic dosing.
To further explore the HPA axis effect of the combination, the individual effects of dipyridamole were characterized in the unique signaling network of AtT20 corticotroph cells. Dipyridamole treatment has been shown to inhibit PDE activity (PDE-2, -4, -10, and -11) and increase cAMP levels [49, 50]. Dipyridamole treatment of AtT-20 cells results in a modest 1.5-fold increase in cAMP relative to CRF stimulation alone, but was less effective than the threefold increase observed with the potent PDE-4 inhibitor rolipram (Figure 6a). Increased cAMP was previously shown to increase ACTH secretion in AtT20 cells , and dipyridamole treatment was found to induce a small but significant increase in CRF-dependent ACTH secretion (Figure 6b), which may be due to increased cAMP. Interestingly, dipyridamole does not increase cAMP in mouse bone-derived macrophages (Fraser CC, unpublished data), suggesting a differential effect of dipyridamole in the immune cell network context as compared with the corticotroph network of the AtT20 cells. Activated GR is also known to activate a negative GRE and to antagonize the effect of cAMP-induced transcription factors to suppress POMC expression . We observed a prednisolone dose-dependent decrease in both POMC expression and ACTH secretion from the AtT20 cells in vitro, and this response was unaffected by the addition of dipyridamole (Figure 6c,d). These experiments demonstrate a lack of synergy between prednisolone and dipyridamole on POMC expression or ACTH secretion in corticotroph cells, unlike the synergistic anti-inflammatory activity of the combination observed in human PBMCs, mouse macrophages, and in rat arthritis models. The differential response to combination treatment is probably due to differences in cellular network context, including expression levels and/or spatial relationships of molecular targets, activation state of relevant signaling pathways, and cell type-specific differences in transcription factor expression or activation. Defining the set of targets responsible for mediating the selectivity of the combination for immune cell networks over alternative cell contexts is the focus of ongoing research.
The success of the SEGRAs and selective GR modulators developed to date is impressive, given the extreme complexity of the GR system. SEGRAs have been identified that potently suppress irritant dermatitis [10, 11, 13], carrageenan-induced and adjuvant-induced arthritis , and CIA , with improved tolerability relative to body weight loss and elevation of blood glucose. For example, the dissociated ligand ZK 216348 exhibited no increase in serum glucose in fasted rats 6 hours after dosing at doses up to 30 mg/kg, as compared with an approximately 60% increase for prednisolone at 10 mg/kg . TAT activity in these rats was minimally increased by acute dosing of ZK 216348, compared with a six-fold increase by prednisolone at an equivalent dose of 10 mg/kg . Similarly, the dissociated GR ligand LGD5552 was demonstrated to be about ten-fold less potent than prednisolone in suppressing bone formation rate in a 4-week study . Unfortunately, the complexity of the GR signaling network that controls transcriptional repression and activation, and the post-transcriptional effects of activated GR make it extremely challenging to develop low-molecular-weight compounds that retain the desirable activities of the native ligand while selectively eliminating undesirable effects.
Dissociation of anti-inflammatory effects and adrenal insufficiency has been especially problematic for the dissociated GR ligand approach. Glucocorticoid release from the adrenal is regulated by both DNA-binding-dependent and -independent (transrepressive) effects of the GR . It may be that all dissociated GR ligands with transrepressive effects equivalent to glucocorticoids will display some degree of undesirable effect on the HPA axis. For example, acute subcutaneous treatment of rats with ZK 216348 resulted in ACTH suppression with potency equivalent to that of prednisolone . Indeed, glucocorticoid activity and safety studies in GRdim mice suggest that not all anti-inflammatory actions of activated GR are retained in the absence of dimerization, and neither are all glucocorticoid-induced adverse effects eliminated . Dipyridamole was not found to amplify corticosterone suppression by low-dose prednisolone in the rat repeat dosing study (Figure 4b), and did not alter the prednisolone dose response for suppression of POMC and ACTH in the corticotroph cell line (Figure 6c,d). These results suggest that cellular network selective amplification of glucocorticoid activity by the multi-target mechanism of low-dose prednisolone and dipyridamole may provide a solution to the challenging problem of dissociating glucocorticoid-induced HPA axis suppression.
There is an increased awareness in the drug discovery industry of the need for novel therapeutics that address the systems biology of disease, and a nascent trend exists that is moving away from the one-drug, one-target paradigm toward a more pathway-focused or multi-target approach to drug discovery [22, 55–57]. Glucocorticoid dissociation efforts to date have focused on modulating GR dimerization or interaction with co-activators and co-repressors to separate the desirable and undesirable effects of glucocorticoids. The systems biology approach presented here exploits multi-target action to amplify glucocorticoid activity selectively in the unique network context of inflammatory cells, rather than attempting to dissect various aspects of GR biology. This approach is advantageous because the anti-inflammatory activity of the combination is therefore derived from amplification of native glucocorticoid action, be it transrepressive or transactivating. Cellular networks that mediate traditional glucocorticoid adverse effects do not support amplification of the low-dose glucocorticoid effect due to the absence of nodes or differential signaling pathway interactions, resulting in the selective action and an increased therapeutic window. The multi-target action of combination drugs may provide a general approach to achieve cell-type specific therapeutic effects.
The authors would like to acknowledge the contributions of many past and present colleagues at CombinatoRx. In particular, the efforts of ER Price, C Keith, J Lehár, J Nichols, M Keegan, P Elliot, M Slavonic, G Nolan, K Kelleher, J Gonzalo, Y Wang, D Crowe, J Luterman, J Randle, and L Baird are gratefully acknowledged.
The authors are employed by CombinatoRx, Incorporated.
GRZ, WA, ALF, MF, CCF, and AAB designed the research and experiments. WA, ALF, and MF performed the experiments. GRZ, WA, ALF, MF, CCF, and AAB analyzed the data. GRZ wrote the manuscript.


Any gluten-free Pita Bread Out There?
My husband and I were in England for six months and loved the gluten-free pita bread we could buy in all the grocery stores there. I've searched gluten-free Mall, the web, and all my regular San Diego stores...Whole Foods, Henry's, Jimbo's, etc. and can't find it anywhere. Europe had great gluten-free baked goods, which we really miss--like stem ginger cookies! However, it isn't as easy to eat out as it is here, so there's always a trade-off. Thanks!
Does anyone know a source for gluten-free pita bread? I just returned from six months in the UK, and they have wonderful pita bread in their supermarkets...I really miss it. I couldn't find any in the online gluten-free stores. Thanks.
Thanks, everyone, for the great information. I will be coming back for more help as we need it. And yes, I do need to remember to tone it down . . . my English coworkers remind me of that as well!
I live in the US - San Diego - and am being sent by my company to London for six months on a temporary assignment. My husband is coeliac (diagnosed last July) and will be going with me (he's retired.) We are very excited! We've never been to the UK and have wanted to go for a long time. I'm not sure where in London we'll be living. Are gluten-free products readily available in stores? What about restaurants? In the US there are certain "chain" restaurants (like PF Changs, Outback) that you can always count on for a gluten-free menu. Is that true in London? Thanks in advance for any help you can give us.
Thanks for such great suggestions, everyone--now I'm going to have my own party to use all these recipes.
Hope I spelled that right! We're going to a birthday party tomorrow for a family member and I was asked to bring an hors duorve. I always try to take something my husband can eat, and all my recipes are pre-celiac! Furthermore, he doesn't like any gluten-free crackers. I'd like it to be something substantial, since he may not be able to eat much else.
OK, let me try this again--and actually send the message this time! My husband was diagnosed with Celiac on July 21, 2006--I remember because it was my birthday! He, too, was "wasted"--skinny as a rail, and his face was the same color as his white beard. He couldn't walk 100 feet without chest pain and he thought he must be dying from something. He was very depressed and "foggy." He sat in his chair all day and watched TV. He had almost no other symptoms--but he was very anemic, which was what got his doctor's attention--finally. The lack of blood was straining his heart--probably would have had a heart attack had we kept on going. Believe me, I didn't know whether to feel sorrier for him or me that day! I am still working at 66, the kids are gone, and I had gradually pretty much given up cooking. I was too busy to change my life--it wasn't fair. And what about the traveling we had been dreaming about for years? I might as well have had celiac myself. Today he is pink-cheeked, gained his weight back, walks five miles up and down hills with no problems, and is full of vim and vinegar--what a handful he is! It's like being married all over again to the man I knew 20 years ago. Sometimes I listen to him laughing and am astonished that I hadn't noticed that he'd stopped. I went through all the trauma you're experiencing--what to eat and not eat, what to throw out and buy new, and scared to death to eat in a restaurant. I made the world's hardest loaf of brick bread. We now have a gluten-free house and eat very well. I remembered how to cook and I bake a lot. Our good friends and family try to accommodate us. We are very, very careful because his symptoms are "silent" and we wouldn't know if he was glutened--except for what it does to his insides. I take our own food to potlucks, parties, and sometimes even to a restaurant. And this summer we are going to Africa. (We're still figuring out how all that will work.) The forum has been a life-saver to me. I've learned something every day. I am so grateful for the people on here who share their struggles and answer the same questions over and over. Sorry for the long post--I've not replied before, but I just wanted you to know there's light at the end of the tunnel. You'll get there and it won't seem so difficult someday.
Thanks, Rachel--we are very excited about our trip. My husband is determined that we are going to do the things we planned to do and not let the celiac rule our lives. He's just grateful to be feeling well again and is very adaptable about what he eats. A Lara bar works for him if there's nothing else around. Yes, I am gluten-free as well, except perhaps when I'm out eating on my own. It makes things much simpler. And I need simple--who can keep track otherwise? BTW this is the first time I've posted--been reading the board since July--so I'm not sure just how this works. Hopefully this is in the right place!
My husband was diagnosed with celiac disease last July, and we both have been very carefully gluten-free ever since--and he's feeling great. We haven't traveled since then, but we have scheduled a trip with a small tour group to Tasmania and Uganda. He's planning on living on the things we pack with us! I'm hoping that it can be better than that. Has anyone done an African safari? Any advice for us?


Baker Beach has long been an eclectic place for gatherings. As early as the 1890s, when the Presidio was still a military post, people began having bonfires at Baker Beach. In 1933, a fire believed to be caused by a careless fisherman camping at the beach spread all the way up the cliffs to within 100 feet of the coastal gun batteries, covering a distance of a half a mile (it took 50 Presidio firemen, 100 coast artillerymen, and three SF engine companies to extinguish the fire). In 1997, Baker Beach became part of the national park system, and today beach fires are no longer permitted.
In June 1986, while the Presidio was still an active Army post, a new tradition started at the northern end of Baker Beach – Burning Man. Initially, the founders of Burning Man were drawn to Baker Beach because of the bevvy of driftwood found there. The northern portion is notoriously known as a "clothing optional" spot for sun bathing and one of the more isolated beaches in San Francisco. Thus it was the perfect place for a group of 20 friends to gather for a bonfire where Burning Man founder Larry Harvey set aflame the eight foot tall wooden structure he called "the Man."
Though Larry was the first to burn a structure of a man, he wasn't necessarily the first to start this effigy burning tradition at Baker Beach. Several years before burning a man was even a twinkle in Larry's eye, Mary Grauberger, a friend of Larry's girlfriend, held annual bonfires at the beach where she'd assemble driftwood statues to burn in honor of Summer Solstice.
Picture this: it's pre-tech boom San Francisco and the City is overflowing with artists, hippies, and free-thinkers. Into this comes the Cacophony Society, "a randomly gathered network of free spirits united in the pursuit of experiences beyond the pale of mainstream society," according to Michael Mikel, one of the Cacophony Society's founders.
Influenced by the Dadaists, Surrealists, and Situationists, the Cacophony Society was a proponent of the "free university." They also had an extensive mailing list and each month they'd invite members to participate in fun and unusual activities – like exploring architecture, putting on plays, or dressing up and having adventures around San Francisco.
Larry Harvey was a member of the Cacophony Society, and in 1988 he invited Michael to attend his first effigy burning at the as yet-unnamed event at Baker Beach. The following year Larry asked Michael to help publicize the solstice gathering through the Cacophony Society's mailing list, and thus the event became the newly minted "Burning Man" in their newsletter. Attendance shot up to 300.
"San Francisco was ripe for something like Burning Man," Michael shared. The following year attendance grew to 350 before local law enforcement shut down the burning of "the Man" due to the group's lack of a fire permit.
In 1990, Burning Man moved to Nevada, far away from the Presidio and the Golden Gate Bridge. And the event has certainly grown since – last year 65,564 people watched a 60-foot structure of a man burn on the playa in the heat of Black Rock Desert.
Michael now lives in Reno so he's closer to Black Rock Desert, but he comes to San Francisco every few months, and he's looking forward to speaking at the Presidio Officers' Club in August 2016. "The Presidio is such a beautiful place," he says. "I came out to Crissy Field recently and gave friends a tour of the whole area. We had a barbecue." When asked if he burned anything while there, he laughed, "No. Surprisingly, I managed to not burn the hot dogs."
Then: June 2012 — Michael Mikel (left) and Larry Harvey (right), founders of Burning Man, at Baker Beach, Summer Solstice, June 20, 2012, for the 25th Anniversary of Burning Man.
Now: Summer 2016 – Baker Beach is still a popular public beach, and one of the warmest sandy spots on San Francisco's Pacific Coast. It's an excellent place to sunbathe, whale watch, or have a picnic. Though bonfires are no longer allowed on the beach here, check out the National Park Service Beach Fire Regulations for information on beaches in the area where fires are permitted.


text:
Амурла́г (Амурский исправительно-трудовой лагерь) — подразделение, действовавшее в системе исправительно-трудовых учреждений СССР.

История

Амурлаг создан в 1947 году. Управление Амурлага располагалось в городе Свободный (ныне Амурская область). В оперативном командовании он подчинялся первоначально Специальному главному управлению Главспеццветмета (СГУ) при Управлении исправительно-трудовых лагерей и колоний Управления внутренних дел Хабаровского края, в 1949 году перешёл в состав такой же структуры Амурской области и в 1953 году вошёл в состав ГУЛАГ Министерства юстиции.

Максимальное единовременное количество заключённых могло составлять более 3000 человек.

Амурлаг прекратил своё существование в 1953 году.

Производство
Основными видами производственной деятельности заключённых Амурлага были железнодорожное строительство, золотодобыча, горные работы и заготовка леса.

Примечания
В период с 1938 по 1941 года в системе НКВД существовал Амурский железнодорожный исправительно-трудовой лагерь, также известный как Амурлаг.

Ссылки
 Амурский исправительно-трудовой лагерь

Лагеря ГУЛАГа
XX век в Амурской области
Появились в 1947 году в СССР
Исчезли в 1953 году

redpajama_set_name: RedPajamaWikipedia


text:
CommentTuesday 14 December 2021
Holy Matrimony? Recognition of same-sex marriage in the Methodist Church
Holy Matrimony? Recognition of same-sex marriage in the Methodist church Mark Rowland is a PhD student at Leeds researching a queer theology of holiness. He has been a Methodist minister since 2009 and currently also serves as Free Church Chaplain at the University of Warwick. He is a founder of...
Researcher of the MonthWednesday 8 December 2021
Researcher of the Month – December 2021, Dr Simon Hewitt
Simon Hewitt is a lecturer in the School of Philosophy, Religion, and the History of Science at Leeds. He is particularly interested in the doctrine of God and political theology, and his latest book is Negative Theology and Philosophical Analysis: Only the Splendour of Light, published by Palgrave Macmillan. Tell...
Event RecordingsThursday 4 November 2021
Inaugural lecture Adriaan van Klinken: Reimagining Christianity and Sexual Diversity in Africa
On 28 October 2021, Adriaan van Klinken delivered his inaugural lecture as Professor of Religion and African Studies at the University of Leeds. In this lecture, loosely based on his recently-published book co-authored with Professor Ezra Chitando, Reimagining Christianity and Sexual Diversity in Africa, he explored how African thinkers, writers, activists,...
Event AnnouncementsThursday 4 November 2021
Research seminar jointly hosted by the Desmond Tutu Centre for Religion & Social Justice, University of the Western Cape the Leeds University Centre for African Studies the Leeds University Centre for Religion and Public Life 10 November 2021, 2:00-3:30pm UK time Registration essential: please email DTC@UWC.AC.ZA In a context...
AnalysisWednesday 20 October 2021
Christian and Jewish Charities in Leeds after WW1: The Response to Disabled Ex-Servicemen
In this blog post, historian Dr Bethany Rowley writes about an often-overlooked part of Leeds' religious history: the response of Christian and Jewish charities to the needs of disabled veterans in the aftermath of the First World War. Based on her archival research, she demonstrates that Leeds was a picture...
Event AnnouncementsMonday 18 October 2021
On 28 October 2021, Adriaan van Klinken will deliver his inaugural lecture as Professor of Religion and African Studies at the University of Leeds, under the title Reimagining Christianity and Sexual Diversity in Africa. The School of Philosophy, Religion and History of Science (PRHS) and the Leeds University Centre for...
Researcher of the MonthMonday 11 October 2021
Researcher of the Month – October 2021: Chrissie Thwaites
Chrissie Thwaites is a PhD student in theology and religious studies, and a member of the Centre for Religion and Public Life. Her researches focuses on the impact of purity culture in UK Christianity, and is funded by the AHRC through the White Rose College of Arts and Humanities (WRoCAH)....
Event AnnouncementsMonday 4 October 2021
We are hosting the inaugural Buddhism in Britain Research Symposium on Wednesday 15th December 2021, 10-4pm, via Zoom, and we are seeking contributions from academics, scholar-practitioners, and research students who are interested in sharing insights into this subject area via 25-30 minute papers and round-table discussions. This new research symposium...
Event RecordingsWednesday 22 September 2021
Religious Communities and Pastoral Care in Times of COVID
On 22 June 2021, the Centre for Religion and Public Life, University of Leeds, held a research day about "Religious Engagement and the COVID Response". Please find here the recording of the second panel of speakers, focused on "Religious Communities and Pastoral Care in Times of COVID". Speakers below are...
Researcher of the MonthMonday 20 September 2021
Researcher of the Month – September 2021, Laura Wallace
Laura Wallace is a feminist researcher undertaking a PhD in the school of Philosophy, Religion and History of Science at the University of Leeds, funded by the White Rose College of the Arts and Humanities (WRoCAH). Originally from Dublin, Ireland, her research interests are in Irish studies of Catholicism, sex...

redpajama_set_name: RedPajamaCommonCrawl


秋冬了!5種適合乾燥室內的植物 | 5 Plants for Home in Autumn and Winter!
In Hong Kong, it is always very dry indoor when autumn/winter comes; an easy way to tackle this problem is to add some plants at home that humidifies as well as making your home more lively.
But choosing the right plant is very important too. Some plants love a humid environment, and some love it warm. To keep your plants happy in this dry winter, you need to know which type to get.
Many people know that aloe is good for our skin, in fact, aloe can also purify air, remove formaldehyde and absorb odor. To keep this useful plant happy, we should keep it in a warm and dry environment. The indoor atmosphere in winter time Hong Kong is perfect for aloe to thrive. As a decoration, aloe has a very strong and distinct appearance that easily adds characteristics to your home. Simply put an aloe in a silver pot and place it on a wooden desktop, and your home instantly becomes chic and classy.
If you have a big enough dinner table, a little pot of aloe acts as an ideal table top decoration. An even more creative way is to add an used food tray at its bottom, like the above photo!
Ivy is a gracious looking evergreen. There are many possibilities to use ivy as a decoration, according to the design of your homes. Through all the micro stoma on its leaves ,ivy effectively filters out any carcinogens from nicotine, absorbs harmful substances, and transform them into sugar and amino acids that are both harmless to our bodies. Ivy doesn't need a lot of humidity for its growth, and since the air is usually more polluted in autumn and winter, keeping an ivy at home can be a very good idea.
Cactus loves sunshine, it can also endures great heat and dryness. Its extreme vitality is a perfect choice of plants for autumn and winter in Hong Kong. Cactus is also known as the 'night time oxygen bar', in which it effectively absorbs carbon dioxide while releasing a good amount of oxygen. Cactus can also absorb dusts and purify air.
Cactus appeals to many people and comes with lots of different shapes and forms; a bigger pot of cactus on its own, by the window or your favorite furniture makes a very eye-catching statement decoration.
Another idea to decorate your homes would be to gather a few smaller cacti and a couple other green plants, and keep them all in the same kind of pots which adds consistency.
Snake plant originates from tropical climates in Africa and India, therefore they don't rely on humidity to grow. Sufficient sunlight is enough to keep it thrive. In winter, create a nice environment for snake plants by keeping them in a warm room facing the sun.
Insufficient sun would cause snake plant's leaves to turn dull, but you shouldn't immediately move a snake plant to the sun after it's been hiding in the shade for a while. Instead, first place the sun-deprived snake plant in a somewhat brighter spot, let it adapt before moving it to the sun; this can prevent the plant's leaves from getting sunburnt.
Snake plants are very low-mainatence, too. You don't have to often trim its leaves. Putting 2-3 pots of snake plants together create an interesting view at home!
Umbella plant can grow under low light conditions, therefore it makes a very good plant choice for indoors. They are highly adaptive, perform great in winter as long as they get enough water.
Because of its strong vitality and gracious appearance, umbella plant gives off a lively feeling. It can also absorb nicotine and other harmful substances, making it one of the world's most popular indoor plants.
All of the above plants have one thing in common: they are low maintenance with a strong vitality. Give these plants sufficient sunlight and care, and they will thrive to be the perfect plant for your homes.
We hope to make design furniture accessible and affordable. Feel free to visit our showroom with your family or friends.


text:
Carbon Black Shipments up 6% for March on Strong Tire Sector Demand
on May 12, 2017 in Market Background, Materials
Shipment volume of carbon black rose 6.1 percent year-on-year to 52,479 tons in March, the third straight month of positive growth, according to records compiled by the Japanese Carbon Black Association. Total shipment for rubber use rose 6.8 percent year-on-year, up for the third straight month, at 49,270 tons. Although shipments for tire use were up 11 percent, shipments for...
Driven by Tires, Japan's Carbon Black Shipments Rise for February
on April 18, 2017 in Market Background
Shipment volume of carbon black in February was up 2.3 percent to 46,531 tons from the same period last year, according to data compiled by the Japanese Carbon Black Association. It marks the second consecutive month of a year-on-year increase this year. Of carbon black used for rubber, shipments for tire purposes increased 6.3 percent, and shipments for general rubber...
Carbon Black Shipments Down 1.3% for October
on December 19, 2016 in Market Background, Materials
Carbon black shipments totalled 50,624 tons in October, down 1.3 percent year-on-year, according to data compiled by the Japanese Carbon Black Association. This marked the 11th consecutive month below last year's shipments. Shipments of carbon black used for rubber totaled 47,456 tons, down 1.4 percent year-on-year. Apart from the shift to negative growth for tires with a 1.1 percent decline...
Carbon Black Shipments Down 0.9% for September
on November 11, 2016 in Market Background
According to data collated by the Japanese Carbon Black Association, 49,729 tons of carbon black was shipped in September. This is a 0.9 percent year-on-year decrease and the tenth consecutive month of a year-on-year decline. Of carbon black used for rubber, shipments of that used in tires turned positive, seeing a 3.3 percent increase. At the same time, carbon black...
Tokai Carbon Increases Price for Carbon Black as Oil Continues to Rise
on November 3, 2016 in Materials
Tokai Carbon Co. Ltd. (TYO:5301) has increased the price of its carbon black products by 3 yen per kilogram. The change was implemented for deliveries beginning Nov. 1, applying to all varieties of carbon black. A release from the company stated that the price of raw oil used in carbon black has been rising incrementally since bottoming out at the...
Carbon Black Shipments Down 7.3% to 49,900 Tons for July
on October 6, 2016 in Market Background
Shipment volume of carbon black in July was down 7.3 percent to 49,900 tons from the same period last year, marking the eighth consecutive year-on-year decline, according to the Japanese Carbon Black Association. Carbon black shipments for rubber products dropped by 6.5 percent for tires and 10 percent for general rubber products. The total shipment volume for rubber products was...


*This paper written by Michael J. Werner, JD, and was developed for the Health and Public Policy Committee: Whitney W. Addington, MD, Chair; Phillip D. Bertram, MD, Vice-Chair; Derrick L. Latos, MD; Philip Altus, MD; Angela McLean, MD; Risa J. Lavizzo-Mourey, MD; William M. Fogarty, MD; David J. Gullen, MD; Nancy E. Gary, MD; Robert A. Berenson, MD; Janice Herbert-Carter, MD; James Webster Jr., MD; and Richard Honsinger Jr., MD. Approved by the Board of Regents on 27 October 1995. Requests for Reprints: Michael J. Werner, JD, American College of Physicians, 700 13th Street, NW, Suite 250, Washington, DC 20005.
As the health care system changes and large managed care entities gain greater control in some markets, proponents of antitrust reform have expressed concern that physicians could lose their autonomy.To respond to this concern, the American College of Physicians has consistently argued that physicians should be allowed to establish their own health plans and networks to provide high-quality and cost-effective care. Moreover, the College has advocated utilization review reform and due process protections to empower physicians in their dealings with insurers.
Under current antitrust law, as interpreted by the federal enforcement agencies, physicians already have the legal authority to form their own health plans and networks, and many state medical societies are sponsoring such plans.The law also allows physicians to operate the clinical components of a health plan, regardless of who owns it. Moreover, physicians can share information about quality, utilization, and, in some circumstances, fees. An examination of federal enforcement agency actions since the mid-1970s shows that physician networks have rarely been challenged. In light of market developments, however, the College has urged the federal antitrust agencies to analyze the effect of their current enforcement policies on physician activities and adopt a more flexible approach. Further monitoring and analysis of the changing health care marketplace are necessary to ensure that physicians are being treated fairly and to determine which factors spur or inhibit the development of physician-run health plans and networks.
Published: Ann Intern Med. 1996;125(1):59-65.
Migrate your mind: the role of palliative care in transcultural cancer treatment : AÂ qualitative analysis.


text:
Ксанторе́я (Xanthorrhoea Sm.) — єдиний рід однодольних ксерофітних вічнозелених рослин підродини ксантореєвих (Xanthorrhoeoideae), типовий рід родини ксантореєвих (Xanthorrhoeaceae). Раніше їх об'єднували з лілієвими.

Назва

Назва роду ксанторея, яка може бути перекладена як «жовтосмолка», походить від кольору смоли, що виділяє ксанторея смолиста (Xanthorrhoea resinosa), яка стала відомою європейцям раніше за інші види роду.

Крім широко поширеної в Австралії назви деревоподібних ксанторей — «злакові дерева», місцеве населення використовує для них ще одну назву — «Чорний хлопчик» («Black boy»), підставою для якої послужила деяка схожість цих рослин з темношкірими людьми, що несуть на голові пучок трави.

Біологічна характеристика

Зазвичай деревоподібні прямостоячі рослини з товстим стовбуром-каудексом, іноді злегка розгалужені, що несуть на верхівці розетку (або розетки) довгого (до 1 м завдовжки) жорсткого лінійного листя і спідницю з відмерлих листків. Пожежі стимулюють появу на верхівці стовбура довгої, позбавленої листя, квіткової стрілки, вкритої густими циліндричними білими або кремовими ароматними суцвіттями, що нагадує качан рогозу. Тичинок 6, розташовані 2 колами. Пиляки прикріплені до тичинкових ниток спинкою. Насіння ксантореї мають блискучу чорну оболонку, майже горизонтально розташований зародок і невеликий білуватий рубчик на одному з кінців. Якщо вони утворюються по кілька в одному гнізді, то вони мають різну величину і форму.

Всі види ксанторей мають цікаву особливість, що ймовірно є засобом захисту від перегріву або механічних пошкоджень: їх каудекс, основи листя, а іноді й суцвіття виділяють велику кількість клейкої смоли, шар якої на стовбурах деревоподібних видів може досягати завтовшки 2 — 4 см. У більшості видів вона має коричневе або темно-червоне забарвлення, у ксантореї смолистої (Xanthorrhoea resinosa) смола жовта. У ксанторей смола дуже густа і застигає прямо біля основи листя. Коли рослина викидає нові листя, вони піднімаються над колишніми, але ростуть також з серцевини рослини. Навколо їх основи у свою чергу застигає шар смоли. Так триває з року в рік, і затверділа смола піднімається все вище, поки по суті, не починає виконувати роль стовбура. Це не стовбур в справжньому сенсі слова, так як всередині він складається з волокнистих основ листя, залитих смолою. Кора відсутня, і поживні речовини досягають вершини по серцевині.

У найбільшого виду цього роду — ксантореї австралійської (Xanthorrhoea australis) — стовбур може досягати заввишки 6 — 7 м і канделяброподібно гілкуватися у верхній частині, утворюючи кілька листових розеток з суцвіттями або без них. У інших деревоподібних ксанторей: ксантореї чотиригранної (Xanthorrhoea quadrangulata), ксантореї Прейса (Xanthorrhoea preissii) і ксантореї деревоподібної (Xanthorrhoea arborea) стовбури-каудекси не перевищують заввишки 2 — 3 м і зазвичай не гілкуються. Деякі інші види, прикладом яких може служити найвідоміший вид роду — ксанторея смолиста (Xanthorrhoea resinosa), мають каудекс, що лише трохи підноситься над поверхнею землі (дуже рідко він досягає заввишки 1 — 1,5 м). У широко поширеної в Південній Австралії і на Тасманії ксантореї малої (Xanthorrhoea minor) каудекс вже повністю прихований в землі, а її пагони утворюють більш-менш густі дерновини з суцвіттями заввишки до 80 см. Ще дрібніше ксанторея карликова (Xanthorrhoea pumilio) з Квінсленда, суцвіття якою підносяться над поверхнею землі всього на 20 — 40 см.

У роді ксанторея відзначається досить широке поширення міжвидової гібридизації, що частково визначається присутністю у всіх вивчених видів цього роду одного і того ж числа хромосом (2п = 22).

Поширення

Ксанторея — ендемік Австралії, характерний для сухих саван цього континенту.

Екологія

За формою росту ксанторею відносять до так званих трав'яних або злакових дерев, через схожість їхнього довгого і вузького листя з листям злаків. Представники цього роду ростуть дуже повільно; щоб вирости на 1 см, деякі рослини повинні утворити 550 листків. Деякі види утворюють 5 листків за 2 роки. Ксантореї живуть до 600 років. Австралійський ботанік Ч. Л'юїс, спираючись на власний метод розрахунку, встановив вік деяких ксанторей, який перевищив 6 тисяч років.

Рослини цього роду адаптовані до чагарникових пожеж, які можуть стимулювати цвітіння і спалюють відмерле листя уздовж стовбура.

Зазвичай ксантореї зустрічаються групами на відкритих ділянках австралійської савани, що іноді нагадують вересові пустки північної півкулі, або серед заростей ксерофільних чагарників, так званого скреба, рідше на відслоненнях пісковика та інших кам'яних порід по схилах пагорбів і гряд. Менші види — ксантореї смолиста і мала — ростуть у вологіших місцях, часто на піщаних і торф'янистих пустках, а іноді навіть на болотах.

Використання

Деревина кванторей служить будівельним матеріалом і йде на різні вироби. Смолу, що місцеве населення виділяє з цих рослин, використовують для виготовлення лаку, але не дуже високої якості. Особливо відома в цьому відношенні жовта смола ксантореї смолистої. Вона має гострий смак і горить з приємним бензойним запахом. У меншій кількості використовують коричневу або червону смолу інших ксанторей. Їхню смолу застосовують також у вигляді клею. У аборигенів Австралії ніжки суцвіть ксанторей йшли на виготовлення списів, наконечники до яких приклеювали цією смолою.

Деревоподібні види ксанторей дуже декоративні і заслуговують широкого введення їх в культуру в садово-паркових господарствах поряд з юками, драценами, агавами і доріантесом.

Види

За даними сайту «The Plant List» рід Xanthorrhoea включає 28 видів:
 Xanthorrhoea acanthostachya D.J.Bedford
 Xanthorrhoea acaulis (A.T.Lee) D.J.Bedford
 Xanthorrhoea arborea R.Br.
 Xanthorrhoea arenaria D.J.Bedford
 Xanthorrhoea australis R.Br.
 Xanthorrhoea bracteata R.Br.
 Xanthorrhoea brevistyla D.A.Herb.
 Xanthorrhoea brunonis Endl.
 Xanthorrhoea caespitosa D.J.Bedford
 Xanthorrhoea concava (A.T.Lee) D.J.Bedford
 Xanthorrhoea drummondii Harv.
 Xanthorrhoea fulva (A.T.Lee) D.J.Bedford
 Xanthorrhoea glauca D.J.Bedford
 Xanthorrhoea gracilis Endl.
 Xanthorrhoea johnsonii A.T.Lee
 Xanthorrhoea latifolia (A.T.Lee) D.J.Bedford
 Xanthorrhoea macronema F.Muell. ex Benth.
 Xanthorrhoea malacophylla D.J.Bedford
 Xanthorrhoea media R.Br.
 Xanthorrhoea minor R.Br.
 Xanthorrhoea nana D.A.Herb.
 Xanthorrhoea platyphylla D.J.Bedford
 Xanthorrhoea preissii Endl.
 Xanthorrhoea pumilio R.Br.
 Xanthorrhoea quadrangulata F.Muell.
 Xanthorrhoea resinosa Pers.
 Xanthorrhoea semiplana F.Muell.
 Xanthorrhoea thorntonii Tate

Примітки

Література

 Brummitt, R. K. 1996. Report of the Committee for Spermatophyta: 44. Taxon 45:674. [conservation proposal considered unnecessary by Committee for Spermatophyta].
 Nelson, E. C. & D. Bedford. 1992. (1059) Proposal to conserve 1042 Xanthorrhoea J.E. Smith over Acoroides C. Kite (Xanthorrhoeaceae).

Джерела

 Семейство ксанторреевые (Xanthorrhoeaceae) // Жизнь растений: в 6-ти томах. — М.: Просвещение. Под редакцией А. Л. Тахтаджяна, главный редактор чл.-кор. АН СССР, проф. А. А. Федоров. 1974.

 Меннинджер Э. Причудливые деревья. Пер. с англ. И. Г. Гуровой. Под ред. и с предисл. П. И. Лапина. М., «Мир», 1970. 371 с. с илл. (В мире науки и техники).
 Xanthorrhoea на сайті Germplasm Resources Information Network (GRIN) Міністерства сільського господарства США
 http://www.tropicos.org/Name/40027049
 Xanthorrhoea на сайті «Агбина»

Посилання

 http://www.desert-tropicals.com/Plants/Xanthorrhoeaceae/Xanthorrhoea.html
 http://www.plantsystematics.org/taxpage/0/genus/Xanthorrhoea.html

Ксантореєві
Ендемічна флора Австралії
Роди холодкоцвітих


text:
/*
 *               In the name of Allah
 * This file is part of The "Quran Teacher or Learn Arabic" Project. Use is subject to
 * license terms.
 *
 * @author:         Fazle Rabbi Rahat
 *
 */
package QuranTeacher;

import java.awt.EventQueue;

import javax.swing.JFrame;

public class Main extends JFrame {
	/**
	 * Contains the main function
	 */
	private static final long serialVersionUID = 1L;

	public Main() {
	}

	/**
	 * Launch the application.
	 */
	public static void main(String[] args) {
		EventQueue.invokeLater(new Runnable() {
			public void run() {
				try {
					//setDesign("com.sun.java.swing.plaf.nimbus.NimbusLookAndFeel");
					MainFrame frame = new MainFrame();
					frame.pack();
					frame.setVisible(true);
				} catch (Exception e) {
					e.printStackTrace();
				}
			}
		});
	}

/*    private static void setDesign(String newLookAndFeel)
	{
		try
		{
			UIManager.setLookAndFeel(newLookAndFeel);
		}
		catch(Exception e)
		{
			System.out.println("Unable to load look and feel");
		}
	}*/

}

redpajama_set_name: RedPajamaGithub


text:
Wrongly Left Out of a Loved One's Will? See a Solicitor Straight Away!
Posted January 13, 2020 by Schofield and Associates & filed under Uncategorized.

If you feel that you have been wrongly left out of a loved one's will, you should see a solicitor straight away. The wisdom of acting promptly was emphasised by a case in which a pensioner who delayed taking legal action almost lost the roof over her head.

The woman had, for over 25 years, shared a home with a man who she claimed was her husband. The property was held in his sole name and, by his final will, he left it and everything else he owned to his daughter. Following his death, the daughter took steps to evict the woman from the property.

The woman launched proceedings under the Inheritance (Provision for Family and Dependants) Act 1975, seeking reasonable provision from the man's estate on the basis that she was financially dependent on him. The claim was, however, lodged long after the expiry of the six-month time limit that applies to such proceedings. A judge refused to extend that deadline and, after the woman failed in a first appeal against that decision, her claim was dismissed.

In upholding her challenge to that outcome, the Court of Appeal found that the judge's decision was plainly wrong. He had completely discounted the fact that the woman risked losing her home if the time limit were not extended. The merits of her claim were strong and it was a plain case for such an extension to be granted.

The Court noted that one of the reasons for the delay in taking legal action was that the woman had, during a crucial period, dispensed with professional advice, relying instead on the guidance of a friend who had no legal qualifications. The Court's ruling opened the way for the woman to proceed with her claim.

Schofield & Associates
The Corn Exchange
Email: eileen.schofield@schofieldandassociates.co.uk
Web: www.schofieldandassociates.co.uk
2 Snow Hill,
Schofield HR
Email: angela.richards@schofieldhr.co.uk
Consent & Personal Data
© 2020 Schofield & Associates. Website by Newman Marketing


text:
Samsung Believes That Millennials Are Looking For Vertical TVs
By Natasha Mathur 2 months, 2 weeks
Samsung knows the millennial generation is obsessed with smartphones. Samsung is also aware that most of this time is spent on Instagram and Snapchat, scrolling through people's stories and posts. Keeping that in mind, Samsung has come up with a new convenient and innovative way to further fuel that obsession with its new "vertical TV", named Sero.
Sero, a new 43.5-inch TV, equipped with 4.1 -channel and 60-watt high-end speaker, can be rotated vertically to stream mobile content (Instagram/Snapchat stories, livestream, et al), providing larger and better display access.
Priced at $1600 (approximately INR 1,11,490), Sero works by synchronizing the mobile screen with the vertical TV screen using a simple function based on NFC. However, this doesn't mean that you can't operate it as a conventional TV. Sero comes with an added ability to flip to landscape mode, giving you the best of both worlds. However, its use-cases can be extended to a lot more than just using it for Instagram and Snapchat.
The TV can also display photos along with clocks and sound walls that can be displayed to create a unique interior at offices and homes. Moreover, there's also a built-in microphone that can be used in conjunction with Samsung's AI virtual assistant platform, Bixby, that can help control various functions in the TV. It can also be used to play music with Samsung Music and the music stored on your phone.
Now, Samsung has targeted the TV towards the millennials, however, it also sounds like an absolutely genius idea for content marketers and advertisers, wanting to promote their brand. We see different brand stories pop up as advertisements on Google. However, the same stories can be broadcast to the larger audience, even to the ones who don't necessarily have an Instagram or Snapchat account to check out these stories. This is because it's a TV at the end of the day (a futuristic one at that), which can be hung within an office or even in the mall, and catch people's attention.
It also sounds like a great idea for businesses and retail industry, wanting to drive more customer interaction and engagement. Organizations can use it in meetings to make better business decisions. Some might argue that using a laptop or a phone to notify people on public feedback to a certain product will do the same thing. However, where Sero can mark the distinction is that it is said to offer a larger display, and better sound, thereby, leading to enhanced interaction among the team.
It can help with showing how well a certain business product is performing, with the help of comments under stories being displayed on a full screen. Overall, it can help enhance the business decisions, which in turn will help with larger and wider audience reach.
Also, the TV sounds like a dream for South Koreans, where live streaming is carried out extensively across online platforms such as Facebook, YouTube, Instagram, et al. Although the livestreaming trend is all the rage in Korea, it has started to pick up all across the world.
Now, many people are labeling the Sero as an idea unwanted and unneeded by millennials, but given the whole hype around Instagram, YouTube, and Snapchat, we might just come across instances in the future where this whole idea catches up with even the millennials!
Cover image: Dhawal Bhanushali / Mashable India
TOPICS: Tech, TV, look of the day, YouTube, Snapchat, Genesis by Mashable India, Samsung, Tv Streaming, Millennials
Tablet. Laptop. Doodlepad. New Samsung Galaxy Tab S4 is bringing it all together
Intergalactic shock: Casio collaborates with Gorillaz to bring back a classic!
Apple acquires one of our favorite apps: Is it bye bye Shazam for Android users?
Ultimate Ears has launched the Wonderboom Freestyle collection and boy are we ready to party
Let's talk about NEX baby: Vivo's new super phone ditches the notch. Here's the review


text:
Thakeham to build 30 new zero carbon homes in next phase of Ockford Ridge redevelopment
New Zero Carbon homes will ease cost of living pressures through homes that are more resilient against the energy crisis
Waverley Borough Council has appointed South East based community creator Thakeham to build 30 new social and affordable homes on a site at Ockford Ridge in Godalming.
Further focus on increasing renewable technology to help local people with the cost of living
The range of 18 new houses and 12 new flats will be made available for shared ownership purchase, social and affordable rent. Work will begin in the Autumn of 2022 and is expected to be completed in October 2023. The £7 million contract is Thakeham's fourth for Waverley Borough Council and the third scheme appointment at Ockford Ridge.
These 30 homes at Ockford Ridge will be Nett Zero Carbon in lifetime operation. This means that the amount of carbon emissions associated with the building's energy on an annual basis is zero or negative. A net zero carbon building is highly energy efficient and powered from renewable energy sources. In a new change in this next phase at Ockford Ridge, each home at Ockford Ridge will feature an air source heat pump on top of the features Thakeham built into previous phases including; a pre-insulated frame, waste water heat recovery, solar panels and EV car charging for most plots.
For Thakeham, Rob Boughton, CEO said:
We have already completed 54 affordable homes at Ockford Ridge – delivering much-needed zero carbon in operation, social and affordable housing. A number of former residents have now returned to the area in new homes.
We are especially proud of the relationship we have built with the local community. We appointed a site dedicated Resident Liaison Officer to manage communications with the community. We are working with Waverley Borough Council and the local community through the school and community centre to ensure that our time at Ockford Ridge creates a legacy for generations to come.
Improvements at Ockford Ridge are being made over six sites; redeveloped in phases up until 2025. The original estate dates to the 1930s and Waverley Borough Council has delivered a modern community at Ockford Ridge. Thakeham have been proud to have worked with Waverley Borough Council delivering 37 and 17 new homes in previous phases.
The broader social impact of the new homes at Ockford Ridge has been carefully considered. Thakeham has collaborated with Waverley and the Ockford Ridge community to foster initiatives such as school visits and learning workshops, led by local charity Skillway. Together, they also supported a breakfast club, run for the community by St Mark's Community Centre, providing free breakfasts for workers from the centre once a week.
The Community Benefit Programme will continue to be delivered over the next 15 – 18 months as they continue to work with the community and WBC to deliver the next phase. Other initiatives have seen Thakeham work with the community centre and Green Oak Primary School on an estate-wide clean up event. Volunteers removed litter, dead leaves and other rubbish to keep the appearance of the estate clean and tidy.
Councillor Nick Palmer, Co Portfolio Holder for Housing (Delivery) said:
The homes built at Ockford Ridge are a mark of pride for the council. We have delivered modern sustainable homes for the community with enhanced environmental standards. The next phase will see the use of renewable technology and homes which will be net zero carbon in operation and we are pleased to continue working in partnership with Thakeham to bring our aims to fruition.
You can stay up to date with all Thakeham news here


I spent 2.25 on a pepsi today, way too much but it came from my 10/week spending budget. I still have 6.20 left to spend.
Tomorrow I turn the big 40, and I'm having a low key day, work and I may take myself out for supper ( there are places you can eat for free on your birthday, so i think I'll go there). On the weekend i'll celebrate with family and friends.
Plus my travel time and gas expenses have dropped too. I used abut 1/4 tank to get here this week, so that will be a huge savings! If I can cut my gas budget from 300/month to 220/month that will be amazing!
Happy birthday!! I hope you have a great day tomorrow!
Happy Birthday!!! Enjoy the big 4-0, make it a great year for yourself!!


Former Meath minor star Conor Nash will make his AFL debut for Hawthorn Hawks on Saturday.
The 20-year-old has been included on the Hawks' bench for their Round 21 clash against Geelong, whom have Zach Tuohy (Laois) and Mark O'Connor (Kerry) in their ranks, at the Melbourne Cricket Ground (MCG).
Nash signed with the Melbourne club as a rookie at the end of 2016 and was elevated to Hawthorn's senior list last month after having shown tremendous progress over the last 12 months.
"I'm absolutely ecstatic," Nash told hawthornfc.com.au after learning the news.
"When I found out yesterday in front of the family in the car, it was just pure joy. I can't wait, and I look forward to contributing to the team."
Having visited on holiday recently, Nash's younger brother and sister were planning to return back home before learning the news this week and rearranged their flights to see their brother make his bow for the Hawks.
"Clarko (Alastair Clarkson, Head Coach) asked them if they could prolong their flights until Sunday or Monday. They'll get to see me play which is a very special moment for them, and for myself as well," Nash added.
Hawthorn will unveil Irish debutant Conor Nash this Saturday against the Cats.


1 John 4:4, You are of God, little children, and have overcome them, because He who is in you is greater than he who is in the world.
This is the "them" that we have overcome. We have overcome lesser spirits in order to know the Spirit of God. In this message, you will learn about the three keys to living the overcomer's lifestyle!
Mark 4:23, "If anyone has ears to hear, let him hear." 24] And He said to them, "Take heed what you hear. With the same measure you use, it will be measured to you; and to you who hear, more will be given. 25 For whoever has, to him more will be given; but whoever does not have, even what he has will be taken away from him."
Hearing is a function of our will to understanding what is being said. We listen with our ears, but we hear with our minds and hearts.
1. Passive Listening • Passive means, "not active, but acted upon." Passive people are generally sluggish and disengaged. Passive listeners only "purk up" when something is said that relates to what they want. In the face of responsibility, a passive listener may pretend to be deaf.
Passive listening produces frustration in any communication or relationship because it creates uncertainly, slow progress, and shortcuts excellence.
2. Convenient Listening • Convenient is defined as, "Situated within easy reach." The convenient listener only applies his heart to hear when the information or task is within easy reach. As believers, we have a dilemma: We live in a world that promotes convenience above everything; but we also live in a Kingdom of which Jesus said, "the violent [or aggressive] take it by force" (Matt 11:12).
4. Active Listening • This should become our goal: Active listeners "hear" the communication accurately the first time. "Active" is defined: "Not waiting to be acted upon. Prepared to apply what is heard and act on it—regardless of circumstances.
James 1:25, But he who looks into the perfect law of liberty and continues in it, and is not a forgetful hearer but a doer of the work, this one will be blessed in what he does.
Always serve God, even when doing so brings difficulty, like sour lemons. Through the power of Jesus Christ, you can bring the water of the word and sweetness of the Holy Spirit to every bitter circumstance. This will produce something good, from something bad, like refreshing lemonade. Then, you will be able to sip and rejoice as God shows you, and others, his love, power, mercy and favor.


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