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Drp1-Dependent Mitochondrial Autophagy Plays a Protective Role Against Pressure Overload–Induced Mitochondrial Dysfunction and Heart FailureCLINICAL PERSPECTIVE
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Background—Mitochondrial autophagy is an important mediator of mitochondrial quality control in cardiomyocytes. The occurrence of mitochondrial autophagy and its significance during cardiac hypertrophy are not well understood.
Methods and Results—Mice were subjected to transverse aortic constriction (TAC) and observed at multiple time points up to 30 days. Cardiac hypertrophy developed after 5 days, the ejection fraction was reduced after 14 days, and heart failure was observed 30 days after TAC. General autophagy was upregulated between 1 and 12 hours after TAC but was downregulated below physiological levels 5 days after TAC. Mitochondrial autophagy, evaluated by electron microscopy, mitochondrial content, and Keima with mitochondrial localization signal, was transiently activated at ≈3 to 7 days post-TAC, coinciding with mitochondrial translocation of Drp1. However, it was downregulated thereafter, followed by mitochondrial dysfunction. Haploinsufficiency of Drp1 abolished mitochondrial autophagy and exacerbated the development of both mitochondrial dysfunction and heart failure after TAC. Injection of Tat-Beclin 1, a potent inducer of autophagy, but not control peptide, on day 7 after TAC, partially rescued mitochondrial autophagy and attenuated mitochondrial dysfunction and heart failure induced by overload. Haploinsufficiency of either drp1 or beclin 1 prevented the rescue by Tat-Beclin 1, suggesting that its effect is mediated in part through autophagy, including mitochondrial autophagy.
Conclusions—Mitochondrial autophagy is transiently activated and then downregulated in the mouse heart in response to pressure overload. Downregulation of mitochondrial autophagy plays an important role in mediating the development of mitochondrial dysfunction and heart failure, whereas restoration of mitochondrial autophagy attenuates dysfunction in the heart during pressure overload.
Autophagy is an important mechanism for degradation of intracellular components, in which cytosolic proteins and organelles are sequestered in double-membrane vesicles called autophagosomes and delivered to lysosomes for degradation.1 Autophagy helps maintain the quality of the intracellular environment by eliminating misfolded proteins and malfunctioning organelles, and preserves the ATP level by recycling extracted amino acids and fatty acids for ATP synthesis. Increasing evidence suggests that autophagy plays an important role in protecting the heart during heart failure (HF).2,3
Editorial, see p 1225
Clinical Perspective on p 1263
Although autophagy can degrade bulk intracellular materials in a relatively nonspecific manner, it can also specifically target malfunctioning organelles. For example, activation of a series of events, including stabilization of Pink1 on depolarized mitochondria, phosphorylation of Mfn2, recruitment of Parkin to mitochondria, and recognition of depolarized mitochondria by autophagosomes, specifically eliminates damaged mitochondria; this process is termed mitochondrial autophagy or mitophagy.4,5 Mitochondria are the primary source of cellular ATP, but when they malfunction, they become a major source of oxidative stress and trigger both apoptosis and necrosis. Mitochondria are therefore normally regulated by quality control mechanisms, including fission and fusion, biogenesis, and autophagy.6 Impairment of any of these mitochondrial quality control processes leads to mitochondrial dysfunction and cell death.
The heart undergoes hypertrophy in response to hemodynamic overload, such as pressure overload (PO), initially with the purpose of reducing wall stress. The heart also initiates various adaptive mechanisms, including autophagy, to cope with energetic stress, increased oxidative stress, and cell death. Previous studies have shown that general autophagy is activated during cardiac hypertrophy and HF, and plays both protective and detrimental roles in the heart.3,7 However, although cardiac hypertrophy and HF are commonly accompanied by general and nonspecific autophagy,3,7 the presence of organelle-specific mitochondrial autophagy and its role during pathological hypertrophy have not been well characterized.
Drp1 is a small GTPase that mediates mitochondrial fission. Drp1 also plays an important role in mitochondrial autophagy in cardiomyocytes (CMs) at baseline and in response to energy stress.6 Although Drp1 is localized primarily in the cytosol, it is recruited to mitochondria in response to stress, thereby inducing mitochondrial fission and mitochondrial autophagy in the heart.6 However, whether Drp1 is involved in mitochondrial autophagy or the maintenance of mitochondrial function during PO-induced cardiac hypertrophy is currently unknown.
Using mice with transverse aortic constriction (TAC) as a model, we investigated the role of mitochondrial autophagy during PO-induced hypertrophy and HF. The goals in this study were to (1) demonstrate activation of mitochondrial autophagy, (2) clarify the role of Drp1 in mediating mitochondrial autophagy, and (3) elucidate the functional significance of mitochondrial autophagy in the development of mitochondrial dysfunction and HF in response to PO.
An expanded Methods section is available in the online-only Data Supplement.
Mouse Models
Cardiac-specific heterozygous Drp1 knockout (Drp1-hetCKO) mice were generated by crossing Drp1 flox heterozygous (fl/+) mice with αMHC-Cre mice.6 Transgenic mice expressing GFP-LC3 (Tg-GFP-LC3) or mRFP-GFP-LC3 (Tg-tf-LC3),8 beclin 1 heterozygous knockout mice (beclin 1-hetKO),9 and cardiac-specific mammalian sterile 20-like kinase 1 (Mst1) homozygous knockout (Mst1-CKO) mice2 have been described. All experiments involving animals were approved by the Rutgers–New Jersey Medical School's Institutional Animal Care and Use Committee.
Keima With Mitochondrial Localization Signal
Keima with mitochondrial localization signal (Mito-Keima) is a mitochondrially localized pH-indicator protein.10 The method used to detect lysosomal delivery of Mito-Keima has been described.6
Tat-Beclin 1
The retro-inverso Tat-Beclin 1 (TB1) d-amino acid sequence was RRRQRRKKRGYGGTGFEGDHWIEFTANFVNT.11 The control peptide, Tat-scrambled (TS), consisted of the Tat protein transduction domain, a GG linker, and a scrambled version of the C-terminal 18 amino acids of TB1 (RRRQRRKKRGYGGWETAFGTTEHIFFDNGV). For in vivo experiments, d-amino acid peptides were dissolved in H2O and stored at –80°C until use. For induction of autophagy in vitro, CMs were washed with PBS(–) and treated for 3 hours with peptides (25 μmol/L) dissolved in OPTI-MEM acidified with (0.15% v/v) 6 N HCl.11 For in vivo induction, mice were injected intraperitoneally with TS or TB1 at 20 mg/kg daily for 2 weeks, beginning 1 week after TAC, or daily for 1 week, beginning immediately after TAC. To evaluate autophagic flux in vivo, chloroquine was injected (10 mg/kg) intraperitoneally12 30 days after TAC. Four hours later, animals were euthanized for immunoblot detection of autophagy markers.
Electron Microscopy
Conventional electron microscopy (EM) was performed as described previously.9 Mitochondrial mass was analyzed using ImageJ.6 The average mitochondrial mass was calculated from 250 mitochondria per slide on 4 different slides.
Real-Time Quantitative Polymerase Chain Reaction for Mitochondrial DNA
Total DNA was extracted from mouse hearts using the DNeasy Blood & Tissue kit (QIAGEN). The mtDNA content was quantified by real-time quantitative polymerase chain reaction of cytochrome b and β-actin as described.6
AAV Transduction
Adeno-associated virus (AAV) vectors were generated from AAV9 vectors. Doses of 1×1011 particles of each vector were administered intravenously through the jugular vein.13
Mitochondrial Assays
The mitochondrial fraction was prepared from mouse hearts (40–50 mg) as described.6 Mitochondrial electron transport chain complex activity was evaluated with the MitoCheck Complex Activity Assay Kit (Cayman Chemical Company). ATP production was measured with an ATP Bioluminescent Assay (Sigma) in which 25 μg of mitochondrial protein was incubated with ATP assay mix and mannitol/sucrose/Hepes buffer containing 625 μmol/L ADP and substrate (10 mmol/L pyruvate and 10 mmol/L malate). A Seahorse XF96 Extracellular Flux Analyzer (Seahorse Bioscience) was used to measure the rate of oxidative phosphorylation in intact CMs.14 The data are expressed as relative experimental values per heart.
Immunoblot Analyses
The methods used for preparation of cell lysates from in vitro and in vivo samples and for immunoblot analyses have been described previously.15 The antibodies used include Drp1 (BD Transduction, 611112), phospho-Drp1 (Ser616; Cell Signaling, 4494S), phospho-Drp1 (Ser637; Cell Signaling, 4867S), LC3 (BML, M186-3), p62 (ORIGENE, TA307334), cyclooxygenase (Cox) I (abcam, ab110413), Beclin 1 (Cell Signaling, 3738S), PGC-1α (Santa Cruz, sc-13067), Cox IV (Cell Signaling, 4844S), Mst1 (BD Transduction, 611052), phospho-Mst1 (Cell Signaling, 3681S), and GAPDH (Cell Signaling, 2118S).
Human Heart Samples
Heart lysates from explanted hearts were obtained from patients who received heart transplants and from age-matched donors. The study was approved by the Institutional Ethics Committee (VGHIRB no. 2012-06-028D). All patients or their families expressed their willingness to participate through an informed consent form. Detailed information regarding the human samples was provided previously.16
Statistical Analysis
Statistical analyses were conducted with the SPSS 22.0 J software program (SPSS Japan Institute, Tokyo, Japan) or the JMP software program (SAS Japan Institute, Tokyo, Japan). Data are expressed as mean±standard error of the mean for the indicated number of experiments or mice. The difference in means between 2 groups and multiple groups was evaluated using the Welch t test and the Welch analysis of variance, respectively. Post-test comparisons for multiple analyses were performed by using the Tukey test. P values were 2-sided and P values of <0.05 were considered statistically significant.
Basal Characterization of C57BL/6J Mice After PO
To evaluate the effects of PO, C57BL/6J mice were subjected to either TAC or sham operation and followed up for 1 hour to 30 days. There was no significant difference in cardiac hypertrophy or function between mice subjected to sham operation and mice that had not undergone any operation at any time point (not shown). Therefore, sham refers to the combined data from the sham-operated mice at all time points. The heart is enlarged in response to TAC in a time-dependent manner (Figure 1A). Postmortem assessment showed that left ventricular (LV) weight/tibia length (TL) was significantly greater 5 days after TAC and thereafter than in sham (Figure 1B). Wheat germ agglutinin staining showed that CM cross-sectional area was significantly greater 7, 14, and 30 days after TAC than in sham (Figure IA in the online-only Data Supplement). Myocardial fibrosis, evaluated with Picric Acid Sirius Red staining, was also significantly greater 7, 14, and 30 days after TAC than in sham (Figure IB in the online-only Data Supplement). Echocardiographic measurements showed that, although the LV ejection fraction (LVEF) was maintained until 7 days after TAC, it was decreased significantly 14 and 30 days after TAC in comparison with sham (Figure 1C and 1D). Lung weight/TL was significantly elevated 1, 3, 6, and 12 hours after TAC than after sham. This index was then normalized at 24 hours of TAC but was again elevated at 14 and 30 days (Figure 1E). LV end-diastolic pressure (LVEDP) was significantly elevated 6 hours after TAC in comparison with after sham. Although the LVEDP declined at 24 hours of TAC to a level slightly higher than that in sham, it rose again and remained elevated in comparison with sham until 30 days (Figure 1F and 1G). The pressure gradient was between 70 and 80 mm Hg, except at 6 hours, and 14 and 30 days (Figure IC in the online-only Data Supplement). We speculate that constriction is still evolving at 6 hours, whereas LV dysfunction decreases the pressure gradient at 14 and 30 days. CM apoptosis, evaluated with terminal deoxynucleotidyl transferase dUTP nick end labeling staining, was increased significantly 14 and 30 days after TAC than after sham (Figure ID in the online-only Data Supplement). These results suggest that TAC induces cardiac hypertrophy within 5 days, LV dysfunction within 14 days, and HF at 14 to 30 days.
Autophagy Is Transiently Upregulated in the Early Phase but Downregulated in the Chronic Phase of PO
We next investigated how TAC affects autophagy in the heart. LC3-II was significantly increased 3, 6, and 12 hours after TAC, was returned to sham level by 24 hours, and then was significantly decreased from 5 days on (Figure 2A). p62/SQSTM1, a protein known to be degraded by autophagy, was significantly decreased 1 and 3 hours after TAC and increased at 6 hours and 3, 5, and 7 days after TAC in comparison with sham (Figure 2B). These results suggest that autophagy is transiently activated by TAC during the initial 12 hours but is suppressed after 5 days. To further validate our results, autophagic flux was evaluated with tandem fluorescent LC3 (tf-LC3). At 6 hours, both green fluorescent protein/red fluorescent protein double-positive autophagosomes (yellow) and red fluorescent protein–positive autolysosomes (red) were significantly increased in the TAC group in comparison with in the sham group. Furthermore, chloroquine induced a significantly greater increase in the number of yellow dots at 6 hours after TAC than after sham operation. However, after 7 days of TAC, both yellow and red dots were significantly decreased in the absence of chloroquine in comparison with sham, and the chloroquine-induced increase in the number of yellow dots was significantly smaller, consistent with the notion that autophagic flux was decreased 7 days after TAC (Figure 2C, Figure II in the online-only Data Supplement). Together, these data suggest that autophagic flux is transiently activated after TAC, peaking within 3 to 12 hours, but inactivated after 7 days.
Activation of Mst1 suppresses autophagy in CMs.2 Mst1-CKO mice were used to test the involvement of Mst1 in the suppression of autophagy after TAC. In control hearts, Mst1 protein and Thr183-phosphorylated Mst1 were increased 7 days after TAC (Figure IIIA and IIIB in the online-only Data Supplement). Although LC3-II was decreased and p62 was increased 7 days after TAC in control hearts, LC3-II was significantly increased and p62 was decreased in Mst1-CKO mice (Figure IIIC and IIID in the online-only Data Supplement). These results suggest that activation of endogenous Mst1, in part, mediates the suppression of autophagy in response to PO.
Mitochondrial Autophagy Is Stimulated After Transient Activation of General Autophagy Ceases
EM analyses showed that autophagosomes containing only mitochondria were observed after 3, 5, and 7 days of TAC (Figure 3A), but not after 6 or 24 hours or 14 or 30 days of TAC or after sham operation (data not shown). Quantitative analysis indicated that mitochondrial autophagy was increased significantly after 3, 5, and 7 days of TAC in comparison with sham (Figure 3B). Mitochondrial DNA content, evaluated through real-time polymerase chain reaction of cytochrome b, was significantly greater 1, 3, 6, and 24 hours and 2 days after TAC than in sham (Figure 3C). However, it transiently decreased below control levels 3 and 5 days after TAC, and returned to control levels after 7 days (Figure 3C). The level of Cox I, a mitochondrial matrix protein, normalized by the level of GAPDH, a cytosolic protein, decreased transiently 5 to 7 days after TAC in comparison with sham (Figure 3D). Cardiac PGC-1α protein expression did not change significantly at any time point, suggesting that PO may not affect mitochondrial biogenesis (Figure 3E). To further evaluate mitochondrial autophagy, we expressed Mito-Keima6 in the heart by using AAV-mediated gene delivery. To confirm the lysosomal localization of Mito-Keima, we coinjected AAV-YFP-Lamp1. Puncta with a high 560/440 nm excitation ratio, reflecting increased lysosomal localization of Mito-Keima, were increased 3 to 7 days after TAC in comparison with sham operation. The high-ratio Mito-Keima dots were colocalized with YFP-Lamp1 dots, suggesting lysosomal localization (Figures IVA and VIIB in the online-only Data Supplement). These results further suggest that lysosomal degradation of mitochondria is stimulated 3 to 7 days after TAC. Although mitochondrial localization of Parkin and polyubiquitination of mitochondrial proteins, indicators of Parkin-mediated mitophagy,17 were both increased 6 hours after TAC, they were downregulated at 7 days (Figure IVB and IVC in the online-only Data Supplement). Beclin 1 was increased significantly 1 and 6 hours after TAC, preceding the initial peak in autophagy, briefly significantly decreased below baseline at 12 hours, and then increased in comparison with sham again after 5 to 7 days, coincident with the occurrence of mitochondrial autophagy (Figure V in the online-only Data Supplement). These results suggest that mitochondrial autophagy is stimulated transiently between 3 and 7 days after TAC, accompanied by transient decreases in mitochondrial content.
Mitochondrial Dysfunction Develops After Mitochondrial Autophagy Is Suppressed
Mitochondrial ATP production and ATP content were both attenuated significantly 5 days after TAC and thereafter (Figure 4A and 4B). The activities of mitochondrial complexes I, II + III, and IV were also significantly attenuated 5 to 7 days after TAC in comparison with sham (Figure 4C through 4E). We also evaluated the rate of oxidative phosphorylation in isolated mitochondria by using a Seahorse analyzer. The basal oxygen consumption rate and the maximum oxygen consumption rate were significantly lower 14 days after TAC than in sham (Figure 4F and 4G), as was the level of proton leak, determined by subtracting the oxygen consumption rate–linked ATP synthesis from basal oxygen consumption rate (Figure 4H). These data indicate that TAC induces mitochondrial dysfunction in the mouse heart after 5 to 7 days of TAC.
Foreshortening of Mitochondria and Mitochondrial Translocation of Drp1 Coincide With Activation of Mitochondrial Autophagy
EM analyses showed that mitochondria in control hearts were primarily rectangular or spherical at baseline, with tubular mitochondria observed less frequently. Mitochondria were enlarged transiently ≈24 hours after TAC, shrunk to a spherical shape again 3 to 5 days after TAC, and then elongated/enlarged again 30 days after TAC (Figure 5A, Figure VIA in the online-only Data Supplement). Quantitative analyses revealed that the mitochondrial mass was significantly greater 24 hours and 30 days after TAC, but was significantly smaller 3 and 5 days after TAC, than in sham (Figure 5A and 5B). The longitudinal length of individual mitochondria in comparison with that of sarcomeres was evaluated as described previously.6 Although the proportion of elongated mitochondria was significantly greater 24 hours and 30 days after TAC than in sham, the proportion of foreshortened mitochondria was significantly greater 7 days after TAC than in sham (Figure VIB in the online-only Data Supplement). Drp1 plays an important role in mediating mitochondrial fission and autophagy in the heart.6 Drp1 is primarily cytosolic at baseline but translocates to mitochondria in response to stress.6 Total Drp1 in the heart did not change significantly after TAC (Figure VIC in the online-only Data Supplement). However, mitochondrial accumulation of Drp1 was significantly increased 2 and 3 days after TAC, whereas it was significantly suppressed 14 and 30 days after TAC in comparison with sham (Figure 5C). Conversely, cytosolic Drp1 was significantly decreased 2 to 5 days after TAC, whereas it was significantly increased 14 and 30 days after TAC in comparison with sham (Figure 5D). Phosphorylation of Drp1 at S616 and at S637 positively and negatively correlates with mitochondrial translocation of Drp1, respectively.18,19 Drp1 S616 phosphorylation was increased significantly 1, 2, 3, and 5 days after TAC in comparison with sham (Figure 5E). In contrast, Drp1 S637 phosphorylation was decreased significantly 12 hours and 2 days after TAC in comparison with sham (Figure 5F). Together, these results suggest that mitochondrial translocation of Drp1 is induced between 2 and 5 days, peaking 3 days after TAC, but that Drp1 is located primarily in the cytosol again thereafter. Furthermore, the mitochondrial translocation of Drp1 after TAC at least partially coincided with S616 phosphorylation/S637 dephosphorylation of Drp1. In myocardial sections obtained from human HF patients, phosphorylation of Drp1 at S616 was significantly lower than in healthy controls (Figure VID in the online-only Data Supplement), whereas phosphorylation of Drp1 at S637 did not differ significantly. Because significant mitochondrial dysfunction and HF only developed in mice after mitochondrial foreshortening and mitochondrial translocation of Drp1 returned to normal levels, we next investigated the functional significance of Drp1 in the heart during PO-induced cardiac hypertrophy and HF.6
Downregulation of Drp1 Exacerbates PO-Induced Cardiac Dysfunction
We used 16-week-old cardiac-specific conditional Drp1 heterozygous knockout (Drp1-hetCKO) mice6 to evaluate the role of endogenous Drp1 during PO-induced cardiac hypertrophy. Immunoblot analyses confirmed that there was significantly less Drp1 protein in Drp1-hetCKO mice than in control mice (Figure VIIA in the online-only Data Supplement). Drp1 fl/fl mice or Drp1 flox heterozygous (fl/+) mice (without Cre) were used as controls. These mice were subjected to either TAC or sham operation for 1, 3, or 24 hours, or 3 or 7 days, but not longer because of the high mortality rate observed in Drp1-hetCKO mice after TAC at later time points. Neither control nor Drp1-hetCKO mice showed any significant difference in cardiac parameters after sham operation. Drp1-hetCKO mouse hearts were enlarged in comparison with control mouse hearts after TAC (Figure 6A). LVEF was significantly lower in Drp1-hetCKO mice than in control mice 3 and 7 days after TAC (Figure 6B and 6C). LV weight/TL, lung weight/TL, and LVEDP were significantly greater in Drp1-hetCKO mice than in control mice 3 and 7 days after TAC (Figure 6D through 6F). Mitochondrial ATP production was significantly attenuated 3 and 24 hours and 7 days after TAC in Drp1-hetCKO mice in comparison with in control mice (Figure 6G). These data indicate that TAC-induced cardiac hypertrophy, dysfunction, HF, and mitochondrial dysfunction were rapidly developed in Drp1-hetCKO mice in comparison with in control mice.
There was significantly less Drp1 in mitochondria in Drp1-hetCKO hearts than in control hearts 3 days after TAC and sham (Figure 6H). Mitochondrial DNA content and mitochondrial mass were significantly greater in Drp1-hetCKO hearts 7 days after TAC than in control hearts or sham (Figure 6I and 6J). The number of high-ratio Mito-Keima dots and their colocalization with YFP-Lamp1 dots were significantly greater in control hearts than in Drp1-hetCKO mice after 7 days of TAC, suggesting that TAC-induced lysosomal translocation of Mito-Keima is Drp1-dependent (Figure VIIB in the online-only Data Supplement). Myocardial fibrosis, CM cross-sectional area, and the number of terminal deoxynucleotidyl transferase dUTP nick end labeling–positive nuclei in CMs were also significantly greater in Drp1-hetCKO hearts than in control hearts 7 days after TAC (Figure 6K through 6M, Figure VIIC in the online-only Data Supplement). The TAC-induced increases in the number of cells with foreshortened mitochondria observed in control mice at 7 days were significantly attenuated in Drp1-hetCKO mice, suggesting that endogenous Drp1 is involved in mitochondrial fission during PO (Figure VIID in the online-only Data Supplement). The activity of mitochondrial complex I was significantly attenuated and that of complexes II + III and IV also tended to be attenuated in Drp1-hetCKO mouse hearts in comparison with in control mouse hearts 7 days after TAC (Figure VIIE through VIIG in the online-only Data Supplement). These results are consistent with the notion that the downregulation of endogenous Drp1 induces accumulation of damaged mitochondria and mitochondrial dysfunction and exacerbates the development of HF after TAC.
TB1 Peptide Treatment Restores Mitochondrial Autophagy and Attenuates Cardiac Dysfunction After PO
To elucidate whether downregulation of autophagy and mitochondrial autophagy contributes to the development of HF, we conducted rescue experiments by using TB1.11 TB1 contains 18 amino acids derived from Beclin 1 (267–284) and potently stimulates autophagy in HeLa and other cells by mobilizing endogenous Beclin 1 from the Golgi apparatus where it is tethered through Golgi-associated plant pathogenesis–related protein 1.11 To evaluate the effect of TB1 on autophagic flux, CMs were transduced with Ad-tf-LC3. Treatment of CMs with 25 μmol/L TB1 for 3 hours significantly increased the numbers of both green fluorescent protein and red fluorescent protein puncta in comparison with treatment with Tat-scrambled peptide (TS; Figure 7A, Figure VIIIA in the online-only Data Supplement). Similarly, daily treatment of GFP-LC3 mice with TB1 for 2 weeks significantly increased the number of GFP-LC3 dots in the heart in comparison with treatment with TS, both in the absence and presence of chloroquine (Figure 7B, Figure VIIIB in the online-only Data Supplement). The chloroquine-induced increase in the number of GFP-LC3 dots in the heart was greater after TB1 treatment than after TS treatment, suggesting that TB1 increases autophagic flux in the mouse heart. We evaluated whether lysosomal clearance of mitochondria is stimulated by TB1 using Mito-Keima.10 High-ratio puncta were significantly increased after 3-hour treatment with 25 μmol/L TB1 in comparison with after treatment with TS, suggesting that TB1 stimulates lysosomal translocation of Mito-Keima in CMs (Figure 7C, Figure VIIIC in the online-only Data Supplement). Treatment of cardiac fibroblasts with TB1 also significantly increased the number of GFP-LC3 puncta in comparison with treatment with TS in the presence or absence of chloroquine treatment (Figure VIIID in the online-only Data Supplement). To normalize the cardiac autophagy level, which is downregulated after 5 days of TAC (Figure 2), we treated mice with TB1 or TS once daily on days 7 to 21 (Figure IXA in the online-only Data Supplement). After 30 days of TAC, LVEF was significantly higher, lung weight/TL and LVEDP were significantly lower, and the pressure gradient was significantly higher in TB1-treated mice than in TS-treated mice (Figure 7E through 7I). Myocardial fibrosis, CM cross-sectional area, and CM apoptosis were all significantly reduced in TB1-treated mice in comparison with in TS-treated mice (Figure 7J through 7L, IXB and IXC in the online-only Data Supplement). TB1 significantly increased LC3-II and decreased p62 in sham-operated mice, suggesting that TB1 stimulates general autophagy at baseline (Figure 7M and 7N). In contrast, TB1 significantly increased LC3-II but did not decrease p62 in the presence of TAC in comparison with TS, suggesting that TB1 failed to increase the level of general autophagy above control in the presence of TAC. Importantly, however, DNA content and Cox I in mitochondria were significantly lower in TB1-treated mice than in TS-treated mice (Figure 7O and 7P). Furthermore, EM analyses of the mouse heart showed that autophagosomes containing mitochondria were observed in TB1-treated but not in TS-treated hearts (Figure 7Q). Quantitative analysis indicated that mitochondrial autophagy was increased significantly in TB1-treated hearts in comparison with in TS-treated hearts (Figure 7R). Mitochondrial ATP production after TAC was greater in TB1-treated than in TS-treated mice (Figure 7S). These results suggest that TB1 treatment stimulates mitochondrial autophagy, thereby preserving overall mitochondrial health in the presence of PO.
The Protective Effect of TB1 Against PO-Induced Cardiac Hypertrophy and HF Is Abolished in the Presence of Drp1 or Beclin 1 Downregulation
To test whether the aforementioned effect of TB1 is mediated through general autophagy or mitochondrial autophagy, we treated Drp1-hetCKO, beclin 1-hetKO, and respective control mice with TB1 or TS. Because of the high susceptibility of these mice to TAC-induced cardiac dysfunction, these mice were subjected to only 7 days of TAC. The mice were treated daily with either TB1 or TS on days 1 to 7 and euthanized on day 7 (Figure XA in the online-only Data Supplement). Neither TB1 nor TS affected cardiac function, mitochondrial content, or mitochondrial function in sham-operated Drp1-hetCKO or beclin 1-hetKO mice. TAC induced cardiac dysfunction and signs of HF in both Drp1-hetCKO and beclin 1-hetKO mice treated with either TB1 or TS. Importantly, LVEF, lung weight/TL, pressure gradient, and LVEDP did not significantly differ between TB1-treated and TS-treated mice after TAC when these peptides were injected into Drp1-hetCKO (Figure 8A through 8E) or beclin 1-hetKO mice (Figure 8I through 8M). Furthermore, TB1 failed to restore mitochondrial ATP production or reduce mitochondrial content or Cox I in Drp1-hetCKO mice (Figure 8F through 8H). Thus, although TB1 prevented exacerbation of PO-induced cardiac dysfunction in control mice, it failed to do so in Drp1-hetCKO or beclin 1-hetKO mice. Taken together, these data suggest that TB1 attenuates the development of HF by stimulating Drp1- or Beclin 1-dependent activation of autophagy and mitochondrial autophagy in response to PO.
The major findings of the present study are: (1) PO initially activates general autophagy but suppresses it thereafter, (2) the time course of mitochondrial autophagy is distinct from that of general autophagy, (3) PO-induced suppression of general and mitochondrial autophagy is followed by the development of mitochondrial dysfunction and HF, (4) endogenous Drp1 plays an essential role in mediating mitochondrial autophagy and maintaining both mitochondrial and cardiac function during PO, and (5) restoration of mitochondrial autophagy attenuates progression of HF during PO (Figure XB in the online-only Data Supplement).
We here show that PO activates autophagy 3 to 12 hours after TAC but suppresses autophagy below baseline after 5 days. One report showed that autophagy is upregulated,7 whereas others have demonstrated that autophagy is downregulated 7 days after PO.20 However, the time-dependent changes in cardiac autophagic activity during PO that were observed in this study have not been shown previously. In the acute phase, PO elicited transient LV dysfunction, as indicated by an increase in lung weight/TL and LVEDP, but the mice recovered within 24 hours. During this period, the heart appeared to retain the ability to activate autophagy. However, autophagy was suppressed below physiological levels after 3 to 5 days, and the flux was significantly suppressed at 7 days. Acute mechanical stress activates Nox2-dependent reactive oxygen species production at the junctional sarcoplasmic reticulum,21 whereas energy stress stimulates Nox4 at the endoplasmic reticulum,22 and we have shown previously that oxidative stress stimulates autophagy.8,22 In contrast, chronic activation of either Mst1 or mTOR suppresses autophagy.2,23 PO chronically activates mTOR and Mst1, both of which are mechanistically involved in the pathogenesis of HF.2,24 We here showed that activation of Mst1 partly mediates the suppression of general autophagy 7 days after PO, but further investigation is required to elucidate the signaling mechanisms mediating the phase-dependent changes in autophagic activity in response to PO.
Mitochondrial autophagy was upregulated 3 to 7 days after TAC and downregulated thereafter. Autophagosomes containing only mitochondria were observed 3 to 7 days after TAC but not at other time points. This was accompanied by decreases in mitochondrial content. This time course of mitochondrial autophagy is distinct from that of general autophagy. Dissociation between mitophagy and conventional autophagy was also observed in mouse embryonic fibroblasts subjected to starvation or hypoxia.25 Because mitochondrial autophagy acts only to eliminate damaged mitochondria, mitochondrial autophagy likely occurs on a much smaller scale than general autophagy. Thus, mitochondrial autophagic activity may not be faithfully reflected by changes in markers of general autophagy. Alternatively, mitochondrial autophagy may be regulated by mechanisms distinct from those regulating general autophagy, such as an Atg5- or Atg7-independent form of autophagy (alternative autophagy), that cannot be evaluated with conventional markers such as LC3-II.25
Activation of mitochondrial autophagy in response to PO coincides with mitochondrial translocation of Drp1, which was also accompanied by phosphorylation of Ser616 and dephosphorylation of Ser637 of Drp1. Furthermore, mitochondrial autophagy is abrogated in Drp1-hetCKO mice. These results suggest that Drp1 plays an important role in mediating mitochondrial autophagy in response to PO. We also found that Beclin 1, a key mediator of autophagy, is upregulated at days 5 to 7 of TAC, and during the initial 24 hours, as well, despite downregulation of general autophagy at days 3 to 7. We have shown previously that both Drp1 and Beclin 1 are important mediators of mitochondrial autophagy in the heart.6 Thus, Beclin 1 may also be critical in mediating mitochondrial autophagy during PO. In contrast, Parkin, a well-known mediator of autophagy, may not be involved in mitochondrial autophagy in response to PO, because neither mitochondrial localization of Parkin nor increased ubiquitination of mitochondrial proteins was observed when mitochondrial autophagy was induced after PO.
Previous reports suggested that mitochondrial autophagy is stimulated rather than inhibited when Drp1 or Miff, a major receptor for Drp1, is downregulated.26,27 Our study used Mito-Keima, EM, and evaluation of mitochondrial content, whereas the other study used mitochondrial accumulation of p62 and LC3-II, aggregation of Parkin, and colocalization of mitochondrial proteins and lysosomes as indicators of mitophagy.26 Our methods broadly detect mitochondrial degradation by both conventional and nonconventional28 autophagy, whereas the methods used by the other study may be more specific to conventional (LC3- and Parkin-dependent) autophagy. We found that mitochondrial autophagy is upregulated only after general autophagy is downregulated in response to TAC. Thus, we speculate that our methods may detect clearance of mitochondria by nonconventional autophagy.
Currently, the molecular mechanisms by which PO leads to Ser616 phosphorylation and Ser637 dephosphorylation of Drp1 are unknown. Cdks, ERKs, and PKCdelta have been shown to be involved in Ser616 phosphorylation,19,29 whereas PKA, ROCK1, calcineurin, and PGAM5 are involved in Ser637 phosphorylation/dephosphorylation.30,31 We predict that the PO-regulated endogenous kinases/phosphatases responsible for Drp1 phosphorylation/dephosphorylation play an important role in mediating mitochondrial autophagy in the heart. Thus, future studies should test whether manipulating the responsible kinase or phosphatase is sufficient to control mitochondrial autophagy in the heart during PO.
Understanding whether stress-induced autophagy is protective or detrimental in the heart is important but complex, because it depends on the type of stress. Here, we show that both general and mitochondrial autophagy are downregulated in the chronic phase of PO, when mice subjected to TAC develop LV dysfunction. In particular, both mitochondrial and LV dysfunction developed only after mitochondrial autophagy was downregulated. Drp1 haploinsufficiency exacerbated PO-induced mitochondrial dysfunction and HF in the mouse heart, whereas stimulation of mitochondrial autophagy with TB1 partially normalized cardiac function in the presence of PO. Importantly, the beneficial effect of TB1 was abrogated when either Drp1 or Beclin 1, molecules shown to be involved in glucose deprivation–induced mitochondrial autophagy in CMs,6 was downregulated. These results suggest that downregulation of mitochondrial autophagy plays an essential role in mediating cardiac dysfunction during PO. The decrease in mitochondrial autophagy is accompanied by increases in mitochondrial mass and mitochondrial dysfunction, suggesting that it causes accumulation of damaged mitochondria and consequent increases in oxidative stress and cell death.
Because the acute cardiac dysfunction during the initial 24 hours of TAC was alleviated after transient activation of general autophagy, peaking at 3 to 6 hours, early activation of general autophagy may also be protective. However, given that the functional significance of general autophagy during PO remains controversial,7 more studies are needed to clarify this issue. In any case, it would be important to conduct loss-of-function studies by selectively inhibiting general autophagy without affecting mitochondrial autophagy. A better understanding of the molecular mechanisms mediating general and mitochondrial autophagy is essential for addressing this issue.
Treatment with TB1, a potent inducer of autophagy,11 attenuated the progression of HF in the presence of PO. Although excessive autophagy can be detrimental in the presence of PO,7 we here show that an intervention that stimulates autophagy can be used to rescue PO-induced cardiac dysfunction if given when mitochondrial autophagy is suppressed below physiological levels. Because TB1 was not effective in Drp1-hetCKO and beclin 1-hetKO mice, we believe that TB1 is particularly effective in stimulating a Drp1- and Beclin 1-dependent form of autophagy such as mitochondrial autophagy.6 In fact, TB1 given on days 7 to 21 of TAC promoted mitochondrial degradation by stimulating mitochondrial autophagy. TB1 also increased general autophagy, but its effect on general autophagy was not as prominent as on mitochondrial autophagy. Although it is unclear why TB1's effect on general autophagy in the presence of TAC was less prominent, it is possible that other mediators of general autophagy are actively suppressed in the presence of TAC. Alternatively, endogenous Beclin 1 released from the Golgi apparatus may be used more effectively for mitochondrial autophagy. Because downregulation of autophagy and consequent mitochondrial dysfunction underlie the pathogenesis of HF, autophagy-inducing agents with proven effects on mitochondrial quality control mechanisms, such as TB1, may have potential for the prevention of HF. It will be interesting to test whether other interventions known to stimulate autophagy equally stimulate mitochondrial autophagy.
It should be noted that TB1 also stimulates autophagy in cardiac fibroblasts. However, the functional significance of fibroblasts in mediating the salutary effect of TB1 remains to be elucidated.
Downregulation of endogenous Drp1 inhibited increases in the number of CMs with fragmented mitochondria 7 days after TAC, suggesting that Drp1 regulates mitochondrial fission during PO. It has been shown recently that cardiac and mitochondrial dysfunction caused by downregulation of Miff can be rescued by concomitant downregulation of Mfn1,27 suggesting that balancing mitochondrial fusion and fission is critical for the maintenance of mitochondrial function in the heart. Aside from the effect on mitochondrial autophagy, whether or not the lack of appropriate fission in Drp1-hetCKO mice also contributes to the enhancement of HF in these mice during PO remains to be elucidated.
In summary, our results suggest that mitochondrial autophagy is chronically suppressed following transient activation 3 to 5 days after TAC, and that suppression of mitochondrial autophagy plays an important role in mediating mitochondrial dysfunction and the development of HF in response to PO.
We thank Daniela Zablocki and Christopher D. Brady (Rutgers New Jersey Medical School) for critical reading of the manuscript. We also thank Dr Toshiaki Otsuka (Department of Hygiene and Public Health, Nippon Medical School, Tokyo) for statistical advice.
Sources of Funding
This work was supported in part by US Public Health Service grants HL67724, HL91469, HL102738, HL112330, and AG23039 and by the Leducq Foundation Transatlantic Network of Excellence (to Drs Levine and Sadoshima). Dr Shirakabe was supported by a Postdoctoral Fellowship from the Uehara Memorial Foundation.
The online-only Data Supplement is available with this article at http://circ.ahajournals.org/lookup/suppl/doi:10.1161/CIRCULATIONAHA.115.020502/-/DC1.
- Received August 16, 2015.
- Accepted January 29, 2016.
- © 2016 American Heart Association, Inc.
- 1.↵
- Gatica D,
- Chiong M,
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- Klionsky DJ
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- Nakai A,
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Mitochondria are intracellular organelles involved in metabolism and ATP production that are vitally important in cardiomyocytes. Heart failure is commonly accompanied by mitochondrial dysfunction. Dysfunctional mitochondria are the major source of reactive oxygen species, and trigger both the apoptosis and necrosis of cardiomyocytes. Mitochondrial damage spreads rapidly through depolarization of the mitochondrial membrane potential and causes deterioration of the function of healthy mitochondria. Thus, cardiomyocytes possess robust quality control mechanisms, and autophagy, which degrades protein and organelles through lysosomes, is a major mechanism for the elimination of dysfunctional mitochondria. In this study, we show that both general autophagy and a mitochondria-specific form of autophagy (mitochondrial autophagy) are downregulated after pressure overload and that mitochondrial dysfunction develops only after mitochondrial autophagy is downregulated. Mitochondrial autophagy was induced in response to pressure overload through a mechanism dependent on Drp1, a protein known to mediate mitochondrial fission. Treatment of animals with Tat-Beclin 1 peptide, known to stimulate autophagy through mobilization of endogenous Beclin 1, a protein essential for autophagy, increased the level of mitochondrial autophagy in the heart and partially rescued the animals from the development of pressure overload–induced heart failure. These results suggest that interventions to restore the level of mitochondrial autophagy should be considered for potential treatment for patients with heart failure.
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- Drp1-Dependent Mitochondrial Autophagy Plays a Protective Role Against Pressure Overload–Induced Mitochondrial Dysfunction and Heart FailureCLINICAL PERSPECTIVEAkihiro Shirakabe, Peiyong Zhai, Yoshiyuki Ikeda, Toshiro Saito, Yasuhiro Maejima, Chiao-Po Hsu, Masatoshi Nomura, Kensuke Egashira, Beth Levine and Junichi SadoshimaCirculation. 2016;133:1249-1263, originally published February 25, 2016https://doi.org/10.1161/CIRCULATIONAHA.115.020502
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- Drp1-Dependent Mitochondrial Autophagy Plays a Protective Role Against Pressure Overload–Induced Mitochondrial Dysfunction and Heart FailureCLINICAL PERSPECTIVEAkihiro Shirakabe, Peiyong Zhai, Yoshiyuki Ikeda, Toshiro Saito, Yasuhiro Maejima, Chiao-Po Hsu, Masatoshi Nomura, Kensuke Egashira, Beth Levine and Junichi SadoshimaCirculation. 2016;133:1249-1263, originally published February 25, 2016https://doi.org/10.1161/CIRCULATIONAHA.115.020502


text:
Here is the club's announcement about their upcoming luncheon:
We are pleased to announce our next event "MetroDolls...Celebrating 10 Years" will be held on October 4, 2015 in Iselin, New Jersey.
Robert Tonner is creating a beautiful 16" fashion doll souvenir for the event and we will be holding an OOAK auction to benefit St Jude Children's Research Hospital.
Hope to see you there!

subset: commoncrawl


text:
Foreigners have been collecting the bulk of royalties paid over by local radio stations for playing music.
This is according to Jamaica Association of Composers, Authors and Publishers Limited (JACAP).
The entity's general manager Lydia Rose said JACAP collected $94.6 million in royalties in 2015. She said that after deducting a $32 million administrative and other costs, a total of $39 million remained for distribution among rights holders.
Of this sum, a mere $7 million was paid to JACAP members while $32 million was paid to overseas societies. Read More via InfoSysTV.


text:
Ceci Cela Gingerbread Man
• Filmmaker Mark Fisher •
New York Promo
A Short Filmed At Eight Turn Crepe In New York
Filmed By Photographer Mark Fisher With Director Mark Fisher,
Filmmaker Mark Fisher. Editor Mark Fisher And Producer Brighton Hughes.
A Mark Fisher Creative Project.
Released By First Stamp Multimedia
Just Be Creative™
No Second Usage Without Permission
All Rights Reserved • Models,Actors, Actresses,Dancers,
Musicians Used In The Web Posts Are Professionals
and Require Fee For Any Publication or Usage.
All Have Management.
Removal Of The Image Or Blog May Violate U.S. Laws.
Photographer Mark Fisher™ Is
A Well Accomplished Published Photographer
In Beauty, Fashion, and Music Photography.
New York City
Based Image And Filmmaker
Has A Worldwide Following.
Is A Member Of The Press.
There Is Fan Page On Facebook.
Certified And Licensed Image Maker
Commissions Accepted Through Contract
• New York • • Paris • • Milan • • London •
Private Client Request Accepted.
Website www.americanphotographernyc1.com
For More Info Contact
Green Key Management
New York, New York
Mark Fisher Is A Member Of The Press.
These Are Official Press Images Of Mark Fisher
Any Attempt To Spam Or Remove Will Be Investigated
As A Violation Of The First Amendment and Freedom Of The Press.
Thank You


text:
Listed at IMDb.com.
|Michael Broderick
|Narrator / Adult Ankar
|Canaan Tai Estes
|Son / Ankar
|Yasmine Marquez
|David Scott Heck
|David L.J. George
|Uncle / Terry
|Shondra Marie
|Joe Grisaffi
|Joshua Michael Gennari II
|E.C. McMullen Jr.
|Writer / Producer / Director
|Luz McMullen
|Still Photographer / Set Designer / Producer /Executive Producer
|Joshua "Déous" Gennari
|AD / Director of Photography
|Kelly Parks
|Associate Producer
|Sean Fernald
|Music Supervisor
|Michael Barela
|2nd AD, Grip
|Christopher M Ross
|Sound Engineer, Grip
|Miguel Aguirre
|Boom! Operator
Movie & Sound Equipment by
Area 51 Productions Shot on Panasonic LUMIX GH2K, OSG Hack Pack Modified
Audio recorded on H4N ZOOM
Additional Movie, Sound, & Still Photography equipment by
Foleco, llc. Shot on Canon 60D. Audio-Technica ATR 6550 microphone
Editing equipment by
Archaic Media. Adobe CS6, Asus computer
The Producers Sincerely Thank,
Trish Avery, SAG-AFTRA – Dallas,
Texas Alfred Cervantes, Houston Film Commission
Alicia Downard, Texas Film Commission
Jerry Ramos, Texas Child Labor Board
Pat Skiles, Aplus Acting – Houston, Texas
Molly Thompson, SAG-AFTRA – Los Angeles, California
We also wish to thank
Drew Beeson, Metro-National
Deepest Thank you and Appreciation to our Contributors
Leticia Ballinas
Randy Feeney
Darren Gold
Made in Cooperation with SAG-AFTRA, New Media
The Night My Monster Died, Copyright 2013, E.C. McMullen Jr.


text: A question that turned into a discussion at work tonight: What is the minimum lateral separation from a thunderstorm while in VFR? After several opinions, including my own, "As far way as I can frikken get...", we consulted a FAAST circular, which stated 5 miles, but 20 miles is recommended. Now for the REAL question, which wasn't clear, and caused much chatter on a SLOWWW evening: Is that in miles, or nautical miles?


text:
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Seiko SQ70 Digital Metronome
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- 473637 000000000 473637000000000
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The SQ70 has all of the features a musician needs . . . and then some. The tempo range is a wide 40 - 208 beats per minute. The large speaker featu...Click To Read More About This Product
6-Month* financing available.
The SQ70 has all of the features a musician needs . . . and then some. The tempo range is a wide 40 - 208 beats per minute. The large speaker features volume control and the lights make keeping time that much easier.
Musician's Friend Gold Coverage
Gold Coverage goes above and beyond the manufacturer's warranty to protect your gear from unexpected breakdowns, accidental damage from handling and failures. This plan covers your product for one, two, three or up to five years from your date of purchase, costs just pennies per day and gives you a complete "no-worry" solution for protecting your investment.
Gold Coverage Includes:
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Other plan features include:
- For products over $200, No Lemon Policy applies. If it fails for the same defect 3 times, it is replaced on the 4th breakdown. (Does not apply to failures due to drops, spills, and cracks)
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*Limitations and exclusions apply. See terms and conditions (Opens in new window) for program details
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- Easy to use (3)
Best Uses
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Reviewed by 4 customers
- Easy To Use
- Facilitates practicing!
- Functional
- Good Quality
- Improves Sound
Best Uses
- Practicing
Comments about Seiko SQ70 Digital Metronome:
The Seiko SR 70 Digital Metronome is a wonderful for assisting me with practicing my piano selections and technique: Excellent sound, EASY to use, wonderful Power pack to use, superior quality: (1) Controls are ideally suitable; (2) Sound Quality is excellent (Volume, tone quality); (3) quick selections of time signatures (i.e., 2/4, 3/4, 4/4, 6/8, other); utility - GETS THE JOB DONE!
Comments about Seiko SQ70 Digital Metronome:
MF emailed me a % off code, so I went shopping for a new metronome. Been using a digital recreation on my PCs and wasn't crazy about it or having to fire up a computer just to run that one program. Even with the laptop, it was just a pain.
Found this, liked the specs. Seemed like it would work well for a guitar player. Saw the one negative review but went elsewhere to find more reviews. Most of them were extremely positive, those that were not tended to reflect issues not important to guitarists.
Ordered it. Been using it for about a month now, every other day or so. Love it. It is super easy to use. It keeps perfect time. It is loud enough that you don't have to use earphones during practice drills. The stand is well executed. Earphone jack. Battery or power supply, the former included the latter not included. It looks like what it is, a quality piece of equipment.
Perfect for may guitar practice needs and I'm betting your in home practice needs as well. One word of caution, check around, the pricing on this thing varies quite a bit.
- Easy To Use
- Functional
- Poor Quality
Best Uses
- Practicing
Comments about Seiko SQ70 Digital Metronome:
A very nice unit and the only reason I gave it 3 stars if for the battery door. Its a friction fit type that snaps into place and feels cheap. For what it cost it should be like on a stomp box where it has screws to get to the battery or a heavier door. Just waiting for it to break and than I am sure you can just buy the battery door.
- Easy To Use
- Good Quality
Best Uses
- Practicing
Comments about Seiko SQ70 Digital Metronome:
In my trumpet practice I always had a hard time hearing the "clicks" with the multitude of little "pocket metronomes" I have owned. The wimpy beep, beeps just got lost. With this SEIKO the CLICKS are louder and, more importantly, have a cutting quality like a woodblock. The unit is a little larger - it won't fit in your pocket, but it is durable. I have used this model in a school setting and it took a lot of abuse. It does not have a lot of bells and whistles like some programable metronomes. Get it for it's sound and basic durability.


text:
Yeah, dangerous thing I know. But I've been doing some studying on the web and in some references and I think there are some things that people aren't paying attention to when it comes to firearms. I'll try and stay both clear and on track (can't guarantee either.)
First off, the genie is out of the bottle for America. Guns are not only a large portion of our heritage, they are a massive portion of our society. Go see an action movie in America. Guns. Guns. Guns. Guns. Guns. And action movies where the hero doesn't use them still glamorize them. *&^%$ You Hollywood! You produce these movies, you actors get paid buttloads of money, you writers have to drop them into your stories, you viewers push them for more, higher kicks. Oh, and don't get me started on the friggin' production companies. So, if you do movies with guns, recently. Or do one in the future, and you stamp your feet about "gun violence", STFU. You are a hypocritical part of the system and your opinions have no value to the citizen with a quarter of a brain.
History wise, our forebears made guns part of their lives; the romanticized old West. The settlers fighting Injuns. The Revolutionary War. Don't fire till you see the whites of their eyes. Sure lots of people didn't have guns. But those that did were part of a system that emphasized the responsibility of people to take of each other. Not how to deal with snowflake's hurt wittle fewwings. When people worried about their neighbors more than their own feelings, suddenly the neighbors were more interested in them! Now we close ourselves off, disconnect from the world and people around us, claiming no responsibility to our neighbors. You didn't do what was right because you had to (and in some cases you had to) but because it was what was best for all involved.
Between history and culture, guns are part of our lives. Maybe in a hundred years you can talk about banning them, but with my generation still here, it just ain't happening.
Do you realize how easy it is to make a gun? About 10 simple parts that anyone with any minimum of tools can make, and you've got a shotgun or a single shot pistol. Add about 15 more parts and you have a semi-automatic. Take away one or two, you've got a fully automatic gun. From any shop. From any shed. If you say 'They can make all the guns they want, if they can't buy ammo, they aren't worth anything", you are sort of right. How's that 'War on Drugs' coming? Other countries will make it and sell it at elevated costs. "Great, less people will get it then." Nope. People will just break more laws, hurt their neighbor to get the money for the ammo they used to be able to afford. Besides, gun powder isn't the only fuel that can project lead at high speeds. Ever hear of propane? Butane? Gasoline? Hydrogen? Rail guns? Take away the bullets and watch people carry around flamethrowers. Crossbows. Explosives. Thermite is so very easy and inexpensive to make, and burns with over 3000 degrees F.
At least as it is you can control what is out there. By banning semi-auto rifles you are just begging for FULL AUTO rifles. Did you know that you can put a longer barrel and a buttstock, along with a slight adjustment in parts on a legal semi-automatic pistol and you have semi-auto rifle? And if you are going to put me in jail for the same amount of time as a full auto, why would I stop at semi-auto? Might as well go out with a bang. Take away too much from people, they'll decide they don't just want a tomato, they want the full garden. Look at lever action rifles. They generally don't have removable magazines, yet can carry 10 bullets lots of times. Or adjust the gun a bit and you've got removable magazines. I've tried quick firing with a lever action. I can fire 10 bullets in about 7 seconds. Add an extended tube and we can move some from 10 to 20 rounds. Or how about we change the design of the lever action to allow quick replacement of an empty tube (weird magazine) – lever action still gets you 80-90 shots per minute. Include tube changes, maybe 5o shots per minute. About half of present semi-automatic's speed. But still a lot of bullets.
And this bull%$#& about "you need to give up your toys for the good of others." Sure. When you donate all of your income to the poor. Include any and all future income. Oh, and get the hell off our backs and get your own pensions, not from us! Think about it Senators and Reps. Your income could save 4, 5 even six families living below poverty. You don't need that money, other people need it more. Let me see the checks.
Didn't think so.


text:
Thinking About Dedicating Yourself to the Service of Your Patron God?
This is not a step to be taken in haste.
It may be, after having worked with and studied a particular Deity for a length of time, you may decide you want to dedicate
yourself to the service of your patron God or Goddess.
This is not a decision to be made in haste. It is also not something that one must do. While some Deities may ask for a dedication from you, it is always your decision.
During my 20 plus years as a Pagan, I have worked with many Gods and been privileged to call several a Patron God or Goddess. I have never been led to take the step in dedicating myself to the service of any One God or Goddess though.
However, if you have thought long and hard, and know this is the right thing for you to do, then this is a very simple Dedication Ritual which you can use. Feel free to make changes and embellish it to suit your own purpose.
Step 1
Several days, or weeks, in advance
Sit quietly and meditate on the following questions. Write your answers as they come to you in your personal journal or Book of Shadows.
- What gifts can I bring to (your chosen God/dess)?
- Why do I want to dedicate myself to (your chosen God/dess)?
- What can I pledge to (God/dess)?
Create your own Book of Shadows, if you have not done so already. Within it, record spells, rituals, prayers, and any other relevant magical material. This book will support you, serving as a platform for exploring your thoughts and feelings over time.
If during your meditation and seeking why you wish you dedicate yourself, your answers include desires to impress your friends, scare your neighbours, freak out your parents, or other words to this effect then you are not ready to dedicate yourself to anything. Continue your studies and learning, but leave the dedications for another time.
If you are doing it as a joke, you are not ready. You may take it lightly, but the Gods tend to take these things very serious.
Step 2
On the Day
Take a special bath, meditate, abstain from eating specific foods, or do whatever you are drawn to do to prepare yourself for your dedication.
Gather together the following representations of the elements:
- Salt
- If all you have is store bought table salt, then by all means use it. If you can get a larger crystal sea salt such as for salt grinders, then use that.
- Incense
- Chose an incense which appeals to you, or one associated with your chosen Deity.
- Holy water
- Candle
This is a simple blessing to create holy water:
Hold your hand over the water and state,
"I cast out from you, all hatred, all fear, all anger
In the Name of (your chosen Deity) So Mote (Must) it Be."
Pause briefly, then holding your hand over the water again, state,
"I fill you with love, with hope, and care and understanding
In the Name of (your chosen Deity) So Mote (Must) it Be."
If the words used here are not appropriate, substitute your own.
Chose a candle type and colour which best represents the Deity you are dedicating yourself to. Inscribe it with the Name of the Deity, or anoint with oils to further forge a link between that Deity and yourself.
Step 3
The Dedication Ritual
Create a sacred space by whatever means feels most comfortable
Sitting within the space, say as follows:
"It is my desire, of my own free will, to dedicate myself to (your chosen God/dess) this night.
I bring with me the gifts of … and speak aloud of the gifts can bring as you have meditated upon them previously.
I put these gifts now to the service of (God/dess)."
Take up the salt and sprinkle a few grains on yourself.
"This is the blessing of Earth, may it strengthen me as I dedicate myself to the service of (chosen God/dess). Through the element of Earth I will walk in balance. "
Take up the incense and left the smoke drift around you.
"This is the blessing of Air, may it inspire me as I dedicate myself to the service of (chosen God/dess). Through the element of Air I go forth seeking knowledge and the wisdom to use that knowledge wisely and with love for all things."
Take up the candle and hold the flame in front of you.
"This is the blessing of Fire, may it purify me as I dedicate myself to the service of (chosen God/dess). Through the element of Fire, I forge new things, giving expression to creative energies."
Take up the water and sprinkle a few drops on yourself.
"This is the blessing of Water, may it cleanse and renew me as I dedicate myself to the service of (chosen God/dess). Through the element of Water I live in love, trusting my emotions and intuition."
Sit quietly for a moment,
I, (your name), do dedicate myself to the service of (chosen God/dess), by my own free will. This is my pledge towards that service…" speak aloud of the pledge you are willing to make as you have meditated upon it
Sit quietly and reflect for a few minutes, then finish in whatever manner seems right to you.
Some may be led to take on a new name at this time, of you wish to do so, add this to the dedication ritual.
"I have received the blessings of the elements. I have pledged myself to the service of (chosen God/dess) and S/He has received me as Her/His own. In this sacred space and time, I am changed. (Your birth name) has been transformed. I ask to be given a new name to reflect that change."
A name may come to you immediately, or it may appear to you gradually over time. Let yourself be open to receiving a new name, from where ever it may come.
If you have any questions about Patron Deities, or dedicating yourself to a Patron Deity, feel free to comment here, or send me a message. I do try to answer questions within a few days.


text:
Wednesday, November 30, 2011
Songs of the year.
January: Runs in the family- Amanda Palmer
Febuary: Untitled- Social Distortion
March: 100 Times Fuckeder- NoFX
April: One Way Ticket To Fuckneckville- NoFX
May:Rise- Eddie Vedder
June: Heart Attack/Handle This- Sum 41
July: Extraordinary- Suicide Machines
August: Where Is My Mind?- the Pixies (I also re-read Horribleville, so that explains a lot)
September: White People for PEace- Against Me!
October: She's Like A Rainbow- The Rolling Stones
November:Rain Wizard- Black Stone Cherry
December:Seeing Double At The Triple Rock- NoFX
Awww what? No Dubstep? Maybe thats because dubstep lacks any literary substance. Go read a book, kids.
Saturday, November 26, 2011
It´s hard enough to even fake a smile, Disillusion? - I´m in denial!
"Happy" weekend to you all (my weekend pretty much rocked, save for right now)
I'm unreasonably pissed off at things that don't even register as problems, and you guys get to deal with it. But first: Heres a long-awaited shout to my two friends Kit Tizzy Bombtastic and Nodjkqr, whomofwhich both have blogs, but for some reason these blogs arent as popular as mine, even though they are way better written!
But anyway, on with this shit.
Yeah, I dont really know what to call this feeling properly, but 'disgruntled' fits too well. Im angry at my facebook feed, Im angry at school, im angry at my friends, im angry at minecraft, skyrim, my brother, my dog, and the fucking pizza place. but I have no reason to be. My facebook feed can be closed, the only person that can make school less shitty is me, my friends did nothing wrong, minecraft is really fun, even moreso when you run it, skyrim can be left alone, my brother has his own life to live, my dog will bark, and its not my fault that the people who work at the pizza place cant drive yet. So what the fuck?
Why am I so unsettled?
I need to get into an old fashioned fistfight with someone, methinks...
Thursday, November 24, 2011
This is just silly.
...But it makes more sense now. Somehow.
Saturday, November 19, 2011
WHAT I THINK OF #Occupy PROTESTS.
Happy Saturday, everyone. Wow. I crawl from the shadowy depths of my well-lit basement for one second, and the entire world is in uproar about how broke they are, and now they're being all homeless on purpose, occupying the streets, bitching about this "1%". Me, having little to no knowledge about whats going on, am deciding to share my opinions about it because I have a small audience and an internet connection. Here I go. (BE WARNED: THIS ENTIRE POST WILL BE THE PRODUCT OF NOTHING BUT MY OWN IGNORANCE. I HAVE DONE ABSOLUTELY NO RESEARCH. DONT QUOTE ME OR USE ME AS A SOURCE FOR ANYTHING. YOU WILL FAIL WHATEVER PROJECT YOUD BE USING ME FOR AND YOUR TEACHER WOULD HATE US BOTH IF THEY DONT HATE ME ALREADY.)
So, here's what I see happening. There's a metric fucktonne of people. Everywhere. And they all live in countries. These countries have governments, and these governments need lots of money from the people that live in these countries. And then something happened in like, 2007, and the entire world suddenly didn't have money. Greece flipped the fuck out, and now they're in a complete state of anarchy, but everyone's content with just stowing that under the rug for now. And then in 2009-2010, North America had some money, but that didn't last long for some reason, and now we have another worldwide economic fuckpile. There were some vicious riots in London, but they're always mad about something or other, be it that university is too expensive, or FUCK THE ECONOMY,(they're the birthplace of punk rock, so what do we expect) or something but I digress... Anyway, so one thing led to another, and now some schmucks in America found out that there's 1% of the American population that doesn't pay taxes because they're super crazy rich or something. That made America very mad. And the rest of the world apparantly, So a bunch of people went to wall street, and started peacefully protesting. The rest of the world slowly followed suit, occupying their own streets, in cities worldwide, and now the European Union (Europe's trading bloc, thank-you social 10-1) is deteriorating because Europe's run out of money AND THEN CALGARY DID IT TO. WHY? WE HAVEN'T HAD AN ORIGINAL IDEA SINCE CHALLENGER FUCKING BLEW UP THATS WHY. So I don't know why the entire worlds protesting, but I like to think they're trying to send the fat cats in wall street a message that theyre fucking dump and should disperse taxes among all 100% of the population, so that the US can have more money, resume trading, and everyone sle can have money, too. But I don't know. That's wishful thinking. Also, memebase has tried to talk about it a lot. Writing things in sharpie about how theyre the 1% of something or other. Heres what I have to say (Aside from all that hogshit up there^):
The protests are going to do no good if you're protesting by any means that follow the rules. The reason there are laws about protesting are to keep it as ineffective as possible. If you're not willing to die for a cause, you're not willing to fight for it at all.
This message I taped to my door is a response to how fucking stupid I think the Occupy protests are and the 0% shows my abhorring indifference to the entire concept.
My dad saw it and took a picture to show people at work, and he thought it was me saying that I wasn't gonna clean my room. I didn't even think my room was messy. the people he showed did at least side with me, somehow. Oh well, I cleaned my room up a little anyway because that wasn't the point.
Sunday, November 13, 2011
Saturday, November 12, 2011
VIC LEWIS 2012! ^o^
Wha-ho! Hey everyone, It's been awhile! There have been many reasons for this, but I don't wanna talk about it right now, these last few weeks were too interesting. What I wanna talk about most though is last weekend, which I spent in Canmore at (yup, you guessed it) THE VIC LEWIS FESTIVAL! I already made a post about this, though I didn't explain much! Well it is a yearly thing, happens every November, and its a thing where a few bands from Calgary get together and... Ignore each-other! Yeah! Anyway, this was a weekend, so lemme break it down for ya like this:
Friday was a normal school day for us (considering fridays end at like noon this isnt that big a deal) so this shit just flew by. I had a math quiz that I bombed because seriously if I gave less of a fuck id be taking one.
So anyway, school overed, but apparently the roads were really bad on the way to Canmore so we almost didn't go, but HEY WE DID ANYWAY because FUCK YOU, NATURE. So we all packed our shit up, drove on a big ass bus to Canmore, unpacked, and not a lot happened that night, except for a big ol game of apples to apples where we actually managed to run out of green cards somehow. Yup. (Helen Keller and Adolf Hitler remain the greatest cards, By the Way)
the night was long and kind of cumbersome, the people in my room were kind of dicks, but more on that later...
The day started way too early, and with nothing to sustain me but the shitty hotel coffee that SOMEONE drank ALL OF the night before because APPARANTLY IT HELPS THEM SLEEP, it was kind of a stressful start. Oh yeah, and we were up and out about fourty-five minutes early. Way the fuck to go, everyone -slowclap-
anyway, some other random bullshit happened, we ended up where we were supposed to go, we played our music, and quite beautifully I may add, our adjudicator loved us, ad awesominum.
Other than that, it was a pretty relaxed day of dickin' around an empty high school with nothing to do but watch happy people shove their happiness in my face. Me, being quite fed up with the circumstances, totally ditched and went and got some poutine: Proof that there is indeed some kind of god, and they're lookin' out for me. It was greasy concession stand poutine, too. Salty, warm, and filthier than what you'd find under Louise's bed. JUST THE WAY I LIKE IT.
Anyway, on with moar Saturday talk. There was a coalition for a tim's run, then it was back to the hotel for a few quick games of Prez, and then back to the school fer the best thing ever. T3H DANCE! LOLZOMGWTFBBQTROMPZORZAGARFZZZZFFGHASDFKLAGKLRASDFZZZSHSHZSHZSBZSHZSHHHHH-*dies*- but k seriously ive never had more fun at a dance than that one. THERE WERE GLOWSTICKS, AND DEADMAU5, AND AND AND THERE WAS KIDS DOING THE MACERENA TO DUBSTEP AND AND AND IT WAS FUCKING RAD! :D Seriously, I had a fucking blast. AND THEN IT GOT RUINED. Remember how I said the people in my room were kind of dicks? Yeah I was speaking in hyperbole. They were superdicks, not even kidding. Fuck, by the end of the night I felt as helpless and unwanted as Piggy from Lord Of The Flies...
You know in Alien, when that guy had the chest burster alien in his stomach? Thats how I felt that morning. And the bacon smell didnt help. THE BACON SMELL. DIDNT FUCKING HELP.
This feeling didnt really go away for quite some time. When I was on the bus I felt like shit. When I was shambling up the steps in the school I felt like shit. When I was trying not to throw up while listening to Aberhart concert Band's set I felt like shit. Eventually I just gave up, pulled my jacket over my head and passed out for a little while. I woke up every so often, cautiously checking my phone to see when my clinic was. When it came about time I sprung to my feet and flailed down the steps through the hallway, and apparated a pair of drumsticks because I fucking could. I didnt question this burst of energy, but accepted it as retribution for the shitty morning I had. It wasnt until after lunch when I bounced back fully. But the more I obtained in vitality I lost in emotional state. I continued to get sadder and sadder, and more and more jealopus of those around me. AND THEN FLETCHER CMAE AND MADE EVERYTHING BETTER BCAUSE THAT KIDS A FUCKING CHAMP. But then I got really lonely and sad again. And then we went home, I bitched about everything, and I went to sleep, not to return to school until after lunch, because band was cancelled, and I had spare second period.
Look who the cat dragged back from the dead.
Me? Is it me? Yeah, its me. HAve I gotten over my depression? Eh, it comes and goes. But who cares?
Wanna see some new posts?
Dont answer that, I KNOW YOU DO!
Expect one tomorrow?


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Baby Anna First Thanksgiving
It was the first Thanksgiving of her daughter Anna, the beautiful and cute girl. Baby Anna was so happy. Time for the ceremony have also come, help them by dressing them both with some special clothes from their closet and after you can enjoy a great image with a mom and her daughter together and well dressed and glittering for the one of the most greatest celebration from the year, Thanksgiving Day.
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Rental Woes in Valley City
Affordable rental housing has become a tenuous issue in Valley City. The loss of dozens of apartments in the name of permanent flood protection coupled with a growing population has left potential renters struggling to find good, affordable housing, especially for families.
Over the course of the next several days the Times-Record will explore Valley City rental issues and what renters can expect for their money alpong with what struggles landlords have finding good tenants. Read more in upcoming issues of the Valley City Times-Record.


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This Halloween, parents have a new and improved tool to help their children stay safe. The Trick or Tracker App was created to ease the minds of parents while traveling in the dark of night for trick-or-treating. Compatible with many smart phones on the market, the GPS enabled app works as a virtual compass between parents and children.
While parents can download the app regardless of what smart phone they own, their children must have an Android OS phone in order to be tracked. The app is easy to navigate for both parent and child, where simple buttons like "Locate my parent" or "Where's my child?" keep each person informed of their surroundings. Additionally, the app enables parents to establish a geo-fence that their children are allowed to travel within- if they wander outside of that zone the GPS will sense their movement and send an alert to parents.
For parents, this tech savvy app is a savior to their worrisome thoughts on All Hallows Eve. From a notable spike in fall and winter crimes to managing your disobedient children, this app certainly utilizes every scare tactic in the book to reel parents in.
It can be argued, however, that this app also leaves parents solely reliant on technology to look after their children. What happened to the old fashioned days when parents would accompany their kids while trick-or-treating? Or maybe, just allowing them to trick-or-treat alone with a well-communicated curfew? Although technology and social media have changed our lives for the better in many instances, this app leans towards minimizing the effort in parenting, which can be argued to have negative or positive effects. What are your thoughts on the matter?


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And after a month in Jogja, it rained for the first time two days ago at 2am. I didnt realise it until I saw most of my friends updating their status about jogja's current weather. Here isnt as hot as at medan but still, when there's no rain after a month you can feel something weird. Luckily it dont rain when Im at campus or outside my boarding house.
Posting another selca = im bored.


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Saturday, August 13, 2011
Directed by John Carpenter
Kurt Russell - R.J. MacReady
Wilford Brimley - Dr. Blair
Keith David - Childs
Scientists in the Antarctic are confronted by a shape-shifting alien that assumes the appearance of the people that it kills.
One of the fun things about doing reviews is I get to go back to older movies I haven't watched in awhile. Then there's movies I go back and watch every year, just because they're personal favorites. The Thing was one of those movies my dad rented for me to watch on a spring break one year. Well, the movie did a good job completely freaking me out. Now this was when I was about 12 or 13 years old, but happily that night after watching it, I didn't have nightmares (that was saved for when I saw Alien and Aliens a couple years later). Anyway, I really didn't connect this movie with being a Mythos type till later on, yet here we are at the movies almost 30th anniversary and a "prequel" coming out later this year so this is a good time to go back and re-watch it as well as review it.
I'm going to start off with the acting in this film. As all the characters aren't giving a back story where they came from, it was up to the actors to make the audience care for them through their actions and personalities and each one pulled it off brilliantly. What's even more striking is that everyone isn't talking twenty four seven. When something is odd or they can't explain it, the characters just keep quiet instead. This adds a lot more to the characters than one might imagine as you don't know what they are thinking. Kurt Russell, who plays R.J. MacReady, to me gives the best performance of his career in this film. Nothing he does is hammed up, he speaks more softly, yet with authority, plus he can't stand computer chess. Kieth David, who portrays Childs is very effective as the pissed off black guy. Yes, I know that's basically type casting him, but that is exactly what David is playing and does surprising well.
One of the main reasons this movie works so well while others built around the same premise is because of the setting of the film. The isolation that is present in the film is palatable to the audience as there really is nowhere to run. All the scientific team can do is stay in one place and rely on each other to help them stay sane in their environment. So what happens when an outside presence is introduced that can take the shape of the living around it is really no surprise as they're trapped. As the paranoia sets in as the team finds out that there is a alien presence among them that is killing and taking on that said dead form. No one is what they seem and that helps feed the unease among the characters as well as drive some to madness. Even in scenes you know will go bad, the shock of watching it happen doesn't really fade over time, no matter how many times this movie is watched. Part of the reason for the tension that is built up during these scenes is the score by Ennio Morricone. I say this because with most horror movies when something bad is about to happen, the music swells to a forceful level which basically lets the audience know something is about to happen, and in return kills part of the surprise. Morricone though went the opposite way and killed the music so where all that you here is the background noises and characters talking. This might not seem that big of a thing, yet in reality it just adds to the tension of the screen.
Another aspect of the film is the special effects. While CG creatures is what is mostly shown on movie screens now, nothing will ever replace practical in camera effects. The reaction of the cast is real when they can actually see and interact with something in front of them. I'm not saying CG effects are terrible, in fact I love what is possible with CG now, but it can never replace practical in my mind. The closest I can put the creature in the film is a shoggoth from Mythos stories made real as it is always taking on a new form. Thank you Rob Bottin for this nightmare creature. But I would be remiss to not mention Carpenter himself who brought all these elements together and put it on screen for all to see, as this is his movie and unlike other movies during that time it has withstood the test of time to become a classic not in just horror, but also in films in general that still has the power to scare, shock,and gross out viewers after all these years. Besides, I love the nihilistic ending where you don't know if Childs is really himself or the Thing.
Dr. Cooper's forearm mini-meal.
You gotta be fucking kidding.
Kurt Russell was almost injured in the scene where he blows up the alien Palmer with a stick of dynamite. Apparently, he had no idea exactly how big of an explosion it would produce, and the reaction that he has in the movie is genuine.
Both Nick Nolte and Jeff Bridges turned down the role of MacReady.
This is the first film in what Carpenter calls his "Apocalypse Trilogy". In order they are:
The Thing (1982)
Prince Of Darkness (1987)
In The Mouth Of Madness (1994)
For a scene where Dr. Copper arms are severed, a real-life double amputee stand-in was used wearing a mask in the likeness of Dysart. The audience focuses on the bloody stumps while the mask goes unnoticed.
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nowell family from Dewsbury in Yorkshire
George Nowell married Frances Rhodes 17th Mar 1794,John Nowell chris.3rd June 1805,John Nowell married Rachel Bullows 11th Dec 1825,Jane Nowell married George Ovens May 21st 1857 in Birmingham. does anyone recognise this if so I would love to talk to you.
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Ray Turner 12/06/02


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Haskin's Road to the Kentucky Derby (Cont.)
From Preceding Page
From Preceding Page
If you think this year's road to the Kentucky Derby has been a bizarre one, imagine how scrambled your brain would be right now if Bobby Frankel hadn't brought some semblance of normalcy to the proceedings.
Kentucky Oaks (gr. I) hopeful Lady Tak, winner of the Fair Grounds Oaks (gr. II) and runner-up in the Ashland, breezed five furlongs in 1:01 Monday for trainer Steve Asmussen.
Edmund Gann's Toyota Blue Grass Stakes winner Peace Rules arrived at Churchill Downs on Monday to prepare for the 129th running of the Kentucky Derby on Saturday, May 3.
Trainer Jeff Mullins sent Santa Anita Derby (gr. I) hero Buddy Gil for a half-mile work in :46 4/6, handily, over the Arcadia oval's muddy training track Monday morning.
Tom Durant's Most Feared tuned up for a run in Sunday's Coolmore Lexington at Keeneland with a sharp five-furlong work on Monday at Churchill Downs.
Hall of Fame trainer D. Wayne Lukas has started more horses in the Kentucky Derby than any trainer in history and has won the "Run for the Roses" four times, but he is down to his final candidate for Derby 129 in Robert and Beverly Lewis' Scrimshaw.
Les Benton, chairman of the Dubai World Cup and chief executive of the Emirates Racing Association, submitted his resignation last week and will leave for Australia in mid-June.
Lockhart Spears, who campaigned homebred grade II winner Miss Baja, died April 11 in a Paris, Ky., nursing home after a lengthy illness
Monday's final under tack show for Keeneland's April sale of 2-year-olds in training was marred by a fatal injury to a Hennessy colt.
The Ohio Racing Commission has conditionally approved the sale of Scioto Downs, a Columbus Standardbred track, to MTR Gaming, owner of Mountaineer Race Track & Gaming Resort in West Virginia.
Though racetrack slot machines still haven't been approved by the Pennsylvania legislature, the Bensalem Township Council is scheduled to vote April 14 on Philadelphia Park's plans for a building to house the machines.
A Prairie Meadows board member said that chairman Jim Rasmussen should temporarily step down until an investigation is completed regarding his possible involvement in an alleged illegal sports betting operation.
Directed Energy, a 15-year-old son of Fit to Fight out of the Buckpasser mare Turnabuck, has been moved to Blackwater Ranch near Fort Thomas, Ariz.
Grade I winner Tactical Cat was represented by his first winner April 12 when Fashion Girl won at 4 1/2 furlongs at Keeneland


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Seasonal Offers
Each Season Brigitte crafts a unique selection of delightful chocolate confections ... These are limited offers, hand made in limited quantities.
Be sure to sign up for our newsletter to received advanced notification for our seasonal offers.
Chocolate as a complement to enhance your Easter brunch or dinner table ... the unique host/hostess gift ... the perfect basket ...
Mother's Day
On May 9th share your love of chocolate with Cocoa Pod's finest confections ...
Let Cocoa Pod Chocolates help you with the perfect gift for Mom this year. Please be sure you get your orders in by April 24, 2010.
Dads and Grads
Congratulations to all the Graduates & Happy Father's Day to all the Dads!
Happy Thanksgiving!
Take your Thanksgiving Table of Desserts up a notch! Bring a memorable host/hostess gift
Enjoy an exquisite Chocolate Experience with this holiday show stopper!
Holiday Season
Chocolate is the perfect gift ... for Family ... Friends ... Employees ... Teachers ... YOU !
We're pleased to be offering our 2011 Valentine's Day Collection of fine chocolates. Everyone loves chocolate ... so what better way to show you care than with a box of handmade chocolate confections from Cocoa Pod
St. Patrick's Day
You don't have to be Irish to enjoy the exclusive Irish Chocolate Confections offered by Cocoa Pod Chocolates ... You and your friends will think you found the "Pot O' Chocolate-Gold" with these mouth watering treats! They also make a unique host/hostess and great way to welcome in Spring!


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The Opportunity to Live Without Knee Pain
VQ OrthoCare® Offers National Football League Players Association (NFLPA) Former Player Members the Opportunity to Live Without Knee Pain
Medical Device Company Offers a Special Discount Program on BioniCare, the Only Device that can Claim to Provide an Overall Improvement to an Osteoarthritic Knee
SAN DIEGO – April XX, 2011 – VQ OrthoCare®, creator of the award-winning BioniCare® Knee System, is pleased to announce its BioniCare discount program for National Football League Players Association (NFPLA) former player members and their families. VQ OrthoCare initiated this discount program based on the need for a non-invasive, non-pharmaceutical treatment option for retired professional athletes living with osteoarthritis of the knee. Professional athletes move their knees up to 20 million times per year. In contrast, the average person uses their knees only three million times per year. The constant knee movements wear down the knee joints, increasing the risk of osteoarthritis of the knee (Overtime Magazine, 2007). In conjunction with the BioniCare discount program, VQ OrthoCare is also offering a financial aid program for former player members needing assistance.
"More than 27 million Americans are living with osteoarthritis of the knee and National Football League players are three times more likely to develop osteoarthritis of the knee than the general population," said VQ OrthoCare president and founder, Jim Knape. "We are thrilled to be offering former player members of the NFLPA and their families the opportunity to utilize this life-changing treatment option that will enable them to live pain-free and regain their active lifestyles."
The BioniCare Knee System delivers a low-level pulsed electrical signal, developed specifically to treat osteoarthritis of the knee by mimicking the naturally occurring signal present in a healthy knee joint. BioniCare is used with an unloading brace or with a night wrap. More than 80,000 patients nationwide have been prescribed the BioniCare Knee System. In a study led by 2 Johns Hopkins Orthopedic surgeons, 62% of total knee replacement (TKR) candidates with clinical and radiographic evidence of severe osteoarthritis avoided a TKR for at least four-years after an average of only 11 months of BioniCare therapy. Only 7% of matched control patients treated at Johns Hopkins, who received therapy but did not use the BioniCare Knee System, avoided total knee replacement surgery in the four-year period. Significant improvement in pain, range of motion activities of daily living and overall improvement in the knee over and above that achieved by other available therapies have been reported in published studies.¹
For more information on VQ OrthoCare or the BioniCare Knee System, visit http://vqorthocare.com or call (800) 266-6969.
About VQ OrthoCare:
Founded in 1989 and based in Irvine, California, VQ OrthoCare® is a leading manufacturer and provider of non-invasive medical solutions focused on bone, joint and soft-tissue conditions. VQ OrthoCare's commitment to treating patients with respect and supporting them in achieving optimum outcomes is unmatched in the industry. Services include in-home patient fitting of braces and other medical devices, and technology-enabled compliance monitoring. VQ OrthoCare's products include: Surgi-Stim4, Ortho-Stim4, Eagle, Catalyst and OActive knee bracing, Fast Start EMS, Fast Start HVPC, VQ Vector, and the BioniCare system. VQ OrthoCare is registered with the U.S. Food and Drug Administration as a medical.
1 Mont MM, Hungerford DS, et al., "Pulsed Electrical Stimulation to Defer TKA in Patients with Knee Osteoarthritis," Orthopedics, October 2006 (10): 887-889.


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Oh my ! Summer seems to be flying by. I find myself reluctant to sit inside and type a blog entry when the weather is so fine and there is summer fun to be had.
We've been going to the beach, vacationing, and having a blast!
The girls and I made up a couple of cute summer dresses a couple weeks ago. It was their first sewing project. They were naturals! I think I may be raising a couple of budding quilters, yeaaaaah! They used my old starter machine first of all because it's simple and second of all my silly machine has been at Baby Lock headquarters being repaired all summer.
Here is how they look!
I finally finished the QUILT FROM HELL for my husbands old boss. I blogged about it in the past. I am just happy to be done with it and all those tiny black lines.
I must admit I was a bit sad to part with it after all the work.
It's a Frank Lloyd Wright quilt.
Here's the BG holding it up for me:
I hope you all are enjoying the summer as much as we are.
Later Gators!


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Undertakers and Bad Gas
I might have entered the ministry if certain clergymen I knew had not looked and acted so much like undertakers.
-- Oliver Wendell Holmes (legendary U. S. Supreme Court Justice)
A man approached a fellow passenger on an airplane and inquired, "Excuse me sir, but as I passed you just now I noticed that you look like a minister. Am I right?"
"No," came the reply, "I just have bad gas.


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As a parent, I don't think you can help projecting things onto your child's personality. And new evidence always upsets the image.
Breaking news: Madge hates gym class.
Or so she said this morning.
I'm working on it.
But see, you can't help doing it either, can you? Now, to you, she's the kind of girl who loves to read, wants to have long hair, loves all things "beautiful" (read: Barbie-esque), and hates gym.
Like I said, I'm working on it.
Dear Frustration,
It appears you and I have been set up on a blind date. What will you be wearing? I will have a crushed pansy in my hatband. I have a feeling we've met before, but I'm willing to give this relationship a fresh chance.
Cautiously yours,
Tuesday, September 12, 2006
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Whatever or wherever your shopping preference, you will have easy access to lots of great eateries. For the those who prefer to just sit and relax, they will find benches and chairs at the Five Oaks Outlet Mall, rocking chairs at The Island in PIgeon Forge, and benches throughout the Old Mill HIstoric District.
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