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Aβ deposits represent a neuropathological hallmark of Alzheimer's disease (AD). Both soluble and insoluble Aβ species are considered to be responsible for initiating the pathological cascade that eventually leads to AD. Therefore, the identification of therapeutic approaches that can lower Aβ production or accumulation remains a priority. NFκB has been shown to regulate BACE-1 expression level, the rate limiting enzyme responsible for the production of Aβ. We therefore explored whether the known NFκB inhibitor celastrol could represent a suitable compound for decreasing Aβ production and accumulation in vivo.
The effect of celastrol on amyloid precursor protein (APP) processing, Aβ production and NFκB activation was investigated by western blotting and ELISAs using a cell line overexpressing APP. The impact of celastrol on brain Aβ accumulation was tested in a transgenic mouse model of AD overexpressing the human APP695sw mutation and the presenilin-1 mutation M146L (Tg PS1/APPsw) by immunostaining and ELISAs. An acute treatment with celastrol was investigated by administering celastrol intraperitoneally at a dosage of 1 mg/Kg in 35 week-old Tg PS1/APPsw for 4 consecutive days. In addition, a chronic treatment (32 days) with celastrol was tested using a matrix-driven delivery pellet system implanted subcutaneously in 5 month-old Tg PS1/APPsw to ensure a continuous daily release of 2.5 mg/Kg of celastrol.
In vitro, celastrol dose dependently prevented NFκB activation and inhibited BACE-1 expression. Celastrol potently inhibited Aβ1-40 and Aβ1-42 production by reducing the β-cleavage of APP, leading to decreased levels of APP-CTFβ and APPsβ. In vivo, celastrol appeared to reduce the levels of both soluble and insoluble Aβ1-38, Aβ1-40 and Aβ1-42. In addition, a reduction in Aβ plaque burden and microglial activation was observed in the brains of Tg PS1/APPsw following a chronic administration of celastrol.
Overall our data suggest that celastrol is a potent Aβ lowering compound that acts as an indirect BACE-1 inhibitor possibly by regulating BACE-1 expression level via an NFκB dependent mechanism. Additional work is required to determine whether chronic administration of celastrol can be safely achieved with cognitive benefits in a transgenic mouse model of AD.
Alzheimer's disease (AD) is an ever-increasing health concern among the aging population and is the most common form of dementia affecting more than 25 million individuals worldwide. While the cause of the disease is uncertain, there are two major neuropathological hallmarks present in the brains of AD patients: the extracellular senile plaques and the intracellular neurofibrillary tangles. Neurofibrillary tangles contain hyperphosphorylated microtubule-associated protein tau, while senile plaques contain a core of β-amyloid (Aβ) peptide. Current treatments for AD include cholinesterase inhibitors and glutamate antagonists. Although useful, these symptomatic treatments do not stop the disease process or prevent neuronal degeneration. There is an on-going need for the development of new treatments for AD. Although the central role of Aβ remains to be proven in clinical trials, data accumulated during the past two decades place Aβ peptides and in particular soluble forms of the peptide as being the main molecule initiating the pathological cascade that eventually leads to AD [1]. Consequently, significant resources have been allocated to identify and develop treatment strategies that alter the metabolism of Aβ. To this end, the discovery of new pharmaceutical entities that have Aβ-lowering ability remains a priority.
Aβ peptides are derived from the sequential proteolysis of the β-amyloid precursor protein (APP) by β- and γ-secretases. The major β-secretase is an aspartyl protease termed BACE-1 (β-site APP cleaving enzyme) [2-4]. BACE-1 cleaves APP within the extracellular domain of APP, resulting in the secretion of the large ectodomain (APPsβ) and generating a membrane-tethered C-terminal fragment CTFβ or C99 which serves as a substrate for γ-secretase [5]. The multimeric γ-secretase complex cleaves at multiple sites within the transmembranous CTFβ generating C-terminally heterogeneous Aβ peptides ranging between 38 to 43 amino-acid residues in length that are secreted [6]. In addition to BACE-1 and γ-secretase, APP can be cleaved by α-secretase within the Aβ domain between Lys16 and Leu17, releasing APPsα and generating CTFα or C83 which is further cleaved by γ-secretase to generate an N-terminally truncated Aβ termed p3. Genetic ablation of BACE-1 completely abolishes Aβ production, establishing BACE-1 as the major neuronal enzyme responsible for initiating the amyloidogenic processing of APP [7].
Given that BACE-1 is the initiating enzyme in Aβ generation, it is considered a prime target for lowering Aβ levels in the treatment and/or prevention of AD. BACE-1 expression is tightly regulated at both the transcriptional and translational level [8]. The BACE-1 gene spans approximately 30 kilobases on human chromosome 11q23.2 and includes 9 exons. The BACE-1 gene promoter lacks the typical CAAT and TATA boxes but contains a variety of transcription factor binding sites, including those for Sp1, GATA-1, AP1, CREB, HSF-1, STAT1, and NFκB, among others. It is likely that these sites influence transcriptional activity of the BACE-1 promoter. Interestingly, both the expression and activity of BACE-1 appear to be elevated in the cerebrospinal fluid and brains of AD patients [9,10] suggesting that BACE-1 expression is altered in AD and that therapeutic approaches aimed at regulating BACE-1 expression may be successful.
Neurons are believed to be the major source of Aβ and BACE-1. However, evidence is mounting that glia, and astrocytes in particular may produce significant levels of BACE-1 and Aβ, especially during inflammation. Glia out-number neurons by a factor of 10, so even a slight increase in glial BACE-1 expression might contribute substantially to cerebral Aβ and exacerbate AD pathology. NFκB is increased in AD and particularly in astrocytes, and it has been suggested that Aβ itself can trigger BACE-1 expression in glial cells via NFκB activation [11-15]. BACE-1 expression has been shown to be particularly elevated both in neurons and astrocytes around Aβ deposits in AD brains [16,17,9]. Thus, pathophysiological conditions leading to the activation of astrocytes may increase BACE-1 expression resulting in increased Aβ production which could exacerbate AD pathogenesis. Small increases in BACE-1 expression have been shown to result in sharply increased Aβ production [18]. Altogether these data suggest that lowering BACE-1 expression levels may have therapeutical potential for AD. We have therefore explored in vitro the effects of various known NFκB inhibitors as a way to reduce BACE-1 expression and shown that NFκB inhibitors reduce Aβ production by targeting the β-cleavage of APP [19]. In the present study, we investigated acute and chronic effects of celastrol, a potent natural triterpene NFκB inhibitor extracted from the roots of the Chinese "Thunder of God vine" (Tripterygenium wilfordii) on Aβ production and Alzheimer-like pathology in a transgenic mouse model of AD. Root extracts of the "Thunder of God vine" have been used as an anti-inflammatory remedy for centuries in China. Celastrol was selected over other known NFκB inhibitors for its ability to cross the blood brain barrier and offer neuroprotection in animal models of Parkinson's disease and Huntington's disease [20]. Previous work has revealed that celastrol can prevent neurodegeneration and extend the life span of a transgenic mouse model of amyotrophic lateral sclerosis [21]. Celastrol displays potent anti-inflammatory activities in vivo and has been shown, for example, to be beneficial against allergy-induced asthma [22] as well as rheumatoid arthritis [23] and to inhibit tumor cell invasion through suppression of NFκB signaling [24].
Cell culture and western-blotting experiments
NFκB activation was quantified using a stable NFκB luciferase reporter cell line of HEK293 cells with chromosomal integration of a luciferase reporter construct regulated by 6 copies of the NFκB response element (Panomics, CA, USA). Cells were grown in DMEM containing 10% serum, 1% penicillin/streptomycin/fungizone and 100 μg/ml of hygromycin B. Confluent cells were treated with 20 pg/ml of TNFα (Sigma, MO, USA) to induce NFκB activation and with a dose range of celastrol for 3 hours. Celastrol was purchased by Gaia Chemical Corporation (CT, USA). Luciferase activity was detected with the Luc-Screen Extended-Glow from Applied Biosystem (CA, USA) and a Synergy HT Biotek chemoluminescent reader (VT, USA). The medium surrounding the cells was collected and used to assess cytotoxicity using a Lactate Dehydrogenase (LDH) assay (Roche Diagnostics, Germany) according to the manufacturer's protocol. Similar experiments were also conducted with 100 ng/ml of phorbol 12-myristate 13 acetate (PMA) purchased from Sigma (MO, USA) instead of TNFa to induce NFκB activation (data not shown). HEK293 cells were used to assess the effect of celastrol on the activation of canonical members of the NFkB signaling pathway following PMA stimulation and on BACE-1 expression levels. Briefly, HEK293 cells were treated 100 ng/ml of PMA or a combination of PMA with celastrol (5 mM) for 15 minutes. Celastrol effects on RAF-1, MEK1/2, p44/42 MAPK and IKBa phosphorylation were monitored by western-blots as detailed below.
7 W CHO cells overexpressing wild-type human APP [25] were grown in DMEM (ATCC, VA, USA) containing 10% fetal bovine serum (Invitrogen, CA, USA), 1% penicillin/streptomycin/fungizone (Cambrex, ME, USA) and 0.3% Geneticin (Invitrogen, CA, USA) and used to determine the impact of celastrol on APP processing and Aβ production. Confluent cells were treated for 24 hours with different doses of celastrol.
Cellular proteins were extracted with 150 μL of ice-cold M-PER Reagent (Pierce Biotechnology, Rockford, IL, USA) containing 1 mM phenylmethanesulfonyl fluoride, 1× of protease cocktail inhibitor (Roche, Inc., USA) and 1 mM sodium orthovanadate. Samples were sonicated, denatured by boiling in Laemmli buffer (Bio-Rad, Hercules, CA, USA) and resolved onto 4-20% gradient polyacrylamide gels (Bio-Rad, Hercules, CA, USA). After electrotransfering onto polyvinylidene difluoride membranes, western-blots were immunoprobed with an anti-APP C-terminal (751-770) antibody (EMD Biosciences Inc., San Diego, CA, USA), with an anti-actin antibody (Chemicon, Temecula, CA, USA) used as a reference antibody to quantify the amount of proteins electrotransferred, with a BACE-1 antibody (Invitrogen, CA, USA), with a phospho-IKBα (ser32) antibody, with a phospho-NFκB p65 (ser 536) antibody, with a phospho-Raf-1 (Ser338) antibody, with a phospho-MEK1/2 and a phospho-p44/42 MAPK antibody using dilutions recommended by the manufacturers. All phospho-specific antibodies were purchased from Cell Signaling Technology Inc (MA, USA). Additionally, sAPPα was detected by Western-blot in the culture medium surrounding 7 W CHO cells using the antibody 6E10 (Signet Laboratories Inc., MA, USA) which recognizes the amino-acids 1-17 of Aβ and sAPPβ was detected in the culture medium using an anti-human sAPPβ antibody (Immuno-Biological Laboratories Co. Ltd., Gunma, Japan). APP CTF/actin, sAPPβ/sAPPα signals, phospho-IKBα/actin, phospho-NFκB p65/actin, phpspho-RAF-1/actin, phospho-MEK/actin, phospho-p44/42 MAPK/actin, BACE-1/actin signal intensity ratios were quantified by chemiluminescence imaging with the ChemiDocTM XRS (Bio-Rad, Hercules, CA, USA). Aβ 1-40 and Aβ 1-42 were quantified in the culture media of 7 W CHO cells using commercially available ELISAs following the manufacturer's recommendations (Invitrogen, CA, USA).
Short hairpin RNAs (shRNAs) were used to stably knock-down the cdc37 gene in HEK293 cells overexpressing APPsw cells. Three different cdc37 shRNAs and a scrambled control shRNA (control cells) were used for transfection. Cdc37 shRNAs cloned into the pRs vector were purchased from Origene (Rockville, MD, USA). Approximately one million HEK/APPsw cells detached with TripLE (Invitrogen, IL, USA) were mixed together with 10 μg of shRNA vectors in Bio-Rad (Hercules, CA, USA) gene pulser cuvettes and electroporated using a square wave protocol (110 V, one 25 ms pulse length using a Bio-Rad gene pulser). Forty-eight hours after, the culture media were replaced by selective media containing 6 μg/ml of puromycin for stable selection of transfected cells. Single colonies resistant to puromycin were isolated. Western-blots were run with cell lysates from these clones to confirm silencing of the cdc37 gene using a 1:1000 dilution of an anti-cdc37 (V367) antibody (Cell Signaling Technology Inc, MA, USA). BACE-1 expression and APP-CTF levels were monitored in cdc37 knock-down HEK293 APPsw cells and HEK293 APPsw cells transfected with a scrambled control shRNA vector as described above.
In order to inhibit HSP90, HEK293 APPsw overexpressing cells were treated for 24 hours with 10 and 20 μM of gedunin (Tocris, MI, USA), a known HSP90 inhibitor which has been shown to inhibit HSP90 with an IC50 ranging from 3 μM to 8 μM in function of the cell type used [26]. Following 24 hours treatment with gedunin, cellular lysates were prepared and assayed for BACE-1 expression and APP-CTF levels by western blotting as described above.
The effect of celastrol on the disruption of the cdc37-HSP90 complex was tested in HEK293 cells overexpressing APPsw. Briefly, HEK293 APPsw cells were treated with PMA 100 ng/ml and celastrol 5 μM for 3 hours. Cellular lysates were prepared as described above and immunoprecipited overnight at 4°C with a 1:100 dilution of an HSP90 antibody (Cell Signaling Technology Inc, MA, USA). Protein A sepharose (GE Healthcare, NJ, USA) was used to pull down the immunoprecipitated complex which was denatured and analyzed by westernblotting for the presence of cdc37.
Animals and treatments
All the experimentations involving mice were approved by the Institutional Animal Care and Use Committee of the Roskamp Institute. Transgenic mice overexpressing the human APP695sw mutation and the presenilin-1 mutation M146L (Tg PS1/APPsw) resulting in over production of human APP and Aβ [27] were used to assess the effect of celastrol on brain Aβ levels. These transgenic mice typically start to develop Aβ deposits by 4 month of age and display a significant amount of Aβ deposits by 6 months [27,28]. An acute treatment regimen was first tested by injecting intraperitoneally 35 week-old Tg PS1/APPsw (post plaque formation) with 1 mg/Kg body weight of celastrol (n = 7) or with 100 μL of the vehicle (50% DMSO in PBS, n = 7) daily for 4 days. One hour after the last injection, brains of the animals were collected, snap frozen in liquid nitrogen and stored at -80°C.
Custom made biodegradable pellets ensuring a continuous release of celastrol at 2.5 mg/Kg of body weight/day and placebo pellets were obtained from Innovative Research of America (FL, USA). Pellets were implanted subcutaneously in 5 month-old Tg PS1/APPsw mice (placebo n = 7; celastrol n = 7) when Aβ deposits are forming. After one month (32 days) of treatment, one brain hemisphere for each mouse was snap frozen in liquid nitrogen and stored at -80°C whereas the other hemisphere was fixed in 4% paraformaldehyde for 48 hours at 4°C prior to embedding in paraffin using a Tissue-Tek automated embedding system (Sakura, USA).
Brain Aβ quantifications
Mice brains (not including the cerebellum) were homogenized by sonication in ice cold MPER reagent containing 1 mM PMSF, 100 mM sodium orthovanadate and 1× cocktail of protease inhibitors (Pierce protein research product, IL, USA). Brain homogenates were centrifuged at 4°C for 30 min at 15000 rpm. The supernatant containing soluble Aβ (GS) was collected and treated with 5 M guanidine isothiocyanate for 1 hour at room temperature. The pellet containing insoluble Aβ (GI) was dissolved and denatured in 5 M guanidine isothiocyanate. Protein concentrations were estimated in both fractions by the BCA method (Pierce, IL). For the mice that received an acute treatment with celastrol, Aβ 1-40 and Aβ 1-42 were quantified by ELISAs according to the manufacturer's recommendations (Invitrogen, CA, USA). For the mice that received the chronic celastrol treatment, Aβ 1-38, Aβ 1-40 and Aβ 1-42 were quantified by electrochemoluminescence in GI and GS fractions using multiplex Aβ assays according to the manufacturer's recommendations (Meso Scale Discovery, MD, USA). Aβ concentrations were calculated in pg/mg of protein and expressed as a % of Aβ obtained in the vehicle/placebo treated animals.
Paraffin embedded brains were sagitally cut into 5 μm-thick sections with a microtome (2030 Biocut, Reichert/Leica, Germany). Sections were mounted on slides and air-dried. Prior to staining, sections were deparaffinized in xylene (2 × 5 min) and hydrated in graded ethanol (2 × 5 min in 100%, 5 min in 85%, 5 min 70%) to water. Endogeneous peroxidase activity was quenched with a 20-min-H2O2 treatment (0.3% in water) and after being rinsed, sections were incubated with blocking buffer (Protein Block Serum-free, DakoCytomation) for 20 min. The monoclonal antibody 4G8 was used to stain Aβ deposits at a 1:750-dilution and purchased from Signet Laboratories (MA, USA). CD45 was immunodetected using a 1:50-dilution of a rat monoclonal antibody (clone IBL-3/16) from Serotec (NC, USA) and used as a microglial marker as previously described [29]. The diluted antibodies were applied onto the sections overnight at 4°C and were detected using the Vectastain ABC (avidin-biotin-peroxidase complex) Elite kit (Vector Laboratories, CA). The labeling was revealed by incubating sections in 0.05 M Tris-HCl buffer (pH 7.6) containing 3,3'-diaminobenzidine (Sigma, MO) and H2O2. For each brain, 4 to 5 stained sections were used to perform the quantification of 4G8 burden and 3 sections were used for the quantification of CD45 burden. The stained area within particular regions (hippocampus, cortex) was quantified using Image-Pro Plus software (Media Cybernetics, MD). An average value was calculated for each brain area from individual mice and expressed as a percentage of the total brain area examined.
Statistical analyses
For statistical analyses, ANOVA, post-hoc comparisons and t-tests were performed where appropriate using SPSS for Windows release 12.0.1.
In vitro effects of celastrol
Celastrol (3-Hydroxy-24-nor-2-oxo-1(10),3,5,7-friedelatetraen-29-oic acid) is a natural triterpene lactone epoxide compound (Figure 1A) extracted from the root bark of the Chinese medicine "Thunder of God Vine" which has been used for hundreds of years as a natural remedy for inflammatory conditions. Celastrol appears to potently inhibit NFκB activation induced by TNFα as monitored by measuring luciferase activity in a stable NFκB reporter HEK293 cell line with a half maximal inhibitory concentration (IC50) below 1 μM (Figure 1B) and no cytotoxicity was observed for doses of celastrol up to 5 μM (Figure 1C). Similar data were observed with celastrol when phorbol ester (PMA) was used in place of TNFα to induce NFκB signaling (data not shown). We investigated the possible effect of celastrol on some members of the canonical NFκB pathway following 15 minutes of stimulation with PMA. As expected, we observed that PMA rapidly stimulated the phosphorylation of RAF1, MEK1/2, p44/42 MAPK and IKBα (Figure 2). Interestingly, celastrol did not prevent PMA induced phosphorylation of RAF1, MEK1/2 and p44/42 MAPK but inhibited PMA induced IKBα phosphorylation suggesting that celastrol inhibits the activity of the IKK (IKappa B Kinase) signaling complex. The IKK signaling complex is also comprised of the catalytic components IKKα, IKKβ and of the regulatory subunit NEMO (
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Figure 1. A) Chemical structure of celastrol, an active ingredient of the "Thunder of God" medication which has been used as an anti-inflammatory remedy for centuries in China. B) Dose dependent effect of celastrol on NFκB activation induced by TNFα in HEK293 cells stably expressing an NFκB luciferase reporter construct. Cells were co-treated with 20 pg/ml of TNFα and with a dose range of celastrol for 3 hours before measuring NFκB luciferase activity. ANOVA reveals a statistically significant main effect of celastrol dose on NFκB activation (P < 0.002). Post-hoc analysis show significant differences between NFκB activity in TNFα treated cells and cells co-treated with TNFα and celastrol for celastrol doses greater or equal to 10 nM (P < 0.005). C) The histogram represents the amounts of LDH released detected in the culture medium of HEK293 cells treated with a dose range of celastrol and with the MPER lysis buffer as a positive control (lysis buffer) to induce complete cellular lysis. ANOVA reveals no significant main effect of celastrol dose (P = 0.420) and a significant effect of the lysis buffer (P < 0.001) on LDH released.
Figure 2. Effect of celastrol on phorbol ester (PMA) induced phosphorylation of key members of the NFκB signaling pathway. HEK293 cells overexpressing APPsw were treated for 15 minutes with PMA in the presence of 0 μM and 5 μM of celastrol. A) Western blots reveal that celastrol did not inhibit Raf-1 phosphorylation, MEK1/2 phosphorylation or p44/42 MAPK phosphorylation induced by PMA but prevented IKBα phosphorylation induced by PMA suggesting that celastrol is inhibiting IKK activity. B) Histogram representing the quantification of Raf-1 phosphorylation/actin, MEK1/2 phosphorylation/actin, phospho p44/42 MAPK/actin and IKBα phosphorylation/actin chemoluminescent signal. ANOVA shows a significant main effect of PMA on the phosphorylation of these different proteins (P < 0.05). Post-hoc comparisons reveal significant increase in phosphorylation after PMA treatment for Raf-1 (P < 0.04), MEK1/2 (P < 0.01), p44/42 MAPK (P < 0.001) and IKBα (P < 0.001) compared to the control conditions. Celastrol does not appear to prevent PMA induced Raf-1 phosphorylation (P = 0.619) but leads to a significant stimulation of MEK1/2 and p44/42 MAPK compared to PMA treatment alone (P < 0.02) and significantly inhibited IKBα phosphorylation compared to PMA treatment alone (P < 0.001).
We used 7 W CHO cells overexpressing wild-type human APP [25] to assess the effect of celastrol on Aβ production and APP processing. Following a 24 hours treatment, celastrol inhibited Aβ1-42 and Aβ1-40 production with an IC50 of approximately 900 nM (Figure 3A) in 7 W CHO cells overexpressing APP. Analysis of metabolites of the APP pathway reveals that celastrol impairs the cleavage of APP at the β-secretase cleavage site leading to a decreased sAPPβ and concomitant suppression of the APP-CTFβ (C99) fragment (Figure 3B). No significant effect of celastrol was observed on APPsα secretion indicating that celastrol does not affect the α-secretase cleavage of APP despite a reduction in APP-CTFα (C83) at 5 μM, therefore suggesting that celastrol may affect the stability or turn-over of C83 (Figure 3B). We did not find that celastrol directly affects BACE-1 activity in a cell free assay at doses inhibiting Aβ production in whole cells (data not shown). A dose dependent inhibition of IKBα and NFκB p65 phosphorylation was observed with celastrol confirming that celastrol inhibited basal NFκB activity in 7 W CHO cells overexpressing APP at doses affecting APP processing (Figure 3). We investigated the effect of celastrol and NFκB stimulation with PMA on BACE-1 expression in HEK293 cells overexpressing APPsw (Figure 4). Celastrol inhibited basal BACE-1 expression and opposed the stimulation of BACE-1 expression induced by PMA. In parallel with the reduction of BACE-1 expression induced by celastrol, a reduction in APP-CTFβ was observed. Moreover, an increased in APP-CTFβ and BACE-1 levels was observed when NFκB signaling was stimulated by PMA (Figure 4) suggesting that celastrol affects APP processing by regulating BACE-1 expression via an NFκB dependent mechanism. Similarly, using a neuronal like cell line (SHSY), we observed that celastrol can reduce BACE-1 expression and oppose PMA induced BACE-1 expression (data not shown).
Figure 3. A) Effect of celastrol on Aβ production in 7 W CHO cells overexpressing wild-type human APP. Human Aβ was measured by ELISAs in the culture media surrounding the cells following 24 hours of treatment with different doses of celastrol. Dose response curves for both Aβ1-40 and Aβ1-42 were established revealing an IC50 of approximately 900 nM for celastrol. ANOVA reveals a significant main effect of celastrol on Aβ 1-40 (P < 0.001) and Aβ 1-42 production (P < 0.015). Post-hoc comparisons show statistically significant effects of celastrol at 1000 nM, 25000 nM and 5000 nM for both Aβ 1-40 and Aβ 1-42 (P < 0.001). B) Effect of celastrol on APP processing and NFκB in CHO cells overexpressing wild-type human APP. Celastrol dose dependently inhibited APPsβ secretion, APP-CTFβ level as well as NFκB p65 and IKBα phosphorylation. C) Histogram representing the quantification of APPsβ/APPsα, APP-CTFβ/actin as well as phospho-NFκB p65/actin and phospho-IKBα/actin chemoluminescent signals. ANOVA reveals a significant main effect of celastrol on APPsβ secretion, APP-CTFβ level, phospho-NFκB p65 and phospho-IKBα levels (P < 0.001). Post-hoc comparisons show a statistical significance for celastrol at 2 and 5 μM for all the parameters studied (P < 0.05) showing that celastrol dose dependently inhibits the β-cleavage of APP while suppressing NFκB activity. (* P < 0.05; ** P < 0.001).
Figure 4. A) Representative western-blot depicting the effect of celastrol on BACE-1 expression in HEK293 cells overexpressing APPsw. Celastrol inhibited basal BACE-1 expression as well as the stimulation of BACE-1 expression induced by a 24 hours treatment with PMA. APP-CTFβ level is reduced when BACE-1 expression is inhibited by celastrol and APP-CTFβ level is increased when BACE-1 expression is stimulated by PMA. B) Histogram representing the quantification of BACE-1 expression in response to celastrol and PMA treatments. ANOVA reveals a significant main effect of PMA (P < 0.002) and of celastrol (P < 0.001) on BACE-1 expression. Post-hoc comparisons shows that PMA significantly stimulates BACE-1 level (P < 0.002) whereas celastrol significantly reduces BACE-1 expression (P < 0.001) compared to control condition and opposes BACE-1 stimulation induced by PMA (P < 0.001). (** P < 0.002).
Since the formation of a transient complex between cdc37 and HSP90 may be required to allow IKK activation [30] and celastrol may interfere with HSP90 or cdc37 [31,32], we explored the effect of celastrol on the formation of this complex in HEK293 cells overexpressing APPsw. Data show that at a dose (5 μM) that fully inhibits NFκB activation and Aβ production, only a slight reduction of cdc37 was observed in the immunoprecipitated HSP90 complex (Figure 5A and 5B). No effect of the NFκB stimulator PMA was observed on the stability of this complex. We next inhibited HSP90 in HEK293 APPsw cells using gedunin, a known HSP90 inhibitor [26] which induces the degradation of HSP90-dependent client proteins similarly to other HSP90 inhibitors. Gedunin did not appear to affect BACE-1 expression or APP processing (Figure 5C and 5D). Moreover, we stably knock-down cdc37 expression in HEK293 APPsw cells using a shRNA approach and observed that cdc37 inhibition did not impact BACE-1 expression (Figure 5E and 5F) and APP processing (data not shown).
Figure 5. Effect of celastrol and PMA on the formation of the HSP90-cdc37 complex in HEK293 APPsw cells. A) Western-blot depicting the amount of cdc37 recovered in the HSP90 immunoprecipitate following treatment with celastrol and PMA. B) Histogram representing the quantification of cdc37 in the HSP90 immunoprecipitate. No significant effect of PMA was observed but a significant effect of celastrol was detected (P < 0.005) showing a slight reduction in the amount of cdc37 present in the HSP90 immunoprecipitates. C) Western-blots showing the effects of the HSP90 inhibitor gedunin on BACE-1 expression and APP processing in HEK293 APPsw cells. D) Histogram representing the quantification of BACE-1/Actin chemoluminescent signal showing that the gedunin treatment does not affect BACE-1 expression in HEK293 APPsw cells (ANOVA reveals no significant main effect of gedunin on BACE-1 level (P = 0.572)). E) Representative western-blot showing the level of cdc37 and BACE-1 expression in HEK293 APPsw cells that were knock-down for cdc37 using a shRNA approach for 3 different clones. 1) non silencing scrambled shRNA; 2) cdc37 shRNA clone 51G9; 3) cdc37 shRNA clone 97H1; 4) cdc37 shRNA clone 95C4. F) Histogram representing the quantification of cdc37 and BACE-1 expression for 9 different clones of HEK293 APPsw cells stably transfected with a silencing cdc37 shRNA vector (cdc37 shRNA) and 4 different clones of HEK293 APPsw stably transfected with a non silencing scrambled shRNA vector (control shRNA). Statistically significant inhibition of cdc37 expression (P < 0.001) and no effect on BACE-1 expression (P = 0.947) was observed in HEK293 APPsw cells knock-down for cdc37.
Acute effect of celastrol on Aβ production in a transgenic mouse model of AD
We next investigated the effect of celastrol on brain Aβ levels using an acute dosage paradigm in 6-month old Tg PS1/APPsw mice. Tg PS1/APPsw mice were injected intraperitoneally, daily for 4 days, with 1 mg/Kg of body weight of celastrol or 100 μL of the vehicle only. One hour after the last injection, mice were humanely euthanatized and their brains collected. We assessed the β-amyloid lowering properties of celastrol by measuring the pools of soluble and insoluble Aβ species in celastrol and placebo treated mice. Following 4 days of treatment with 1 mg/Kg/day of celastrol, we observed a reduction in brain soluble Aβ 1-40 and Aβ 1-42 by approximately 40% whereas insoluble Aβ was reduced by approximately 50% compared to placebo treated animals (Figure 6).
Figure 6. Acute effects of celastrol on brain Aβ levels in 35 week-old Tg PS1/APPSw. Mice were treated for 4 days with an intraperitoneal injection of celastrol (1 mg/Kg of body weight) or the vehicle only (placebo). Multivariate analysis reveals a significant main effect of celastrol on brain soluble and insoluble Aβ levels (P < 0.05). Statistically significant differences were observed for soluble Aβ 1-42 values (P < 0.03) between vehicle and celastrol treated mice and for insoluble Aβ 1-40 (P < 0.001) and Aβ 1-42 values (P < 0.001) between vehicle and celastrol treated mice. (* P < 0.05; ** P < 0.001).
Chronic effect of celastrol on Alzheimer-like pathology in a transgenic mouse model of AD
To evaluate whether celastrol could affect Alzheimer-like pathology in Tg PS1/APPsw, we investigated the effects of a longer treatment with celastrol. We used a matrix-driven delivery pellet system implanted subcutaneously that was designed to ensure a continuous delivery of celastrol and avoid subjecting the animals to repeated injections and excessive handling. Tg PS1/APPsw mice were treated for one month with this pellet delivery system at a dosage of 2.5 mg of celastrol/Kg of body weight/Day. Analysis of brain Aβ levels by ELISAs revealed that celastrol reduced brain soluble Aβ 1-38, Aβ 1-40 and Aβ 1-42 by approximately 50% and the levels of insoluble Aβ 1-38, Aβ 1-40 and Aβ 1-42 by approximately 60% (Figure 7). To determine whether celastrol treatment affected β-amyloid plaque pathology, brain sections from Tg PS1/APPSw mice implanted with a placebo pellet and Tg PS1/APPSw mice implanted with a celastrol pellet were immunostained with the antibody 4G8 recognizing Aβ. A significant reduction in plaque burden (approximately 50%) in both the hippocampus and the cortex of celastrol treated animals compared to animals receiving the placebo was observed (Figure 8). We also estimated the impact of celastrol on microglial activation by measuring the percentage of CD45 immunostained area [29] on brain sections in placebo and celastrol treated animals. An approximate 50% reduction in microgliosis was observed in the cortex of celastrol treated animals compared to placebo treated animals (Figure 9).
Figure 7. Chronic effects of celastrol on brain Aβ levels. Five month-old Tg PS1/APPsw mice were implanted subcutaneously with biodegradable placebo pellets and celastrol pellets ensuring a constant release of celastrol at a rate of 2.5 mg/Kg of body weight/day. After 32 days of treatment, brain Aβ levels were analyzed by electrochemoluminescence. A) Histogram representing the level of soluble Aβ 1-38, Aβ 1-40 and Aβ 1-42 quantified in the brain of placebo and celastrol treated Tg PS1/APPsw mice. B) Histogram representing the level of insoluble Aβ 1-38, Aβ 1-40 and Aβ 1-42 quantified in the brain of placebo and celastrol treated Tg PS1/APPsw mice. Multivariate analysis reveals a statistical significant differences between celastrol and placebo treated mice for the levels of brain soluble Aβ 1-38 (P < 0.005), Aβ 1-40 (P < 0.03), Aβ 1-42 (P < 0.007) and brain insoluble Aβ 1-38 (P < 0.004), Aβ 1-40 (P < 0.03), Aβ 1-42 (P < 0.02). (* P < 0.05); ** P < 0.01).
Figure 8. Chronic effects of celastrol on Aβ plaque burden in Tg PS1/APPsw mice. A) Representative photomicrographs (200× magnification) showing the extent of Aβ plaque burden detected by 4G8 immunostaining in the cortex (right panel) and hippocampus (left panel) of Tg PS1/APPsw mice (6 month-old) treated with biodegradable pellets of placebo or celastrol for a period of 32 days. B) Histogram representing the quantification of Aβ burden by image analysis in the cortex and hippocampus of Tg PS1/APPsw mice treated with placebo and celastrol pellets. Multivariate analysis reveals a statistically significant effect of celastrol treatment on Aβ plaque burden for the cortex (P < 0.002) and hippocampus (P < 0.004). (**P < 0.005).
Figure 9. Chronic effects of celastrol on microgliosis in Tg PS1/APPsw mice. A) Representative photomicrographs (taken with a 20× and 60× objective providing a respective magnification of 200× and 600× respectively) depicting the presence of CD45 reactive microglia around Aβ deposits in Tg PS1/APPsw treated with a placebo and celastrol. B) Histogram representing the burden of activated microglia (CD45 positive) in the cortex of Tg PS1/APPsw mice treated with placebo and celastrol pellets. Statistically significant difference in microgliosis burden (P < 0.03) was observed between placebo and celastrol treated mice. (* P < 0.05).
NFκB has been shown to regulate BACE-1 expression level [11,17,13-15], the rate limiting enzyme responsible for the production of Aβ. For this reason, we investigated whether NFκB inhibition could lead to an inhibition of Aβ accumulation in a transgenic mouse model of AD. In this study, we selected the natural product celastrol over other known NFκB inhibitors because it is a potent NFκB inhibitor that has been shown to exert anti-inflammatory effects in vivo [33,34] and neuroprotective effects in animal models of Parkinson's disease and Hungtington's disease [20]. We first characterized the in vitro activity of celastrol towards Aβ production using a CHO cell line overexpressing Aβ. Celastrol dose dependently inhibited Aβ1-40 and Aβ1-42 with an IC50 comparable to its IC50 for inhibiting NFκB activation in response to TNFα (around 900 nM). Celastrol dose dependently reduced the production of APP-CTFβ and APPsβ confirming that celastrol impacted the β-cleavage of APP. Moreover, celastrol appeared to reduce BACE-1 expression and to oppose BACE-1 upregulation induced by NFκB stimulation.
We then investigated the impact of celastrol on Aβ production in vivo using a transgenic mouse model of AD. Following an acute treatment with celastrol in Tg PS1/APPsw, we observed a reduction in brain soluble and insoluble Aβ species. We used a biodegradable pellet delivery system implanted subcutaneously to assess the Aβ lowering effects of a chronic treatment with celastrol. After 1 month of treatment, pathological evaluation of the brains of celastrol treated Tg PS1/APPsw mice showed a 50% reduction in Aβ plaque burden in the hippocampus and cortex compared to placebo treated animals. Additionally, evaluation of activated microglia following CD45 immunostaining revealed that microgliosis was significantly reduced in celastrol treated animals compared to placebo treated mice paralleling the reduction of Aβ burden observed. We further characterized the β-amyloid lowering properties of celastrol by measuring the pools of soluble and insoluble Aβ species in celastrol and placebo treated mice. Data showed that this chronic treatment with celastrol in Tg PS1/APPsw mice reduced brain levels of soluble Aβ 1-38, Aβ 1-40 and Aβ 1-42 by approximately 50% and the levels of insoluble Aβ 1-38, Aβ 1-40 and Aβ 1-42 by approximately 60%.
The exact molecular mechanism of action of celastrol remains unclear. Celastrol has been suggested to be an ERK inhibitor and to inhibit of p44/42 MAPK phosphorylation [35], however our data show that celastrol inhibits NFκB activation without preventing the phosphorylation of MEK1/2 and its downstream target p44/42 MAPK when NFκB is stimulated with PMA, suggesting that celastrol is acting downstream of MAPK. Indeed, celastrol appears to oppose both IKBα and NFκB p65 phosphorylation which suggest that celastrol is inhibiting the IKK complex in agreement with previous studies showing that celastrol inhibits IKK activity [36]. It has been recently suggested that celastrol interacts with HSP90 and disrupts the formation of the cdc37-HSP90 complex [31]. However, NMR analyses of celastrol binding to cdc37 and HSP90 have suggested that celastrol directly interacts with ccd37 and not HSP90 [32]. Interestingly, the formation of a transient complex between cdc37 and HSP90 may be required to allow IKK activation [30] suggesting that celastrol may inhibit NFκB activation by preventing the formation of this complex. However, the disruption of the cdc37-HSP90 complex by celastrol requires 10 μM of celastrol [31] and at 5 μM (dose that fully inhibits Aβ production and NFκB activity in vitro), we only observed a modest reduction (20%) in the amount of cdc37 co-immunoprecipited with HSP90 suggesting that the effect of celastrol on Aβ production is unlikely to be mediated via a disruption of this complex. This is further substantiated by our observation that pharmacological inhibition of HSP90 does not significantly affect APP processing and BACE-1 expression, ruling out the possibility that the Aβ lowering properties of celastrol are mediated via HSP90 dependent events. Moreover, inhibition of cdc37 expression does not appear to affect APP processing and BACE-1 expression excluding cdc37 as a molecular target responsible for the Aβ lowering properties of celastrol. Celastrol has also been shown to inhibit lipidoperoxydation in vitro with an IC50 of 7 μM [37], which is 7-fold higher than it's IC50 for inhibiting Aβ production; it is therefore unlikely that the antioxidant properties of celastrol play a major role in the Aβ lowering activity of this compound. Overall, our in vitro data suggest that celastrol inhibits Aβ production by regulating BACE-1 expression level most likely via an NFκB dependent mechanism.
At doses similar to the doses of celastrol that we tested in vivo, celastrol has been shown to significantly inhibit NFκB activity in the brain, to reduce TNFα production and the expression of the CD40 ligand in astrocytes [20]. It remains possible that the in vivo Aβ lowering properties of celastrol may be the result of multiple mechanisms in addition to those affecting APP processing and BACE-1 expression as observed in vitro. For instance, celastrol may also lower brain Aβ levels by opposing inflammation, cytokines production and CD40 ligand expression, all of which are known to modulate the accumulation of Aβ in the brain [38-40]. So far, we have tested the effects of celastrol on brain Aβ accumulation when Aβ deposits are initially forming in a transgenic mouse model of AD, which could be regarded as an early intervention. It remains to be determined whether celastrol can display therapeutic efficacy when Aβ deposits are already highly abundant. Interestingly, celastrol has been shown to display neuroprotective effects in different animal models [20,21] and to improve memory, learning and psychomotor activity in rats [34]. Chronic effects of celastrol on memory impairment and tau pathology remain to be investigated in transgenic mouse models of AD to determine whether this compound may offer therapeutic benefits suitable for the treatment of AD.
Our data suggest that celastrol is a potent Aβ lowering compound that may act in part by decreasing BACE-1 expression levels and Aβ production via an NFκB dependant mechanism. More experimentation is required to determine the exact mechanism of action responsible for the Aβ lowering properties of celastrol. It remains to be determined whether this compound can also improve cognition and tau pathology in transgenic mouse models of AD and whether this molecule displays the pharmacokinetic characteristics and toxicity profile required for a drug candidate.
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
DP conceived of the study, developed the methodology for studying APP processing and NFκB activity, carried out the in vivo treatments with celastrol, participated in animal care, participated in data analysis and drafted the manuscript. NG and VL carried out the immunostaining studies. NP, DB, AM, CB participated in animal care, samples preparation and carried out some of the western blotting and ELISA studies. GA generated the cdc37 knock-down HEK293 APPsw cells and helped with cell culture experiments. MM helped write the manuscript and gave a critical analysis of the manuscript. All authors read and approved the final manuscript.
This work was supported by an Alzheimer's Association grant IIRG-08-91076. The authors are grateful to Mr. and Mrs. Roskamp for their generous support which helped to make this work possible. We thank Dr. Michael Wolfe (Harvard Medical School, Boston, Massachusetts, USA) for providing HEK293 APPsw and 7 W CHO APP overexpressing cells.
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Open & Shut: This week in restaurant openings and closings
With the New Year comes new beginnings, and in this case – one temporary ending.
Hecho: A new Mexican restaurant called Hecho is opening in Phinney Ridge in early March. Hecho in Spanish means "made" – and that's what everything in the restaurant will be: hand-made, that is. From drinks, to salsa and tortillas, and even the tables. Hecho will use local ingredients to serve and make food from every corner of Mexico. Eater Seattle reports that this collaboration of Chris Navarra, Shannon Wilkinson and Chris Girke will be taking over the old Carmelita space on Greenwood Ave N.
Amaro Bistro: Six Oaks Apartments announced yesterday that Amaro Bistro will open inside its downtown Bothell complex in the fall of 2014. The bistro will specialize in traditional and modern Italian cuisine, and is the second restaurant for Nick Wiltz – the owner of Il Bistro in Pike Place Market.
Tutta Bella: Great news if you love this popular Seattle restaurant, but live on the Eastside! Owner Joe Fugere has opened his fifth restaurant, this time in the Crossroads Shopping Center in Bellevue. This 300-seater is the largest of the five, and has imported ovens from Naples, and a private space and patio for Microsoft to use as a meeting place. Sign us up for that meeting!
CLOSED (for now)
Saigon Boat Café was closed Sunday night by a Public Health food inspector for safety violations including hazardous foods at unsafe temperatures, improper cooling of food, poor personal hygiene and improper sanitation of utensils. West Seattle Blog reports the café will be reopened once all issues have been resolved.
Any other openings or closings we missed? Let us know in the comments!

subset: commoncrawl


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Stubborn Love Acoustic chords by The Lumineers, added: March 5th, 2013
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Artist: The Lumineers
Song: Stubborn Love (acoustic on 98.7 FM Penthouse)
Video 1: http://www.youtube.com/watch?v=0_DWlCJIL60
Video 2: http://www.youtube.com/watch?v=2wf2ADGQvcg
Transcription: SCA, email@example.com
Tuning: standard, NO capo
Chords used:
Fmaj7: x.188.8.131.52.x (actual name: F/C)
Fsus2: x.220.127.116.11.x (actual name: Csus4 or Fsus2/C)
Am: x.0.2.2.1.0
G: 22.214.171.124.0.0 (actual name: G6)
Gsus4: 22.214.171.124.1.0 (actual name: G6sus4)
C: x.22.214.171.124.0
Gadd9: 22.214.171.124.3.x
Am/G: 3.x.126.96.36.199
Watch the video, for riffs, hammer ons and
pull offs.
Fmaj7 Am Fsus2 Fmaj7 C
Fmaj7 Am G Gsus4 G C
Fmaj7 Am Fsus2 Fmaj7 C
Fmaj7 Am G Gsus4 G C
Fmaj7 Am Fsus2 Fmaj7 C
She'll lie and steal and cheat, and beg you from her knees
Fmaj7 Am G Gsus4 G C
And make you thinks she means it this time
Fmaj7 Am Fsus2 Fmaj7 C
She'll tear a hole in you, the one you can't repair
Fmaj7 Am G Gsus4 G C
But I still love her, I don't really care
Fmaj7 C Am G Gsus4 G
When we were young, Oh Oh, we did enough
Fmaj7 C Am G Gsus4 G
When it got cold, Ooh Ooh, we bundled up
Fmaj7 C Am G Gsus4 G
I cant be told, Ah Ah It cant be done
It's better to feel pain, than nothing at all
The opposite of love's indifference
Pay attention now, I'm standing on your porch screaming out
And I wont leave until you come downstairs
Fmaj7 C/G Gadd9 Am Am/G
So keep your head up, keep your lo-ove
Fmaj7 C/G Gadd9 Am Am/G
Keep your head up, my lo-ove
Fmaj7 C/G Gadd9 Am Am/G
Keep your head up, my lo-ove
Fmaj7 C/G Gadd9 Am Am/G
So keep your head up, keep your lo-ove
I don't blame ya dear for running like you did, all these years
I would do the same, your best believe
The highway signs say we're close, but I don't read those things anymore
I never trusted my own eyes
When we were young oh oh, we did enough
When it got cold, Ooh ooh we bundled up
I can't be told, Ah ah, can't be done
So keep your head up, keep your love
Keep your head up, my love
Keep your head up, my love
Keep your head up, keep your love
Note II:
If you have any corrections or improvements to the tab write
them on the comments section, or send me a message,
or e-mail, so we can together make this tab even better.
SCA, email@example.com
Doesn't Fmaj7 need to have an E in it? The chord you have seems to just be a F/C
Love the tabs otherwise.
Yes you're right, Fmaj7 must have E in it. And in this song,
as in Hey Ho, he doesn't play Fmaj7, he plays F/C.
I didnt update the tab because if I change the name of the chord,
I will then need to adjust all other chords to be just above the right
lyrics. It seemed easier just to highlight the notes in chords than changing the name.
But you are 100% right


text:
G-Force is being released in Blu-Ray and DVD on December 15 so I wanted to share this article with you in anticipation of its release. It looks like it will be a cute movie! Are you hoping to get some radical, high tech , totally mind- blowing, possibly body transporting, electronic gadgets this Christmas?… [read more]
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A Woman's Right to Choose
Guest Post by Bill Walker
One of the most difficult aspects of writing, in my humble opinion, is dialogue. It is also one of the most important. In fact, I would wager that most people wouldn't care to read a book without it. So, how does one develop an ear for dialogue? After all, we hear people around us speak… [read more]
Testing the Limits on Governmental Intrusion Into Personal Privacy Rights
Testing the Limits on Governmental Intrusion Into Personal Privacy Rights: Cases Raise the Question as to Whether the Police Be Permitted to Install a GPS Device on a Suspects Car Without a Warrant Our society is continually being called upon to evaluate how far our law enforcement officials and other governmental agencies should be permitted… [read more]
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Is it true that gang members do not hurt people that are not gang members or is that just a myth? This is a tricky question because the moral values on human life for organised crime gangs throughout the world are different. In the research I did on organised crime for my book, Purged by… [read more]
Introducing Avon's Winter Fashion Collection
This holiday season, look super chic without spending a bundle! Avon introduces the perfect way to spice up the holiday with new stylish essentials and accessories. Here are their top five holiday picks that will help you save and stay in style all season long. Stay warm with the Cashtastic Ruffle Scarf (On Sale… [read more]
Honouring Selfless Woman Award with Joolwe.com
Selfless Woman Jewelry Award Thanksgiving is coming, and there's no better time to thank people who have touched your life and that of others'. Joolwe.com is proud to announce the launch of its first annual "Joolwe Selfless Woman Jewelry Award"; the program is searching for members of the community who always put the needs… [read more]


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Downhill Area:
Cypress Creek Grill - The Cypress Creek Grill eatery is located on the second level of the magnificent post and beam Cypress Creek Lodge. The open concept food court features hand crafted pizza's, juicy flat grilled burgers, soups and chili's, Mexican burrito's and taco salads, fresh daily baguette sandwiches, pizza oven toasted sandwiches, grab and go salads, fresh baked goods and deserts. New this season is our line-up of Gourmet Grilled Cheese Sandwiches. Check our website cypressmountain.com to keep up to date on our delicious value priced Daily Specials. The Cypress Creek Grill is licensed with a selection of beer, wines and cider.
For more information on the Cypress Creek Lodge and photos please click HERE [2]
Gold Medal Cafe – Named to honour the four Olympic Gold Medal performances by Canadian athletes on Cypress Mountain in 2010, the Gold Medal Café is a favourite spot for a light breakfast or try our full selection of Barista espresso drinks and delicious fresh brewed Coffee. The GMC is located on Level 1 of the Cypress Creek Lodge across from Big Bear Sports. Light lunches and soft drinks are also served during the day.


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Class of 1969
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Robert N. Foley, MB, MSc, FRCPI, FRCPC
Dr. Foley graduated from the National University of Ireland Medical School. He completed his residency training in Medicine at Cork University Hospital, Ireland. He received a nephrology fellowship from Memorial University of Newfoundland, Canada, where he also received a masters of science degree in the field of clinical epidemiology.
Dr. Foley focuses his attention on research at the Chronic Disease Research Group and the United States Renal Data System, with a primary interest in the clinical epidemiology of cardiovascular disease in chronic kidney disease.
Dr. Foley is also an Associate Professor of Medicine at the University of Minnesota School of Medicine.
Medical School
National University of Ireland Medical School, Cork, Ireland
Nephrology, Cork University Hospital, Cork, , Ireland
Nephrology, Memorial University of Newfoundland, Newfoundland, , Canada
Other Specialty Location(s)
Clinical Epidemiology, MB, Memorial University of Newfoundland, Newfoundland, , Canada


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- July 18th, 2013
- Posted In: News
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- Continue Reading
Music has charms to sooth a savage breast…to soften rocks, or bend a knotted oak… poet William Congreve was correct when, in 1697, he wrote to the power of music.
Greetings from Memphis:
The horrific and cowardly events that transpired at Monday's Boston Marathon are painfully difficult for all of us to understand, much less accept – even in the context of today's tumultuous world. Despite our religious, political, and cultural differences here on planet Earth, I find it unfathomable that anyone would strive to kill or maim innocent people in order to illustrate their own barbarism and savagery.
I have worked within the music industry for the past 50 years, and have found time and again that poet William Congreve was correct when, in 1697, he wrote: "Music has charms to sooth a savage breast…to soften rocks, or bend a knotted oak…" When we were making music in Memphis at Stax Records in the 1960s and '70s it was to make people "feel good," and we need more of that "feel good" music today – now more than ever.
A Letter from Al Bell to President Obama:
April 16, 2013
Mr. Barack Obama
President of the United States of America
The White House 1600
Pennsylvania Avenue, NW Washington, DC 20500
Dear Mr. President,
I am Al Bell, the former owner of Stax Records and current Chairman of the Board of the Memphis Music Foundation. I am a Memphis music entertainment entrepreneur born in Arkansas, and I humbly state that I played a major role in the building and global recognition of Stax Records, the Memphis sound, and the careers of many fine and talented artists and musicians that make Memphis music what it is today. With great respect, appreciation, and insight into the phenomenon occurring at Stax Records, I was inspired to brand our Memphis music home as Soulsville USA. Continue Reading


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NEW DELHI: Canadian handset maker BlackBerry in association with Porsche Design today launched a new all-touch Porsche Design P'9982 luxury smartphone.
Though the price of the handset was not disclosed, BlackBerry said the device is fashioned from high-quality materials and features a customised version of its 10.2 operating system.
BlackBerry has also launched a second Crocodile model strictly limited to 500 devices globally, it said in a statement.


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Aaron Briscoe of the CSU Facilities Management Department puts recycling materials into bins Wednesday as part of a trash audit. The audit is part of recycling campaign, held on campus to show students how much trash can be recycled.
- Filed Under
As dozens of CSU students walked by, curious looks on their faces, Aaron Briscoe picked through several tons of trash outside the Morgan Library in Fort Collins on Wednesday, looking for recyclables.
He didn't have to look very hard. In the mound of trash were items ranging from steel soup cans to soda bottles, paper and shoe boxes. All can be recycled but instead were tossed into trash bins outside residence halls at Colorado State University this week.
Wednesday's "trash audit" is part of CSU's efforts to improve recycling rates at what administrators have dubbed "The Green University." ...


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News LA Times - Had Your McYoga Today?
LA Times - Had Your McYoga Today?
Bikram Choudhury looks to franchise his torrid technique.
By HILARY E. MacGREGOR, Times
Staff Writer
His yoga.
It is from here--a nondescript warehouse on La Cienega Boulevard north of the Santa Monica Freeway--that this spiritually driven, trash-talking, Rolls-Royce-driving guru is building his vast yoga empire.
Next to him is a map of the world, with pins for every Bikram yoga studio in America. In 1973 there would have been three pins--one in Honolulu, one in San Francisco and one in Beverly Hills. In January there were 500. Today, 650 pins stud the map. Bikram studios, he says, are opening around the world at a rate of two a day. Some cities--such as San Francisco--are so saturated with Bikram yoga studios that Choudhury may put a moratorium on the area.
"It was like this for years," Choudhury says, drawing a flat line through the air. "Then in the last years it shot up, like this! Like a rocket!" He launched his hand skyward and nearly leaps out of his chair.
Choudhury has taught his trademark brand of sweaty yoga--practiced in 100-plus-degree heat-- in Beverly Hills for nearly three decades. He has always done well. But now, he is riding the crest of the yoga tsunami sweeping the nation, and he is poised to cash in on his "product" the American way. Since January, a San Diego lawyer has copyrighted and trademarked Bikram's name, products, logo, clothing line, his scripted 90-minute teaching dialogue, and his sequence of 26 asanas, or postures.
Choudhury is not the first to copyright or trademark yoga products. But by fall, if all goes as planned, Bikram's Yoga College of India will become what is believed to be the first yoga franchise in the United States. The announcement has caused a stir in the yoga world, where Choudhury is already a controversial figure, dubbed "yoga's bad boy" by Yoga Journal.
By franchising, Choudhury makes tangible uncomfortable links between spirituality and commerce.
"You don't typically think of yoga and business being that intertwined," said Michael Harris, who runs a Bikram yoga studio in Portland, Ore.. "But yoga and business country have become intertwined."
Trisha Lamb Feuerstein of the Yoga Research and Education Center, a Santa Rosa-based nonprofit that tracks yoga trends, said franchising defies the spirit of yoga. "Yoga is huge and infinite. What's being branded are the physical aspects of the practice. You can't brand the spiritual aspects. Yoga is not hamburgers."
Choudhury counters that his popularity has created copycats and diluted his teachings. Franchising, he hopes, will keep his yoga pure and his workout as reliable as a cup of Starbucks coffee.
"Many people think yoga belongs to the world," said Choudhury's attorney, Jacob Reinbolt of San Diego."That is wrong." Bikram's unique method, he says, "is one of the most easily protectable pieces of intellectual property there is."
So will Bikram yoga be as standardized as McDonald's?
For a moment the charismatic yoga-vangelist who preaches the miracles of the discipline in an amped-up patter of Indian-accented hyperbole until he is hoarse, grows reflective.
"You could say that," he finally says. "Only I sell life."
In 21st century America, yoga is big business. There are yoga retreats, yoga weekends, yoga books, yoga clothing lines, yoga calendars, yoga videos, baby yoga, even a yoga cruise.
It's unclear how many millions of Americans are practicing yoga, but a 1998 survey by the Wall Street Journal and NBC pegged the number at more than 18 million (up from 6 million in 1994). Although there are no current figures, the editor of Yoga Journal believes the number is significantly higher.
Nowhere is yoga more popular than in America's largest and most stressed-out urban centers. But with perhaps more brand-name yoga gurus per capita than any other city in the country--and legions of celebrities doing "downward dog" daily--Los Angeles has emerged as a center for yoga innovation. Still, for critics, franchising a form of the 5,000-year-old sacred path to divine realization seems like the biggest, and crudest, step yet in the commodification of spirituality in America.
"We have never heard of this," said Deborah Willoughby, founding editor of Yoga International, a Pennsylvania-based magazine that focuses on the spiritual dimensions of yoga. "A lot of places have branch centers, like Shivananda, or the Himalayan Institute, where students will open a center and work under the guidance of a spiritual director. But it is not like it is owned, or licensed. It is just a desire to spread the spiritual teachings.
Franchising, she said, is "hard to imagine."
"But it's very California," she said. "I think it probably means everyone else will be doing it soon."
On Easter Sunday, 300 giddy wannabes have descended on Bikram's Yoga College of India on La Cienega for an intense, nine-week boot camp that will transform them into yoga teachers. They have come from France and Hungary, from Vancouver and Denver, from San Diego and Silver Lake. They have paid $5,000 each for the chance to study at the feet of the Indian master.
Choudhury offered his first teacher training course eight years ago, to 35 students. Today his courses fill as soon as they are offered. This is his largest class ever. Choudhury earns $3 million annually in teacher training alone.
The training program is the key to Choudhury's breakneck expansion; these fresh arrivals are the foot soldiers of the empire. They will practice yoga twice a day, drill each posture until they get it right and listen to lectures until the wee hours.
They will also have to memorize every word of "the dialogue" verbatim, complete with Bikram similes and turns of phrase. After the course ends, they will be dispatched to their hometowns like missionaries--taking Bikram to the people.
Bikram Choudhury, 56, was born in Calcutta, where yoga is a way of life. Like many Indians, he began doing yoga at the age of 4. His guru, or teacher, was Bishnu Ghosh, brother of Paramahasa Yogananda, who founded the Self-Realization Fellowship. At 13, Choudhury won the National India Yoga Contest--which judges suppleness and strength, knowledge, and presentation--and remained its undefeated champion for three years. Choudhury then became a weightlifter, winning the All India Weight Lifting Competition in 1963 at age 16. A year later he dropped a 380-pound barbell on his knee. He says doctors told him he would never walk again. He turned to his teacher--and yoga. Six months later, he says, his knee was healed.
Bikram's yoga is made up of a sequence of 26 postures (each of which is performed twice), and two breathing exercises, culled from 84 classical postures and more than 10,000 combinations. He heats the room to between 100 and 105 degrees, he says, to work bodies like a blacksmith.
At the urging of his guru, Choudhury came to the West. He opened his first studio in Los Angeles in Beverly Hills in 1973, in the basement of a former bank building. He was young and gorgeous, at a time when the nation was bewitched with all things Indian--from bedspreads and incense to Ravi Shankar and Krishnamurti.
"In those years he was trying to be a pure yogi," recalls Emmy Cleaves, who was Bikram's 15th student in Los Angeles and is now a senior teacher. "He slept on the floor there. That's the kind of sacrifice he was willing to make. He has invested a lot in this, because his guru told him to go out and teach yoga. From an Indian perspective, he was doing the right thing."
In those days Choudhury was celibate. He didn't drink, nor does he now. Some recall that the young Choudhury was so shy he would barely meet people's eyes when he spoke. He ran his yoga school as if it were in Calcutta, rather than Beverly Hills. "Do you know, every yoga school in India is free?" Bikram asked. "My school was free. We don't think to charge money. For us, a yoga school is like a temple. I had a little box, and people could put money in it. Like a church."
Shirley MacLaine, one of his earliest students, took him aside. "You cannot run a yoga school the Indian way," he says she told him. She hired a manager and a security guard, he says, and he began to charge for classes.
"If it's free, only 15 people would come," Choudhury said. "That's the way America is." (A 90-minute class at Bikram's Yoga College of India costs $20, making it one of the most expensive in the city.)
In 1984, by arrangement of his guru's son, Choudhury wed Rajashree, a famed yogini--or yoga practitioner-- and five-time All India Yoga Champion. "That was the main reason I married her," he says. "She beat all the men, 1400 people. The year we got married she was the champion."
They have a daughter, Laju, 12, and a son, Anurag, 9. Rajashree has proved a strong businesswoman and a gentle presence against Choudhury's braggadocio.
On this, their first day of orientation, the students shed their shoes and walk into the 10,000-square-foot room where they will spend more than 12 hours a day for the next two months. This is no ashram, with wafting incense, soft light and chanting in the background. This is a yoga factory.
Mirrors line the walls. A carpet stretches forever. Lines for yoga mats are etched like parking places on the floor. Students trim their mats so everyone can fit in. Fluorescent lights glare down from above. The light, the mirrors, the microphones, give the room a carnival-esque quality. During yoga classes, heaters blow so hot, people grow lightheaded. The stench of sweating bodies hits a visitor like a wall.
On the second day, around noon, Choudhury finally arrives. (The master does not rise early because he stays up late watching Hindi movies.) Applause ripples through the room as he walks across the carpet, touching the hands of students who reach for him like a rock star.
Choudhury defies the stereotype of a soul-soothing, aphorism-spewing bearded yogi. He likes expensive cars and Rolex watches. He likes to drop the names of famous people he claims to have saved (MacLaine, Raquel Welch, Quincy Jones), the athletes whose careers he claims to have has resurrected (Kareem Abdul-Jabbar, John McEnroe), the president he says he cured (Richard Nixon--of phlebitis).
He climbs onto a chair perched like a throne on a large mirrored cube, and dons a headset. His introduction ranges from the compassionate to the profane, from the incomprehensible to the profound. Now, he is a soulful Indian. Now, an abuse-spewing Hollywood executive. (Cleaves calls his tendency to swear and exaggerate "Bikram-isms.") "Sometimes he uses a little profanity. He never did that in the early years," she says. "He learned a lot of that from the show-biz people. He thought that was America."
He welcomes his students and warns them what lies ahead.
"In America, they say, you can take a horse to the pond, but you can't make him drink," he says. "My way is different. I only take the horse to the pond to make him drink. If he doesn't, I'll cut his head off."
He asks the students to introduce themselves.
A man rises.
"I'm from San Diego. I've been doing yoga for 11 months. I love Bikram. He is God."
Some of the students hope to open studios, some want to teach. Some just love the yoga.
Kenny Colgan, 38, was a Northern California software executive who quit his job to study with Bikram. "He is a world-class evangelist," Colgan said. "And I've known a lot of them in the technology world."
Kate Besleme, 30, took up yoga to help her overcome running injuries. Just out of graduate school, she has been hired by the Bush White House to help develop environmental policy. She waffled about disclosing to her new bosses the yoga training, fearing she would be labeled a flake. She said she hopes to bring Bikram Yoga to the White House. Seriously. "Why not bring this gift of healing to the people who are making this country function and run?" she asked.
On the 61st day of training, just two days before graduation, teacher training is winding down. Choudhury will lecture on the postures. Normally, he teaches his yoga classes in a tiny Speedo and lectures in street clothes. But tonight he is in tight white shorts decorated with red and orange flames. He climbs on the mirrored box and dances, as his students cheer. In his day, they say Choudhury ruled the discos. He still loves to dance.
Here, there are no typical Indian greetings of "namaste." No oms. "Welcome to Bikram's Torture Chamber," he announces, and he is not joking.
"This is beyond human imagination," Bikram says of his yoga. "You are not Kareem, McEnroe, or Nixon. You are just a piece of [expletive]." He threatens, in street language, to do damage.
He segues into a discussion of the postures but digresses again. "Your country, your life, they are a joke," he says. "This country is a mental hospital. It is never too late. You are never too old. You will be cured by only doing one thing," he says. "What is that?'"
"Yoga," they cry out.
Students having trouble with postures come to the front of the room to be corrected. A 50-ish man comes to the front of the room and sits. Should he push himself in a posture that really hurts his knee? He tries to lean back. Students stand, craning to see if he will make it, holding their breath. Will he hurt himself?
Bikram stands over him like a Marine drill sergeant.
"Of course it hurts! What do you think this is, cheesecake? When I crushed my knee, I used to bite my finger [in pain]. I had 10 bloody fingers. If you die, call me. American men are like women.
"What do I sell?" Choudhury asks as the man struggles.
"Pain," they scream.
Later, a woman comes up and does a beautiful "camel." She leans back on her knees and grabs her feet. She arches beautifully. Bikram climbs onto her, his feet on her hip bones, surfing her like a circus acrobat.
And then, Choudhury will stun with his compassion. A stiff guy comes up to do the camel. He kneels. Choudhury instantly perceives an old ankle injury and grows gentle. His ability to discern injury and illness is uncanny, say his students. From afar, he can spot a twisted spine, an injured knee, a weak shoulder.
Some wonder if the guru has lost his way in America. Those who know Choudhury say beneath the trappings of wealth and glamour, he is still a man committed to protecting his special form of yoga, and--as his guru ordered--to spreading it as far as possible whatever it takes.
"Being in L.A., having all his cars, having all those people around him, has changed him," said Tony Sanchez, a yogi in San Francisco who studied with Choudhury and has a copyrighted yoga system of his own. "Underneath all that, though, I think he is a very pure, a very simple, and a very beautiful person."
Choudhury wearies of hearing that his brand of yoga is somehow not spiritual enough.
"I teach spirituality," he said. "I use the body as a medium. I use the body to control your mind, to make your spirit happy."
But before he can expand on the thought, his inner salesman emerges, touting ambitious new projects that seem to sprout from his head by the minute. He is building five new studios in L.A., have you heard? He is expanding the La Cienega studio. He is making an infomercial that will reach millions. He is working with partners to build a "health city" in India.
A few weeks later, he was on the phone from Paris.
"I was just at the opening of a studio here today," he said. "This is unbelievable. This has become a multimillion-dollar corporation.
"Did I tell you about the first world yoga contest in Los Angeles, from Sept. 25-28? There will be judges from all over the world.... "


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Villa Sorriso, Napa, California
Happy Friday! Feast your eyes on some sophisticated property porn that's worth a pretty penny.
$35,000,000 (yes, that's right) gets you 20,000 square feet of living space: 5 suites, library, theater, elevator, cellars for art and wine, bridge gallery and viewing tower overlooking the valley but not to exclude the pools, sculpture gardens, tennis court, stables and about 600 acres worth of vineyards where olives and world class Cabernet Sauvignon grapes are grown.
Don't have a green thumb in grape growing? Not to worry, the "fabulous expanse" is carefully cared for by an on-site manager.
Yea, the house is incredible but it's the ground we're after. What do you think?
More info here via Sotheby's Realty.


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- Filed Under
On Tuesday, four new members of the Pensacola City Council are scheduled to take office and begin four-year terms.
They are:
» Charles Bare, who will represent the the city's "at-large B seat." He defeated Council President Sam Hall in a three-way race in the August primary. Bare ran unsuccessfully for mayor in 2010 but regrouped to win this citywide seat on the nine-member council. He promised to focus on improving city neighborhoods.
» Jewel Cannada-Wynn, who had served for six years until a defeat in 2010, will replace Ron Townsend, who decided not to run for re-election in District 7. During her campaign, she talked of the importance of the mayor and council working as a team.
» Andy Terhaar, who was elected by default in November when District 3 incumbent Maren DeWeese withdrew. She plans to run for mayor in 2014. During his campaign, Terhaar said he would work for district residents - including a new fire station - and improving the city as a whole.
» Gerald Wingate, who defeated incumbent John Jerralds in District 5. Wingate expressed optimism about the city's future during his campaign. He also said any friction between the council and Mayor Ashton Hayward could be resolved with better communication.
Three new council members with a veteran returning. They each will bring ideas to the council. We wish them luck.
They will join Larry Johnson, Sherri Myers, Megan Pratt, Brian Spencer and P.C. Wu on the council. (Wu was re-elected without opposition this year.) The other four are in the middle of four-year terms.
Even casual observers of the City Council know the last two years have been fraught with problems. Operating under a new city charter, the council and mayor were at odds over a number of issues. That enmity prompted two lawsuits, a failed attempt at a budget veto override and the threat of subpoenas, which, when sanity finally prevailed, was dropped earlier this month.
The mayor has seized on the new role and working to move the city forward. His plan to stem the city's declining population through annexation and a first-time home buyers program is one example. He promotes Pensacola and is a positive ambassador for the city.
As the city government evolves, the council's role is changing. Members, however, still remain vital if their tenure is marked by improving the neighborhoods they represent; working with the mayor to attract employers; and finding ways to reduce crime and poverty. Those may not be glamorous, headline-grabbing tasks, but they will improve the city.
As we have stated, healthy debate on issues is needed and encouraged, especially between the branches of government. We daresay no one has cornered the market on good ideas for Pensacola's future. But for ideas to become reality, there has to be mature discussion that includes listening.
Positive momentum is building in Pensacola and we encourage the four new members to help lead the city without the rancor and political gamesmanship that has plagued the current council.


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|Basic Information
|Biotope classification
|Habitat preferences and distribution
|Species composition
SS.SSa.CMuSa.AalbNuc recorded () and expected () distribution in Britain and Ireland (see below)
A hydrodynamic regime of weak tidal streams and shelter from waves (owing to the depth of the seabed offshore) creates conditions for the formation of circalittoral muddy sands. Sediments in areas of seabed largely undisturbed by water movement are less well sorted, with substantial amounts of silt and organic matter, which favours deposit and suspension feeders of all types.
The CMS.AbrNucCor biotope is dominated by bivalve molluscs. Bivalves that inhabit muddy low energy environments are typically deposit feeders, although suspension feeders, e.g. Corbula gibba may also be abundant, or species may be able to switch between feeding methods, e.g. Abra alba. When deposit feeding, bivalves remove, microzooplankton, organic and inorganic particles, and microbes including bacteria, fungi and microalgae from the sediment. They also probably absorb dissolved organic materials in much the same manner as when filter feeding (Dame, 1996). Deposit feeding bivalves adopt two approaches to feeding: bulk feeding and particle sorting. Some may ingest large amounts of sediment in a relatively nonselective manner, or may sort particles before they are ingested and reject the majority as pseudofaeces. Deposit feeding bivalves may process up to 20 times their body weight in sediments per hour with as much as 90 % of the sediment egested as pseudofaeces (Lopez & Levinton, 1987). Suspension feeders also demonstrate some selection of particles ingested, the efficiency of which is related to palp size rather than gill type or turbidity in the bivalve's environment. For instance, Corbula gibba demonstrated a 10% selection efficiency in feeding experiments by Kiørboe & Møhlenberg (1981).
Polychaetes are also characteristic of the infauna of the biotope. Members of the families Spionidae (e.g. Prionospio spp., Spiophanes bombyx, Spio filicornis) and Cirratulidae (e.g. Chaetozone setosa, Tharyx spp.) are small slender worms which burrow through the sediment and use their long anterior palps or tentacles to collect organic particles. Nephtys hombergii is carnivorous and captures molluscs, crustaceans and other polychaetes with its eversible, papillated proboscis. Other carnivorous polychaetes include glycerid polychaetes such as Goniada maculata, and Glycera alba. The flabelligerid worm, Diplocirrus glaucus is a commensal of sea urchins, e.g. Echinocardium cordatum and feeds on its faecal material (Hayward & Ryland, 1996). Some polychaetes, however, are less mobile and construct tubes or burrows in the sediment. Lagis koreni constructs a tapered tube of sand grains, open at both ends, but orientated so that the worm's head is down in the sediment, drawing water and food into its burrow below the surface. Echiuran worms, e.g. Echiurus echiurus, also create burrows within the sediment and Thomsen & Altenbach (1993) found that the numbers and biomass of bacteria and foraminifera were up to three times higher around burrows of Echiurus echiurus than in surrounding sediment.
The heart urchin, Echinocardium cordatum occurs in both muddy and clean sands, although it grows at a considerably slower rate in the former than the latter (Buchanan, 1966). It is a relatively large infaunal species whose burrowing activity may serve to enhance oxygenation of the sediments and make them less compact.
The burrowing and feeding activities of deposit feeding macrofauna, are likely to modify the fabric and increase the mean particle size of the upper layers of the substrata by aggregation of fine particles into faecal pellets. Such actions create a more open sediment fabric with a higher water content which affects the rigidity of the seabed (Rowden et al., 1998). Such alteration of the substratum surface can affect rates of particle resuspension.
Bioturbation by the infauna on a variety of scales is also likely to be of particular importance in controlling chemical, physical and biological processes in marine sediments, especially when the influences of physical disturbances such as wave action or strong currents are minimized (Widdicombe & Austen, 1999).
In summary, a mix of infaunal burrowers (bivalves, polychaetes and echinoderms) in a sedimentary biotope such as this will generate a complex and continually changing 'mosaic' of habitat patches experiencing different types and levels of disturbance. The differing responses of individual species to such patchiness are likely to be a factor in the maintenance of local species diversity. The depth of penetration into the sediment by infaunal species is also likely to be enhanced by the physical and chemical consequences of infaunal activity (Hughes, 1998).
Epifaunal species include brittlestars, Ophiura albida and Ophiura ophiura, these species compete with neighbours for space, as they are surface deposit feeders. Like other echinoderms inhabiting soft sediments, they have pointed rather than suckered tube feet, the latter being of little use for attachment to soft sediment (Wood, 1988). Other epifaunal organisms associated with muddy sands are predominantly mobile species, including the crabs Liocarcinus depurator, Atelecyclus rotundatus and Macropodia spp. Predatory fish are also likely to frequent the biotope to feed upon bivalves, polychaetes and brittlestar arms, and include Dover sole, Solea solea and members of the cod family. The infaunal, tube-building, polychaete Lagis koreni is a significant food-source for commercially important demersal fish, especially dab and plaice, e.g. Macer (1967), Lockwood (1980) and Basimi & Grove (1985).
This review can be cited as follows:
Budd, G.C. 2006. Abra alba, Nucula nitida and Corbula gibba in circalittoral muddy sand or slightly mixed sediment. Marine Life Information Network: Biology and Sensitivity Key Information Sub-programme [on-line]. Plymouth: Marine Biological Association of the United Kingdom. [cited 28/11/2015]. Available from: <http://www.marlin.ac.uk/habitatecology.php?habitatid=62&code=2004>


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Now you can buy the new album at Aorheaven.com
Press Release - Sandviken, Sweden, November, 2013 - Speechless Music Production proudly announces the release of GRAVITY, the highly anticipated fifth studio album by the Swedish band HIGHER GROUND. The album GRAVITY was produced, recorded and mixed by HIGHER GROUND at their own studio.
With GRAVITY the band breaks into new musical territory, though still preserving the borders around theunique sound of HIGHER GROUND. The title is a free interpretation of the term gravity, as well as a bold statement from a band that has evolved for 19 years with its original lineup: HIGHER GROUND being the force that holds the different songs of the album - as well as the different band members - together!
From the dramatic opening of "Fate", the heaviest rock song on offer – To the dark bluesy end of "Favorite Friend", HIGHER GROUND delivers a variety of expressions. "Missing You" is a guitar based pop/rock song with sweet melodies, which also brought the band attention in the semi-finals of Swedish chart competition "Svensktoppen Nästa", in 2012. The sweet pop/rocksong "Glory" proved equally successful in the same event 2013. "Supernatural" is a catchy, retro style up-tempo song that holds a clue to the album title in its lyrics.The slow ballad "Remember me" is a rare occasion."Again" takes us into modern mid-tempo pop/rock. The rock song "Waste of Time" is a time well spent."Tightrope" is a melancholic melodic rock song. GRAVITY simply proves HIGHER GROUND at their best – A force to be reckoned with!
The album GRAVITY was released in November 2013 and can be ordered at www.higherground.nu and also on iTunes, Amazon, Spotify and several other digital music distribution services.
Speechless Music Production


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The soccer team Brooklyn United, posing with Police Commissioner Ray Kelly, wins the NYPD Commissioner's League Cup. Courtesy: Picasa user Arab AmericanNY. © All rights reserved
In 2009, the Arab American Association of New York sponsored the Brooklyn United, a team in the New York Police Department's youth soccer league. "We were trying to engage with law enforcement, get kids off the street and it was kind of putting out our hand to the NYPD," said the organization's executive director, Linda Sarsour. That first year, the Brooklyn United won the tournament trophy and even posed for the above photo with Police Commissioner Ray Kelly. But by 2011, the AAANY withdrew its sponsorship after learning that the league was also being used as a way to monitor the Arab, Muslim and South Asian players and their families.
The question now hangs in the air: Were the NYPD youth soccer leagues as well as the teams that compete for the "NYPD Cricket Cup"—yes, there is such a thing—set up explicitly for the purposes of surveillance? Was the trust of hundreds of families who signed up their children for these leagues violated in the name of intelligence gathering? Were these leagues just a way to practice a more effective form of racial and ethnic profiling? Sarsour certainly thinks so. "The NYPD created these spaces," she said. "When I think about it I get goosebumps. It is so outrageous. What parent would think if you were part of a Little League or Police Athletic League that the police would be tracking your kids on the basis of their ethnicity? When the leagues started we thought they were trying to engage our community through sports. We were wrong."
These families have the right to know whether the NYPD specifically set up these leagues for the purposes of keeping tabs on a sports-loving community or if it just found a rich opportunity for surveillance once everyone was organized to play. Its community outreach and media divisions have still not returned my requests for comment. If and when they do, we will share their response.
I was able to speak with Matt Apuzzo, co-author of the new book Enemies Within, which has blown the lid off of the full extent of NYPD's surveillance of Arab, Muslim and South Asian communities.
"What we know is that they did set up soccer and cricket leagues from youth to adults," said Apuzzo. "We also know that they encouraged their detectives to join the adult cricket and soccer leagues. I don't know if we can say they created the leagues for the express purpose of surveillance as opposed to outreach. But we do know from their own documents that they do see these sports leagues as an opportunity to keep tabs on conversations. Either way, we certainly can say that any effort at actual legitimate community outreach can be undermined by the surveillance aspect because it makes people suspicious of motives."
Whether the leagues themselves were part of a master plan or clumsy happenstance doesn't really matter, of course, to the communities that feel their trust was betrayed. Rinku Sen, President of the Applied Research Center, described the using of sports leagues to spy on kids as "abusive." She also made the point to me that the NYPD's unwelcomed entry into this space exacts a particularly serious price. "Coming out of a regional history fraught with religious and national conflict, sports are one arena in which Hindus, Sikhs, Christians, Muslims and others have been able to come together, especially in the diaspora. The NYPD spying brings a layer of suspicion into this world that has otherwise been an important place to build trust and camaraderie."


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Filth and Wisdom
Dir: Madonna. UK. 2008. 81 minsContrary to billing, there's not much filth and precious little wisdom in Madonna's directing debut. Even so, you have to applaud her for daring to get behind a camera, given that her career in front of it has been strewn with clunkers. While Filth And Wisdom may not quite inhabit the same Hall of Shame as Shanghai Surprise, Body of Evidence and (God save us) Swept ...
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An inspector calls
Who could be more quintessentially British than Max Hutchinson, former RIBA president, founder of Architects for Aid, media star and introduction agent for Stewart McColl? Obviously keen to preserve Blighty's values in Cannes, he was spotted on the Croisette in bowler hat and trenchcoat, towing a compact suitcase. Had he got the wrong festival? He looked less like UK architecture at MIPIM, rather more the reincarnation of Peter Sellers at the film fest.
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