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Bungalow Bar hits heights with top atmosphere
FROM East to West, the pub-mobile headed for Connacht last week and it was the lovely town of Castlebar that we settled on.
We last visited this area in the early parts of 2012, but as the Old Commando rightly pointed out, there are so many pubs in Castlebar that we would have no problem visiting twice a year.
After a three-hour journey, we finally arrived and our designated driver had to wake some of our crew who had drifted off during the trip.
"Sorry about that," the Young Commando said as our designated driver parked the pub-mobile just off Rathbawn Street. "But there's only so many stories I can listen to from your man in the passenger seat," he moaned.
He was referring to the Old Commando, who had been nattering since the moment we left the Pale and was giving us all the background information on the town of Castlebar.
As we've said on numerous occasions before, Pub Spy and his team never get involved in political talk in pubs. We don't think it's the right place to debate politics and, ultimately, it will lead to an argument.
However, those rules do not apply to the pub-mobile and the Old Commando was only too willing to discuss Castlebar's political exports for the duration of the journey – exports that include two Taoisigh in Charles Haughey and Enda Kenny and a former EU Commissioner in Padraig Flynn. No wonder some of us drifted off.
With our wagon safely parked, we headed for Main Street and the first pub we came across that looked lively was the Bungalow Bar.
The bright red exterior stood out and it was in good condition. We arrived just in time for a live soccer match and the Young Commando was happy to sit at the bar and enjoy the game. We placed our order with a very engaging young barman and we really enjoyed his company.
The Young Commando quickly established that there was a big soccer feel to this bar, with framed Arsenal jerseys, scarves and flags hanging on the wall. Even the barman was togged out in an Arsenal jersey.
Some banter ensued, but it was all in a friendly nature. The pints of stout were very reasonably priced at just €4, while our designated driver paid just €1.50 for his mineral.
There are pints of Carling available for just €3.50, but we all stuck with the stout.
There are two sections, with the first containing a pool table just inside the front door, a large-screen TV and a roaring open fire where we huddled for a few moments to warm our toes.
The second section is located up a small set of steps and it too has a large-screen TV, a jukebox (which thankfully was turned off) and some gaming machines, although nobody was using them.
We noticed the calendar was still showing December 2015, and while we wish it was still Christmas, it's not, so it's time to confine it to the skip.
The bar was well stocked and it was clean too. The timber floor wasn't laid today nor yesterday; it's quite worn, but it was clean. There was an all-male clientele and while it wasn't overly busy, there was a good atmosphere, which was helped by the chatty and polite young barman.
The Old Commando ran an eye over the toilets and he reported back to us that they were in good condition.
A sign behind the bar advises that 'alcohol doesn't solve your problems, but neither does milk' and we can safely say we had no problems with this pub in Castlebar. The pints were cheap and cheerful and the barman was most welcoming. We'd definitely call again some day.


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June 30, 2010 BY John Kim
A release reminder in the middle of the week? Quite an oddity for Jordan Brand, but we can anticipate slower-than-usual sales this weekend due to the Independence Day holiday. Dropping tomorrow, July 1st, is the Air Jordan IX (9) Retro in the Black/Citrus colorway; the Citrus IX was one of the first colorways Sneaker News gave you a heads up on in our Air Jordan IX (9) Fall 2010 Preview from January, and it surely held its own next to the two OG colorways. While some spots have released the Citrus IX early, tomorrow will be the official released date of this great colorway. Check out the sickly sweet display from Extra Butter after the jump and remember you can still pick up a pair on eBay.
Filed under: Air Jordan Just Released

subset: commoncrawl


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How did your life change?
Most Helpful Girl
Haha OMG I was an ugleeeey pre teen!
Like I was chubby between the ages 13 and 15, and extra weight (although it wasn't drastic, I wasn't like obese) doesn't look good on me because I gain fat easily in my face and it makes me look reallyyyyyy bad (my face doesn't carry extra pounds really well)! I lost the weight at the end of 15-16 and looked so much better and started getting attention from guys in High School! I was treated better from both guys and girls and became more popular.20|20|0
And I had nerdy glasses, and was the teachers' pet... didn't help haha
Aww I like chubby faces? :( yea same I gain weight on my face fist.
Most Helpful Guy
I personally changed hugely from 16 to 20 and I'm still changing I suppose.
I was never really ugly but I didn't like the way I used to look.
Now, I get things at cafes or restaurants for free once in a while and even have the modeling industry interested in me.
I even had a girl "run away" from me because she must have thought I am a "jerk" or that I am "out of her league"
On the other hand, I have tons of "Likes" , "Super-Likes" and "Matches" on those online dating apps, but fewest even respond. Others believe I'm not real - they also don't believe me when I'm telling them what my career path is.
That 1 girl chickening out on me in real life explains why those girls online do not believe I'm real or that "I'd ever be genuinely interested in them"
I don't believe in leagues... and I still sometimes see myself as my 16-year-old-self.
So, I rather doubt myself in this hindsight...00|00|0


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Tire TPM Sensor; 07-11 Jeep Wrangler JK
This replacement tire pressure monitor sensor from Omix-ADA fits 07-11 Jeep Wrangler (JK). One needed per tire.
Pricing: $150.99
Direct OE replacement Jeep parts and accessories built to the original specifications by Omix-ADA. Limited five year Manufacturer's warranty.
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- Item Requires Shipping
- 0.1 lbs.
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janetd0987 Review
Very disappointing. I've ordered quite a few items from Overstock, and this is the first time that I have been disappointed. The quality is very poor, and the packaging was horrible. There was absolutely no padding or cushioning to protect the product, and the larger basket arrived with one of the woven pieces broken right in the front of the basket. I'm pretty handy, but there is no way to fix this, as the material is stretched so tight that it can't be repaired. The only reason why I didn't return the item was that I needed this to use at work, and didn't want to bother with the return. So I just used the smaller basket, which was okay. Also, the lids on these don't really sit right, they aren't aligned very well. Next time, I will take the time to go to Target and get a similar item. Very disappointing. My advice? Don't purchase this item.
- How did the color on site compare with the actual product? The actual color was the same as shown on the site.
- Please tell us about the quality of the product. Very poor quality.
- How accurate was the on site description of the product? The description was accurate.


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2004 P Keel Boat Jefferson Nickel PCGS MS67 !!!
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Pro Era- School High (Instrumental) (Loop) MP3
RIP Capital Steez. This instrumental is looped. Subscribe and check out my channel for more beats/instrumentals. Follow me on Twitter: ...
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Oxygen homeostasis represents an essential organizing principle of metazoan evolution and biology. Hypoxia-inducible factor 1 (HIF-1) is a master regulator of transcriptional responses to changes in O2 concentration. HIF-1 is a heterodimer of HIF-1α and HIF-1β subunits. O2-dependent degradation of the HIF-1α subunit is mediated by prolyl hydroxylase, von Hippel-Lindau protein (VHL)/Elongin-C E3 ubiquitin ligase, and the proteasome. O2-independent degradation of HIF-1α is regulated by the competition of RACK1 and HSP90 for binding to HIF-1α. RACK1 binding results in the recruitment of the Elongin-C E3 ubiquitin ligase, leading to VHL-independent ubiquitination and degradation of HIF-1α. In this report, we show that calcineurin inhibits the ubiquitination and proteasomal degradation of HIF-1α. Calcineurin is a serine/threonine phosphatase that is activated by calcium and calmodulin. The phosphatase activity of calcineurin is required for its regulation of HIF-1α. RACK1 binds to the catalytic domain of calcineurin and is required for HIF-1α degradation induced by the calcineurin inhibitor cyclosporine A. Elongin-C and HIF-1α each bind to RACK1 and dimerization of RACK1 is required to recruit Elongin-C to HIF-1α. Phosphorylation of RACK1 promotes its dimerization and dephosphorylation by calcineurin inhibits dimerization. Serine 146 within the dimerization domain is phosphorylated and mutation of serine 146 impairs RACK1 dimerization and HIF-1α degradation. These results indicate that intracellular calcium levels can regulate HIF-1α expression by modulating calcineurin activity and RACK1 dimerization.
All metazoan organisms possess physiological systems to regulate the delivery and/or utilization of O2. Hypoxia-inducible factor 1 (HIF-1)3 is a critical mediator of adaptive responses to reduced oxygen availability in many developmental, physiological, and pathological contexts through its transcriptional regulation of genes that encode proteins required for tissue oxygen delivery and energy metabolism (1–4). HIF-1 is required for embryonic vascularization. Mouse embryos with complete HIF-1α deficiency or expression of a dominant negative form of HIF-1 in endothelial cells fail to develop proper vascularization and die at embryonic days 10–11 (4–6). HIF-1-dependent induction of angiogenic factors promotes vascularization and cardiac function in the initial compensated phase of left ventricular hypertrophy caused by pressure overload (7). In contrast to this adaptive response, HIF-1 contributes to the pathogenesis of hypoxia-induced pulmonary hypertension and right ventricular hypertrophy (8). HIF-1 also plays an important pathogenic role in many critical aspects of cancer biology, including cell immortalization, energy metabolism, vascularization, invasion, metastasis, and resistance to radiation and chemotherapy (9).
HIF-1 is a heterodimeric transcription factor composed of a HIF-1β subunit, which is constitutively expressed, and a HIF-1α subunit, the expression and transcriptional activity of which are regulated by the cellular O2 concentration (10). Both subunits contain basic helix-loop-helix domains that mediate dimerization and DNA binding, as well as a second dimerization motif referred to as the PAS domain (10, 11). HIF-1α protein accumulates in response to hypoxia and is rapidly degraded upon reoxygenation (10, 12, 13). Under hypoxic conditions, HIF-1α dimerizes with HIF-1β and translocates into the nucleus. HIF-1 binds to its target DNA sequence within hypoxia response elements, recruits the coactivators p300 and CBP, and activates transcription (14, 15). An increasing number of HIF-1 target genes are known, including those encoding metabolic enzymes, cytokines, growth factors, receptors, and other signaling proteins (1–4).
The HIF-1α transactivation domain located at the carboxyl terminus is regulated by the binding of factor inhibiting HIF-1 (16). Under non-hypoxic conditions, factor inhibiting HIF-1 hydroxylates asparagine 803 of the 826-amino acid human HIF-1α using O2 and α-ketoglutarate. Asparagine hydroxylation blocks the binding of HIF-1α to coactivators p300 and CBP, thus disrupting its ability to activate target gene transcription (17). O2-dependent regulation of HIF-1α protein degradation is mediated by a 200-amino acid oxygen-dependent degradation domain that is located in the middle of the protein (18). HIF-1α prolyl hydroxylases (PHDs) also utilize O2 and α-ketoglutarate as substrates to hydroxylate proline residues 402 and/or 564 of human HIF-1α (19, 20). Osteosarcoma protein-9 binds to both PHDs and HIF-1α, leading to increased prolyl hydroxylation and proteasomal degradation (21). The von Hippel-Lindau tumor suppressor protein (VHL) is the substrate recognition subunit of an E3 ubiquitin ligase complex. Undernon-hypoxic conditions, VHL binds to both hydroxylated HIF-1α and to Elongin-C, which recruits Elongin-B and other subunits of the E3 ubiquitin ligase that targets HIF-1α for ubiquitination and subsequent degradation by the 26 S proteasome (22–25). Renal carcinoma cells lacking functional VHL have high HIF-1α expression even under non-hypoxic conditions (22).
HIF-1α protein stability is also regulated through an O2-independent pathway involving the interaction of HSP90 and RACK1 with the PAS domain (26–28). HSP90 and RACK1 compete for binding to the PAS-A subdomain of HIF-1α. When the interaction of HSP90 and HIF-1α is disrupted by the HSP90 inhibitor 17-allylaminogeldanamycin, RACK1 binds to HIF-1α and recruits an Elongin-C E3 ubiquitin ligase complex to HIF-1α, leading to HIF-1α ubiquitination and degradation. Thus, VHL and RACK1 recruit Elongin-C E3 ubiquitin ligase complexes to HIF-1α via O2-dependent and O2-independent pathways, respectively.
Calcineurin (also known as protein phosphatase 2B) is a calcium/calmodulin-dependent and serine/threonine-specific protein phosphatase that is conserved in all eukaryotes. It is composed of a catalytic subunit A (CnA) and a regulatory subunit B (CnB) (29, 30). The best characterized substrate of calcineurin is nuclear factor of activated T-cells (NFAT). Upon activation by calcium, calcineurin dephosphorylates NFAT at multiple residues, leading to its nuclear translocation and activation of gene expression (31, 32). The immunosuppressants cyclosporine A and FK506 bind to calcineurin and inhibit its phosphatase activity (33, 34). Calcineurin is involved in a number of physiological and pathological processes including T-cell activation and cardiac hypertrophy (35). In this study, we report that calcineurin promotes HIF-1α expression by dephosphorylating RACK1, blocking RACK1 dimerization, and inhibiting RACK1-mediated HIF-1α degradation.
Human HEK293T embryonic kidney and RCC4 renal carcinoma cells were cultured as described (1, 13). Cells were maintained at 37 °C in a 5% CO2, 95% air incubator. For hypoxic exposures, cells were placed in a modulator incubator chamber (Billups-Rothenberg) flushed with 1% O2, 5% CO2/balance N2 and incubated at 37 °C.
Ionomycin and cyclosporine A were from Sigma. [γ-32P]ATP was from GE Healthcare.
Construction of the FLAG epitope-tagged HIF-1α (27) and hemagglutinin (HA) epitope-tagged calcineurin (51) expression vectors was previously described. HEK293T cells were seeded onto 48-well plates and transfected with plasmid DNA using FuGENE-6 (Roche). Reporter plasmids pSV-Renilla (1 ng) and p2.1 (10 ng), empty vector (EV), and/or expression vectors were cotransfected (14). Cells were lysed and luciferase activities were determined in a Wallac Victor3 multiwell luminescence reader (PerkinElmer Life Sciences), using the Dual Luciferase Reporter Assay System (Pro-mega). Three independent transfections were performed for each experiment. For immunoblot (IB) and immunoprecipitation (IP) assays, HEK293T cells were seeded onto 6- or 10-cm culture dishes. The following day, the cells were cotransfected with expression vectors using FuGENE-6. 24 h after transfection, the cells were exposed to 20 or 1% O2 for 4 h and lysed in RIPA buffer with protease inhibitor mixture (Roche). For RNA interference assay, the expression vectors encoding short hairpin RNA for RACK1 and scrambled negative control were described previously (27).
Cells were lysed in RIPA buffer (IB assays) or buffer Na150NP0.1 (50 mM Tris-Cl (pH 7.5), 150 mM NaCl, 0.1% Nonidet P-40) with 1 mM dithiothreitol and protease inhibitor mixture. 2 μg of antibody was incubated with 1 mg of whole cell lysates (WCL) overnight at 4 °C. 20 μl of protein G-Sepharose (GE Healthcare) was added to the samples and incubated for 2 h at 4 °C to capture IgG. Beads were washed 3 times with buffer Na150NP0.1. Proteins were eluted in Laemmli sample buffer and analyzed by SDS-PAGE. Antibodies used in IB assays were: FLAG and HA (Sigma), RACK1 (BD Bioscience), glutathione S-transferase (GST) (GE Healthcare), His (Santa Cruz), β-actin (Santa Cruz), Myc (Roche), and HIF-1α and CnA (Novus Biologicals).
Cells were separately transfected with vectors encoding: (i) FLAG-HIF-1α-(P402A/P564A), (ii) His-tagged ubiquitin, (iii) HA-CnA-(1–401), or (iv) empty vector. The cells were lysed in phosphate-buffered saline, 1 mM CaCl2, 1 mM MgCl2, 0.1% Tween 20. Lysates were mixed, incubated, and precipitated with anti-FLAG antibody, followed by immunoblot analysis using anti-His antibody. The levels of ubiquitinated FLAG-HIF-1α were quantified by densitometric analysis.
GST fusion protein was purified as described (21). [35S]Methionine-labeled proteins were generated in reticulocyte lysates using a T7-coupled in vitro transcription/translation (IVTT) system (Promega). 3 μg of GST-RACK1 fusion protein and 10 μl of IVTT product were incubated in 500 μl of buffer Na150NP0.1 at 4 °C for 2 h. 20 μl of glutathione-Sepharose 4B beads were added to the samples and incubated at 4 °C for 1 h to capture the GST fusion proteins. After washing 3 times with 1 ml of buffer Na150NP0.1, the proteins were eluted in Laemmli sample buffer and analyzed by SDS-PAGE followed by anti-GST IB assay and autoradiography.
3 μg of GST-RACK1 fusion proteins were incubated with 1 mg of HEK293T WCL transfected with EV or CnA for 30 min at 4 or 37 °C for the phosphorylation reaction. ATP, [γ-32P]ATP, ionomycin, or CaCl2 was added to the reaction as indicated. The reaction volume was brought up to 500 μl with buffer Na150NP0.1 and the samples were incubated for 2 h at 4 °C for dimerization. 20 μl of glutathione-Sepharose 4B beads were added to the samples and incubated at 4 °C for 1 h to capture the GST fusion proteins. After washing 3 times with 1 ml of buffer Na150NP0.1, the proteins were eluted in Laemmli buffer and analyzed by SDS-PAGE. IB assay was used to detect dimerization of GST-RACK1 and endogenous RACK1. The gel was also stained with Coomassie Blue (Simple Blue Stain, Invitrogen) and ProQ-Diamond (Pierce), followed by autoradiography.
After incubation with WCL, GST-RACK1 was captured by glutathione-Sepharose 4B beads and eluted in 30 mM Tris-Cl (pH 8.5), 7 M urea, 2 M thiourea, 4% CHAPS. The proteins were loaded onto a 7-cm, pH 4–7 IPG strip (GE Healthcare), which was rehydrated for 10 h actively at 50 V. Isoelectric focusing was performed for 10 kV-h. The IPG strip was reduced, alkylated, equilibrated in Laemmli buffer, and second dimension SDS-PAGE (4–12% NuPAGE BisTris precast gel, Invitrogen) was performed. The gel was stained with Coomassie Blue.
Data are presented as mean ± S.D. Differences between experimental conditions were analyzed for statistical significance (p <0.05) by Student's t test. The NetPhos program was accessed at www.cbs.dtu.dk/services/NetPhos/.
The calcineurin inhibitor cyclosporine A was previously shown to inhibit HIF-1α protein expression, but the mechanism was not determined. D'Angelo et al. (36) reported that cyclosporine A activated HIF-1α proline 564 hydroxylation, whereas Kong et al. (37) concluded that cyclosporine A inhibited HIF-1α in a VHL- and hydroxylation-independent manner. The involvement of calcineurin was not extensively studied in these reports. We hypothesized that the calcineurin catalytic subunit CnA induces increased HIF-1α protein expression, and thus, inhibition of calcineurin by cyclosporine A would reduce HIF-1α levels. To test this hypothesis, we transfected HEK293T cells with an expression vector encoding HA-tagged full-length CnA. Overexpression of CnA modestly increases calcineurin phosphatase activity, which can be further increased by stimuli that increase intracellular calcium levels (50). Co-transfection of expression vectors encoding FLAG-tagged HIF-1α and HA-CnA led to increased FLAG-HIF-1α protein levels in cells cultured under non-hypoxic (20% O2) or hypoxic (1% O2) conditions (Fig. 1A).
To determine whether HIF-1-dependent transcription was induced by CnA, cells were co-transfected with reporter plasmid p2.1, in which expression of firefly luciferase was driven by a HIF-1-dependent hypoxia response element upstream of an SV40 promoter; control reporter pSV-Renilla, in which expression of Renilla luciferase was driven by the SV40 promoter alone; and expression vector encoding HA-CnA and/or FLAG-HIF-1α. HIF-1-dependent reporter gene transcription mediated by FLAG-HIF-1α was increased by HA-CnA in cells incubated at 20 or 1% O2 (Fig. 1B).
We next investigated whether endogenous HIF-1α protein expression and endogenous HIF-1 transcriptional activity were also increased by HA-CnA under hypoxic conditions. The effect of HA-CnA on the levels of HIF-1α protein in cells cultured at 1% O2 was striking (Fig. 1C, compare lanes 3 and 4). HA-CnA had a similar effect on transcription of the HIF-1-dependent reporter gene in cells cultured at 1% O2 (Fig. 1D). Although endogenous HIF-1α protein levels in non-hypoxic cells were below the limits of detection by IB assay (Fig. 1C), in the more sensitive luciferase reporter gene assay, HA-CnA was shown to significantly increase HIF-1-dependent gene transcription 2-fold under non-hypoxic conditions (Fig. 1D, inset). Taken together, the results presented in Fig. 1 suggest that overexpression of CnA is sufficient to induce HIF-1α protein expression and HIF-1-dependent transcription in transiently transfected HEK293T cells.
Calcineurin activity requires expression of both calcineurin A and B subunits and the binding of Ca2+/calmodulin. We compared the effect of transiently co-transfecting various calcineurin expression vectors (51) into 293 cells along with the HIF-1-dependent reporter plasmid p2.1 or the reporter plasmid pNFAT-Luc, the transcription of which is dependent upon calcineurin-mediated dephosphorylation of NFAT. In each condition tested, the responses of the HIF-1-dependent reporter p2.1 were remarkably similar to those of the calcineurin-dependent reporter pNFAT-Luc (Fig. 2A). These results indicate that endogenous CnB levels in HEK293T cells were sufficient to form heterodimers with ectopically expressed CnA or its truncated, constitutively active form to reconstitute calcineurin activity; and that transient transfection was a sufficient stimulus for calcineurin activation.
Transactivation of p2.1 by FLAG-HIF-1α was stimulated by overexpression of HA-CnA alone (column 1) or in combination with CnB (column 6) and by expression of a constitutively active form of CnA consisting of residues 1–401 (column 3). In contrast, reporter activity was eliminated by a dominant negative form of CnA containing the missense mutation H160N (column 4) and by cyclosporine A treatment (column 5). The effects of co-transfected CnA expression vectors on FLAG-HIF-1α protein levels (Fig. 2B) were in complete agreement with the results of the reporter assay (Fig. 2A), indicating that calcineurin activity stimulates HIF-1 transcriptional activity by increasing HIF-1α protein levels.
To determine the effect of inducing endogenous calcineurin, cells were treated with the calcium ionophore ionomycin, which induces calcineurin phosphatase activity by increasing the binding of calcium to calmodulin, which in turn binds to the carboxyl-terminal region of CnA, thereby disrupting the interaction of the autoinhibitory and catalytic domains; in addition, binding of calcium to CnB also occurs during calcineurin activation (31). Ionomycin increased FLAG-HIF-1α protein levels in a dose-dependent manner, whereas the endogenous CnA protein levels were unchanged (Fig. 3A). The data suggest that calcineurin phosphatase activity, rather than CnA protein levels, are the determining factor for HIF-1α regulation. In further support of this conclusion, cyclosporine A, which selectively inhibits calcineurin phosphatase activity, blocked the increase in HIF-1α levels that was induced by HA-CnA without affecting HA-CnA protein levels (Fig. 3B).
The increased HIF-1α protein levels that were induced by calcineurin may result from increased synthesis or decreased degradation. The proteasome inhibitor MG132 equalized FLAG-HIF-1α levels in cells cotransfected with HA-CnA or empty vector (Fig. 4A, compare lanes 1 and 2 to lanes 3 and 4). These data suggest that CnA regulates HIF-1α levels by inhibiting proteasomal degradation. The proline to alanine mutation of residues 402 and 564 of human HIF-1α renders the protein resistant to PHD-dependent hydroxylation and subsequent VHL-dependent ubiquitination and degradation (21). HA-CnA also increased the levels of FLAG-HIF-1α-(P402A/P564A) (Fig. 4A, lanes 5 and 6), an effect that was mimicked by MG132 treatment in the absence of HA-CnA (lane 7). HA-CnA also potentiated the activation of reporter gene transcription by wild type or mutant FLAG-HIF-1α (Fig. 4B). The finding that CnA had similar effects on the wild type and mutant forms of FLAG-HIF-1α indicates that CnA regulates HIF-1α independently of the PHD pathway.
RCC4 renal carcinoma cells do not express functional VHL and have high endogenous HIF-1α levels even under non-hypoxic conditions (22). Ionomycin dose-dependently increased HIF-1α protein levels in RCC4 cells, whereas cyclosporine A decreased HIF-1α protein levels (Fig. 4C). The data indicate that CnA regulates HIF-1α independently of VHL.
To determine whether CnA affected the ubiquitination of HIF-1α, cells were transfected with expression vectors encoding FLAG-HIF-1α-(P402A/P564A), His-tagged ubiquitin, and either empty vector or vector encoding HA-CnA-(1–401). Analysis of anti-FLAG immunoprecipitates for ubiquitination using anti-His antibodies revealed that HA-CnA-(1–401) decreased the ubiquitination of HIF-1α-(P402A/P564A) 2-fold (Fig. 4D). Taken together, the data presented in Fig. 4 demonstrate that CnA inhibits HIF-1α ubiquitination and proteasomal degradation by an O2/PHD/VHL-independent pathway.
Previously we identified RACK1 as a critical factor in an O2/PHD/VHL-independent HIF-1α degradation pathway. RACK1 competes with HSP90 for binding to the PAS domain of HIF-1α. RACK1 recruits an Elongin-C-containing E3 ubiquitin ligase complex to HIF-1α, thereby increasing HIF-1α ubiquitination and proteasomal degradation (27). RACK1 was reported to bind to PP2A, a protein phosphatase that has high sequence similarity to CnA (38). We therefore investigated the interaction of RACK1 and CnA. Purified GST-RACK1 bound to in vitro translated full-length CnA and amino acids 1– 401, which encompass the phosphatase catalytic domain of CnA (Fig. 5A). HA-CnA co-immunoprecipitated with endogenous RACK1 in HEK293T cells (Fig. 5B).
RACK1 consists of seven tryptophan-aspartate-rich WD40 domains and the fourth of these domains (WD4) mediates homodimerization. The isolated WD4 domain bound to both full-length CnA and amino acids 1–401 (Fig. 5C). Based on this finding of physical interaction between RACK1 and CnA, we investigated whether RACK1 was required for the reduction in HIF-1α protein levels in cells treated with the calcineurin inhibitor cyclosporine A. RACK1 knockdown by RNA interference increased HIF-1α protein levels by 2-fold and blocked the degradation of HIF-1α in cyclosporine-treated cells (Fig. 5D). The data in Fig. 5 demonstrate that RACK1 binds to CnA both in vitro and in human cells and is required for cyclosporine A-induced HIF-1α degradation.
We have previously shown that RACK1 recruits Elongin-C to HIF-1α and that they both bind to the 48-amino acid WD6 domain of RACK1 (27). Two models were proposed for the mechanism by which RACK1 promotes the ubiquitination of HIF-1α: HIF-1α and Elongin-C may bind to two separate subdomains within the WD6 domain of monomeric RACK1; alternatively, RACK1 may form a homodimer in which one monomer binds to HIF-1α and the other binds to Elongin-C. RACK1 was shown to dimerize within its WD4 domain both in vitro and in vivo (39). We constructed a T7-tagged RACK1 expression vector, which only contained the carboxyl-terminal WD5–7 domains of RACK1. If the first model were correct, WD5–7 would have the same effect as full-length RACK1 to degrade HIF-1α. If the second model were correct, WD5–7 would not be able to degrade HIF-1α because it lacks the WD4 dimerization domain. The data in Fig. 6A support the dimerization model because RACK1-WD5–7 failed to induce HIF-1α degradation.
RACK1 dimerization is based on non-covalent interactions, which are disrupted by the denaturing conditions used for SDS-PAGE. To study RACK1 dimerization, we developed an in vitro assay to measure the ability of purified GST-RACK1 to pull down endogenous RACK1 from cell lysates. Because GST-RACK1 and RACK1 have different molecular masses, we could analyze them individually by immunoblot assay. GST did not bind to RACK1 (Fig. 5C and data not shown), indicating that GST-RACK1 pulled down endogenous RACK1 by dimerization. This conclusion was supported by the finding that GST-RACK1-WD56, which lacked the WD4 dimerization domain, did not pull down endogenous RACK1 (Fig. 6B, lanes 1 and 2). Full-length GST-RACK1 (WD1–7) purified from E. coli pulled down endogenous RACK1 when incubated with cell lysates at 37 °C but not at 4 °C (Fig. 6B, lanes 3 and 4).
These results suggested that an enzymatic reaction, which occurs at 37 °C but not at 4 °C, is required to modify GST-RACK1 prior to dimerization with endogenous RACK1. Two-dimensional gel electrophoresis revealed that incubation of GST-RACK1 with cell lysates and ATP at 37 °C reduced the pI of the protein without changing its apparent molecular mass (Fig. 6C), which is consistent with a post-translational modification, such as phosphorylation, that introduces a negative charge. The ability of GST-RACK1 to pull down endogenous RACK1 through dimerization was promoted in the presence of ATP and inhibited in the presence of ionomycin (Fig. 6D).
The data suggested that phosphorylation promotes RACK1 dimerization and that ionomycin activates a calcium-dependent phosphatase such as calcineurin, which dephosphorylates RACK1, and inhibits RACK1 dimerization. In support of these conclusions, RACK1 dimerization was reduced in the presence of overexpressed HA-CnA and/or added calcium (Fig. 6E). ProQ-Diamond phosphoprotein staining revealed that phosphorylation of GST-RACK1 was also reduced by HA-CnA or calcium. Thus, GST-RACK1 phosphorylation correlated with competence for dimerization. The data suggest that phosphorylation of RACK1, perhaps within the WD4 dimerization domain, is required for the RACK1 dimer formation and that calcineurin inhibits this process by dephosphorylating RACK1. Furthermore, activation of calcineurin by ionomycin decreased the interaction between HIF-1α and Elongin-C (Fig. 6F). Thus, calcineurin blocks RACK1 dimerization, which is required for HIF-1α/Elongin-C interaction.
We used the NetPhos phosphorylation prediction software (40) to identify potential phosphorylated residues within the WD4 dimerization domain of RACK1. Scores >0.5 indicate residues that are possibly phosphorylated, with 1.0 representing the highest possible score. Serine 146 (Ser146) and Ser157 were assigned scores of 0.99 and 0.97, respectively (Fig. 7A), and were therefore strong candidate sites for phosphorylation. We investigated the effect of serine to glycine substitution at amino acid residue 146 of RACK1. In HEK293T cells, the T7 epitope-tagged RACK1(S146G) mutant protein demonstrated impaired ability to reduce the levels of co-expressed FLAG-HIF-1α compared with wild type T7-RACK1 or T7-RACK1 with other serine mutations (Fig. 7B). Remarkably, GST-RACK1(S146G) was also less effective in pulling down endogenous RACK1 compared with wild type GST-RACK1 in the dimerization assay (Fig. 7C). Furthermore, when GST-RACK1 was incubated with cell lysates in the presence of [γ-32P]ATP, the incorporation of radioactive ATP was decreased in the S146G mutant compared with the wild type protein (Fig. 7D). The data in Fig. 7 indicate that Ser146 is a phosphorylation site and that mutation of this residue results in reduced RACK1 phosphorylation, reduced RACK1 dimerization, and reduced RACK1-mediated degradation of HIF-1α.
In this study, we further investigated the previously reported observation that cyclosporine A inhibits HIF-1α protein expression (36, 37). We have demonstrated that promoting calcineurin phosphatase activity through ionomycin treatment or overexpression of CnA is sufficient to increase HIF-1α protein levels and HIF-1 transcriptional activity. Calcineurin promotes HIF-1α expression by inhibiting its proteasomal degradation through a ubiquitin-dependent and O2/PHD/VHL-independent pathway.
We investigated the involvement of RACK1 in this process based on our previous discovery that RACK1 is an important mediator of O2/PHD/VHL-independent HIF-1α degradation (27) and a recent report that RACK1 interacts with PP2A, a protein phosphatase with high sequence similarity to calcineurin (38). We found that RACK1 binds to CnA both in vitro and in human cells. Furthermore, RACK1 is required for cyclosporine A-induced degradation of HIF-1α protein. These results provide a molecular basis for the recent demonstration that cyclosporine administration blunted the development of hypoxia-induced pulmonary hypertension (41), which has previously been shown to involve HIF-1 (8).
We focused on the effect of calcineurin on RACK1 dimerization after determining that RACK1 dimer formation is required for HIF-1α degradation. Enhancing calcineurin activity through calcium addition and/or CnA overexpression decreased RACK1 phosphorylation and dimerization, leading to reduced Elongin-C E3 ubiquitin ligase recruitment and subsequent HIF-1α degradation (Fig. 8). We identified Ser146 in the WD4 dimerization domain as a phosphorylation site and demonstrated that mutation of this residue results in impaired RACK1 dimerization and impaired ability to induce HIF-1α degradation.
Our data indicate that phosphorylated Ser146 is likely to be the direct target for calcineurin-mediated dephosphorylation. An antibody that specifically recognizes RACK1 phosphorylated at Ser146 would provide a means to definitively prove this hypothesis. Further studies are required to determine which kinase is responsible for the phosphorylation of RACK1 that is required for its dimerization. Although the experimental approach to answer this question is beyond the scope of the current study, we note that Ser146 is present within a context that matches the consensus sequence (SXXE/D) for phosphorylation by casein kinase II.
RACK1 dimer-mediated HIF-1α degradation requires that each monomer bind to either HIF-1α or Elongin-C. This strategy may seem inefficient because by chance only half of the RACK1 dimers will co-recruit HIF-1α and Elongin-C, whereas the other half will bind two molecules of HIF-1α or Elongin-C. However, this process provides a mechanism for regulation by controlling RACK1 dimer formation. In this report, we have shown that calcineurin inhibits RACK1 dimerization, thereby linking the regulation of HIF-1α expression to intracellular calcium levels and calcium-activated signal transduction cascades. These findings complement previous studies, which demonstrated that HIF-1α translation is calcium-regulated by protein kinase C-α signaling (42) and HIF-1α transactivation function is regulated by calcium/calmodulin kinase-mediated phosphorylation of p300 (43). In some cell types, hypoxia induces increased intracellular calcium levels (42), which may amplify the induction of HIF-1α in these cells.
RACK1 was originally identified as an anchoring protein for activated protein kinase C (44). However, RACK1 is now recognized as a multifunctional scaffold protein that plays an important role in diverse biological processes including intra-cellular signal transduction (45) and assembly of the 80 S ribosome from 40 S and 60 S subunits (46). Some processes that are mediated by RACK1, such as HIF-1α regulation, require RACK1 dimerization, whereas other processes, such as ribosome assembly, involve RACK1 monomers (47). For most of the processes in which RACK1 has been implicated, it is not known whether dimers or monomers are utilized. Calcium-activated calcineurin signaling is likely to regulate many physiological pathways that involve RACK1 dimerization. Furthermore, RACK1 dimers increase calcium release by binding to inositol 1,4,5-triphosphate receptors (48). Thus, a feedback loop involving RACK1 dimerization and calcium influx may serve to tightly control the RACK1 dimer:monomer equilibrium. HIF-1 may contribute to the complexity of this regulatory circuit by mediating transcriptional activation of the TRPC1 and TRPC6 genes, which encode store-operated calcium channels (49). Calcineurin plays a key role in physiological responses such as T lymphocyte activation and cardiac hypertrophy. Further studies are required to determine whether these responses are dependent upon modulation of the RACK1-HIF-1α regulatory pathway by calcineurin.
We thank Karen Padgett (Novus Biologicals Inc.) for providing CnA antibody.
*This work was supported by National Institutes of Health NHLBI Grant N01-HV28180.
3The abbreviations used are: HIF-1, hypoxia-inducible factor 1; RACK, receptor for activated C kinase; PHD, prolyl hydroxylase domain protein; VHL, von Hippel-Lindau protein; HSP, heat shock protein; CBP, CREB-binding protein; CREB, cyclic AMP response element-binding protein; PAS, PER-ARNT-SIM homology domain; PAS-A, subdomain A of PAS; CnA, calcineurin A; NFAT, nuclear factor of activated T cells; GST, glutathione S-transferase; IVTT, in vitro transcription/translation; WD, tryptophanaspartate-rich; IB, immunoblot; EV, empty vector; WCL, whole cell lysates; SV40, simian virus 40; IP, immunoprecipitation; sh, short hairpin; HA, hemagglutinin; CHAPS, 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid; BisTris, 2-[bis(2-hydroxyethyl)amino]-2-(hydroxy-methyl)propane-1,3-diol.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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text: London (ANI): While James McAvoy might be thanking his stars to have landed a role opposite Angelina Jolie, the Hollywood beauty made him see stars in broad daylight by beating him up on the sets of their new movie. The Atonement star himself revealed his awful experience of starring with the Tomb Raider beauty. Angelina kicked seven colours of poo out of me but, thankfully, Im still in one piece, British tabloid Daily Mirror quoted him as saying. In Wanted, James plays Wesley Gibson, a 25-year-old employee who works in a cubicle. His character is offered the opportunity to avenge the death of his father, who was an assassin. He is then trained by his fathers partner, Sloan (Morgan Freeman), and by Sloans second-in-command, played by Angelina Jolie. The film is set to be released on June 27.


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Forums > MM Market > Photo Gear > FS: Quantum T5d-r with power pack and wireless
Posts: 447
Tacoma, Washington, US
Due to switching largely to continuous lighting, I'm looking to sell my Quantum strobe. It's a lightly used T5d-r with a Turbo SC power pack, plus a FreeXWire receiver and transmitter pair. Asking price for the set $600, including a PC sync cord and power adapter for charging the power pack.
Jan 26 13 10:40 am Link
Posts: 8833
Las Vegas, Nevada, US
Moderator Note!
moved to photo gear
Jan 26 13 12:51 pm Link


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"I wasn't sure if I really had the skills to help someone else, but I now understand that what matters the most is that I am there."
Adrian B, OzChild Volunteer
How can I help?
Volunteers are crucial to the work OzChild provides helping support children and families and are essential in allowing us to deliver our Volunteer Family Support Program. Volunteers offer assistance to families in practical and emotional ways often helping them through difficult times.
Volunteers of the Family Support Program are asked to commit two – four hours per fortnight to support children and families with a range of activities such as;
- Supporting a parent with household routines and positive parenting.
- Simply spending time listening to a child
- Sharing hobbies and skills.
- Cooking, arts and crafts
- Sharing activities such as kicking a football, visiting the park, shooting hoops or a trip to the beach.
- Helping with school work
- Providing emotional support and company to parents and children.
- Helping families' access libraries play groups and the wider community.
- Providing transport to activities and appointments.
Why Volunteer?
Joining OzChild's volunteering services can be incredibly rewarding experience. By volunteering with OzChild you provide vital support to children helping them access new opportunities – enabling them to thrive. Your support can make a huge difference by benefiting not only the child but also their families and in turn the communities in which they live.
Volunteering can also provide opportunities to:
- Share your skills and life experiences
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How do I volunteer?
You do not need any special qualifications or have previous experience to volunteer with OzChild. The only qualities you need to possess are compassion and the desire to improve the lives of children and families. To be eligible as a volunteer you must be:
- Over the age of 18
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- Attend an interview
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Enquire to become a Volunteer by completing our online Application Form here.
Volunteers are needed in the following regions:
Enquire to become a volunteer
Complete an application to become a Volunteer here or contact our Volunteer Recruitment, Retention and Training Coordinator on (03) 8796 0000 or email firstname.lastname@example.com
OzChild disability volunteers support our clients on recreational activities, camps and school holiday activities. Our Flexible Respite Programs offer activities as wide ranging as arts and craft lessons to a trip to the zoo. As a volunteer you may provide assistance helping a child to learn karate, dancing lessons, attend a movie night, access a yoga class or participate in an African drumming workshop. The list is endless and ensures our children and young people are engaged, learning and having fun. Volunteers will be asked to support the clients and to enhance their experience on the day. A disability support worker is always with you for additional support and guidance.
The time commitment of volunteers within the disability program is determined by the volunteer themselves from one day on a weekend to everyday during school holidays, 3 nights at camps or 5 hours at karate. As a volunteer we encourage you to choose the acitivites which most suit you and your skills and abilities.
Flexibility is just one of the benefits when people volunteer with the Disability program. It's a great way to make a difference, to gain new skills and experiences and to get involved within your community.
Volunteers are accepted from 16 years up, and will need to be willing to undertake a police check, working with children's check and DWES (Disability Workers Exclusion Scheme) once they turn 18.
Complete an application to become a disability volunteer here or call our Volunteer Co-ordinator on (03) 5975 7644
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Former Australian coach John Buchanan, who is the Director of Cricket for New Zealand Cricket (NZC), has defended the decision to appoint another Australian for a key role in New Zealand Cricket.
It comes after Kim Littlejohn was appointed as the national selector manager, who will be responsible for the selection of men's and women's squads from under-19s through to the Black Caps and White Ferns.
Littlejohn previously played and coached club cricket in Australia, then had high-performance roles in baseball and bowls. He joins other Aussie- assistant Black Caps coach Trent Woodhill and Black Caps bowling coach Damien Wright.
"Like any business you scout around your opposition and if you see something working well, that is where you go to get their IP (intellectual property) or borrow their people," stuff.co.nz quoted Buchanan, as saying.
"Australian cricket has for some time been doing some things well, not just in cricket, but in high performance, so naturally we gravitate there. I don't think the bias is Australian, the bias is looking where the things we need can be best located," he added.
Littlejohn's initial task will be to work with Wright to select the New Zealand squad for the upcoming tour of Zimbabwe. (ANI)


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The location was perfect for a short trip to DC! Carols the day manager was excellent! All of the staff was great. They were very friendly and more than happy to help us. This was our first time to DC and didn't know where to start, and the entire staff was great in helping us; from metro maps to shuttle to and fro to nearby metro station. We would definetly stay at this hotel again. It saved us alot of headaches touring DC; money and time. The free shuttle service to and from the metro station was great. We didn't have to worry about our car and parking. You could do all of the things you want in DC and you can relax at the end of your day and catch the shuttle back to the hotel. Don't have to worry about fighting the traffice in unfamiliar place!! It was a great experience!!! You could always say that hotel could be better, but for the price and the location it was worth it. TV could use an update model and the controller was sticking, but the room was clean and bedding was too. Overall experience of this hotel was great!
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Sharepoint Web Part #1
Budget $30-5000 USD
We need a web part which would do the following:
From within a particular view of a contact list there is a checkbox named "send email". Records which are selected will be sent an email via the BCC field. When a contact is emailed a note will be automatically added to a "email history" field which will list the date/time and subject of the email.
## Deliverables
Rent A Coder requirements notice: As originally posted, this bid request does not have complete details. Should a dispute arise and this project go into arbitration "as is", the contract's vagueness might cause it to be interpreted against you, even though you were acting in good-faith. So for your protection, if you are interested in this project, please work-out and document the requirements onsite.
1) Complete and fully-functional working program(s) in executable form as well as complete source code of all work done.
2) Deliverables must be in ready-to-run condition, as follows? (depending on the nature? of the deliverables):
a)? For web sites or? other server-side deliverables intended to only ever exist in one place in the Buyer's environment--Deliverables must be installed by the Seller in ready-to-run condition in the Buyer's environment.
b) For all others including desktop software or software the buyer intends to distribute: A software? installation package that will install the software in ready-to-run condition on the platform(s) specified in this bid request.
3) All deliverables will be considered "work made for hire" under U.S. Copyright law. Buyer will receive exclusive and complete copyrights to all work purchased. (No GPL, GNU, 3rd party components, etc. unless all copyright ramifications are explained AND AGREED TO by the buyer on the site per the coder's Seller Legal Agreement).
## Platform
Windows 2003, Windows Sharepoint Services


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Kart's in an established food brand in Malaysia specializing in Malaysian ethnic food and finger food snacks as well. Kart's wide product range is processed and packed in two manufacturing centres in Shah Alam and Kelantan
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|Overall Rating:
I've only bought their frozen pizza.
Kart's is a brand that I am familiar with as I tend to accompany my mother to the supermarket. In Giant Supermarket at the frozen food section, they sell Kart's Pizzas in different varieties. I bought their beef pepperoni pizza to try it out at home. The box definitely did make it seem delicious, that's for sure.
As I tried it at home, I was highly disappointed. It didn't taste anywhere close to a regular beef pepperoni pizza. If anything, it was beef slices with regular tomato sauce on bread. Don't worry, I didn't eat it while it was frozen as I did put it in the oven first. It's definitely not worth the money.


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The Pope John Paul II volleyball team got a sample of the Pontchartrain Center last season, reaching the Division IV playoff quarterfinals.
The Jaguars are hoping to take the next step and win the school's first volleyball state title. They have steadily climbed in the power rankings, starting at the fourth sport , then moving to No. 2 before landing at the top.
"All season long, we have strived to be the No. 1 seed heading into the playoffs," coach Rachel Battistella said. "The girls have been working so hard to get there and to stay there. I don't think they feel like it's any pressure. They just want to keep working hard to win. The girls are pushing really hard to bring home the trophy. I'm glad they have been in the Pontchartrain Center before because they know what to expect now."
Pope John Paul II enters the playoffs with a 36-5 record. Their losses have come against Mount Carmel, St. Joseph's, St. Thomas More, Barbe and Country Day. They also boast a win against Country Day, the No. 1 seed in Division V. The Jaguars won a district title for a second straight season, going undefeated in district for a second straight year. The season also included a 17-match winning streak.
Battistella, in her first season as head coach after serving as an assistant in 2013, was expecting this type of success from a team that returned most of its starters. And while the returning experience has been a key factor, the team's unity has also played a major role.
"This team is very close- knit," she said. "Our four seniors are great leaders, and the younger kids are great players. It's a family- oriented team, and they all look out for each other, even at practice. They are always encouraging each other, so you couldn't ask for a better group of girls."
The seniors are: Caitlyn Battistella (the coach's daughter), Allison Hartmann, Christian Cutura and Sarah Worchel. Other key players are: outside hitters Marlena Cutura and Camille Dedeaux, setters Milayne Danna and Bailee Maillet, Sarah Trosclair, Aixa Crespo, and libero Tyler Sanderson.
Christian Cutura and sister Marlena are seeking their second state title of the year. The two helped lead PJP to a soccer state title in the spring.
Danny Tullis, who won a state title at Fontainebleau in 2011, is in his first season as an assistant at Pope John Paul II. Like Battistella, he was expecting the Jaguars to have a splendid season.
"We had a really good preseason and did really well in the camps," he said. "So we thought we had a chance. We lost two starters, but we felt really good about the kids who would be taking their places."
He also hopes last year's experience, losing to Riverside in the quarterfinals, will carry over to this year.
"Last year was the first time this group had been to the Pontchartrain Center, so they played a little tight," he said. "They played a little tight. Any one of the top 8 to 10 teams has a chance to win it, so getting the No. 1 seed is good, but it's not the end all, be all."
PJP earned a bye in the bidistrict round and awaits the winner of Thursday's match between Catholic-New Iberia and Delcambre.
"We will use this week to practice hard to try to reach our goal," Battistella said. "The girls are handling themselves well, They know what to expect. They are ready to go."


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So, I was out dancing a few nights ago and met a girl with whom I had amazing dance chemistry. I danced better with her than anyone else there. I LOVED it.
I told her I loved the dance chemistry, and that it was the best I'd had all night. She agreed with me, enthusiastically. The style we were dancing in was pretty sensual, close, and so on.
I told her I thought she was damned cute and asked her if she was single...
She then said it was complicated at the moment and she wasn't sure how to answer. She actually kind of started to explain, but I cut her a bit short and said,
"Let me ask you an easier question... would you like to get some hot chocolate with me some time?"
She said without missing a beat "Yes!"
And then added a second later,
"But no strings attached."
I agreed.
My first impression was that she was saying "I'm not looking for a 'relationship'."
I don't know. What's your impression, in this context, of the "no strings attached" comment?
A girl said "no strings attached" in an ambiguous way?
So, I was out dancing a few nights ago and met a girl with whom I had amazing dance chemistry. I danced better with her than anyone else there. I LOVED it.
Have an opinion?
What Girls Said 1
In that context it kind of sounds like "don't expect anything from me, in exchange for this hot chocolate".
I would include sexual, emotional, and romantic in the "no strings attached" comment until other wise clarified.
in my opinion anyone under "its complicated" status should be left alone, they're messy people.00|00|0
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text:
The Essay is a daily personal piece submitted by readers. Have a story to tell? See our guidelines at tgam.ca/essayguide.
The last time I saw my grandparents was 35 years ago. Today, I am repairing their tomb in rural Jamaica.
Much has changed. Grandmother's prized garden, filled with countless species of plants and flowers – where she would kneel and pull stubborn blades of grass from the dark soil – is dead. The fidgety, chirping, yellow-chested bananaquits that used to suck the juice from the red and mauve hibiscus blooms are gone. That scruffy-trunk mango tree on the edge of grandma's garden, from which I once fell on barbed wire that ripped my chest, is barely alive. Finally, I can forgive that withered old fruit tree.
The L-shaped, paint-faded, three-bedroom brick house is smaller than I remembered. The bluish-green hills of Spring Garden in the Trelawny region are dotted with houses and farms, but the land around my grandparents' house is fruitless – a sign of how long it has been since Hutch, my grandfather, died.
I hold my breath and walk slowly up the seven weather-beaten red steps onto the veranda, lingering briefly between the two old empty chairs where he and grandma spent afternoons reliving their youth until the sun set behind the mountains.
I grin – smirk, perhaps – as I recall how Gwendolyn Fluxby, the district's gossip queen, would force her wide bottom into those narrow veranda chairs. Under the heavy load, the metal frame would screech against the tile floor. Gwendolyn would lean back, kick off her shoes, remove her purple head wrap and mop her forehead before fanning her face with a red-and-white handkerchief and complaining about the scorching heat.
Grandma would make lemonade with stout chunks of ice in the glass. After taking a few hasty, loud gulps, Gwendolyn would press her palm against her belly and burp like a deflating truck tire for many seconds. After begging pardon, she would settle down and release the latest windy hearsay into grandma's right ear – the one that still listened to the world. Once, I heard her counselling my grandparents not to buy the condensed milk imported from Cuba because it was spiked with a potion that converted hard-working citizens into idle socialists.
I slip through the front door into the faded scent of peppermint and a flood of memories: Hutch's lanky frame, his starched and ironed khaki pants and spotless, white, short-sleeved shirt as he sat beside his Telefunken radio listening to the BBC news.
My nine-year-old eyes shone brightly beside him on my first visit to his farm in Cockpit Country. Prime lands had been set aside for primary citizens of the British Empire, so Hutch's land was cratered with sinkholes and strewn with boulders. But it was the place where he sowed the seeds of his character, and reaped an iron will through hard work and rivers of sweat.
Dense clusters of century-old trees, their fat, crusty trunks reaching for the sun, swallowed us. Mist from whitewater streams gushing over limestone rocks soaked my short pants. A sawtooth of greenish-blue hills rose in the distance, and creatures called in haunting voices I had never heard. I stomped my feet like a marching soldier to prevent the buzzing soprano choir of mosquitoes feeding on me until Hutch's fire could drive them away.
Suddenly, it was quiet and I could almost hear the barking voices of British military men, "Load your musket and fire!" To which Maroon Captain Cudjoe rallied his own troops. "Destroy those Redcoats!"
Grandfather's stories of ancient battles hung trapped in the humid air, and the peaceful hills roared like thunder. Musket balls ripped flesh apart. Projectiles whistled as they ricocheted off rocks and trees. Arrows and spears fired by Maroon warriors swished through the air, punching holes in the Redcoats' jackets. It was after these guerrilla wars that the Empire negotiated a peace treaty, recognizing the Windward Maroons of Trelawny Town as an independent nation, but many were later deported to Nova Scotia.
A shot from my grandfather's rifle removed a swift-flying white-crowned pigeon from the sky, cutting short my day-dream.
Now, looking out the car's window, my eyes travel with the quiet Caribbean Sea. Its turquoise waters incline gently into the cloudless sky, forming a seamless bond. That's when the old man splashes into my mind: the sound of his crackling laughter as he played cricket with me and his other four grandsons. The uncommon wisdom he always gave us: "Pick sense out of nonsense."
The sky was cloudy on our final hunting trip. Age had wounded his health and a walking stick helped as a third leg. The rifle was heavy, and I carried it most of the time. I pointed to a wild boar in a thicket. He took aim, his hands trembling. He handed the rifle to me, as though tapping my shoulders with a sword to complete my knighthood. I grabbed the weapon, raised it and fired. It growled, a promise of spring to a wintered man under tropical skies.
Montego Bay Airport was crowded when I left him the last time, but my 3 p.m. flight to Toronto was on time. I whispered to grandma, "Tell Hutch I miss him already when you get home." He died one year after that, and grandma eight months later. But today, in the laughter of his sons, daughters, grandchildren and faded black-and-white photographs I see him, alive again.
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The look of leather for less makes this glider rocker with ottoman a must for any - or every - room in your home. A uniquely shaped back is accentuated with accent stitching and paired with a rounded front seat for supportive comfort. A matching ottoman offers cushioned support for your legs and invites you to put up your feet and stay awhile. Pillow armrests top the metal base, which features glide and recline mechanisms for total relaxation. Your choice of black or bone leatherette lets you dress this recliner with ottoman set up as much or as little as you like for a beautiful addition to your den or living room decor.Includes
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Carnival Girl: Searching for God in the Aftermath of War by Sonja Herbert
Posted by Krystal Larson at 11:44 PM Saturday, July 14, 2012
The only life little Sonja Francesco has ever known is traveling the carnival circuit and living with her five siblings in a tiny caravan home. The family never stays anywhere long enough for Sonja to make friends or develop roots. The only one in her family, Sonja always believed in God and wants to belong to a church.
At fourteen, Sonja meets the Mormon missionaries and develops a strong testimony of the truth of the Gospel. But can she live the commandments while traveling with the carnival and running one of the attractions every Sunday? Will it be possible for her to leave her family's life behind and live the life she has always dreamed of?
Goodreads Summary
How many times have we wished we could be part of a carnival? Sonja, the main character, travels in a caravan home and helps "build" carnivals. Her life may seem exciting, but it is apparent that it may be less than ideal. Margot, Sonja's mother, first started travelling with the carnival to hide during WWII. Margot is not a religious woman, can be a little crass, and rarely fawns over her children. Whether or not Margot knows it, she plays a large part in shaping Sonja's character. Sonja mainly wants her family to appreciate and love her. She doesn't always feel noticed or care about and simply wants to do her best. It could be said that Sonja's loneliness led her to God. Will Sonja's deepening faith lessen or cause trouble with her family?
This memoir is not exactly up-lifting. The readers will most likely find themselves wanting to hug Sonja. Sonja speaks plainly and succinctly in the memoir; the only subject the reader may want to know more about would be Sonja's parents-how did they meet? What happened to Margot during the War? Getting to know Sonja made this book worth reading for me. The book isn't fast-paced, but this isn't a book a reader would probably want to skim through. This book is set in a very topsy-turvy, unsettled time in history, a setting that adds to the novel. Overall, this book is recommended to adult readers.
3 1/2 Stars
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3 comments:
Sonja Herbert said...:
Thanks for your very nice review!
V (BookBorne) said...:
I love books with a carnival or circus element. with that and the time period, this book sounds very interesting.
erin said...:
thanks for a great review! Gotta go check this one out :)
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PRAGUE – Every year, it seems, starts with the return of a noxious tradition: the annual dispute between Russia and Ukraine over energy, a dispute that left millions of Europeans shivering last winter. That dispute, in turn, exposes the European Union's fragmented energy-supply system and lack of political cohesion, which undermines its ability to forge a long-term energy strategy.
Diversification of energy sources is the first step needed to build a viable energy-security strategy. This requires long-term planning and coordination, because the necessary infrastructural decisions will take decades to implement.
Calls for supply diversification stem largely from the EU's reliance on Russian gas, and Russia's willingness to use energy for political purposes. Moreover, Russia controls the vast majority of the gas pipeline system, and has negotiated individually with European countries, with the result that it holds a disproportionate degree of leverage over the European gas market.
This system is inherently unbalanced, and must be corrected. New and alternative pipelines such as Nabucco must be built, and closer relationships with other suppliers encouraged. It is unrealistic to call for an end to Russian gas imports, or even an end to Europe's major reliance on them, but the EU can increase its own leverage by developing alternative sources of supply.


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Our desire is to study God's Word together and to meet with each other with a sense of authentic community. We feel like having Community Groups mainly on Sunday mornings between our services gives us an intentional way of caring for each other's need and growing in Christ
Our mission is to see every member, attender and follower be part of a Community Group that unifies us as a body of believers that are 'one hin heart and mind' (Acts 4:32). We desire for these groups to be led by people that want to study God in the Word and in the Spirit. When we study together we are becoming more connected to each other and able to be a stronger body of followers of Christ.
These Community Groups are:
- Open to All and Welcoming
- Consistent and Accountable
- Equipping and Discipling
- Bible-based, Gospel-centered Studies
- Community-centered and Multi-generational
We envision a ministry that effectively cares for people, accelerates spiritual growth within people, and shifts the work of community to the people (Eph 4:12).
DO I NEED TO SIGN-UP? Nope. We just want you to be present and coming. All groups are upstairs in the new classrooms (Jerusalem, Bethlehem, ect) so head there in-between the services and join a group. Or you can contact Scott and April Tappe to know what group is available.


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Earlier this week, Google introduced a feature that lets you
Google's "Call Phone" feature has so far made a splash, inspiring
A favorable response is good news for Google; the last opt-out Gmail feature,
Google's Gmail call feature is undoubtedly a dangling carrot that can boost Google's formidable bottom line and reinforce some of its other services. If it's heavily adopted, as we suspect its convenient placement in the inbox will warrant, the company's dominance as a trusted, go-to brand will surely continue to grow.
Watch our video above for a step-by-step tutorial on how to make calls from Gmail to get started.This story originally appeared on CNET


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Parking: Street parking (most meters nearby are free after 8pm) / There are parking garages and outdoor lots off of Market Street that are in walking distance. Most of them charge $1 an hour or have a flat nighttime rate.
If you are taking the Trolley: Closest stops are " Fifth" off of the Blue Line and "Convention Center" or " Gaslamp Quarter" if you are taking the Orange Line.


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The true story of a resilient circle of shrimp boat captains who faced and withstood the wreckage of Katrina but now find their courage tested by a greater threat: the disappearance of their livelihood and their centuries-old bayou culture.
"Vivid prose, first-hand testimony and solid, heartbreaking reportage make this disaster debrief hard to put down, and worth the attention of every U.S. citizen." (Publishers Weekly)
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It Puts St Bernard on the Katrina Map
- Martha A. Murray
Chris Andrews should have consulted with people who live in that area how to pronounce the names of places. For instance the city Houma is NOT pronounced WHO mah. It's HOE ma. Lafitte is laFEET, NOT La FIT. And you pronounce the S in Ysclosky!I found his constant mispronunciations extremely annoying.
I lived in New Orleans from 1977 to 1986. For the first two years I ran crew boats in the Kerr-McGee production field in Breton Sound. Our base was in Hopedale, so every other week (we worked 7 days on/7 off) I drove up and down those roads and I ran boats around in those bayous. The last three years I lived on a shanty boat at the Gulf Outlet Marina on Bayou Bienvenue in Chalmette, so I have a real feeling for the area.New Orleans got all the media attention, and it's understandable. People in Detroit, Denver, and even Düsseldorf, Germany know about New Orleans, but none of them ever heard of Shell Beach of Ysclosky. I saw videos of the flooding in St. Bernard and couldn't believe my eyes. 95% of the structures in the Parish were flooded! The Gulf Outlet Marina, which sat INSIDE the levee system, the story says, was "obliterated." I could, literally, visualize the narrative as it unfolded, knowing Violet and the canal and what those houses and camps along the bayou all the way down to Hopedale were going through.Thank you for this book.
- Richard


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Is he interested or is it too soon to say? So there's a coffee shop I've been going to for the past 5 months now and I come across the same barista guy. Once when I walked in, he was cashing out his register and when he saw me, he kept staring at me while he was taking the money out. About a month later in December when he was making my drink he put my drink at the edge of the counter, smiled reallyyy big and said "Have a nice day." At the beginning of this month I started to study there on Fridays and would sometimes catch him looking. I've been there like a total of 4 times to study there and I stay for several hours. Once when I was about to walk out I asked him what time they open and close. I asked this while I was walking and he started walking with me but he was behind the counter if that makes sense. I came with my friend late Saturday night and he was there. While I was waiting for my drink he kept looking up every now and then to look at me. When I went up to grab my drink he smiled super bigg, waved, and said "Hiiiii!!!" I got shy and just said thank you. My friend said "Omg he totally wanted to talk to you!" I went back today and he was looking at me a lot. I told him I was sorry for not speaking to him last week and he made a funny sound with a hand motion that basically said "Why didn't you talk?' I said I was really shy. Then he introduced himself and asked what my name was and shook my hand. We asked each other questions and I told him about this big grad school exam Im taking soon and he said "Oh so you're too busy to hang out with friends?" But I said "It depends on the day but I can always make time." We talked for 15 mins and I told him that he can always come by my table and talk to me even if Im studying and he said he will. Is it too early to tell?
Is he interested or is it too soon to say?
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The mystery over whether ConnectiCare would stay on or exit the state's Access Health CT insurance exchange ended late on Sept. 13, when the insurer said it would remain.
At issue was a disagreement between ConnectiCare and the Connecticut Insurance Department over a proposed rate hike. Earlier this year, the insurer sought an increase of 14.3 percent, later revised to 17.4 percent and ultimately climbing to 27.1 percent. When the Insurance Department approved the 17.4 percent increase, ConnectiCare sought a court injunction to invalidate the decision. It was denied by a Superior Court judge in New Britain on Sept. 9.
That same day. ConnectiCare sent a letter to Access Health CT saying it was terminating its agreement. Although AHCT announced its receipt of the letter and seemed to acquiesce to the termination, ConnectiCare President and CEO Michael Wise insisted the company was still "actively working" to find a mutually acceptable arrangement allowing it to remain on the exchange.
late on Sept. 13, Wise released another statement announcing that the insurer would remain with AHCT with the 17.4 percent increase.
'Throughout the rate-setting process, ConnectiCare has emphasized its goal of remaining on the Connecticut health insurance exchange in a manner that is fiscally responsible," Wise said in the statement. "After hearing from state officials, providers and beneficiaries about the importance of our plan to Connecticut, we have decided to move forward into 2017 as a plan on the exchange at the rates approved by the department."
ConnectiCare has withdrawn its legal appeals and rescinded its Sept. 9 termination notice in a letter to the exchange. Wise said.
The statement left open the possibility that the rate-hike issue may arise again. "We look forward to continuing to partner with the department and the exchange to continue to meet this promise and to address the broader policy issues, such as risk adjustment, that affect the sustainability of the health insurance market place," Wise said.
For now, however, it appears both sides are heaving sighs of relief.
Gov. Dannel Malloy said ConnectiCare's decision "is welcome news, as many Connecticut residents who depend on the carrier for their health care services will continue to benefit from their inclusion in the exchange."
"Recent data show that Access Health CT has brought the number of uninsured people in our state to its lowest rate in history, and among the lowest rates in the country," Malloy said. "Our administration will continue working to ensure that Connecticut remains a leader in implementing an exchange that provides quality, secure, and affordable health care to all of our state's families."
ConnectiCare covers 47,597 people through the exchange. Had it exited, only Anthem would have remained with AHCT, following United Healthcare's decision in April to leave most Obamacare-created insurance exchanges and the Insurance Department's July decision to prevent the financially strapped HealthyCT from continuing to do business.


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Bangkok Seashell Museum
The type of search By District | By Place
Map Have Map No Map
Bangkok attractions
Attractions in Thailand
Operating day: Daily
Operating time: 10.00 - 18.30
A place with a large collection of beautiful Shell around the world, Selected and obtained to show their never-ending range of colors and forms. More than 3000 specimens of 624 selected species from the world are exhibited here!
Our legacy from many generation to show the visitors "Knowledge for little creatures" to understand the world and its background history through enormous fossils, cute vivid snails, colorful pectens, poisonous cones and much more.
This Museum will be another colorful experience for your Bangkok destination. A place where knowledge and art are combined in harmony.
Buildings where huge outdoor Ammonite-like decorate are seen just a zoometer walk from Surasak sky train station, at the corner of Silom 23 Road, near Leardsin Hospital.
Website: www.bkkseashellmuseum.co.th
Tel: 022340291
Mobile: 0896813814
Category: ●Art, Culture and Heritage
Group: ●Museums
Tag: ●Bangkok Seashell Museum
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Auschwitz Concentration Camp by: Allen Sharpe
Auschwitz also known as Auschwitz opened in 1940 and was the largest of the Nazi concentration and death camps. Located in southern Poland. Auschwitz initially served as a detention center for political prisoners. However it evolved into a network of camps where Jewish people and other perceived enemies of the Nazi state were exterminated often in gas chambers, or used as slave labor. The gas chambers were numbered 2 and 3, and 4 and 5. The first opened on March 31 1943, the last on April 4 1943. The total area of the gas chambers was 2,255 square meters; the capacity of these crematoria was 4,420 people. Those selected to die were undressed in the undressing room and then pushed into the gas chambers. It took about 20 minutes for all the people to death.
10 interesting facts
1.Auschwitz concentration camp was very big and large because it had three parts. It was located on the 25 square miles of land.
2.There were smaller satellites located inside the Auschwitz. It also contained an exclusion zone. That's why this camp was called as the largest one when all the construction was completed.
3.The Jewish people who become the prisoners in the Auschwitz were murdered in gas chambers. They will be sent directly to the gas chamber with naked body. They tell them that it was a shower.
4.Arbeit macht frei was seen on the front gate of Auschwitz camp. It means "work sets you free". When people entered the camp, the Nazi gave them false hope by thinking that they would be free if they worked in the camp.
5.Auschwitz III was also called as Monowitz. It was owned by IG Farben. It was the palce where many prisoners had to work to create many types of chemicals
6.Sonderkoandom was the prisoners whose job was to assist the German Nazi in the crematorium. To keep everything secret, German Nazi killed Sonderkommando randomly.
7.Josef Mengele was the famous doctor who made an experiment on the prisoner while in Auschwitz camp. He was interested more to conduct the experiment on twins and kids.
8.e first commandant in Commandant Camp was Rudolf Hoess. In March 1946, he was caught and faced the trial. In April 1947, he was hanged on the entrance gate of Auschwitz camp.
9.There was a revolt occurred in Auschwitz II. Some prisoners who helped the German Nazi on the crematorium damaged the Crematorium IV. The women who worked outside camp used the explosive to destroy the gas chamber
10.The allied force which liberated Auschwitz was the Soviet Union on January 27, 1945. Most prisoners had been killed on the chamber. There were only 7,500 people saved from the camp.
In September 1935 the Nuremberg law were enacted. These laws prohibited marriages between Jews and people of Germanic extraction. and the employment of German women under the age of 45 as domestic servants in Jewish households. A supplementary decree outlining the definition of who was Jewish was passed on 14 November and the Reich Citizenship Law officially came into force on that date
5 People involved in Auschwitz
1.Adolf Hitler: started WW II
2.Nazis: Guarded Prisoners
3.Gypsies: Where prisoners
4.Polish:Where prisoners
5. Jews:Where prisoners
American Soldiers Are Heros
The American Soldiers are heroes because they came to all of the labor cams and death camps and released all of the prisoners.
This is a picture of Auschwitz Dethcamp
Why is Auschwitz Important
Auschwitz is important because it has poisoned people have been killed their.
- "Auschwitz." History.com. A&E Television Networks, n.d. Web. 02 June 2015.
"Auschwitz-Birkenau Extermination Camp (Poland)." Auschwitz-Birkenau Extermination Camp (Poland). N.p., n.d. Web. 02 June 2015.
- "Auschwitz." United States Holocaust Memorial Museum. United States Holocaust Memorial Council, 20 June 2014. Web. 02 June 2015.


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New year's shineCategory: Photo frames >Events > New Year
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