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getting rid of oberon media
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All In One Heat Pump For Hot Water - Chinaprice: contact company for price
This Series of Eco-friendly Water Heater is desighed to make mestic and overseas sanitary hot water. As a lovely equipment, it can be put in storage garage or near the kitchen or the washing room. A lot of attention has been put to its appearance design. To make the whole unit small, the structure inside is also very compact and organized well.
2)All in one heat pump
1. Stainless steel tank inside, plus 50mm insulation.
2. Extremely compact structure, good looking face, and easy demountable for access.
3. High efficiency compressor with R410A refrigerant.
5. Touch design LCD control panel, and the control signals are easier to be transferred to the control system;
6. Automatic defrosting function included.
7. Clock & Timing on/off function.
8. Electrical heating element inside as back up for emergent request.
9. 65°C high temperature weekly auto antisepsis and changeable Magnesium stick inside tank to ensure good quality water.
10. Careful protection functions such as: high pressure, low pressure, and anti-freezing, phase order, discharge temp etc.
11. Optional extra solar coil can be added inside tank to combine using with solar heaters
12. General testing and operational test carried out for every unit before package in the fact
Company Contact:
- Posted By: Power World Manchinery Equipment Co., Ltd
- Phone: 86075533850784
- Address: BaoAn District,ShenZhen China., Shenzhen, Guangdon, shenzhen, China
- Email:
- Website: http://www.heatpumpcn.com
Published date: September 12, 2013 -
- Business Description: Power World Machinery Equipment Co., Ltd was established in 1998,has
been OEM for 10 years.Our mainly products are includes air source heat
pumps, diesel generations and so on. When you work with us, you will
always find a competitive price for the products because you are
dealing with a professional organization backed by ten years of
experience. As one of the leading manufacturers in heat pump industry
in China, we pride ourselves on our reputation for marketing high performance
price products.
In the field of the diesel generator,we mainly focus is on small and large diesel powered generators from 8kW to 3000kW. We specialize in designing with famous brand engine like Cummins, Detuz, Deawoo, Deere, Perkins, Volvo, Shangchai and Weichai.
In the field of heat pump,we have launch three series:commercial heat pump series, swimming pool heat pump series,household heat pump series.Most of our products asopt famous brand like Copeland,SAGINOMIYA and so on.
We possessed international advanced whole set equipment of manufacture and test, our factory was operated strictly according to the standard of ISO 9001 2000, and our products have been widely proved by CCC Certificate and CE Certificate. We have established nationwide sales and service network, and we sincerely welcome your coming to create mutual benefits together.
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Swimming Pool Heat Pump For SalesGeneral Manufacturing - Power World Manchinery Equipment Co., Ltd - China - September 12, 2013 - contact company for price
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Commercial Heat Pump Water Heater For House Supply 24 Hours HotGeneral Manufacturing - Power World Manchinery Equipment Co., Ltd - China - September 12, 2013 - contact company for price
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Pool Heat Pump China Top Ten Professional ManufacturerGeneral Manufacturing - Power World Manchinery Equipment Co., Ltd - China - September 12, 2013 - contact company for price
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text: I had my 2nd set of tube surgically removed after having it in my ear for 11 years ago.I am curious was having the tube removed the right thing to do? I had middle ear infection in the summer that causes some pain as well as drainage. I am also curious from the time of the surgery to get the ear tube removed, how should should I wait until I get back to physical activity? Some of the activity I am refeering to are the following: running, cycling, and tennis.


text:
All right, it's 10:00 AM and no one has had a mid-morning nosh yet? Don't tell me that the Beltway is devoid of coffee and bagel shops? Ah, the non-existent nourishment… a staple of 24. That and the lack of exciting costume changes. I'm already getting tired of seeing Jack in that trench coat. But if I wanted fun clothes I'd watch Sex and the City (which I do – in case there was a bet going or something).
And on with the show.
Taylor's Secretary of State comes to see her with the unfortunate news of Colonel Dubaku's demands. As we expected, he's threatening more disastrous events like planes crashing and whatnot if Taylor does not move the US troops out of Sangala. And we've noticed that at every turn, her Chief of Staff Ethan Kanan suggests that a U.S. military presence in Sangala might not be so peachy. And now, with the line clearly drawn in the sand, Kanan keeps pushing for withdrawal. I'd agree with him, except for his most interesting and disturbing conversation with poor First Gentleman Henry Taylor.
Ethan told Henry that he needs to forget this conspiracy theory regarding his son's death. According to Ethan, Roger was about to be investigated by the SEC for some unsavory money dealings. Again, this goes back to what we've learned earlier during Redemption — it was Chris, Roger's friend who was involved in something troublesome. Roger was ready to try and help Chris before Nichols (Chris's boss) and baddie Jon Voight's "Jonah Hodges" had Chris killed. And we knew there was a connection between Nichols, Hodges, and Dubaku as well.
So, can we, and more importantly Taylor, trust Hodges? More questions of trust and betrayal come up when Jack gets the chance to interrogate Tony. Stoic Jack must still be struggling to understand that Tony is not only still alive, but not a very nice guy. Certainly Boss Moss (that does just roll off the tongue nicely, doesn't' it?) is not thrilled to have Jack anywhere near Tony, but Renee convinces him that it just might work. Oh, and it does work, just not the way that anyone expected, least of all Jack.
Tony really does not say much at first, but he finally talks about how he became embittered after Michelle died, and held the government responsible. Although he didn't use words like "embittered." And he hopes Jack will commiserate, so he reminds him about Audrey, Kim, Teri – that his personal life has been all kinds of f**ked up since working for the government. Jack gets appropriately pissed and does a trademark slam-against-the-wall move on his old pal. Just what Tony was hoping for.
No, he didn't work a fancy self defense technique, instead he mutters, "I really can't lie", or maybe it was "eat more pie." But when Moss and Renee burst into the interrogation room, Jack leaves and makes a phone call. Ah, it was "deep sky", an old CTU code phrase. So, Jack responded the way he was trained by punching in a number and giving the code.
And this was a sweet moment; Jack is connected to Bill! I didn't realize I was holding my breath until I heard Bill's voice. Didn't know exactly when he and Chloe were due back, just that they would be part of the show. Nice that they didn't keep us waiting for too long. So, the thing is, Tony has actually been working undercover. Now he's not all sweetness and light and puppy dogs, he actually was recruited by Emerson, and partook in some nasty deeds, because of his deep-seated resentments. But when he realized what Emerson was planning do with the CIP, he decided to contact Bill Buchanan and "turn."
Elsewhere in the FBI office, Renee had put Janis on a security task. She told her about the suspected breach, and while Janis is working computer magic, she finds Sean Hillinger's name come up where it didn't belong. Cue dramatic music! But this, for now, is one of 24's famous red herrings; we think Sean is bad because he poised his eyebrows just so, stared for a fraction longer at someone, or stabbed his computer keys with too much vigor.
But this vigor – as it turns out – was induced because his wife is actually on board one of the many planes that have been put in holding patterns in the sky. Sean has been trying to get more information on the flight's status. Come on, wouldn't you?
And rounding up the hour is the exciting escape from the building, Jack has to temporarily knock out Renee with a sleeper hold, and he purloins the key-card. Poor Renee is all confused now, she thought she really could trust Jack. (Psst, Renee, you can – but you're not supposed to know that yet). Anyway, Bill picks them up and brings them back to his lair – I mean his headquarters. (Did I hear right that it was an old, closed branch of CTU?)
And on we continue, back to the Oval Office. Poor President Taylor. She certainly does seem to have a good sense of command, and she's got a fierceness tempered with humanity. She's got to make an impossible decision, but even in her indecision she's got a lot more class than Charles Logan did when he was faced with the choice of letting US citizens die from Sentox gas poisoning, or let his important ally – the Russian President Subarov – die from assassination. Allison Taylor thankfully does not express herself with those old nasty grimaces.
And Ethan is still pushing for withdrawal. What's the matter Ethan, do you own a diamond company over there or something? Taylor meets again with the former Sangalan Prime Minister, Matobo and explains the situation. He empathizes but has to push for the end of Juma's regime, with American aid of course.
After Jack dutifully admires Bill and Chloe's setup, Bill explains that they don't report to anyone. They've found evidence of corruption very, VERY high up in the government (no shit Sherlock!) and need to bypass certain folk in order to bring down Dubaku – and then Juma. They also need to get Tony back into Emerson's group, and decide to bring Jack along as a bonus spy.
Emerson is no fool though, he initially welcomes Jack, but then decides that he's not necessary. Jack, who has been waiting down in the rumpus room with a couple of goons, senses that things might not go well, and brings down both goons. Emerson then decides that Jack just might be someone to have on your side in a tight spot. He also gives them the next part of the plan. Kidnap Matobo. Cue dramatic music again.
While they head to nap Matobo, Renee and Janis have gone to the hospital. Our old pal Tanner (the sniper) has regained consciousness. My goodness it's like watching Jack and Chloe. Renee is a quick study, and soon enough she's waving her gun around, pointing it at Tanner, pointing it at the mirror, at bad art on the wall… No, of course, not, but she does threaten Tanner with her gun and then she blocks his ventilator. Meanwhile Janis is outside trying to stall Tanner's lawyers with a song and dance. No Chloe scowling, but she does make geek girls proud everywhere.
Before Jack, Tony and Emerson reach Matobo, the FBI has been able to warn him of trouble. He and his wife are hustled off to a safe room, with Jack and Co. close behind. Luckily no Chinese consulates were harmed during the filming of that scene. Next week could be another story though. Stay tuned.
Best quote:
"Who ever it is, they're good!" (Janis Gold, referring to Chloe's busting in on the FBI's surveillance systems)Powered by Sidelines


text:
St. Ann Center will begin its second annual Winter Farmers Market on November 5. With over 20 seasonal vendors there is something for everyone. End of the season produce is available usually through the beginning of December. Many of the vendors from the South Shore Farmer's market move indoors to St. Ann Center in November.
There will be seasonal, natural and organic produce, preserves, canned goods, homemade items, jams, jellies, soups, soaps, lotions, makeup, jewelry, personal items and much more.
There will be free door busters for the first 50 people, amazing raffles, live musical
entertainment, and product demonstrations. There will even be added crafts for the children on certain days and special events/ classes during the season.
St. Ann Center is located at 2801 East Morgan Avenue, Milwaukee, 53207. The hours for the market are 9:00a.m. to 1:00 p.m. and will happen every Saturday from November 5 through April 7, 2012.
On November 5, 2011 is our Market Kickoff - Doorbusters for the first 50 people and a $1.00 Magic Show for the Kids featuring "Rainbow Randa"
November 12 is our Veteran's Day Celebration. We will have a salute to our Veterans and a Veteran's Day Craft for the kids.
On November 19 Join us as we remember to be thankful for the little things in life. Don't forget to look for the fixings for your Thanksgiving dinner. We will also have a Thanksgiving craft for the kids.
On November 26 is Black (Friday) Saturday at the Market - Come shop till you drop for your loved ones. Stop by for our special coupon flyer from you favorite vendors.
There is something for everyone. Stay warm and stay indoors while you enjoy all that a traditional farmer's market has to offer!
For more information go to www.stanncenter.org/Farmersmarket
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- St Francis School District Rummage Sale
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Gift Premium Subscription To acsimmo2
Join Date:02-16-2011
Last Activity:03-09-2011 1:09 AM
1) Crisp Amber Caramel Ale:
recipe found here:http://beerrecipes.org/showrecipe.php?recipeid=1117
Notes: BLAND. not a bad beer, definitely drinkable but i will never make this beer again.
2)24 Black Silk:
recipe found here:http://beerrecipes.org/showrecipe.php?recipeid=465
Notes: Made with French Vanilla coffee. Best beer i ever had. I might be tasting a since of self confidence no one else can taste, but to me, i love it.
3) Tamalpais Wit
recipe found here: http://beerrecipes.org/showrecipe.php?recipeid=706
Notes: way to much coriander. Overpowers been and gives it a strong spice flavor, next time try only 1/2 ounce coriander.
4)24 Black Silk:
recipe found here:http://beerrecipes.org/showrecipe.php?recipeid=465
Notes: Made with Hazelnut coffee. not as good as french vanilla. Hazelnut is overpowering, use different coffee next time. otherwise good.
- Blackberry Stout; recipe found here: http://beerrecipes.org/showrecipe.php?recipeid=578
- In a Relationship
- Raliegh
Posted in thread: OG question on 02-17-2011 at 08:57 PM
is that 2-3 weeks in secondary fermention or 2-3 weeks after the yeast has been pitched? also,
with this new style of fermentation i am going to try, how long should it sit in primary
Posted in thread: OG question on 02-17-2011 at 06:05 PM
Thanks, i get i read the OG right before pitching the yeast, but im confused when you say i
need to take multiple readings. is the easiest way to just leave the hydrometer in the carboy
when i switch ...
Posted in thread: OG question on 02-17-2011 at 03:37 PM
I have brewed a couple of times and always went by the rule:1 week bucket2 weeks carboy2 weeks
bottlesand been getting good results, but now i am really starting to get into it and realized
that the a...


text:
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Not so known battery saving tips
Hi everyone,
The other day i saw 30% drop in battery overnight. I had already researched battery saving ideas and had the usual settings (i.e., google now off, dim screen, etc.)
I came across 3 different tools that helped me big time...i am currently sitting at 80% after 16 hours
1) application manager -> swipe right to all - go through the list and disable "show notifications" for pretty much every app you don't need a notification for. also unistall updates/diasable/force stop any app you dont need (since there are a lot of random apps, only do this for obvious ones)
2) download (all free) cpu spy (confirm your phone is hitting deep sleep), my data manager (determine which apps are using data), and gsam battery monitor (determine which apps are both keeping your phone awake, and waking your phone). cross reference my data manager and gsm battery with the below.
3) the holy grail as someone referred to it - settings -> data usage -> scroll down to bottom -> manually disable background data for all obvious apps. i personally left my weather, email, and the android os ones alone. it is important to note that if you open your browser (or any other app) it is no longer a background app. you must do the hold home button clear open apps to close it out.
i am pretty good with tech, but am no expert and this worked for me (so far), but if anyone knows better please let me know
nice to see my battery finally in beast mode
my original post (i have a verizon galaxy note 2) - Not so known battery saving tips
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Re: Not so known battery saving tips
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Re: Not so known battery saving tips
I have no issue with my battery for now but maybe these tips will help me in the future.
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Facebook programmer or iphone app programmer
This project received 13 bids from talented freelancers with an average bid price of $815 USD.Get free quotes for a project like this
Project Budget$250 - $750 USD
Total Bids13
Project Description
for my own social website im looking for someone who can add some facebook features in my website
and im looking for a iphone app
please send a private message for details
Skills Required
Looking to make some money?
- Set your budget and the timeframe
- Outline your proposal
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- Forbes
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- Times Online


text:
Objectives: β-Adrenergic receptor kinase (βARK) phosphorylates and thereby inactivates agonist-occupied β-adrenergic receptors (βAR). βARK is thought to play an important role in the regulation of cardiac function. Therefore, we studied βARK activation and its inhibition in intact smooth muscle cells and in cardiomyoblasts. Methods and Results: βAR agonist-stimulated translocation of βARK was monitored by immunofluorescence labelling with specific antibodies and confocal laser scanning microscopy in DDT-MF 2 hamster smooth muscle cells and in H9c2 rat cardiomyoblasts. In unstimulated cells, βARK was mainly located in the cytosol. After βAR agonist stimulation, the βARK signal was partially translocated to the membranes. Liposomal gene transfer of the COOH-terminus of βARK ('βARKmini') as a βARK inhibitor led to functional expression of this protein in both cell lines with high efficiency. Western blots with βARK antibodies showed a gene concentration-dependent immunoreactivity of the 'βARKmini' protein. 'βARKmini'-transfected myocytes demonstrated reduced membrane targeting of the βARK immuno-fluorescence signal. Additionally, the effect of 'βARKmini' on βAR-induced desensitization of myocytic cAMP accumulation was investigated. In control cells, desensitization with isoproterenol led to a subsequent reduction of βAR-induced cAMP accumulation. In 'βARKmini'-transfected myocytes, this βAR-induced desensitization was significantly diminished, whereas normal βAR-induced cAMP accumulation was unaffected. A gene concentration of 2 μg 'βARKmini' DNA/100 000 cardiomyoblasts, and of 0.7 μg 'βARKmini' DNA/100 000 DDT-MF2 smooth muscle cells led to ≈5.9- and ≈5.6-fold overexpressions of 'βARKmini' vs. native βARK, respectively. These gene doses proved sufficient to attenuate β-adrenergic desensitization significantly. Conclusions: (1) βARK translocation was evidenced in DDT-MF2 smooth muscle cells and in cardiomyoblasts by confocal laser scanning microscopy. (2) Feasibility of 'βARKmini' gene transfer to myocytes was demonstrated, and necessary gene doses for βARK inhibition were titered. (3) Overexpression of 'βARKmini' functionally interacted with endogenous β-adrenergic signal transduction, leading to sustained cAMP accumulation after prolonged β-adrenergic stimulation.
Time for primary review 38 days.
1 Introduction
Several biochemical mechanisms contribute to β-adrenergic receptor desensitization [1]. Most importantly, agonist-occupied active receptors become phosphorylated by the specific β-adrenergic receptor kinase (βARK) [1, 2], facilitating the subsequent binding of the inhibitor protein, β-arrestin, to the receptors [3]. Thereby, receptor function is inhibited by up to 70% [4, 5]. Phosphorylated β-adrenergic receptors lose their capacity to activate subsequent second-messenger systems, such as adenylyl cyclase [1, 2]. Alternative mechanisms, such as the reduction of receptor number by down-regulation, become operative only after 4–12 h [1]. βARK is thought to play an important role in the regulation of cardiac function. In a previous study, we found a significant increase in cardiac βARK activity and mRNA levels in human heart failure [6]. This activation specifically involved the mRNA for βARK-1, but not for βARK-2 or for β-arrestin [7]. The activation of βARK seems to be a very early event during the development of heart failure, since it preceded the alterations of β-receptor density or of G-proteins in the model of pacing-induced heart failure in the pig [8]. Chronic exposure to β-blockers, in turn, decreased cardiac βARK activity in cytosol and membranes [9]. Also, cardiac ischemia increases βARK activity in cardiac membranes [10].
Pivotal work by Lefkowitz and collaborators has shown that activation of βARK is closely related to a translocation of the enzyme from the cytosol to the membranes. βARK membrane targeting is specifically mediated by G-protein βγ-subunits. βγ-Subunits can bind to the pleckstrin-homology (PH) domain, which forms part of the COOH-terminus of βARK [11, 12]. Overexpression of the COOH-terminus of βARK-1 (amino acids 495–689, 'βARKmini') has subsequently been shown to exert a 'scavenging' effect on a variety of mechanisms mediated by βγ-subunits, such as α2-adrenergic receptor-mediated stimulation of adenylyl cyclase II [13], or muscarinic M2-stimulated inositol phosphate turnover [13]. 'βARKmini' also inhibits Gβγ-stimulated phosphorylation of rhodopsin in vitro [14]. Most importantly, overexpression of 'βARKmini' in the hearts of transgenic mice also improved cardiac contractility, as demonstrated by heart catheterization in vivo [14]. Up to now, however, direct evidence for the role of 'βARKmini' on βARK translocation in intact cells and in myocytes has not been provided. Generally, βARK translocation has been investigated in isolated, prepared cell fractions, instead of whole cells. Moreover, the inhibitory effect of 'βARKmini' on adenylyl cyclase activity has only been shown in reconstituted systems of recombinant proteins [15]. In contrast, direct effects of 'βARKmini' on endogenous β-adrenergic desensitization in myocytes have not been demonstrated, since adenylyl cyclase activity in myocardial membranes was not altered by 'βARKmini' [14].
Therefore, we set out to characterize βARK translocation directly in intact myocytes. Similarly, we intended to investigate more precisely the effects of 'βARKmini' on the endogenous β-adrenergic system in cardiomyocytes. Thereby, we sought to determine adequate gene concentrations of 'βARKmini' needed for relevant inhibition of β-adrenergic desensitization in smooth muscle cells and in cardiomyocytes.
2 Methods
2.1 Cell culture
DDT-MF-2 smooth muscle cells derived from hamster ductus deferens leiomyosarcoma were a kind gift of M. Lohse (Würzburg). The cells were grown as monolayers in Dulbecco's modified Eagle's medium (DMEM), supplemented with 10% fetal calf serum (FCS), non-essential amino acids, 2 mmol/l glutamine, penicillin (100 IU), and streptomycin (100 μg/ml) in 5% CO2 in water-saturated air at 37°C.
H9c2 cardiomyoblasts (ATCC CRL 1446, cardiac myoblasts from rat) were purchased from American Type Culture Collection (Rockville, MD, USA) and cultured as monolayers in DMEM, 10% FCS, 2 mmol/l glutamine, penicillin (100 IU/ml), and streptomycin (100 μg/ml) in 7% CO2 in water-saturated air at 37°C. Both cell lines were transfected transiently by liposomal transfer, using 20 μg/ml lipofectamine (Gibco/BRL, Gaithersburg, MD, USA) and varying concentrations of pβARKmini (as indicated), a kind gift of R.J. Lefkowitz (Duke University, Durham, NC, USA). Alternatively, the myocytes were transfected with control plasmid pRK which did not contain the specific insert. For transfer, cells were freshly plated and grown for 24 h. Then, medium was exchanged for preincubated liposome/plasmid mix in Optimem medium (Gibco/BRL, Gaithersburg, MD, USA) for 6 h. Cells were harvested 36 h later. The study conforms with the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23, revised in 1985).
2.2 Western blot
For immunoblotting, cells were scraped from dishes, placed in 500 μl of TE buffer (20 mmol/l Tris-HCl, pH 7.5, 1 mmol/l EDTA) and sonicated for 3 min. Where indicated, cytosolic and particulate fractions were prepared as follows: Cells were homogenized for 30 s in a Polytron tissue mincer and centrifuged for 30 min at 100 000×g. The supernatant represented the cytosolic fraction. The pellet was resuspended in TE buffer to be used as membrane fraction, and homogenized again. The volume of both fractions was reduced by filtration in a microconcentrator Centricon 30 (Amicon, Düsseldorf, Germany) to yield 100 μl. Total protein levels in the concentrates were determined according to Bradford, as described [6]. Samples containing 30 μg of protein were boiled with Laemmli buffer for 5 min, and electrophoresed on 12% SDS-polyacrylamide gels. Proteins were then blotted onto Amersham Hybond-C membranes. Efficiency of transfer was verified by Ponceau red staining of the blots. The blots were blocked with 2% ovalbumin in TBS. Immunogenic signals of native βARK or 'βARKmini' were detected by probing with rabbit GRK-2 antibody directed against the COOH-terminus of βARK-1, purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), followed by peroxidase-coupled secondary antibodies (from Sigma, Deisenhofen, Germany) using chemoluminiscent substrate (ECL, Amersham, Braunschweig, Germany) and Kodak XAR films. Densitometric analysis of the Western blots was carried out using NIH graphics software.
2.3 Confocal laser scanning microscopy
pβARKmini- or pRK-transfected myocytes were grown in 6-well tissue culture plates on microscope cover slips for 24 h. Stimulation of the cells was carried out with 1 μmol/l (−)-isoproterenol in PBS. After rinsing 3 times with PBS, cells were fixed for 15 min with 4% para-formaldehyde in PBS. Cells were then permeabilized with 0.1% saponin for 10 min. To avoid non-specific antibody binding, cells were treated with 10% FCS in DMEM for 30 min prior to labelling with GRK-2 antibody (1μg/ml). After incubation with goat anti-rabbit IgG FITC-conjugate (Sigma, Deisenhofen, Germany), the coverslips were mounted upside-down on microscope slides and sealed with nail polish. For microscopy, samples were visualized by differential interference contrast microscopy (Nomarski optics) and by excitation with a low-power argon laser at 488 nm.
Digitalized microscopic images of fluorescent myocytes were analyzed densitometrically. The signal intensity in representative membrane areas was quantified as arbitrary units.
Transfected myocytes were grown on 6-well tissue culture plates for 24 h. Cells to be used for fluorescence analysis were treated as described above for immunofluorescence microscopy and removed from the plates with trypsin-EDTA prior to FACS analysis. Samples were analyzed on a FACScan cytometer (Becton Dickinson) equipped with an argon laser. Cell signals were plotted in a forward versus side scatter, and 50 000 particles were analyzed at a rate of less than 100 events per second. A gate was defined in order to exclude all debris and aggregates. Logarithmic amplification was used for both, the fluorescence and light scatter signals. The mean intensity of immunofluorescence was used as a quantitative measure of antigen representation on cells.
2.5 cAMP accumulation in intact myocytes
pβARKmini- or pRK-transfected myocytes were grown in 6-well Falcon tissue culture plates to 80–90% confluence. They were incubated in PBS containing 1 mmol/l of the phosphodiesterase inhibitor, IBMX, for 30 min. For desensitization, cells were incubated in 1 μmol/l (−)-isoproterenol in PBS and IBMX for 15 min to cause maximal stimulation of the cells' β2-adrenergic receptors. Controls were treated equally without isoproterenol. Following aspiration of the buffer, the cells were rinsed 4 times with PBS and incubated with various concentrations of (−)-isoproterenol for 15 min. The incubation was stopped by addition of 0.5 ml of boiling TE buffer at pH 7.5. The cells were then scraped off the plates and were separated from soluble material by centrifugation at 5000×g for 10 min. The pellet was kept for protein determination.
The cAMP content in the supernatant was determined by a [3H]cAMP assay system purchased from Amersham (Braunschweig, Germany) according to instructions supplied by the manufacturer.
3 Results
3.1 Western blots
Western blotting of whole cell extracts of both DDT-MF2 smooth muscle cells and H9c2 cardiomyoblasts showed a weak, but clear signal at 80 kDa, the expected size of native βARK (Fig. 1A,B, lane 7). A second, non-specific band at a size of ≈60 kDa was obvious on cell extract blots of DDT-MF 2 cells, and was also present on blots exposed to preimmune serum instead of specific βARK antibody (Fig. 1A, lane 8). After liposomal transfection of 0.25–5 μg pβARKmini to 100 000 DDT-MF2 smooth muscle cells or of 0.3–3 μg pβARKmini to 100 000 cardiomyoblasts, a gene dose-dependent band appeared at a size of 27 kDa (Fig. 1A,B, lanes 2–6). Both native βARK signals and 'βARKmini' signals were competitively inhibited by increasing concentrations of the GRK-2 peptide. Lane 9 of Fig. 1A,B demonstrates the effect of 10 μg/mL GRK-2 peptide on the signal of the extracts shown in lane 4. Analysis of signal intensity by laser densitometry revealed that the transfer of 0.7 μg pβARKmini to 100 000 DDT-MF 2 smooth muscle cells led to ≈5.6-fold overexpression of 'βARKmini', compared to native βARK. Similarly, 2 μg pβARKmini transferred to 100 000 cardiomyoblasts led to ≈5.9-fold overexpression of 'βARKmini'.
Western blotting of membrane extracts of both cell lines demonstrated a strong signal of the 'βARKmini' transgene in cytosol and a weaker signal in the membranes. Fig. 2A shows that in membranes from DDT-MF 2 smooth muscle cells transfected with control vector pRK, the signal for native βARK was increased after stimulation with 1 μmol/l isoproterenol for 30 min (lanes 3 and 4). In contrast, this increase was clearly diminished in smooth muscle cells transfected with 0.7 μg pβARKmini/100 000 cells (lanes 1 and 2). No detectable differences in the cytosolic immunosignal of native βARK occurred under any condition (lanes 5–8). Similarly, cardiomyoblasts transfected with 2 μg control vector pRK/100 000 cells showed an increased native βARK membrane signal after agonist exposure (Fig. 2B, lanes 3 and 4). This increase was markedly attenuated in cardiomyoblasts transfected with 2 μg pβARKmini/100 000 cells (lanes 1 and 2). Again, the cytosolic signal for native βARK was not altered under any condition (lanes 5–8).
3.2 Flow cytometry
Flow cytometric analysis was performed on 50 000 cells after being detached from culture flasks. Fig. 3A demonstrates the analysis of the fluorescence intensity of DDT-MF2 smooth muscle cells which had been incubated with the secondary antibody only, or with specific GRK-2 antibody, and of myocytes which had been transfected with 0.7 μg pβARKmini/100 000 cells. The smooth muscle cells which had been incubated with the specific GRK-2 antibody showed significantly higher fluorescence intensity than cells exposed to FITC antibody only. After gene transfer with pβARKmini, a second peak of nearly 10 scale units higher intensity appeared. The areas under the two curves were estimated by geometric analysis. Based upon this approximation, ≈40% of the DDT-MF2 smooth muscle cells exposed to the 'βARKmini' plasmid had gained additional immuno-fluorescence intensity, corresponding to the cytoplasmic expression of 'βARKmini' in addition to native βARK.
Similarly, cardiomyoblasts showed specific immunolabelling with the GRK-2 antibody (Fig. 3B). After gene transfer with 2 μg pβARKmini/100 000 cardiomyoblasts, ≈25% of total cells gained significantly higher specific fluorescence intensity, as estimated by geometric analysis of the surfaces under the two curves.
3.3 Confocal laser scanning microscopy
FITC-labelled cells were also subjected to confocal laser scanning microscopy. Fig. 4 shows representative light transmission and fluorescence images of DDT-MF2 smooth muscle cells transfected with either control plasmid pRK or pβARKmini and stimulated with 1 μmol/l isoproterenol or vehicle. Fig. 4A shows the absence of specific signals in cells exposed to FITC antibody only. Fig. 4B shows the largely homologous cytosolic distribution of native βARK in unstimulated cells. After exposure to isoproterenol, the βARK signal was significantly translocated to the membranes, whereas less signal was found in the cytosol (Fig. 4C). Translocation reached its maximum within 10 min and did not increase thereafter. After transfection with 0.7 μg pβARKmini/100 000 cells, approximately 30% of the cells showed a markedly higher fluorescence signal. Such a cluster of positive cells is shown in Fig. 4D. After stimulation of 'βARKmini'-expressing cells with isoproterenol, markedly less translocation of the native βARK signal to the membranes occurred (Fig. 4E). Densitometric analysis of the membrane areas of 'βARKmini'-expressing and 'βARKmini'-negative cells after agonist stimulation showed significantly higher signal intensity in cells which did not express 'βARKmini' (Fig. 6A). DDT-MF2 smooth muscle cells transfected with more than 1.5 μg pβARKmini/100 000 cells changed their morphology, and a significant number did not survive the gene transfer. Lower concentrations of pβARKmini, however, did not change cell morphology detectably.
Also in cardiomyoblasts, native βARK was translocated after agonist stimulation (Fig. 5B,C). Fig. 5C demonstrates that stimulation with isoproterenol led to a somewhat asymmetrical increase in the βARK signal to one side of the membrane. This was a typical finding in almost all stimulated cardiomyoblasts investigated. Additionally, the cytosolic βARK signal appeared more granular after agonist stimulation, possibly corresponding to an internal translocation of βARK to intracellular microsomal structures, as suggested by some authors [16, 17]. Again, translocation reached its maximum within 10 min. After transfection with 2 μg pβARKmini/100 000 cells, nearly 25% of the transfected cells gained markedly higher βARK immunofluorescence, due to the additional expression of 'βARKmini' (Fig. 5D). 'βARKmini'-expressing cardiomyoblasts showed no detectable translocation of the native βARK signal after agonist stimulation (Fig. 5E). Densitometric analysis of the membrane areas of 'βARKmini'-expressing and 'βARKmini'-negative cells after agonist stimulation showed significantly higher signal intensity in cells which did not express 'βARKmini' (Fig. 6B). Higher doses of pβARKmini >5 μg/100 000 cardiomyoblasts were cytotoxic.
3.4 Adenylyl cyclase desensitization
cAMP concentrations were measured in intact myocytes after stimulation with isoproterenol. Fig. 7 demonstrates that isoproterenol in concentrations ranging from 0.001 to 10 μmol/l led to intracellular accumulation of cAMP. On average, cAMP levels were raised from 1 to 38 pmol/mg protein in DDT-MF2 smooth muscle cells, and from 13 to 50 pmol/mg protein in cardiomyoblasts. In DDT-MF2 smooth muscle cells transfected with 0.7 μg pβARKmini/100 000 cells (corresponding to ≈5.6-fold overexpression of 'βARKmini' vs. native βARK, as estimated from Western blots), basal and isoproterenol-induced maximal cAMP levels were unaltered, with EC50 values ranging from 100 to 150 nmol/l. When the cells were desensitized by preincubation with isoproterenol for 15 min, the response to a subsequent, repeated exposure to the β-agonist was markedly blunted. This desensitization reached significance only in pRK-transfected smooth muscle cells (P<0.02), whereas cAMP accumulation after agonist exposure did not differ significantly in pβARKmini-transfected cells. After prior exposure to isoproterenol, 'βARKmini'-transfected cells accumulated significantly more cAMP (Fig. 7A, P<0.05). When cells were transfected with lower gene concentrations of pβARKmini (0.1 μg/100 000 cells, corresponding to equal expression of 'βARKmini' and βARK), they could still be subjected to significant desensitization, accumulating significantly less cAMP after pre-exposure to isoproterenol (P<0.05, Fig. 8A). pβARKmini doses exceeding 1.5 μg/100 000 cells (≈15-fold overexpression of 'βARKmini') led to a change in cell morphology, and to a decrease in cAMP response to isoproterenol (Fig. 8C).
Similarly, cAMP accumulation was identical in cardiomyoblasts transfected with 2 μg pβARKmini/100 000 cells (corresponding to ≈5.9-fold overexpression of 'βARKmini'), and in cells transfected with 2 μg of pRK. Also in cardiomyoblasts, expression of 'βARKmini' did not affect regular β-adrenergic stimulation. After pre-exposure to isoproterenol, however, the cAMP response elicited by isoproterenol was significantly higher in pβARKmini-transfected cardiomyoblasts than in pRK-transfected cardiomyoblasts (Fig. 7B). Expression of 'βARKmini' led to a virtually complete reversion of desensitization. In contrast, cardiomyoblasts transfected with 0.1 or 0.3 μg of pβARKmini (corresponding to ≈0.7- and ≈1.1-fold overexpression of 'βARKmini', respectively) showed a pattern of desensitization which did not differ significantly from that of control cells (not shown).
4 Discussion
The present study characterizes βARK translocation in intact myocytes, as detected by immunofluorescence labelling. βARK translocation and β-adrenergic desensitization could be inhibited effectively by a plasmid-induced expression of the inhibitor protein, 'βARKmini', in intact myocytes and cardiomyoblasts. Transfection of more than 0.7 μg of pβARKmini to 100 000 DDT-MF 2 smooth muscle cells, and of 2 μg of pβARKmini to 100 000 cardiomyoblasts led to sufficient expression of this protein to significantly attenuate endogenous β-adrenergic desensitization in these myocytes.
βARK selectively phosphorylates and thereby inactivates agonist-occupied β-adrenergic receptors. This process has been linked to translocation of the enzyme from a cytosolic compartment to the membranes [1]. We have demonstrated that specific antibodies can be used to effectively label βARK in permeabilized smooth muscle cells and cardiomyoblasts. Confocal laser images of these cells suggest that βARK is actually translocated rapidly to the myocytic membrane after β-agonist stimulation. In DDT-MF2 smooth muscle cells, which display a narrow cytoplasm, translocation seemed to be directed to all sides of the myocyte, and the cytosol appeared to clear rapidly of the βARK immunosignal after agonist exposure. Since small cytoplasmic margins compromise the microscopic differentiation between cytosol and membranes, we compared these effects with another myocytic cell line with larger cytoplasm. Our data corroborate the results of an earlier publication on the translocation of βARK activity in DDT-MF2 smooth muscle cells after β-adrenergic stimulation [18].
In cardiomyoblasts, which possess a pronouncedly larger cytoplasm, translocation appeared asymmetrical in all cells investigated. The homogeneous cytosolic distribution of the βARK immunosignal seemed to assume a more granular appearance after the agonist challenge, possibly corresponding to an internal translocation to a microsomal compartment, as suggested by others [16, 17]. This issue can, however, not be decided with certainty, since the spatial resolution of fluorescence imaging is limited. Translocation reached a maximum within 10 min and was not further increased thereafter. This time course corresponds quite well with previous studies in which the capacity of cell extracts to phosphorylate rhodopsin or β2-adrenergic receptors was used to determine βARK kinetics [19].
Considerable interest has evolved in inhibiting cardiac βARK activation since overexpression of the inhibitor protein 'βARKmini' has been shown to increase cardiac contractility in transgenic mice [14]. These animals, however, developed high constitutive expression of 'βARKmini', since the transgene was coupled to the α-myosin heavy chain promotor which was active during the entire life of the animals. Our study shows that also external plasmid-derived gene transfer of 'βARKmini', which leads to less expression of the inhibitor protein, can effectively inhibit β-adrenergic desensitization in intact myocytes. Our results demonstrate that the effect of 'βARKmini' can be tested biochemically by estimating the amount of immunolabelled βARK which translocates after agonist stimulation, and by quantifying intramyocytic cAMP accumulation after previous desensitization. This finding is of some relevance, since measurement of adenylyl cyclase activity in membranes prepared from 'βARKmini'-transgenic and control hearts did not reveal any differences [14]. Normal cAMP accumulation was not affected by 'βARKmini', but sustained cAMP formation after prolonged agonist stimulation occurred. Therefore, 'βARKmini' might interact specifically with the mechanisms which lead to rapid β-adrenergic refractoriness in heart failure. We conclude that the analysis of single myocytes contributes to the understanding of the biological effects of 'βARKmini' in cardiomyocytes.
The amount of plasmid needed for the expression of 'βARKmini' in biologically relevant concentrations was titered to be 0.7 μg for 100 000 DDT myocytes, and 2 μg for 100 000 cardiomyoblasts, corresponding to similar overexpressions of 'βARKmini' vs. native βARK (5.6- and 5.9-fold, respectively). Since several reports have described the possibility of cardiac gene expression after direct plasmid injection [20, 21], the direct application of a plasmid by intracardiac injection might suffice to attenuate β-adrenergic desensitization in vivo. This information about gene effects in cardiomyocytes should be useful for further studies, since several investigators have described a gene concentration-dependency of plasmid-derived expression of recombinant proteins in vivo. Such an example was given recently by the successful transfer of a plasmid encoding VEGF into the popliteal artery of a patient, leading to enhanced collateralisation and better clinical outcome [22]. In this study, effects started at a plasmid dose of 2000 μg. An even higher biological effect of 'βARKmini' can be expected from future experiments with viral transfer systems which will allow even more efficient gene transfer to a higher percentage of cells.
4.1 Limitations of the study
In the present study, we distinguished the signals of 'βARKmini' and of native βARK by their sizes on Western blots, and by their localisation by confocal laser microscopy. On Western blots, the bulk of 'βARKmini' was found in the cytosolic fraction. A small amount of 'βARKmini' in the particulate fraction is probably due to massive overexpression of the protein, and to minor contamination during preparation. This is in line with previous studies [13], since 'βARKmini' is known to be a strictly cytosolic protein which lacks all components necessary for membrane translocation, namely the site of isoprenylation. Therefore, the translocation observed by means of immunofluorescence (i.e., the gain in membrane signal after agonist stimulation) can be assumed to be due to translocation of native βARK. In contrast, the cytosolic signal for native βARK could not be studied in 'βARKmini'-expressing cells by confocal microscopy. This signal could, however, be differentiated on Western blots of cytosolic fractions.
5 Note added in proof
During the review process of the present manuscript, an extensive study on the effect of adenoviral transfer of 'βARKmini' to cardiomyocytes was published. This paper corroborates our data on the effects of 'βARKmini' on β-adrenergic desensitization (Drazner et al., J Clin Invest 1997;99:288–296).
We thank Ms. Claudia Rokitta for skilful help with the FACS analysis. This study was supported by a grant of the Deutsche Forschungsgemeinschaft (Un 103/1-1).
Isner JM, Pieczek M, Schainfield R et al. Clinical evidence of angiogenesis after arterial gene transfer of phVEGF165 in patient with ischemic limb. Lancet 1996;348:370–374.
- isoproterenol
- gene transfer techniques
- cytosol
- fluorescence
- fluorescent antibody technique
- genes
- tissue membrane
- phosphotransferases
- beta-adrenergic receptor
- translocation (genetics)
- antibodies
- cyclic amp
- ddt (insecticide)
- agonists
- protein overexpression
- muscle cells
- microscopy, confocal scanning laser
- attenuation
- receptor desensitization
- myocytes, smooth muscle


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Because Everyone Deserves a Great Smile!
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Riad Dar Dmana Guesthouse
20 Rue Salaj Douh Batha, Fez, Morocco
What's Included
- Breakfast
- Linen
- Luggage Room
- Towel
- Air conditioning
- Free Breakfast
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Check-in / Check-out
Earliest check-in: 00:00
Latest Check-out: 02:00
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Group of 15 or more traveling to Fez?
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The Riad Dar Dmana is located in the centre of Fez city.Distance from the airport 15 km the hotelDistance from the Railway station is approx 2 km the hotel


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Every day for the next 3 days, I would go to the log cabin and see Adam. We got to know each other and trust each other and I can now say that I am totally in love with him. Nobody knows what
happened to him and no one's asking me any questions on his whereabouts. The cops got involved and now there's a search team looking for him all over the town. Even his dad is worried about
"I don't believe you," he said when I told him.
"I'm not lying. He loves you and you can't deny it."
"I can and I do. I hate him and he doesn't even care I'm gone. I'll bet he's even happy." His eyes were filled with confusion. He couldn't help but turn away when a tear ran down his pale, snow white skin.
"It's okay to cry, you know. I understand how you feel. You're confused and lonely and you just want your parents to notice you for once. I felt the same way when my dad left for Australia."
He turned around, tears rolling down his face. He took off for the stairs and rushed up to the small room. I slowly went up and opened the door. He was on the bed curled up in a ball.
"You are such a baby," I say as I go sit next to him. I wrap my arm around his shoulders and he leans on mine.
The next day, he was outside throwing rocks into the river. Each toss was hateful, I could tell. He hated the river after what happened with Mr. Gorge. That might have been the reason he stopped coming here a long time ago.
"Your mom thinks you're dead," I tell him, trying to convince him to go back home.
"Why should I care? She's not even my real mom," He said coldly. I looked at him with confusion.
"What?" I asked totally lost.
"They aren't my real parents. My real mom is in mexico. My real dad is dead. I've lived in 23 foster homes and have been to 37 different families. I can't go back and face them. I don't want to. They don't care and they never will. None of them cared. None of them will. My real mom doesn't even care. She left me here on the streets and ran to Guatemala. Yeah, I'm living in a real life hell. It sucks. Get over it like I did. It was hard but still. I had to...." His voice trailed off.
The next day in history class, I was the first one there and was all alone in the room with Mr. Gorge. He was sitting at his desk and there was an awkward silence. I was worried about him coming anywhere near me and was so nervous my palms were sweating.
"What ever happened to Mr. Sullivan?" He practically hissed that at me.
"I don't know who you're talking about," I lie. I hope he can't tell.
"Don't play dumb with me. I saw the way he looked at you and the way you were clinging to him. Don't be so nervous, girl. I paid my dues now all you need to do is get over it."
Did he just say that?
"Okay..." I say, my voice quiet and nervous. He was glaring at me with dark eyes that were cold like winter. That's when everyone finally started to file in, breaking the silence. I still don't understand the reasoning for letting him teach again.....
After school, I decided to hang out with my friends at the skate park.
"Where have you been?" asked my friend, Collin. He's been my friend since third grade.
"None of your business."
"I see how it is. You have a new boyfriend that you don't want me to know about. Is that right?" I just start blushing and shook my head.
"I knew it!" Said Lucy. "Why didn't you tell me? I thought I was your best friend!"
"You are but he's NOT my boyfriend. We just kinda hang out sometimes." I told her.
"You've been 'hangin out' every day for the past two weeks, Zoe," noted Collin. I just gave him a dirty look. Kinda like the one Adam gave me.
"So when are we gonna meet this mystery guy of yours?" Says a voice from behind us. We turn around and it's Ronnie.
"Hey, I thought you were in Italy," says Collin.
"I was. But now I'm back," he says casually.
"Hey guys, I gotta go," I say.
"Okay. Have fun with your boyfriend," Collin teases. I give him another dirty look before I take off for home.
At home I get cleaned up then take Stephanie to see Adam. When we get there, I knock on the door and wait. When he opens the door, he looks like he hasn't slept in days.
"You okay?" I ask, concern filling my voice.
"Fine. Who's this?" He says looking behind me at Stephanie.
"This is my daughter, Stephanie..." He chuckles. I'm nervous and he can tell.
"She's beautiful," he says. She steps out from behind me, already warming up to him. He opens the door wider and lets us in. When we're inside, there's a warm smell of vanilla that fills the air.
"So, what's up?" He asked.
"Nothing," I said.
"You know, you don't have to come here every day. If I were you, I'd be tired of myself already."
"I know, and I'm not tired of you. I actually enjoy coming here. It's nice to have someone that actually seems to care." I say.
"I see..."
We follow him up the stairs and into the bedroom and sit on the bed.
"So how old are you?" He asks Stephanie. She holds up three fingers and he smiles.
"That's how old my nephew is," he says, seeming to be lost in thought. She smiles a smile I haven't seen since she was 6 months old. It was nice to see.
We were there for hours just talking. Sephanie fell asleep in his arms.
"She's so comfortable with you," I tell him.
"I noticed," he said smiling. "I love kids."
I smile back and lean on his shoulder. We stayed like that for what seems like forever.
"You should get home," he whispers. I nod my head. We get up and he carries Stephanie to the redwood cabin door.
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Columbus, Georgia
We specialize in wedding photography, engagement photography and more! We believe we capture your wedding day in a way that cannot be found in Columbus. Photography is much more than a nice camera. We believe it's about chasing light to direct the outcome of your photos and if light isn't available - then we create our own. We use this found or produced light and mold it into an image that's more appealing. Then we like to turn this good photo into a great one! This is done by creating an expression. The expression is what sells the photo. The expression is what makes it into a great photo. Our goal is to bring our craft to your wedding day and create these beautiful photos.
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You're invited!
Join us for an online information session to learn more about our program!
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So as it turns out, I'm a person. Believe it or not. I know, it's pretty hard, innit? It's even hard for me to believe. I've spent my life thinking I wasn't. Oh well. Life goes on. I'm hungry.
I just ruined the moment, didn't I? Damnit. The joys of being a distracted writer.


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Core A, the Administrative Core of this National Institute of Neurological Disorders and Stroke (NINDS) Morris K. Udall Parkinson's Disease Research Center of Excellence (Udall Center) at the University of Pennsylvania (Penn), will facilitate accomplishing the goals of this multidisciplinary research program. Briefly, the goals of this Udall Center are to elucidate mechanisms of progressive neurodegeneration in Parkinson's disease (PD), especially those mechanisms underlying the cognitive and executive impairments in PD patients without and with dementia (PDD) and in dementia with Lewy bodies (DLB) patients. We hypothesize that disease progression leads to neuron dysfunction and death resulting from the differential transmission of pathologic alpha-synuclein species or strains and their neurotoxic effects in patients with PD/PDD/DLB. Thus, the Aims of Core A are to oversee budgetary and fiscal aspects of this Udall Center;promote and foster interactions between Core and Project investigators, as well as interactions of Udall Center investigators with scientists outside the Center at and beyond Penn;monitor the progress of the Cores and Projects;facilitate the sharing of data, reagents, and resources generated by Penn Udall Center investigators with other researchers in accordance with NIH policies;and participate in annual Udall Center meetings to accelerate the pace of advances in understanding PD/PDD/DLB. By utilizing the strategies in these Aims, Core A will play an important administrative role in this Udall Center by fostering the accomplishment of its overall mission..
Core A is highly relevant to the mission of the Penn Udall Center because it supports the efforts of each of the Cores and Projects in the Center to challenge and re-define current understanding of and research on PD/PDD/DLB by implementing the Specific Aims of Core A.
|Kalia, Lorraine V; Lang, Anthony E; Hazrati, Lili-Naz et al. (2015) Clinical correlations with Lewy body pathology in LRRK2-related Parkinson disease. JAMA Neurol 72:100-5
|Swanson, Christine R; Li, Katherine; Unger, Travis L et al. (2015) Lower plasma apolipoprotein A1 levels are found in Parkinson's disease and associate with apolipoprotein A1 genotype. Mov Disord 30:805-12
|Irwin, David J; McMillan, Corey T; Suh, EunRan et al. (2014) Myelin oligodendrocyte basic protein and prognosis in behavioral-variant frontotemporal dementia. Neurology 83:502-9
|Luk, Kelvin C; Lee, Virginia M-Y (2014) Modeling Lewy pathology propagation in Parkinson's disease. Parkinsonism Relat Disord 20 Suppl 1:S85-7
|Avants, Brian B; Libon, David J; Rascovsky, Katya et al. (2014) Sparse canonical correlation analysis relates network-level atrophy to multivariate cognitive measures in a neurodegenerative population. Neuroimage 84:698-711
|Peelle, Jonathan E; Powers, John; Cook, Philip A et al. (2014) Frontotemporal neural systems supporting semantic processing in Alzheimer's disease. Cogn Affect Behav Neurosci 14:37-48
|Cook, Philip A; McMillan, Corey T; Avants, Brian B et al. (2014) Relating brain anatomy and cognitive ability using a multivariate multimodal framework. Neuroimage 99:477-86
|Ash, Sharon; Menaged, Anna; Olm, Christopher et al. (2014) Narrative discourse deficits in amyotrophic lateral sclerosis. Neurology 83:520-8
|McCluskey, Leo; Vandriel, Shannon; Elman, Lauren et al. (2014) ALS-Plus syndrome: non-pyramidal features in a large ALS cohort. J Neurol Sci 345:118-24
|Escobar, Valerie Drews; Kuo, Yien-Ming; Orrison, Bonnie M et al. (2014) Transgenic mice expressing S129 phosphorylation mutations in ?-synuclein. Neurosci Lett 563:96-100
Showing the most recent 10 out of 105 publications


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|Message view
|« Date » · « Thread »
|« Date » · « Thread »
|RE: New directory API...
|Mon, 22 Jan 2001 06:45:56 GMT
> > I dislike combining the readdir and the stat call. We can provide what > > comes for free, anything more should be the result of a separate stat > > call. > > Alrighty then... I will live with that till we are proven wrong. I presume > we mean to 'reuse' the apr_finfo_t struct in the subsequent call, so we > need to watch for any 'oops'. Of course, the user is left testing what > they 'wanted' twice, once to see if apr_readdir picked it up, and again to > see if apr_stat did the second time around. Very sub-optimal. Yeah, but in the other case, they are left grabbing information they didn't want, and that could be expensive. Take mod_autoindex. We do a quick readdir, and then use the filename for a sub_request. If we put the stat in the readdir call, then we end up stating twice. Once in the readdir and once in the sub_request. If we leave the stat out, then we only stat in the sub_request. In my mind that behaviour is non-negotiable, but I am always willing to be convinced. :-) > > > This is a sub-optimal as the original. I'd expect most apps would > > > simply accept this; > > > > > > finfo->fcase = thedir->entry->dname; > > > > That's how I originally had it. The problem is that the testfile program > > then fails miserably. My goal was to have the readdir function return an > > finfo with a name that we could then use for apr_finfo. When I just used > > the thedir->entry->dname field, I couldn't do that. > > Two choices. apr_dir_fileinfo() which deals with it and merges them. > Or preallocate a buffer in the opendir of the fname length (dir name) plus > the length of the longest allowed filename, up to the limit of the total > path length. No sense returning a name >_MAX_PATH, I already have this > exception in Win32 (and just skip over such bogus files.) > > I'll have the Win32 implementation tommorow about noon for all to consider. > I'd like to see this be done so the api continues to reflect reality. I'm > going to sleep - need rest. preallocating a buffer won't work, because we end up with thread-safety issues. Not a problem for Apache, but for other programs... boom! :-) Let me sleep on this tonight. Ryan _______________________________________________________________________________ Ryan Bloom firstname.lastname@example.com 406 29th St. San Francisco, CA 94131 -------------------------------------------------------------------------------
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Closeouts. Step out of the shower and into the thick, absorbent softness of a genuine Turkish cotton terry robe. Fabric is luxurious with a plush, tightly woven surface texture.
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Who Was First? An Examination of an Early Hedonic Study
G. C. Haas produced a hedonic study more than 15 years prior to A. T. Court who first published the term hedonics. Haas's application was to agricultural land prices with a particular focus on distance to the city center and city size. Thus, Haas's work has much of the flavor of contemporary urban economics. A reestimation of Haas's model reveals that he did a respectable job in an age before computational machinery was available. The estimation of a new model reveals a tiny value gradient and shows that some of Haas's adjustments to price, especially his time adjustments, were amazingly accurate.
If you experience problems downloading a file, check if you have the proper application to view it first. In case of further problems read the IDEAS help page. Note that these files are not on the IDEAS site. Please be patient as the files may be large.
As the access to this document is restricted, you may want to look for a different version under "Related research" (further below) or search for a different version of it.
Volume (Year): 75 (1999)
Issue (Month): 4 ()
Pages: 620-626
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The following is an index of Publications for this department available to read online and/or order a printed copy.
The Government of Prince Edward Island has a new website princeedwardisland.ca
The publication you are searching for may have moved. Try searching using key words at www.princeedwardisland.ca/en/publications.
If you still cannot find what you are looking for, please use the Feedback tab on the new site to contact us. Thank you for your patience while we transition to the new site.
Recent Publications
Prince Edward Island Coastal Property Guide
January 25, 2016Communities, Land and Environment
A Water Act for Prince Edward Island - White Paper
July 8, 2015Communities, Land and Environment
Annual Report 2013, Environmental Advisory Council
April 15, 2014Communities, Land and Environment
Annual Report 2012 Environmental Advisory Council
August 28, 2013Communities, Land and Environment
Report of the Action Committee for Sustainable Land Management
February 15, 2013Communities, Land and Environment


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Endogenous Timing of Actions Under Conflict between Two Types of Second Mover Advantage
Young-Ro Yoon
Indiana University Bloomington - Department of Economics
July 30, 2007
CAEPR Working Paper No. 2007-013
In a model, two players, heterogeneous in their information quality, compete with each other with perfect information about the other player's information quality. If they can decide their timings of actions endogenously, the less-informed player has an incentive to delay her action for learning. On the other hand, the more-informed player wants to delay her action to prevent her information from being revealed, not to enable her to learn. The conflict of these two types of second mover advantages yields a war of attrition. Although both players can benefit from acting as the follower, the gain from a delay for learning is greater than that for preventing the other's learning. Therefore, a cost for the delay in action plays an important role in characterizing the equilibrium. In contrast to the literature, in which only informational externalities are considered, this article shows that the introduction of payoff externalities contributes to different procedures and reasoning processes through which the heterogeneous players' timings of actions are decided endogenously.
Number of Pages in PDF File: 24
JEL Classification: D81, D82
Date posted: August 1, 2007
Suggested Citation
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Call (888) 651-9785
Certified photos by travelers like you. What's this?
Borneo_Bibz Malaysia
February 2017
I learned there were 3 types of crow. I can't remember them now.
Borneo_Bibz Malaysia
February 2017
Mini bus to pick and drop you from visitor center to the site
December 2016
hut near the visitor center don't miss this so you have more pix
May 2016
Stonehenge - totally worth it. Amazing in person
August 2013
Kraaien hebben hun vaste stek op de eeuwenoude stenen van Stonehenge. Als oude geesten waren ze er rond.
August 2013
C'est ma photo grace à l'excursion ! Une vraie photo de carte postale. Un merveilleux souvenir.
June 2013
I (James) giving a thumbs up to one of the coolest places on the planet! STONE HENGE
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iPhone apps
- Sale 50% off through April - "Excellent iPhone Game..." -- Kotaku.com- OpenFeint Enabled - Colorblind is a game that challenges your ability to memorize and create colors. Remembering numbers, letters, words, sounds...that's easy.….. Read more
iPhone Screenshots
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Rated 4/5 based on 3 customer reviews.
5/5 stars
Paigez write:
4/5 stars
Good casual game
Toby Delrahim write:
4/5 stars
Good workout
jurassictodd write:
Be the first question this app.
Users Community
The information may be outdated. For actual information go to iTunes
Related to «Colorblind» applications
DanKam: Colorblind Fix
Can your iPhone correct your color blindness?"I used it today in the real world. It was amazing! I was at Target with my girlfriend and saw a blue plaid shirt that I liked. She asked me what… [more..]
Price: $2.99 |
ColorBlind Assistant
ColorBlind AssistantUse your iPhone or iPod Touch's photo library to quickly identify colors.Quickly draw a circle around the test area. The app performs the following:- identifies the hue,… [more..]
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IOS4 now supported!*Update coming with new live view*The perfect tool for anyone who is colorblind. Now you'll be able to identify the color of anything just by taking a picture of it with your iPhone… [more..]
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ColorBlinds Easy is the easy to use tool to help you as a colorblind person to distinguish colors and gain understanding and respect from family and friends for your color deficit by simulation… [more..]
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Government & Organizations, Non-Profit, News & PoliticsFollow
|Government & Organizations
A chance to go inside Council on Foreign Relations events. Listen to world leaders and foreign policy experts discuss and debate the most pressing issues in international affairs.
December 1st, 2014
Episode 100 of 727 episodes
Stanley Fischer, vice chairman of the Federal Reserve System, discusses past financial crises and the future of the Federal Reserve System.
You'll need to sign in to write a comment.
Welcome to the daily audio workout for your head.


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If you're on Windows 10 and are not satisfied with the new Mail app, Mailbird is a great alternative to Microsoft Outlook and worth checking out.
Windows Tips & Tricks
3 Apps to Quickly Configure Privacy Options in Windows 10
Windows 10 collects a lot of data which is considered an invasion of privacy. Here are some of the best apps to manage privacy options in Windows 10.
How to Uninstall Pre-Installed Apps in Windows 10 Using Powershell
Windows 10 comes with pre-installed apps, but there's no direct way to remove them. Luckily, you can uninstall pre-installed apps with the Powershell command.
How to Configure or Disable Automatic Updates in Windows 10
Windows 10 features automatic updates. If you're looking to configure or disable automatic updates in Windows 10, here's how to do that.
How to Disable Action Center in Windows 10
The Action Center is a new Windows 10 feature; notifications stay there until you clear them. If you don't like that, here's how to disable Action Center.
How to Turn Windows 10 into a WiFi Hotspot
Windows 10 allows you to create a WiFi hotspot and share your wireless connection with several devices. Here's how to create it and switch it on.
My 10 Favorite AutoHotKey Scripts to Make Life Easier
AutoHotKey is one of the best Windows automation programs that can perform both simple and complicated tasks. Here are 10 AutoHotKey scripts worth a look.
Add Tabs to the Microsoft Office Interface in Windows
Tabs make everything easier and allow you to open multiple items as once. If you want to add tabs to the Microsoft Office interface, this guide is for you.
Tip to Download Windows 10 ISO from Microsoft
Want to download Windows 10 ISO? The idea is to trick Microsoft servers to think you're not using Windows 7 or above. Here's how to do that.
How to Disable Windows Update Delivery Optimization in Windows 10
If you're on a limited Internet connection, here's how you can disable Windows Update Delivery Optimization in Windows 10 to save Internet bandwidth.
Adjusting Play Count in iTunes [Windows]
Are you interested in adjusting the play count in iTunes manually? If so, this tutorial will show you just how to do that on Windows.
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Not a fan of light colors? Don't like the light color scheme in Windows 10? You're in luck. Here's how to enable Dark Mode to see the dark side of Windows.
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Take your next company picnic or outdoor event to the next level with customizable blankets and picnic supplies! Guarantee that your brand stands out from the competition with merchandise that's sure to impress employees and onlookers. Browse our selection of cushions, chairs, baskets, serving ware, and even full picnic kits. Embellished with your logo, each distinctive item will allow you to widen your customer base in a fun and creative way. If you want to be the hit of your next office outing, bring along supplies and swag attendees will adore!
This promotional beach mat will promote your company or event at the beach, park, or any outdoor event.
Polyvinyl promotional beach mat with inner inflatable pillow.
These beach custom blankets are water and sand resistant and fold up into a handy carrying case for easy storage.


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plant is called Gold heart and I love mine. I went to get a second
this year and they are still expensive. But awesome in the garden
and a
great grower. I have it in a prominent location. I've been
told it is easy
to divide dicentras, but it will be a couple of years before I feel safe
touch it. The Essentially English garden on SR35 in Jamestown
OH has large
ones for $20, but he does not ship..... They are stunning!


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Posted by Sara on Thursday, September 6, 2012 at 6:14am.
s=sqrt(80²+60²) = 100 m.
v=60•1.1/100 = 0.66 m/s.
sin φ= 0.66/1.1 = 0.06
φ=arcsin 0.06 = 3.44º
Elena, i have a little problem with your calculation. I was thinking the velocity of the swimmer should tally with one of the adjacent sides of the triangle and not the hypothenus. the hypothenus should be in line with the resultant velocity of the swimmer.


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Shares of Mears, a little maintenance services group, have enjoyed a splendid run since it reported interim figures at the end of last month. They are now at a peak of 33p. Analyst Roger Brocklebank at the Old Mutual investment group believes there is a case for a price much nearer 50p.
Shares of Mears, a little maintenance services group, have enjoyed a splendid run since it reported interim figures at the end of last month. They are now at a peak of 33p. Analyst Roger Brocklebank at the Old Mutual investment group believes there is a case for a price much nearer 50p.
They arrived in the no pain, no gain portfolio in April at 23p. I must admit I was apprehensive about them and at the last moment almost reversed my decision to include them.
My anxiety stemmed from a sudden surge in the share price. When I examined the company I was impressed by its attitude, management, prospects and record. At the 19p they then stood, I felt, the shares looked an ideal buy.
But the rapid advance to 23p put a different complexion on them. Such a movement by the low-priced share of a small company looked as though I had missed the boat? Luckily, I stayed aboard. And the portfolio is the beneficiary.
When I alighted on Mears it had just produced encouraging figures which, I felt, the stock market had overlooked, as it often does with small companies. An internet tip service apparently came to the same conclusion and its enthusiastic online comments spurred the sudden advance.
Now the Mears half-year figures have underlined the company's undoubted strength. Interim pre-tax profit was £850,000 against £412,000 with the dividend (an often-overlooked factor in modern investment) again increased.
Mr Brocklebank sees year's profit of the AIM-traded group emerging a little below £2m with £2.4m next year. My guess is he is being rather cautious.
Mears is run by Bob Holt, ex-Mitie, the highly successful building services group. It could be argued that Mears closely resembles the much-smaller Mitie of a few years ago. Mr Holt, who is 45, was once chief executive of Tottenham Hotspur and was at Blue Arrow, the controversial recruitment group.
He is keen to expand Mears, and acquisitions, probably in the North, are likely. But with its growing profitability and a strong order book - around £110m - it must look an enticing takeover target. It is thought to have had approaches but talks clearly failed. Some believe it is such a well-run and progressive company that it cannot hope to retain its independence much longer. At the present price it is capitalised at only £16.6m.
A great many successful small companies are, on traditional measurements, undervalued. Mears is no exception. The shares are selling at less than 10 times prospective earnings, roughly half the sector average. A direct comparison with Mitie, probably selling on more than 50 times expected earnings, is unjustified. Even so such a yawning rating gap is too wide.
Another feature in Mears favour is that unlike many small players, it is not shunned by institutional shareholders. Eaglet Investment Trust, admittedly specialising in small companies, has 20.6 per cent. Other declared holdings take the institutional presence to more than 40 per cent. Directors have 20.3 per cent.
The group operates from 30 branches and has extensive links with local and other public authorities. Most of its contracts are long-term.
It is under contract to maintain around 140,000 local authority houses, out of a total of 3.2 million or so. Therefore the potential to win new business on that front is considerable. In fact, Government plans to increase spending on housing and education should ensure Mears continues to prosper.
Certainly it seems, at least on trading grounds, that Mears will be able to keep its followers happy. But the stock market's failure to embrace small companies could inhibit the shares. But they are destined to retain their membership of the no pain, no gain portfolio and even at their present price are still a worthwhile buy.
Dru Edmonston at stockbroker Durlacher is, like Mr Brocklebank, keen on the shares. He thinks they are a buy up to 35p.
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Saturday, February 18, 2017
Pavel Ivanov (Mak) (1891-1967) ... The Princess and the Phoenix, pen and ink , watercolours and gold ink on paper 1926
Monday, November 21, 2016
Maxwell Ashby Armfield (1882-1972)... Aquarius - study for the Signs of the Zodiac or 'Gates of the Makara', pencil and watercolour, 1949
Sunday, August 21, 2016
Tuesday, June 7, 2016
Sunday, May 22, 2016
Wednesday, May 4, 2016
Wednesday, April 20, 2016
Saturday, April 2, 2016
Friday, January 22, 2016
Tuesday, December 8, 2015
Diego Rivera...Human sacrifice before Tohil 1931
Watercolor painted by Diego Rivera in San Francisco, Calif. during the summer of 1931, originally commissioned to illustrate a never-published English translation of the Popol Vuh by John Weatherwax.
Monday, December 7, 2015
Sunday, December 6, 2015
Grace W. Pailthorpe (1883-1971) Sea creatures, watercolour 1967
Monday, October 26, 2015
Born today...Karin Boye,poet, October 26, 1900 Gothenburg, Sweden
Say, where does the world's heart burn,
the world's heart of fire?
It lives on coarse, heavy prehistoric coal:
black darkness, dense night, Chaos.
Seek there!
itself a struggle, glowing struggle -
For thus is the nature of fire:
strong with its foe's struggle -
has no other nature.
And the victory? When the darkness has disappeared in flames?
and fire wants to conquer.
Is the victory death?
Empty question and empty fear!
The world's heart is fire,
Lilith and Faun
Life and Death
Wednesday, January 21, 2015
Dolorosa de la Cruz... works in progress no1 2015
Sunday, January 4, 2015
Rikki Ducornet ...poem and watercolour & gouache ... 1989
The lunatic algebra
of Love.
The frenzied orbits
of Mood.
The malarial temperatures
of Wound.
Symbols of the Cult
of Seizure:
This flesh, this amulet
This hot spoor
of predators.
This zodiac savaged
in the sky.
The Cult of Seizure 1989
Saturday, July 27, 2013
Tuesday, March 20, 2012
Thursday, September 22, 2011
Dolorosa... new watercolours ... the Twins I ...2011
New work exhibited at the annual Esoteric Book Conference in Seattle Sept 2011.
the Twins I ~ 2011 - Dolorosa
I have one print available for sale of a limited edition of 3, A4 size with 1 inch border printed on Hahnemühle FineArt paper ~ e-mail me at email@example.com for more details
photo by Patrick John Larabee
Monday, August 22, 2011
Dolorosa... new watercolours ... 2011
Dolorosa ~ Untitled Watercolour 2011
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After just 22 days on the job, Rich Bisaccia has left Auburn to reportedly take a job as special-teams coach for the Dallas Cowboys. Bisaccia, who spent the last two seasons as the Chargers special teams coach, left San Diego for Auburn after he was hired to become a running backs coach/special-teams coordinator on Jan. 3. AL.com reported that Bisaccia will accept the Cowboys job later Friday.
Report: Bisaccia to Coach Dallas' Special Teams
From ESPN.com
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Anna Netrebko
The star of the highly-anticipated Manon opera at the Royal Opera House
Despite the fact that the opera marketing industry continues to
launch one diva after another, nobody among today's young singers
is as attractive and sensual as
Anna Netrebko. Her already mythical status was
surely consolidated by the story of her career's beginnings -
widely recounted in the art world and the media - in which she's
practically depicted as a modern-day Cinderella
who was cleaning the floors at the Mariinskij Theater of
Saint Petersburg.
Before being discovered by the powerful director Valery Gergiev, who is the man responsible for launching the soprano into the international scene, from Russian operas to the Italian belcanto to the French repertoire. Her breathtaking beauty helped earn her starring roles in operatic films like La Bohème, as well as unconventional contemporary renderings of classics like the famous Traviata in Salzburg. Between the New York Metropolitan, the Royal Opera House in London and Paris's l'Opéra, the singer - who is fully booked until 2013 - took a break of several months two years ago in order to give birth to her son, whom she had with the baritone Erwin Schrott from Uruguay.
As Italy awaits her return to their stages, where her absence has been felt, her fans can find her this month at London's Royal Opera House for the highly-anticipated production of Manon by Massenet, conducted by Antonio Pappano (6/22-7/7), where she will doubtlessly charm and seduce packed audiences.
(Photo by Mat Hennek/Deutsche Grammophon)
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- Details
- Hotel pickup included
- Informative, friendly and professional guide
- Perfect introduction for first-time visitors
After leaving the capital, you'll start the day by heading for the Reykjanes Peninsula. The peninsula is almost completely covered with old lava flows - you will see lava combs and craters of all sizes and shapes, plus Iceland's famous tectonic plates.
You'll then continue to the lighthouse on the westernmost tip of the peninsula, and marvel at this area's rich birdlife. One of the world's biggest north gannet breeding colonies is at Eldey Island, where surf crashes against the beautiful basalt cliffs. It's a simply stunning sight!
From there, you'll continue through the lava fields to the nearby fishing village of Grindavik. After a short stop by the Blue Lagoon, you'll return to Reykjavik to pick up your Gulfoss and Geysir Express half-day tour, to see Iceland's best known historical sites and natural phenomena.
The afternoon tour begins with a visit to Thingvellir, a geologically unique place where the American and Eurasian tectonic plates meet. The Icelandic parliament Alpingi was established here in 930, and in 2004 Thingvellir was added to UNESCO's World Heritage List.
You will also see the majestic and beautiful waterfall Gullfoss, the incredible spouting hot springs of Geysir and Strokkur. After a visit to the village of Hveragerdi, with its large greenhouses, the tour returns to the capital.
Schedule details
Hotel pickups commence 30 minutes prior to this time.
Pricing information
Gulfoss Waterfall, Iceland
- Hotel pickup and drop off
- Professional guide
- Transport by air-conditioned coach
- Food and drinks
- Gratuities (optional)
- Towels and bathing suits
- Confirmation will be received at time of booking
- Bringing a packed lunch is highly recommended
Cancellation Policy
If you cancel between 3 and 6 day(s) in advance of the scheduled departure, there is a 50 percent cancellation fee.
If you cancel within 2 day(s) of the scheduled departure, there is a 100 percent cancellation fee.


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Hermit and the Well, the
€ 17,99
Lieferbar innert 2 Wochen
Januar 2004
Beschreibung"The Hermit and the Well" is a story is based on an actual event in Thich Nhat Hanh's life.
As a young boy, amidst the suffering and destruction that the war had brought to Vietnam, Hanh had a profound transformative experience after seeing a reproduction of a Buddha statue on the cover of a magazine. Moved by the Buddha's serene expression and peaceful appearance, he decided to become a monk. Through his ensuing school years, his interest and commitment deepened and led to visits of temples and other places of worship including a class fieldtrip to a hermit living in the mountains.
Thich Nhat Hanh entered a Buddhist monastery at age 14.
Untertitel: col. Sprache: Englisch.
Verlag: Parallax Press
Erscheinungsdatum: Januar 2004
Seitenanzahl: 32 Seiten
Format: gebunden
Alle Preise inkl. MwSt., ggf. zzgl. Versandkosten


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Total Granny Flats
Accommodation, Cabin, Granny Flats
Total Transportables is a family owned & operated company based in Rockingham building high quality transportable buildings including offices, Granny Flats & ablutions on time & at a competitive price. Being licensed builders, we can offer a complete service from helping you design your building, organising council approval, construction, and delivery, installation & connection to services. We build a large range of transportable buildings for hire & sale, supplying individuals right through to big companies. We build Granny Flats, backpacker accommodation, shower & toilet blocks, site offices, lunch rooms, and family dwellings. Total Transportables also have been regularly supplying mine sites with a large variety of buildings. We build to the customer's needs and, unlike larger companies, don't charge for 'changes' to existing building plans. You tell us what you require and we will endeavor to design & quote a building which is exactly what you are after and which is very competitively priced. We have had excellent feedback from past & existing customers and are receiving many referrals by 'word of mouth' which speaks for itself.
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Quality Transportable Buildings at an affordable price Transportable offices, ablution, accommodation buildings, Granny Flats, lunch rooms for hire & sale. Complete service from planning, construction to delivering & connecting to services.
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