text:
CHICAGO, August 13, 2013 – After announcing the close of their first investment round, Dan Klein & Patrick Tannous, co-founders of Tiesta Tea, offer tips of raising money for young entrepreneurs like themselves. Their success allowed Tiesta Tea, a loose-leaf tea company specializing in exotic flavors and simple functions, to double its original funding goal, close the round within four months and ultimately turn a handful of investors away.
Raising money is all about attitude. There is a big difference between begging for money and showing investors this is the way for them to make money. The latter answers the single greatest concern investors have.
"Don't be afraid to be cocky. A lot of the times it translates to confidence. We spoke with confidence about our product, our vision and our team. This was truly the first step in showing investors we had something special and that they're going to want in on it," stated Tannous.
Raising money is like selling a product. It's hard to bring attraction without a good selling point. Since day one, Tiesta Tea has taken nontraditional approaches to selling that has set them apart from competitors. The success of their unique in-store demo program, "Demo Derby", has allowed the company to refuse slotting fees from grocery chains, which is an unorthodox tactic in the industry and extremely attractive to investors.
"After all investors were in and we closed the round, I asked them why they did it. A lot of investors liked our marketing approach of our Demo Derby and saw room for growth throughout various aspects of our company. Most importantly, they said they believed in us," said Klein.
Raising money is like hosting an event; you need to create a buzz in order to get and maintain interest. On top of sending out applications to angel investment groups and investment firms, Tiesta Tea hosted their own investment presentations in Chicago. The result was a significant turning point for the young start-up.
"We rented a huge ballroom and set up our in-store demo booth with every Tiesta Tea blend on display. This was the only way we could completely convey our vision to investors and ultimately show them what a great opportunity this is. The events were such a success that we actually signed a few on the spot. I also got emails from people that weren't at the event telling me that they were interested in the opportunity," said Klein.
Raising money is about creating scarcity and making it as a limited edition. In the middle of the financial round, Tiesta Tea sent out an email list of investors who had already signed up. This was done to create a feeling of high demand.
"Create a situation where people feel they are lucky to be able to invest. As if they don't answer this email by tomorrow with an offer, they miss out on this amazing offer. If you can do that, you are going to be very successful in raising money," said Klein.
Tiesta Tea took a nontraditional route to raise their money. It was a risky and exciting time for the young company. Through this investment round, Klein and Tannous gained valuable lessons and believe many young entrepreneurs can relate to and even learn from these same challenges.
# # #
Tiesta Tea Company is a loose-leaf tea company based out of Chicago, Illinois. Their all-natural, loose-leaf teas are packed with high quality fruits and herbs from all around the world. Their goal is to take the confusion out of tea by organizing the teas into five functional lines that explain the effects they have on the drinker (Energizer, Slenderizer, Relaxer, Immunity and Eternity). They believe this simple approach, along with unique flavors like Kiwi Cherry Bonanza and Banana Split will appeal to the masses and raise awareness for loose-leaf tea. Check out Tiesta's teas online at www.tiestatea.com or find them in major grocery retailers across the United States. Find out more information about Tiesta Tea Company by going to www.tiestatea.com or calling 312-202-6800. Keep up with Tiesta Tea by following them on Twitter or liking them on Facebook.


text:
No one in their right mind looks forward to doing this. But, unless you're going to self publish, (and we'll get to that soon) you've got to do it.
You're whole book. In one paragraph. One paragraph small enough to fit onto a one page letter with at least three other paragraphs, two address blocks, a formal salutation, and closing. And no, you can't fudge the margins or use an eight point font. Call it less than one hundred words, possibly seventy-five. Eeep!
Now, I mentioned earlier that this paragraph is not the back of the book copy. But why not? After all, you are trying to sell your book to the agent, and the back of the book copy is trying to sell the book to everyone who picks it up.
And that's precisely why not. You are selling your book to one person. One person you can research. You can find that person's likes and dislikes. You know what books he's purchased before. You know what kinds of people he's worked with before.
In the case of my own book, and yours probably, there are a lot of angles you can play up when it comes to convincing your agent-to-be that you've got the book of his dreams. I can work on the fact that my main character is Jewish, the intense religious themes, and questions about theodicy and forgiveness if I aim at an agent who handles religious fiction. I can paint a story of an Epic Fantasy trilogy that will span modern day Urban Fantasy to High Quest Fantasy while weaving a classic love story between worlds, if I was hunting a fantasy agent. If I want I can minimize the fantasy aspect and show how it's dripping in erotic romance if I'm hunting an erotica agent.
So, as you sit there with your list of potential agents, take the time to write down what precisely they've sold, what it is they want in a book, and start to work on why your book is exactly what they want. At the same time (because, after all this is never easy) you need to keep in mind what your book actually is. In my case I may tone down the fantasy aspect, but if I leave it out completely and actually do get a manuscript request from an agent who handles erotic or religious fiction, I'll get a "no thanks" letter awfully fast, and be out the cost of a manuscript.
I'll once again borrow from the car shopping world. If you've ever had someone try to sell you a car that wasn't quite right, say trying to get you into one that cost too much, or had a bunch of things you didn't need, or was in much worse shape than claimed, you know how frustrating and annoying that is. You need to realistically assess you story. You need to know if you've got a Rolls Royce, a Subaru, a Honda or a Dodge and sell it as the best possible version of whatever it is you've got. Spending your hundred words talking about the aching beauty and immense clarity of human experience in your Literary Fiction story is great, assuming all of that is true.
So, how best to go about figuring out what your book is about? I suggest having a few people you know and trust read it and tell you. Seriously, what you think your book is about and what everyone else thinks it is about may be two very different things. Better yet, people you know and trust who you haven't already told what the book is about. (Because we all have talented reader buddies who we haven't talked the ears off of already...Yeah, I know exactly how realistic that proposition is. But assuming you have some people like that in your life, now is the time to ply them with gifts and a copy of your manuscript.)
Once you know it's time to write. And re-write. And re-re-write. And probably do it a few more times. Because here's the crux of this issue: this is the paragraph that makes your chance of ever getting an agent. There are a million ways of losing potential agents, and this is the one way to actually snag one. If you know people who have agents, beg them to read your letter, ask for brutal feedback. If you don't know anyone along those lines try to get people who will give you an honest answer to read your letter and answer this question: "Having read this, do you want to see my book?"
Lastly, good luck!

subset: commoncrawl


text:
From all the gum trees, I think Spotted Gum is the most beautiful. It is also known as Corymbia maculata or Eucalyptus maculata. The large majestic trees are native of Victoria, New South Wales and Queensland. The slow growing trees in average will grow up to 35 m but many will reach up to 45 m high or more. The tallest in record was said to reach 91 m high (according to Wikipedia). The large mottled trunks are colourful with a mixture of greyish green, brown and orange pink. More
Majestic Spotted Gum Tree
6. Wonderful Plants, Street Trees Australian Native Plant 4 Comments
Blooming Sarcochilus Orchid
3. Orchids Australian Native Plant, My garden, Sarcochilus 2 Comments
It seems that this year only one Sarcochilus orchid has flowers (2 spikes). It would be more beautiful if all of them are having flowers now to fill the pretty basket pot. However, I just have to be happy with what I have 🙂
Xanthorrhoea, si Bujang Hitam
6. Wonderful Plants Australian Native Plant, Indonesian Language 2 Comments
Ingat waktu menyaksikan kejadian yang cukup langka dibulan October 2006, yaitu disaat tanaman Blackboy Grass Tree secara serentak berbunga di Brisbane Ranges National Park yang letaknya sekitar 80 km sebelah barat kota Melbourne. Sebetulnya tanaman rumput Blackboy itu lambat sekali tumbuhnya dan jarang sekali terlihat berbunga, jadi kalau semuanya tiba-tiba dengan serentak berbunga, itu agak aneh juga.
Anggrek Sarcochilus
3. Orchids Australian Native Plant, Indonesian Language, My garden, Sarcochilus 2 Comments
Bulan Desember tahun lalu, bersama suami, saya pergi ke Langwarrin (kira-kira 40 km dari rumah kami) untuk beli anggrek Sarcochilus. Kami lihat iklannya di Gumtree dan harganya juga tidak terlalu mahal. Sebelumnya saya tidak tahu seperti apa jenis anggrek ini, jadi penasaran juga untuk melihatnya. Ian yang kira-kira seumuran kami amat ramah dan rupanya dia memiliki 'hobby farm' tanaman anggrek dihalaman rumahnya. Dari pinggir jalan, rumah dan bangunan untuk menanam anggrek tidak kelihatan karena depannya lebat dengan pohon-pohon Eucalyptus yang tinggi.
Akhirnya kami beli dua jenis anggrek Sarcochilus yaitu S. fitzhart dan S. hartmanii. Keduanya berwarna putih tapi lain warna bibirnya. Walau bunga kecil ukurannya, tapi karena tumbuh menggerombol jadi kelihatan cantik juga dan semerbak mewangi. Ian yang murah hati malah memberikan bonus 2 pot Sarcochilus yang masih kecil, yang satu juga S. fitzhart dan satu lagi S. Bernice Klein 'Pink Blush' dan belum berbunga. Waktu saya tanya seperti apa bunganya, jawabannya adalah misteri jadi harus tunggu sampai berbunga. Moga-moga saja daun bunganya akan pink seperti namanya dan bukan putih lagi. Waktu saya mengagumi bunga anggrek Dendrobium nobile yang berbatang lunak, kami juga diberi anakannya yang masih kecil gratis.
Sarcochilus Orchids
3. Orchids Australian Native Plant, My garden, Sarcochilus Leave a comment
My husband and I went to Langwarrin to an Orchid Farm and bought Australian native orchids 'Sarcochilus'. The small and pretty blooms are fragrant. These native orchids species are endemic to Northern, Eastern Australia and Tasmania. These epiphytic (grow on other plants) and lithopythic (grow on rockeries) orchids grow wildly on high altitude mountain ranges. In Victoria, Sarco orchids can be found in certain areas around Mt. Dandenong. There are also quite a few of hybrid varieties.
As this is the very first time for me to have this species of orchid, I have to learn the growing requirements. These are notes that I have made:
- Location: To grow Sarco orchids will need cool and airy shaded areas around 60 % – 70% with high humidity. It is common to place Sarcochilus under tables or benches. Never expose the plants to direct sunlight. Keep them in shadier place during summer months.
- Temperature: The ideal is between 8 deg.C to 28 deg.C. In the summer when the Melbourne temperature can soar well above 30 deg.C, keep the plant in darker place and water down the surrounding area to make the air more humid. When the winter temperature is well under 8 deg.C, keep the plant under cover or if it is outside, the plant can be covered with plastic sheet. Never expose the plant to frost.
- Potting media: consist of orchid barks or coco bark. A little bit of river pebbles or styrofoam pieces can be added into the barks.
- Watering: As Sarco orchids have no Pseudobulb, they will not be able to store water. Watering should be done regularly to keep roots moist. Reduce to about once a week during winter time. Rain water is more suitable to use. Do not let the leaf crown wet and keep flowers dry.
- Fertilizer: During summer and autumn apply weak mix of liquid fertilizer such Aquasol, Fish Emulsion or Seaweed Extract once a week with exception, once a month use water only to flush the fertilizer residue (once a week for three weeks apply weak fertilizer and only water on the fourth week). Apply less often fertilizer in spring. DO NOT apply fertilizer at all during winter time.
- Propagating: Sarcochilus will form a large clump that can be divided over time.
- It can grow mounted on a shady tree trunk, on rockeries or just in ordinary pots. The proprietor of the orchid farm said that Sarco orchids will grow many long aerial roots. In older plant, the aerial roots can creep and cling on the wall, on rocks or on tree trunk. If grown in pots, when the clump is bigger, wide squat pot is better to use.
We bought two pots and both are flowering, the one with pink flowers (S. fitzhart) is larger than the white one (S. hartmannii sp.). We were given two extra in smallest pots. One is another flowering S. fitzhart and the other one is Sarco bernice klein 'Pink Blush' with no flower. So it will be a mystery how the flowers will look like, but I guess it will be pinkish. We were also given a small pot of soft cane Dendrobium (no flower) for free.
Dendrobium delicatum
3. Orchids Australian Native Plant, Dendrobium, My garden Comments Off on Dendrobium delicatum
It is said that Australian native orchid Dendrobium delicatum was the result of natural hybrids between Dendrobium tarberi and Dendrobium kingianum. Though the small and mostly white little flowers are less showy, they are prolific bloomers and very sweet scented. It is very easy to grow in tropical and temperate climates.
D. delicatum is perfect to grow on rocks or up on tree branches. It can also be grown in pots. The plant will quickly grow new shoots. The best growing medium is orchid potting mix together with a bit of compost. Liquid fertilizer should be applied after finished flowering and I also use liquid seaweed to promote healthy growth. Dendrobium delicatum will need reasonably bright sunlight to flower.
We put ours near the front door and the gentle spring breeze is blowing the sweet delicate scent into the house. It is just beautiful and refreshing….
Pretty Scaevola 'Aussie Salute'
6. Wonderful Plants Australian Native Plant, My garden 3 Comments
I spotted a beautiful plant with the pretty purple flowers in the market. The plant name is Scaevola aemula 'Aussie Salute'. The shape of the flowers is quite unusual. It looks like the flower has been ripped into half. As it is half circular, this plant is also known as 'Fan Flower'.
I still have this plant in a pot, but I plan to transplant it in a sunny spot in the garden.


text:
Small in size but big on style, this tiny oxblood color leather pouch bag features porcelain details and long leather cord strap.
Made in the United States. Pouch Bag. Oxblood color leather (unlined) with porcelain details and leather cord straps.
AHAlife is committed to help you live a conscious and inspired life.
Many of the items you discover here are exclusive to AHAlife, which is what makes our community of designers and artisans so special.
If you find one of our products anywhere else for less than it's being sold on our site, we will match it. No questions asked.
Our dedicated Concierge Team is here to help. Reach out any time at:


text:
Australian eyewear brand COLAB continues to present interesting collaborations. Today we show you their work with artist Rockin Jellybean. The capsule collection consists of several models, for both men and women. While all of them keep a clean appearance on the exterior, the interior graphics make them stand out, along with the accessories, such as a custom Rockin Jellybean lense cleaning cloth. The sunglasses are now available from GR8.
Detailed images of the collection follow after the jump.


text:
will it work with my iphone 4?
people have been asking about it working with the new macbook
so i was curious if it would work with my iphone 4!
- Asked by Misbah A
Apple Remote
Product No Longer Available
1 Answer from the Community
Yes it will, but only if you have the universal dock. If you dont have it, just seach for it on the apple store website.
- Answered by Caleb E from Simi Valley


text:
First Choice Auto Service
Review for First Choice Auto Service
JOYCE W. on 04/28/2015
"Great service."
These guys have worked on my vehicles for many years and always do a good job. The shop is clean, employees are polite and knowledgeable.
Vehicle: Pontiac Grand Am
Service Date: 04/27/2015
Review Created: 04/28/2015 06:43 PM


text:
Voting, Fireworks and a Dance-off
Comments Off on Voting, Fireworks and a Dance-off
This time I'm not talking about the US election, but the Australian federal election. It's been called for October 9. Bring it on.
I went to RiverFire at Southbank on Saturday night. The fireworks were pretty good. The F-111's are always a great finale. Unfortunately I was in a spot where I was only going to get one shot at them, and they were too quick for the camera. It was more enjoyable just hanging out and experiencing it with a group of friends, and the other thousands upon thousands of people there.
Still at Southbank, we went to sit down and relax in the Piazza. The teen angst was palpable. This was definitely the place to be if you wanted to see and be seen. We took a seat and found we had chosen the best vantage point to witness the dance-off. That's right folks, a dance-off. I was amused, impressed and privileged all at the same time. It was funny to see these guys facing off to prove themselves to each other and the very sizeable crowd that had gathered to watch. Alternating sides sent out the representatives to challenge… This went on for about 15 minutes. Some of them were really good. Some were fairly average. The whole experience was a little surreal, but kind of cool. Eventually the police came and told everyone they had to leave, but both groups had just been standing around looking at each other for five minutes by then anyway. I think they'd run out of moves.
Just chose one fireworks shot. None of them were that great. I didn't have my tripod.
The Danceoff – dueling dancers
The Danceoff's only clear shot because, yes, he just balanced there.
Parko attempts a backflip after we leave the danceoff.
His feet do hit the ground first…but I can't say he landed on them.
Copy Editor in Cambodia
August 28, 2004 by Dave Earley
Filed under Uncategorized
Comments Off on Copy Editor in Cambodia
This job makes me want to pack my bags. It's not the location. It's the perks: adventure, inconvenience, making a difference in a fragile democracy and furthering a free press. I'm sold. If you think I'm joking, I assure you I'm not. Pity about university…
I've just signed up with Hello from Picasa, so testing it out by posting one of my favorite photos. This is looking up Monument steps, all 345 of them…
To vote or not to vote
Australian readers may say, "But it's compulsory to vote". Yes, but I'm not talking about the Australian federal election which I wholeheartedly look forward to voting in. I'm talking about the election of the next president of the United States.
It may be the US Democrats urging expats to vote, but what if I wanted to vote Republican? I don't, but what if I did? On visiting their site, the Democrats Abroad Australia group asks me (not require, just ask) to fill out a membership form. Hell no! You ain't co-opting me into your database! I don't need any stinkin' democrat knocking on my door the next time I vote republican! I will, however, directly visit OverseasVote2004.com to fill out the form…
Argh, it's run by the Democratic National Committee too! Oh well, they're going to love me – Iowa's a key state in the Electoral College isn't it? Yay, I know not much about US politics, but will participate in my first American election. Unless of course George W. manages to invalidate absentee votes, the military, African Americans and all of Florida… then, well, I tried.
Comments Off on Sellout
Google/Blogger is promoting their Adsense program for blogs. Basically content-based advertising…
Because I have thousands of visitors to this site every day – thousands I tell you! – I'm thinking of selling out. Oh yes, it's the high life for me from now on. Goodbye university. Goodbye career. Goodbye food stamps. Hello no work, lazing around and weekend jaunts to the Ba-HA-mas! Wait, that's already my life. Except the Bahamas of course, I can only get there on long weekends at the moment…
News story written in 90 minutes and handed in this morning was not even worth mentioning really. But I just did. Now to find a real story.
I'm posting from the library at university. I have my Ethics case study outline due in 15 minutes. Yeah it's not happening. I'm looking for three stories from Iraq to analyse. Are there still embed journos in Iraq, or are they all independent of the US Army now that the war is 'over' (I laugh)? What I'm looking for is reporting on the same event by an embed, independent, and Al-Jazeera journalist. Then analyse the ethics of each. Hm, maybe I'll just go with an American, Australian and Al-Jazeera journalist.
Next time I blog, there may be advertising content seen on this site. Then again, that might involve more effort than I feel comfortable expending. hoo-roo
I'm back
August 25, 2004 by Dave Earley
Filed under Uncategorized
Comments Off on I'm back
And what have I learned? That I shouldn't have a computer connected to the internet in my bedroom. It's just too damn easy to waste time. I must admit, I cheated a little. I did check email, and read some blogs, but my computer was never connected to the internet. Anyway, I didn't have withdrawals or anything, and I did actually get some stuff done.
So, five minutes back on the net on the computer in my bedroom (eh, things will change), what do I have for you? RSS Owl, the newsreader I occassionally use, tells me yet another update is available. While downloading that, I check out some of the feeds I never read (because I never use my newsreader), and discovered this link to Transformers, busting out the mad floor routine. Good stuff. I recommend it… and have patience if you have a slow download rate (it's streaming), which I'm experiencing as I type this and try to watch it. Alright, it finished streaming (3min 45sec) and I watched it uninterrupted beginning to end. Truly a masterpiece. Those Transformers can dance!
Otherwise, there's a lot more important things to be done than write in this blog, like
- write at least one news story
- write my case study outline for journalism ethics (I'm thinking 'war' reporting, either Iraq or Sudan)
- other miscellaneous university requirements
Till next time.
My Isaac
Comments Off on My Isaac
I like the new Blogger Navbar up the top. I've enjoyed random blogsurfing with the "next blog" feature. At the same time, however, I've found it difficult to enjoy. It's a guilty pleasure. At the back of my mind is that nagging, insisting, reminding, voice that all I'm doing is avoiding other more important responsibilities.
Nothing should ever consume so much of your time it reduces your quality of life. With this thought in mind, I'm pulling the plug on the internet for a week. A WEEK?!What emails will I miss? How will I keep up with the world? What will I do? The very fact that it's a little daunting confirms to me that I need to cut the cord (not literally).
The one line I remember with crystal clarity from a sermon two weeks ago was, "God knows your Isaac." The reference was to the story of God testing Abraham by asking him to sacrifice his only son Isaac. I always considered sport to be that one thing I was holding in too high regard, but I've gotten over my obsession for the most part. I know I can walk away from it if I have to.
The internet, however, is consuming hour upon hour of time I don't have to waste. And so until 7.30pm next Wednesday night, or later, I bid you adieu.
Michigan sucks
Comments Off on Michigan sucks
Hard work and dedication does pay off! The coveted 'Michigan sucks' shirt from Caliblog.com arrived in the mail the other day. I wore it to university on Tuesday and received some strange looks. That's right, that's what it says!
Michigan may suck, but South Carolina (state of my birth, though I haven't been there in 15 years) rules, as do Volkswagen Kombi vans and Gopher College rugby! Gopher College will take some explaining, but it's not going to happen now. Neither are you going to get the pic that doesn't cut out both the rugby ball and my whole head.
[singing] Lazy blogging, yeah it's for me.
Comments Off on Cartoons
Jaws in 30 seconds – reenacted by bunnies.
I had to blog this. Too funny. Check out the other cartoons there also. Thanks to Dan from Surgical Strikes for the reference.
Another good site for cartoons is Weebls Stuff. Some funny cartoons, some very strange, and some quite disturbing really.
August 17, 2004 by Dave Earley
Filed under Uncategorized
Comments Off on Gmail
By the way, I still have one Gmail account available to anyone who wants it.
That's One Gig of email space… excitement.
Inform Me
August 17, 2004 by Dave Earley
Filed under Uncategorized
Comments Off on Inform Me
Cave possibly used by John the Baptist found
"The discovery, if confirmed, would be among the most significant breakthroughs for biblical scholars in memory".
'Evangelical X'ianity Has Been Hijacked': An Interview with Tony Campolo
"there's a difference between evangelical and being a part of the Religious Right. A significant proportion of the evangelical community is part of the Religious Right. My purpose in writing the book was to communicate loud and clear that I felt that evangelical X'ianity had been hijacked.
When did it become anti-feminist? When did evangelical X'ianity become anti-gay? When did it become supportive of capital punishment? Pro-war? When did it become so negative towards other religious groups?
We ought to get out of the judging business."
You tell 'em Tony.
And then, to turn to a completely different focus… And no, this is not being intemperate or negative towards other religious groups… or is it? You tell me, I'm just presenting the news.
Abdullah urges Muslims to restore Islam's glory
Prime Minister Abdullah Badawi [of Malaysia] has called on Muslims to have a plan of action to restore the glory of Islam, saying that rebuilding the Islamic civilisation is their responsibility.
He added that efforts to develop a respectable lifestyle on earth by seeking out success and building an Islamic civilisation at the same time were actually part of the requirements in Islam.
Islamic expansionism?
Saudi prince declares victory in war against terrorists
'We directly went to the heads of the snakes to cut them off, and we know the tails are a group of our sons who went astray,' Prince Ab- dullah said. 'We know how to deal with them and how to bring them back to the right path.' He did not elaborate.
Perhaps it involves not funding them anymore. Too bad for the US most of the insurgents in Iraq are Iraqis, not foreign nationals, or Saudi Arabia would have the answer to global terrorism… if they chose to elaborate.
Polls in 2000 showed Muslims favored Bush over Mr. Gore by a wide margin – some showed support in the 90 percent range
With a little work he [George W.]may break out of the single digits with Muslim-American voters [in the 2004 election].
Everyone […] knows a wavering former Bush voter […] or, worse for the administration, someone who is sure they aren't voting for Bush this year. In the last four years he has done something that offends them or taken a stand they cannot abide.
What I find surprising is that he hasn't done something that offends everyone in the last four years, but that's just me.


text:
Wednesday, February 1, 2017
dab for the gram
I spent most of my Saturday chatting and taking photos with my co-worker and friend Daquan. We had quite a day and this photo was one of my favorites so I thought it was a perfect post for how I feel about Wednesdays and the weekend.
If you want to see more of my photos, check out my instagram ( instantdork2.0 ). Comment your anagram names bellow so I can check you guys out!
Subscribe to: Post Comments (Atom)


text:
Picture: * PRINCE HARRY 'WILL NOT SERVE IN IRAQ' LATEST: British royal PRINCE HARRY will not be deployed to Iraq, according to senior army officials....
Young British royal Prince Harry celebrated the end of his school exams yesterday (19JUN03) by indulging in a drunken pub crawl.
The 18-year-old prince went on a six-hour tour of some of London's top nightspots with his friends and two detectives, to mark their final A' Level examination that day.
One onlooker remarks, "He certainly seemed to be having a good time, and was definitely in no hurry to head home. Harry really was enjoying himself - he was very jolly."
Harry, whose mother DIANA, Princess Of Wales died in 1997, was seen knocking back bottles of lager, cocktails and wine throughout the evening.
A fellow drinker says, "He was very chatty and keen to mix. There were lots of boys and girls of his own age around.
"Even though he was obviously out late, he didn't seem too much the worse for wear and handled the scene well. He just wanted to get stuck-in and have fun."20/06/2003 13:10


text:
Gregg writes,
Will Grass Out Max work to eliminate grass from a baseball infield? And will it kill weeds to?
Grass Out Maxx is only labeled to control grassy weeds. It will not kill flowers, shrubs, broadleaf crops or other broadleaf plants. Agrisels Gly Pho-Sel Pro with 41% Glyphosate would be a better product to use, just be sure to only apply to the weeds/grasses you are wanting to kill as it can kill anything that it is applied to.
Answer last updated on: 02/09/2015
Was this answer helpful to you? Yes No


text:
32nd Aerospace Sciences Meeting & Exhibit: 94-0640 - 94-0669
American Institute of Aeronautics and Astronautics, 1994 - Aeronautics
From inside the book
72 pages matching structures in this book
Where's the rest of this book?
Results 1-3 of 72
What people are saying - Write a review
We haven't found any reviews in the usual places.
Other editions - View all
Common terms and phrases
3-D TBL accuracy AIAA AIAA Paper Aircraft airfoil algorithm angle angle of attack average axisymmetric boundary layer boundary layer thickness bow shock calculated cell centered expansion centerline coefficient compressible configuration contours convective convergence curvature cylinder delta wing developed diameter dimensional distribution domain downstream duct dynamic dynamic modulus effect exit experimental flat plate flow field Fluid Mechanics flux Formulation freestream frequency grid images implicit incoming boundary layer interaction laminar laser linear Mach number measured mesh method modulus Navier-Stokes equations nozzle obtained parameters particle plane present problem reconstruction region Reynolds number Scalar scheme shear layer shock shown in Figure simulation solution solver spanwise spatial streak streamwise stress structures subsonic supersonic surface temperature tions turbulence model turbulent boundary layer turbulent flow unstructured unstructured grid unstructured-grid upwind USLIP values velocity Velocity Profile computed viscoelastic viscous vortex vorticity wake wall wind tunnel Yaw Angle


text:
You can positively impact lives right in your own community with the Y. Volunteers of all ages help the Y nurture youth, improve health and well-being and assist our neighbors in need.
Youth Development
Our volunteers nurture the potential of thousands of kids year-round. They work with our Child Watch staff, assist with our quarterly Kids' Night Out events, plan, execute and chaperone our Father-Daughter Dance, coach Girls on the Run and encourage athletes at the Youth Triathlon.
Healthy Living
Y volunteers provide support, resources and guidance to kids, adults and families so they can achieve greater health and well-being in spirit, mind and body. Y volunteers work at the annual Robin Run, Girls on the Run, Yoga Day and additional special events.
Social Responsibility
It is the community of YMCA volunteers that help provide Christmas gifts to families in need through our partnership with the Hattie Daniels Day Care Center, help gather backpacks and school supplies for the 1000 Backpacks in 100 Days initiative through the Wilson Public Schools and donate gently used shoes for Soles 4 Souls.


text:
As always, our green season gives clients an opportunity to forget their busy lives and immerse themselves in unique sights, smells and sounds of the African bush. The rains have already made a majestic appearance this year with Impala already having their little calves and Wildebeest doing the same. The recordings so far seem higher than last year's rainfall recordings.
The Hwange National Park has undergone a remarkable transformation and Somalisa is now embedded in a place of many shades of green. Although we are already in the green season, extremely good sightings are still being recorded around Somalisa, and are most vividly described by our guests themselves who wrote:
"We saw three wild dogs being chased from the pan by a big Bull elephant. Two buffalo were killed on the same night about 20 meters away from the pan by two female lion. These same two female lion had a little rumble and tumble and chased off a big male lion the following night… they were protecting their cubs… the females were batting him with their paws etc right next to the guest toilet by the main area." -November 2008
"The male lion has been in and out of the camp and we hear him every night…" – November 2008
"We have had sable at the pan, a big herd of 60 buffalo this morning also at the pan along with Zebra, a crowned crane and many other birds" – November the 17th
The Chobe Enclave on which our Linyanti Bush Camp is located has also received refreshing rains. This is quite a relief as the whole of Southern Africa, mainly Zimbabwe, Botswana, Zambia and Namibia experienced severe field fires this last October. A number of measures initiated by the Camps managers and National park officials have always been in place to contain these natural fires.


text:
JoySpring Celebrates Successful Launch of New Natural Thyroid Support Supplement
JoySpring executives are elated at initial response to their new Thyroid Support Supplement
Charlotte, NC (PRWEB) May 17, 2016
JoySpring, a premium brand of natural health supplements, is proud to announce the US release of their new Thyroid Complex, a complete thyroid support supplement for individuals facing the effects of hypothyroidism.
After years of research, analysis and development, JoySpring's Thyroid Energy is now available over the counter for the very first time.
During pre-launch trials hundreds of satisfied users reported a boost in metabolism and reduced fatigue. The adoration has continued post-launch with hundreds of 5 star consumer reviews in mere weeks. The new product is currently one of the highest rated iodine support supplements available on Amazon.
With a blend of clinically tested ingredients including L-Tyrosine, Iodine, B-Vitamins, copper, magnesium, and traditional Chinese herbs, and the highest potency hypothyroidism support formula available in an all-natural form, JoySpring's Thyroid Complex promises to become one of the most popular thyroid supplements available.
L-Tyrosine plays a key role in Thyroxine and Triiodothyronine (T3) production, two hormones responsible for regulating thyroid hormone (TSH) production.
Clinical tests demonstrate that L-Tyrosine, combined with Iodine and B-Vitamins can increase TSH production and ease the symptoms of hypothyroidism.
JoySpring combines the power of L-Tyrosine, Iodine, and B-Vitamins with natural Schizandra and Aswaganda, two ancient Chinese herbal remedies long used to cure metabolic disorders. Current research indicates that these herbs can also help regulate the metabolism and fight against many thyroid disorder side effects.
Effective thyroid supplementation is increasingly becoming a required part of a healthy diet.
Nearly 20 million Americans currently suffer from a form my thyroid disease and more than 12% of people in the United States will develop a thyroid condition during their lifetime. Current estimates indicate that up to 60% of people with thyroid disease are unaware of their condition.
Low energy levels, food cravings, weight gain, metabolic disorders, and sleep disturbances are just a few of the silent signs of hypothyroidism.
Blood tests are available to determine thyroid hormone levels and medical treatments including hormone therapy are frequently prescribed to treat the myriad of symptoms associated with thyroid disease.
Until now, high-potency clinically-tested natural supplements to support thyroid function were difficult to find in a single source.
Combining over 7 research-backed ingredients, JoySpring's Thyroid Complex offers a single supplement solution to thyroid support. Consumers treating thyroid conditions or combining medical treatment with over the counter supplements no longer need to rely on multiple bottles of expensive supplements.
All production runs of Thyroid Complex are made in a US-Based GMP certified facility following FDA production and supplement safety guidelines.
Due to high quality standards, supply sourcing, and a limited production run, quantities of JoySpring Thyroid Complex are limited. Customers wanting to learn more about the rising health company can learn more on their website at http://www.hellojoyspring.com or on http://www.amazon.com
For the original version on PRWeb visit: http://www.prweb.com/releases/2016/05/prweb13410732.htm


text: So my phone is rooted and I want to know if when I run an antivirus will it show my root or apps as viruses if they have superuser permissions? Because when I run antivirus it shows several viruses. If I clean them idk what it will do. Any help would be greatly appreciated thanks. And when I say several I mean like 50 to 100 files show infections. Gmail is cjohns8792 if anyone would like to help me through it with gmail thanks.


text:
Tadanori Yokoo (born 1936) is one of Japan's most successful and internationally recognized graphic designers and artists. His early work shows the influence of the New York based Push Pin Studio (Milton Glaser and Seymour Chwast in particular) but Yokoo himself cites filmmaker Akira Kurosawa and writer Yukio Mishima as two of his most formative influences. In the late 1960s he became interested in mysticism and psychedelia, deepened by travels in India. Because his work was so attuned to 1960s pop culture, he has often been (unfairly) described as the "Japanese Andy Warhol" or likened to psychedelic poster artist Peter Max, but Yokoo's complex and multi-layered imagery is intensely autobiographical and entirely original.
(These scans are from the ever-inspiring blog "A Journey Round My Skull." I highly recommend taking a gander at his flickr page and blog for more illustration and graphic design sweetness.)
Also of note is the Two LP picture disc "Opera from the works of Tadanori Yokoo" by Toshi Ichiyanagi which Mutant Sound has made available, and describes as:
"...an unique amalgamated mixture of lysergic demented psychedelic assault-like stunt rock, a aural whirlwind filled with acid folk ramblings, tape collages, field recording excerpts, radio commercial snippets, roaring jet engines, electronic music excursions into the vast unexcavated canyons and dungeons of your mind, squealing sound fragments of frogs mating in a nearby pond, drowned out enka escapades, kayokyoku excursions into no mans land, Takakura Ken nasal singing, spoken word fragmentation bombs by Kara Juro amongst others, traffic noises, sonic sound clusters of blistering fuzzed out psychedelic mayhem, stratosphericstatic electronic hissing, radio news broadcast flashes, vintage electronic tape music escapades, chirping cricket orchestras in a distant backgroundand so much more non-adaptive and deranged sonic activity."
(Jordan, I'm going to recommend it to you without even giving it a listen.)
photo of Tadanori Yokoo
I can't gather much about this, but I love this too:
Tadanori Yokoo & Bohemians ビッグバンダナ TATOO FRONT & TATOO BACK
I can't gather much about this, but I love this too:
Tadanori Yokoo & Bohemians ビッグバンダナ TATOO FRONT & TATOO BACK


text:
Richard Strauss: Elektra Trailer (2006)
Elektra, Op. 58, is a one-act opera by Richard Strauss, to a German-language libretto by Hugo von Hofmannsthal, which he adapted from his 1903 drama Elektra. The opera was the first of many collaborations between Strauss and Hofmannsthal. It was first performed at the Dresden State Opera on January 25, 1909. Elektra is a difficult work and is musically complex. A performance requires musicians with great stamina. The role of Elektra in particular is one of the most demanding in the dramatic soprano repertoire.
Watch the official Richard Strauss: Elektra 2006 trailer in HD below or find more Richard Strauss: Elektra videos on Vidimovie.


text:
Hello, brand new here. My wife got great insight here when we did a Pac Northwest trip a few years back.
A few of my good friends and I are planning a trip to Europe in June 2012. We're basing the trip around Amsterdam and trying to figure out where we go from there.
1. Trying to decide to either go 1 of 2 ways. Amsterdam, London, Brussel, Paris -or- Amsterdam, Brussels, Munich, Prague, Amsterdam. Any thoughts or suggestions?
2. Any insight on booking a rail pass? I was looking around and it doesn't look like all the cities/countries are compatible with each other.
Is the Eurostar the only line that runs under the channel to London?
If we get a pass on eurail or raileurope would we have to then get another pass to go to London?
If someone could point me in the right direction on this that would be great! Thank you!
Recent ActivityView all Europe activity »
- 1 Pubs showing NFL football in London?
- 2 Lisbon neighborhoods
- 3 Help With Itinerary By Train: London, Paris, Nice, Florence
- 4 Looking for Good Eating in Valencia
- 5 The Little Cyclades, Santorini, Vienna
- 6 [Crete] Trapped in Skouloufia? Please help!
- 7 What happened to Sundriedtopepo
- 8 Help with Venice Trip?
- 9 Dubrovnik
- 10 Norte Dame light show!
- 11 Rome, Tuscany & Umbria
- 12 Where to Eat in Barcelona
- 13 Adventureseeker returns to Italy! As glorious and detailed as before!
- 14 Help with 11 night Portugal Itinerary
- 15 London - Paris - Amsterdam trip planning help
- 16 10 Days in Austria
- 17 Czech Republic & Germany in Eleven Days
- 18 Sampling Some of Sicily and Bits of Italy Beyond
- 19 Greece in end October
- 20 Champagne Tours
- 21 GTG Paris December 2017
- 22 Restaurant suggestion Bamberg
- 23 Russian-American Relations: safe to travel? Get Visas?
- 24 Mantova, Ravenna, Brisighella, Bologna
- 25 Le Marche Research Help
Trip advice - first time
Hello, brand new here. My wife got great insight here when we did a Pac Northwest trip a few years back.
Report Abuse
I'll jump in with the usual questions.
How long will you be in Europe? Including or excluding your travel days?
What do you like to do? Art and culture, history, outdoors activities like hiking or biking? Drinking, smoking, good food, etc? Do you prefer big cities, small towns, the countryside, mountains, beaches, or a mixture of some/all of those?
On the Amsterdam, London, Brussels, Paris option, it makes the most sense to follow the order of: Amsterdam, Brussels, Paris, London. Or the reverse.
I've never gotten a rail pass, but based on the advice I've seen given here, it's not always the best financial decision - I think you need to price it out vs. point-to-point tickets to your destinations.
Whatever itinerary you end up with, it makes the most sense to travel open jaws - in one city and out another. That way you aren't backtracking.
Report Abuse
mmm - what is it that draws you to Brussels?
ignoring that, Lexm is right - open-jaw and using the eurostar to get from Paris to London, [or vive versa] is the best option.
Report Abuse
And yes, the eurostar is basically the only passenger train to and from London (there is a car train as well I believe). Be sure to book your tickets as early as you can to get the best price.
Report Abuse
Thank you for the quick replies.
We can certainly scratch Brussels.
This trip has been "talked about" for about 12 years - "Let's go to Amsterdam when we're 30" and he we are.
We're going for about 12 days (2 travel days) in early in June. This is a trip with my 3 college buddies so we're looking more at drinking, smoking, eating and site-seeing. In my opinion, we should scratch London and Paris and go west of Amsterdam (Germany, Czech). I'm going to London and Ireland in February with my wife so that's why I'm kind of leaning towards not going to London with the guys.
Round trip flight from Chicago to Amsterdam and back is significantly cheaper and we are on a budget. Plus I don't think we'd mind having to back track to Amsterdam.
Report Abuse
Make sure you're checking the pricing on open jaws flights carefully - in my experience, the flights themselves tend to be the same as a round trip flight. (Including flights from Chicago when we lived there.)
Also, especially if you end up in Prague as your eastern-most location, you'll need to factor in travel time - out of your 10 days of vacation time - and the cost to get back to Amsterdam. Let's say 8 1/2 hours, at least, if you drive, and 10-12 hours by train. Even on a budget, your time is worth money.
Report Abuse
Make sure you're checking the pricing on open jaws flights carefully - in my experience, the flights themselves tend to be the same as a round trip flight. (Including flights from Chicago when we lived there.)
Also, especially if you end up in Prague as your eastern-most location, you'll need to factor in travel time - out of your 10 days of vacation time - and the cost to get back to Amsterdam. Let's say 8 1/2 hours, at least, if you drive, and 10-12 hours by train. Even on a budget, your time is worth money.
Report Abuse
Open-jaws is multi-city on airline web sites. When you compare costs, add in the cost to return to Amsterdam and the time you'll lose backtracking.
Report Abuse
Smoking in Czech is still prevelant, a little less in Germany and not really acceptable in Netherlands. Dutch food has improved in the last few years but you will pay for it, Germany generally is more reasonably priced and Czech even easier (though the meat content goes up as you pass to the East)
While this time is not a good focus for beer festivals there are some good wine festivals. Do not think the gentile wine sniff and slurp of the states, think glasses (up to a pint in size) of wine and lines of roasting pig leg cooked to perfection and stuffed in buns to help the wine go down. German tourist info has a list of most festivals in 2012.
Report Abuse
a useful article http://www.mybikeguide.co.uk/German_Wine_Festivals.php
Report Abuse
Thank you all. Your info is much appreciated. We're kind of all over the place here.
I did more research on flights and we decided to scale it back to 3 major cities with most likely an open-jaw flight.
Chicago, London, Paris, Amsterdam, Chicago
Chicago, Amsterdam, _______ , _______ , Chicago
We're hoping to all sit down on Monday night and figure out what the priorities are.
Report Abuse
I would recommend you look at the trip as a whole. As you already found out, optimizing solely on the basis of transatlantic flight lead to an inefficient routing.
Different people can/want to travel different number of destinations in a given time. Some are very emphatic about their approach. But in the end, it is up to you how you want to spend time.
Play with different open-jaw combinations. For each identify what you can do with them, how to connect the dots, and the cost implications.
In 10 days, some possible open-jaw combinations if you are staying on the continent:
AMS in, back from CDG (+sw Netherlands/Bruges)
AMS in, back from FRA (+Mosel/Rhine)
AMS in, back from MUC (+Neuschwanstein/Salzburg)
AMS in, back from CPH (+northern Germany)
and so on.
I presume the trip had to be in June? It is when the airlines hike prices to catch families with kids in school who cannot travel earlier.
Sign in to comment.


text:
My Nonprofit Reviews
Review for Erowid Center, Grass Valley, CA, USA
I discovered Erowid 10 years ago. Sinse then, Erowid has been my one and only reliable source of info in this field. Here you will find everything you "need", and should"want" to know; concerning the information provided. I always recommend Erowid to my associates, because learning this information will save your life.
More Feedback
If I had to make changes to this organization, I would...
Advertise on other domains to better help those out, who are unaware of the dangers that Erowid has stopped many of people from falling in to.
Would you volunteer for this group again?
For the time you spent, how much of an impact did you feel your work or activity had?
Did the organization use your time wisely?
Very Well
Would you recommend this group to a friend?
When was your last experience with this nonprofit?


text:
The Poly Prep Case
Poly Prep Country Day School (known familiarly as "Poly Prep") is the subject of a federal lawsuit filed in Brooklyn in 2009 centering on the sexual assault of students by Philip Foglietta, the head football coach from 1966 to 1991. A 2004 state suit against the school was dismissed due to the statute of limitations. The 2009 case cites the Racketeer Influenced and Corrupt Organizations (RICO) Act in alleging that past and current administrators engaged in a coverup of the abuse. The school acknowledged in 2002 that it had received "credible allegations that such abuse occurred at Poly Prep."
Legal Documents
- August 28, 2012 - Zimmerman v. Poly Prep Country Day School, Memorandum & Order
For a complete listing of all the articles written about Poly Prep, please do a search for the term from our news section. Below are a few of the more pertinent news reports:
- Sex Abuse Suit Against Poly Prep Survives School's Bid to Dismiss - NY Law Journal, 8/30/12
- Lawyers Say Ruling Allows Abuse Suits - Wall Street Journal, 8/30/12
- Judge Says School Abuse Case May Proceed - NY Times, 8/28/12
- Lawsuit Claims Poly Prep Covered Up Sex Abuse Of Young Boys For Decades - NY Daily News, 12/7/11
- Brooklyn's Poly Prep Covered Up Football Coach's Sex Abuse - NBC, 12/6/11
- Simpson Thacher Partner on Sex Abuse Case Against His High School: "I Have to Do This" - Zack Lowe, 4/12/10


text:
The nucleoside adenosine acts on the nervous and cardiovascular systems via the A2A receptor (A2AR). In response to oxygen level in tissues, adenosine plasma concentration is regulated in particular via its synthesis by CD73 and via its degradation by adenosine deaminase (ADA). The cell-surface endopeptidase CD26 controls the concentration of vasoactive and antioxidant peptides and hence regulates the oxygen supply to tissues and oxidative stress response. Although overexpression of adenosine, CD73, ADA, A2AR, and CD26 in response to hypoxia is well documented, the effects of hyperoxic and hyperbaric conditions on these elements deserve further consideration. Rats and a murine Chem-3 cell line that expresses A2AR were exposed to 0.21 bar O2, 0.79 bar N2 (terrestrial conditions; normoxia); 1 bar O2 (hyperoxia); 2 bar O2 (hyperbaric hyperoxia); 0.21 bar O2, 1.79 bar N2 (hyperbaria). Adenosine plasma concentration, CD73, ADA, A2AR expression, and CD26 activity were addressed in vivo, and cAMP production was addressed in cellulo. For in vivo conditions, 1) hyperoxia decreased adenosine plasma level and T cell surface CD26 activity, whereas it increased CD73 expression and ADA level; 2) hyperbaric hyperoxia tended to amplify the trend; and 3) hyperbaria alone lacked significant influence on these parameters. In the brain and in cellulo, 1) hyperoxia decreased A2AR expression; 2) hyperbaric hyperoxia amplified the trend; and 3) hyperbaria alone exhibited the strongest effect. We found a similar pattern regarding both A2AR mRNA synthesis in the brain and cAMP production in Chem-3 cells. Thus a high oxygen level tended to downregulate the adenosinergic pathway and CD26 activity. Hyperbaria alone affected only A2AR expression and cAMP production. We discuss how such mechanisms triggered by hyperoxygenation can limit, through vasoconstriction, the oxygen supply to tissues and the production of reactive oxygen species.
- adenosine
- adenosine receptor
- CD26 peptidase
- hyperbaric conditions
- hyperoxia
in vivo and in cellulo studies on the nervous system, cardiovascular function, and membrane transport have shown that severe perturbations result from pressures of a few atmospheres, whereas in vitro, at the molecular level, denaturation of molecules requires pressures of hundreds of atmospheres (26, 57). In addition, as early as in the 1970s and with in vivo relevance for terrestrial species, studies showed that low hydrostatic pressure in the range of (patho)physiological conditions modifies major processes such as cell signaling, membrane traffic, and cell proliferation (2, 8, 15, 34). Exerting hydrostatic pressure on cells or living animals is therefore a means to address physiological mechanisms.
Hyperbaric conditions, notably hyperbaric oxygenation, have established medical, mountaineering, and diving applications because of their capacity to increase the oxygen level in tissues, 1-atmosphere oxygen being used for example in the treatment of carbon monoxide poisoning and 2-atmosphere oxygen being used as breathing gas by divers in extreme conditions (31, 62). Although it displays major side effects such as production of reactive oxygen species (3), the use of hyperoxygenation is strongly growing.
Adenosine is a nucleoside produced from ATP at the cell surface by CD39 and the ecto-5′-nucleotidase CD73. Adenosine is also produced in the cell before secretion by membrane transporters (48). Monitoring CD73 production is a useful means to address the regulation of adenosine synthesis (16, 29) because CD73 expression contributes to regulate the concentration of extracellular adenosine as previously reported in a context of adenosine deaminase (ADA) deficiency (51). Adenosine plasma concentration increases during hypoxia, ischemia, inflammation, and oxidative stress (20, 22, 56). Various mechanisms including altered expression of several enzymes involved in ATP metabolism and transcription of genes involved in adenosine metabolism and transport act in concert to modulate the extracellular adenosine concentration in response to the oxygen supply to tissues. For example, hypoxia decreases the intracellular ATP concentration and increases the production of its degradation product adenosine, whereas it inhibits the adenosine kinase-mediated adenosine degradation. The coordination of these processes induces a rise in the concentration of the nucleoside (21, 25, 56).
Adenosine acts via four G protein-coupled receptors that are conserved among vertebrates (22). The A2A receptor (A2AR) is of particular interest because it is expressed in the brain, where it inhibits dopaminergic pathways of the basal ganglia, and in the cardiovascular system, where it controls the heart rate and the blood pressure by its vasodilator capacity (18, 23, 39, 55, 63, 64). It is known that A2AR expression increases in hypoxic conditions (11, 13, 22, 37). Regarding hyperoxia, the adenosinergic response has been already addressed in the lung, and the supplemental use of high oxygen levels to treat the acute respiratory distress syndrome was found to weaken the local tissue hypoxia-driven and adenosine-mediated anti-inflammatory mechanism, hence further exacerbating lung injury (16, 48, 61). Recently, the effects of hyperoxia on the CD39/CD73-driven production of adenosine and A2AR expression were addressed from a cancer therapy perspective. Indeed, there is a need for reversing the immunosuppressive effects mediated by A2AR on T and NK cells in the hypoxic adenosine-rich and tumor-protecting microenvironment and for converting the latter into a normoxic and adenosine-poor environment that induces tumor regression (29, 30). These pioneer studies have evaluated the effects of hyperoxia in the lung or in cancer/inflammation models, and there is therefore a need for addressing the situation in the brain using normal animals/tissues.
ADA and surface CD26 are other major components of the response to blood oxygen level. Plasma ADA degrades adenosine into inosine and therefore controls the extracellular level of the nucleoside that, besides its anti-inflammatory effect, exhibits cytotoxicity (20). In higher mammals, ADA interacts with CD26 at the surface of mononuclear cells and the capacity of the aminohydrolase to bind CD26 was interpreted as a means to protect, via adenosine degradation, cells from cytotoxic effects of high adenosine concentration in the immediate vicinity of the cell surface (1, 19, 33, 46). In rodents, it is considered, however, that ADA does not bind cell-surface CD26 (19).
In mammals, CD26 also exhibits the dipeptidyl peptidase-IV (DPP-IV) activity. DPP-IV cleaves NH2-terminal dipeptides from peptides containing proline or alanine in the penultimate position to control the concentration of their active intact form (7, 41). CD26 substrates include peptides produced in response to oxidative stress or abnormal oxygen supply to tissues (41). These peptides act on the redox stress and energetic metabolism, in particular in high-energy-consuming tissues such as the skeletal muscle, as well as on brain function and oxygen supply to tissue by ino/chronotropic effects and vasodilation in response to oxygen deficiency or inflammation (e.g., substance P, neuropeptide Y, vasoactive intestinal peptide, glucagon-like peptide 1, and brain natriuretic peptide) (7, 41). CD26 is therefore a major player in the systemic response to changes in blood oxygen level. However, whereas changes in CD26/DPP-IV expression are documented following ischemia/reperfusion and hypoxia (20, 41, 46), CD26 activity during hyperoxia and hyperbaria remains unclear.
For these reasons, we sought to address here the effects of high blood oxygen level and hyperbaria on various components of the adenosinergic pathway, from changes in the extracellular adenosine concentration up to cAMP production following A2AR activation. CD26 was also addressed.
Animals and reagents.
Rats (male Sprague-Dawley rats; Charles River, F; 300–350 g) were treated according to the European Guidelines (Brussels, Belgium; 86/609/EEC), the French law (decree 87/848), and the rules established by our Institutional Animal Care and Use Committee (agreement number B13.0005.8) Rats were kept at 22 ± 1°C under a 12-h:12-h light/dark cycle (lights on at 7:00 AM) with food (A03, UAR) and water available ad libitum. Rats (n = 8 per condition) were exposed in a pressure chamber for 4 h (25°C; H2O: 40–60%; CO2 < 300 ppm) as follows: normoxia at atmospheric pressure (210 mbar O2, 790 mbar N2), a condition known to generate a partial pressure of oxygen in the arterial blood (PaO2) of ≈90 mmHg, i.e., the control situation (10, 58) (normoxia thereafter); hyperoxia at atmospheric pressure (1 bar O2), a condition generating a PaO2 of ≈450 mmHg (10, 58) (hyperoxia thereafter); hyperoxia in hyperbaric conditions (2 bar O2), a condition generating a PaO2 of ≈900 mmHg (hyperbaric hyperoxia thereafter); and normoxia in hyperbaric conditions (2 bar with 210 mbar O2, 1,790 mbar N2), a condition generating a PaO2 of ≈90 mmHg and that is often considered to experimentally reproduce authentic normoxia under such hyperbaric conditions (9) (hyperbaria thereafter). Isoflurane was used as an anesthetic agent before brain collection. Other reagents were purchased from Sigma-Aldrich.
Cell line expressing A2AR.
We used a murine Chem-3 cell line expressing the functional A2AR (HTS048C; Millipore). Suspension cells were cultured in RPMI 1640, 10% FCS, 0.8 mg/ml geneticin (G418), 100 IU/ml penicillin/streptomycin, and 1/100 dilution of the Chem-3 growth supplement (Millipore) under various pressure and gas conditions for 4 h before storage of cell pellets at −80°C.
Adenosine plasma level.
Blood samples (1 ml) were obtained following cardiac puncture using a syringe containing 0.25 ml of cold stop aqueous solution [0.2 mM dipyridamole, 4.2 mM Na2-EDTA, 5 mM (9-erythro-3 nonyl) adenine, 79 mM αβ-methylene adenosine 5′diphosphate, 1 UI/ml heparin sulfate, and 0.9% NaCl] to prevent subsequent degradation and uptake of adenosine (28). Samples were then deproteinized using 5% perchloric acid (1 vol:4 vol), and adenosine was identified and quantified using high-performance liquid chromatography coupled with a diode array detector (Chrom System) as described (28). Adenosine was identified by spectrum and elution time (detection threshold: 5 pmol in plasma; coefficient of variation: <5%).
After cardiac puncture and centrifugation, serum ADA was measured as described previously (28); adenosine (750 μl, 28 mM) was incubated with serum (125 μl) in saline (125 μl, 7% BSA) for 37 min at 37°C. The reaction was started by adding adenosine and stopped in ice water. Ammonia resulting from adenosine degradation by ADA was measured using a Synchron LX 20 analyzer (Beckman Coulter) and expressed as international units per liter.
A2AR and CD73 expression.
A2AR expression was assessed using a semiquantitative Western blot procedure (13). Frozen brains or Chem-3 cells were quickly thawed and treated using a 4% SDS aqueous solution supplemented with a protease inhibitor cocktail (Sigma-Aldrich) and a 15-min sonication treatment at 47 kHz. After protein quantification (microBCA assay; Pierce Biotechnology), samples (10 μg) were diluted in loading buffer (65.2 mM Tris·HCl buffer, pH 8.3, containing 10% glycerol, 0.01% bromophenol blue, and 5% 2-mercaptoethanol) and analyzed using 12% SDS-PAGE before electroblotting onto PVDF membrane. Filters were incubated for 20 min with Adonis (1 μg/ml), a mouse monoclonal antibody directed against human A2AR (12) that can readily detect and semiquantify in denaturing conditions A2AR antigens from various origins (e.g., human, mouse, rat, heart, peripheral blood mononuclear cells, kidney, brain). Filters were then incubated with horseradish peroxidase-labeled anti-mouse antibodies and a chemiluminescent substrate (SuperSignal West Femto, Pierce Biotechnology). Densitometry analysis of the 45-kDa species corresponding to the A2AR antigen was performed using the ImageJ software (NIH). CD73 expression was similarly studied in brain samples (100 μg) using Western blot analysis and the H-300 rabbit polyclonal IgG (Santa Cruz Biotechnology; 1:200) before densitometry analysis of the corresponding band (70 kDa).
A2AR mRNA.
Total mRNA was extracted from brain samples (PAXgene, Blood RNA kit, Pre Analytix, Qiagen). Real-time quantitative mRNA expression analysis was performed using 250 ng of cDNA encoding the A2AR sequence (Light cycler, Roche). The forward primer sequence was 5′-GAGGACGTGGTGCCCATGAAT-3′, and the reverse primer sequence was 5′-TGCAGCGTGGACCGAGT-3′. β-Actin was used as housekeeping gene.
cAMP production.
Measurement of total cAMP in Chem-3 cells was performed using the Biotrak kit (GE Healthcare Biosciences) according to the nonacetylation procedure described by the manufacturer. Cells (106 cells/well) were incubated with/without various concentrations of the A2AR agonist CGS-21680 for 60 min at 37°C under various gas and pressure conditions. The incubation period was stopped by adding the dodecyltrimethyl ammonium bromide acetate lysis buffer provided with the kit. Production of cAMP in each well was then determined (n = 3) using the competitive enzyme immunoassay according to the manufacturer's instructions (11). Results were given either as a percentage of cAMP produced in the condition of interest vs. the basal production level or as pg/106 cells.
Blood samples (5–10 ml) were obtained following intracardiac puncture using a syringe containing calciparine. Mononuclear cells were isolated using a separation gradient-based procedure; blood was diluted (vol/vol) using 0.9% NaCl and 0.05% NaN3 and mixed (vol/vol) with the lymphocyte separation medium Lymphosep (MP Biomedicals). Cells were isolated (21°C, 400 g, 40 min) before being washed (n = 3) using RPMI 1640 culture medium. Cells (2 × 106/ml) were then incubated with a colorimetric substrate of DPP-IV, Gly-Pro-P-nitroanilide (3 mM, 2 h in 75 mM glycine buffer, pH 8.7) (46). Signal background was determined using incubation in 1 M acetate buffer, pH 5, a condition in which DPP-IV is inactive. Optical density was measured at 405 nm, and the results were expressed as international units.
Statistical analysis.
Data were expressed as means ± SD. A two-way ANOVA was used for intergroup comparisons. A P value of <0.05 was considered significant.
Adenosine plasma concentration (in vivo).
Hypoxic conditions promote accumulation of adenosine inside and outside the cell, notably in the blood (22). Here, we quantified adenosine in blood sampled from animals submitted to hyperoxia combined, or not, with hyperbaric conditions. Hyperoxia alone (1 bar O2) reduced the adenosine plasma concentration compared with normoxia, the control situation (means ± SD: 0.40 ± 0.18 vs. 0.77 ± 0.15 μM, respectively; Fig. 1A). Hyperbaric hyperoxia (2 bar O2) further affected this parameter (0.15 ± 0.02 μM). No significant difference was found between the adenosine level obtained using hyperbaria alone (0.65 ± 0.25 μM) and normoxia. These data show that the adenosine plasma level was mainly decreased by the oxygen level, hyperbaric treatment lacking significant influence per se under normoxic conditions.
CD73 regulation (in vivo).
CD73 expression is a marker used to address the regulation of adenosine synthesis (29, 52). CD73 expression in brain samples was analyzed by Western blotting. The 70-kDa species corresponding to the CD73 antigen (Fig. 1B) was semiquantified using densitometry analysis. Hyperoxia alone increased CD73 expression compared with normoxia (means ± SD: 3.43 ± 0.64 vs. 1.10 ± 0.19 AU, respectively; Fig. 1C), hyperbaric hyperoxia exhibiting a similar effect (2.95 ± 0.72 AU). Hyperbaric treatment alone had no significant effect vs. normoxia (1.49 ± 0.27 AU). These results show that hyperoxia increased CD73 expression, hyperbaric conditions having no significant influence.
ADA regulation (in vivo).
High adenosine concentration can exhibit cytotoxic effects, and its extracellular level is regulated by an adaptive response that includes degradation into inosine by the serum aminohydrolase ADA. We addressed here the effects of hyperoxic and hyperbaric treatments on serum ADA. We found that hyperoxia alone tended to increase ADA compared with normoxia (means ± SD: 16.3 ± 6.3 vs. 8.8 ± 3.7 IU, respectively) (Fig. 1D). Hyperbaric hyperoxia further increased the activity (28.9 ± 8.5 IU). No significant difference appeared between the hyperbaric condition alone (8.7 ± 6.2 IU) and normoxia. As a control, we quantified the transaminase level in the corresponding blood samples (using a Cobas 8000; Roche Diagnostics) and found no significant differences between conditions (means ± SD; normoxia: 67 ± 14 IU; hyperoxia: 78 ± 21 IU; hyperbaric hyperoxia: 71 ± 13 IU; hyperbaria: 59 ± 11 IU; P > 0.05 compared with normoxia). These data show that hyperoxia increased plasma ADA, hyperbaric treatment having no influence per se under normoxic conditions.
CD26/DPP-IV regulation (in vivo).
CD26 expressed on blood mononuclear cells displays the DPP-IV activity, which controls by endoproteolysis hypoxia-regulated peptide substrates with proline in penultimate position (7). We monitored CD26 expression on blood mononuclear cells sampled from animals using the chromogenic peptide substrate Gly-Pro paranitroanilide. Hyperoxia alone significantly decreased DPP-IV activity compared with normoxia (means ± SD: 0.87 ± 0.16 vs. 1.08 ± 0.11 U) (Fig. 2). Hyperbaric hyperoxia further decreased the activity (0.78 ± 0.16 U). Hyperbaric treatment alone did not significantly modify DPP-IV (1.01 ± 0.17 U) compared with normoxia. These data show that CD26 expression was mainly downregulated by the oxygen level, hyperbaric conditions having no significant influence per se under normoxic conditions.
Regulation of A2AR expression (in vivo).
Adenosine acts following binding to the surface A2AR of numerous cell types, notably in the cardiac and nervous systems (22, 63, 64). We chose here to monitor the effects of hyperoxic and/or hyperbaric treatments on A2AR expression in brain, dissociating heart tissue into its component cells being a long and difficult process that affects the integrity of protein components, hence their biochemical characteristics/characterization. We studied mRNA expression of A2AR (Fig. 3A). Hyperoxia alone decreased mRNA expression compared with normoxia (means ± SD: 1.9 ± 0.6 vs. 3.0 ± 0.6 AU, respectively). Hyperbaric hyperoxia further decreased the expression (1.3 ± 0.8 AU), hyperbaric treatment alone inducing a similar effect (1.5 ± 0.3 AU). The protein expression of A2AR was also semiquantified, and a similar pattern was found (Fig. 3, B and C; normoxia: 9.9 ± 1.5 AU; hyperoxia: 8.2 ± 1.5; hyperbaric hyperoxia: 6.9 ± 1.2 AU; hyperbaria: 6.1 ± 2.3 AU). Overall, these results show that hyperoxic and hyperbaric conditions downregulated A2AR expression, the regulation being exerted at the transcriptional level.
Regulation of A2AR expression and cAMP production (in vitro).
Adenosine binding to A2AR stimulates the adenylyl cyclase pathway and GS proteins, which induces a rise in the intracellular cAMP level and activates a plethora of biological processes (36). We addressed here the influence of hyperbaric and/or hyperoxic treatments on cAMP production using two biological tools, a murine cell line and the potent and selective A2AR agonist CGS-21680. Production of cAMP is difficult to evaluate in samples derived from in vivo experiments because the protocol used to isolate cell samples from animals (e.g., length of procedure, use of chemical reagents, and physical effects) does not comply with the requirements of a reliable cAMP quantitation. The cell line we used readily expresses the functional A2AR, and it is therefore a useful tool to address cAMP production resulting from activation of the receptor (6). To validate this cell line as a model to test the effect of hyperbaric and/or hyperoxic conditions on the cAMP component of the adenosine response, we examined whether A2AR expression responded to hyperbaric and hyperoxic treatments. Changes in A2AR expression observed in the cell line submitted to the various conditions used to treat animals were found to be similar to those observed in brain samples (normoxia: 13.1 ± 0.7 AU; hyperoxia: 11.2 ± 0.1 AU; hyperbaric hyperoxia: 10.1 ± 0.4 AU; hyperbaria: 7.1 ± 0.2 AU). Regarding CGS-21680, its use instead of adenosine was attributable to its stability, the natural ligand being rapidly (few seconds) degraded in the cell environment. First, the concentration of CGS-21680 that induced 50% of the maximal production of cAMP (EC50) in the condition of interest was studied (Fig. 4A). The EC50 value reflects the ability of a ligand to activate a receptor and to subsequently cause transduction (its efficacy). Hyperoxic and/or hyperbaric treatments did not modify the EC50 values found in normoxia. Second, cAMP production was examined (Fig. 4B). Hyperoxia alone decreased the production compared with normoxia (means ± SD: 2,990 ± 130 vs. 3,990 ± 250 pg/106 cells). Hyperbaric hyperoxia had a similar influence (3,200 ± 260 pg/106 cells). Hyperbaric conditions alone exhibited the strongest effect (2,150 ± 180 pg/106 cells). Overall, these results show that hyperoxia and hyperbaria downregulated cAMP production in a context where the ligand efficacy remained unchanged compared with normoxia.
This study adds to the line of research that was started by demonstration that A2AR is critically involved in the downregulation of hypoxia/CD73/A2AR-mediated inflammatory damage (50). The present findings are that increasing the oxygen level in inhaled air 5- to 10-fold, and hence the partial pressure of oxygen in the arterial blood, compared with the terrestrial conditions decreased in a dose-dependent manner adenosine plasma concentration, A2AR expression, and CD26 level, whereas it increased both CD73 expression and ADA activity. The consequence of hyperbaric treatment alone was dual: the treatment lacked significant effect on adenosine, ADA, CD73, and CD26, whereas it downregulated A2AR expression.
These results were obtained using wild-type animals although transgenic mice and rats deficient in A2AR expression or overexpressing A2AR are commonly used to study the adenosinergic pathway. We considered, however, that the use of such models had no interest here because manipulating the expression of A2AR induces pleiotropic effects, including high blood pressure, altered heart rate, changes in blood cell properties, metabolic changes, and chronic inflammation (65), which all display patently obvious influences on the variables we studied, thus precluding valuable conclusions regarding authentic mechanisms.
A large amount of literature demonstrates the importance of high adenosine concentration and high A2AR expression in pathophysiological mechanisms resulting from ischemia/hypoxia. It is also known that adenosine and A2AR are important factors in the hyperoxia-induced lung pathophysiology: on one hand, recent studies further described how pulmonary inflammation and edema resulting from hyperoxia are associated with an increase in adenosine concentration and A2AR expression, a hypothesis being that these changes occur to protect the vascular barrier function (16, 17); on the other hand, it was also shown that hyperoxia alters the tissue-protecting mechanism in acute inflammatory lung injury by decreasing the A2AR signaling pathway, and inhalative application of A2AR agonists during oxygen therapy was proposed as treatment (61). Although the interest of examining the effect of hyperoxia on the lung pathophysiology is obvious, the situation was poorly examined for other organs such as the brain and in normal animals/tissues.
However, besides providing knowledge on the impact of systemic high oxygen level on the brain, addressing brain A2AR is greatly needed because the receptor recently emerged as a potential therapeutic target to implement the treatment of acute ischemic stroke (42, 43). Evidence suggests, however, that A2AR has additional roles during stroke: in the first phase of ischemia, it promotes excitotoxicity mechanisms, whereas, in the second phase, immune blood cell-associated A2AR promotes cell adhesion mechanisms and infiltration in the ischemic parenchyma (14, 27, 42, 43, 67). Thus, in addition to induction of reactive oxygen species production, systemic hyperoxia may be detrimental to the brain integrity and function through A2AR expression, and we addressed here the receptor and its ligand following hyperoxygenation.
Measuring the intracerebral adenosine level faces major difficulties (its half-life <10 s, its compartmentalization by the various cell populations, and its release induced by in situ sampling microprocedures). Although the intravascular adenosine concentration may only imperfectly represent the extracellular, active, concentrations found in the various regions of the brain, we proceeded to study blood adenosine level. In contrast with recent studies in the lung (16, 17) but in agreement with conclusions reached from other studies (29, 61), we observed that hyperoxygenation induced a decrease in both plasma adenosine concentration and A2AR expression in the brain in a concentration-dependent manner. Regarding CD73, the present work and a previous study (16) show that its expression increased following hyperoxia in the brain and in the lung. However, hypoxia upregulates CD73 expression (59), and one can expect that hyperoxia does the reverse. The increased expression of CD73 we found here in hyperoxic conditions suggests that the corresponding low adenosine plasma concentration did not result from a decreased synthesis of adenosine by CD73 but from an increased degradation by ADA. Consequently, the high CD73 expression observed here in hyperoxic conditions where adenosine plasma concentrations were low may result from a feedback mechanism to restore the plasma level of the nucleoside.
With consideration that lack of A2AR activation promotes vasoconstriction (47), the low plasma adenosine concentration as well as the low A2AR expression level we found here in the brain following hyperoxia may also constitute an adaptive response by inducing vasoconstriction to diminish the oxygen supply to tissue and the deleterious consequences of production of reactive oxygen species. In this respect, it is worthwhile to note here that 1) the hyperoxic conditions achieved in our experiments mimic conditions that are used in human, inasmuch as 1-bar O2 is a common treatment of CO poisoning and 2-bar O2 is used notably by combat divers (31), suggesting that the effects we observed here can be found in humans; 2) the way hyperoxia regulates the adenosine response, as shown here, contributes to explain how and why hyperoxia promotes vasoconstriction (24); 3) our data are consistent with the observation that a low oxygen level induces adenosine release and high A2AR expression, in particular in peripheral blood mononuclear cells (13, 22, 35, 56, 63).
The hyperbaric condition alone potently decreased A2AR expression in the brain and in the cell line we used even when the oxygen supply to tissue was maintained at an ambient level (normoxia). The decrease in the surface expression of A2AR following hyperoxic and hyperbaric treatments is consistent with the decrease in mRNA production we observed here. Various receptors are affected by the hydrostatic pressure, notably in brain tissues (54). Pressure alters protein expression via a transcription/translation regulation process (4, 34, 57), and this mechanism may have pathophysiological relevance. For instance, with the use of hydrostatic pressure in the range of glomerular capillary hydraulic pressure observed in experimental glomerulosclerosis (uninephrectomized spontaneously hypertensive rats), it was shown that discrete pressure changes increase cell proliferation via DNA synthesis (34).
A2AR is a G protein-coupled receptor that modulates the production of the second messenger cAMP, a regulator of important cellular functions such as proliferation, differentiation, inflammation, and apoptosis (36, 40, 64). Here, cAMP production decreased during hyperoxia associated, or not, with hyperbaric treatment, the effect being maximum when the hyperbaric condition was used alone. This observation is consistent with the downregulation of A2AR in a hyperoxic or hyperbaric environment as shown here. The modulation of cAMP production we observed here is also in agreement with previous reports that used systems submitted to hyperoxia or (patho)physiological hydrostatic pressure. For example and regarding oxygen level, a decrease of cAMP production was found following hyperoxia in the carotid body of rats, a region that is enriched in A2AR and is involved, through adenosine release, in mechanisms maintaining high cAMP levels during hypoxia (45). Regarding the hydrostatic pressure, the condition decreases accumulation of cAMP in cells isolated from the proliferative region of tibia epiphyseal cartilage, a zone where pressure-dependent cell signaling likely has physiological significance (8). Thus hydrostatic pressure affects per se cAMP production as part of a general regulatory mechanism (8, 15, 45) that was further observed here using hyperbaric treatment alone.
Regarding the effects of the oxygen level on ADA and surface CD26, both catalysts increase during hypoxia in humans where both proteins are interacting at the mononuclear cell surface (20, 46). This increase was interpreted as a means to protect mononuclear cells from cytotoxic effects of high adenosine concentration in the vicinity of the cell surface via binding to CD26 of the aminohydrolase that degrades adenosine, and hence as an evolutionary advantage (46). Here, in the rodent context where ADA does not bind CD26 (19, 53), we observed that hyperoxia increased the serum ADA level. That low adenosinemia and high ADA level coexisted here following hyperoxia is in apparent discrepancy with the idea that high ADA levels observed during hypoxia in humans are induced by high adenosinemia to counteract adenosine cytotoxicity. The high ADA level we measured here following hyperoxia and its consequences in terms of adenosine degradation are fully consistent, however, with the low adenosine level we found in our study. In contrast, hyperbaric conditions did not affect per se adenosine plasma level, CD73, ADA, and CD26.
Finally, we found that a high oxygen level tended to decrease CD26 expression in rat, which is in agreement with previous findings in humans where an inverse relationship was found between blood oxygen level and CD26 expression (46). Our present data also support the idea that downregulation of CD26 expression by high oxygen concentration constitutes an advantage. Besides its role in the immune response, there are various ways by which CD26 may participate in controlling the response to hyperoxia. First, a diminished degradation of CD26 substrate peptides with antioxidant capacity promotes their activity, which represents an obvious advantage in hyperoxic conditions (5, 32, 44, 49, 60). For example, a decreased degradation of the vasoactive intestinal peptide is consistent with the observation that its serum level increases following production of reactive oxygen species, which constitutes an adaptive response because the peptide has per se antioxidative capacities via its capacity to scavenge singlet oxygen and to reduce hydroxyl radical production (5, 32, 41, 44). A similar conclusion can be reached for the glucagon-like peptide 1 that fights against redox-generated cell senescence (49). Second, a diminished degradation of cardio/vasoactive CD26 substrates such as the potent neuropeptide Y results in an increase of the vascular tone, which reduces the oxygen supply to tissues (41, 66). Third, CD26 controls energetic metabolism and energy supply to cells via degradation of peptides such as the glucagon-like peptide 1 (41). Last, but not least, a long-term loss of CD26 activity was reported to increase the capability against the oxidative stress and reactive oxygen species (38). Although addressing CD26 substrates was out of the scope of this work, monitoring them during hyperoxia certainly deserves further study.
In conclusion, we showed that increasing PaO2 by up to 10-fold tended to downregulate the adenosinergic pathway and CD26 activity. This mechanism may participate in an adaptive response to extreme values of oxygen supply to tissues and their deleterious consequences such as reactive oxygen species production through A2AR-mediated vasoconstriction and decreased vasoactive and antioxidant peptide degradation.
No conflicts of interest, financial or otherwise, are declared by the authors.
Author contributions: L.B., L.N., J.C., N.A., L.M., J.-L.B.-L., N.K., M.L., J.-M.D.-G., and E.F. performed experiments; L.B., J.-C.R., L.N., J.C., G.M., L.M., J.-L.B.-L., J.F., N.K., J.-M.D.-G., Y.J., R.G., J.R., and E.F. analyzed data; L.B., Y.J., R.G., J.R., and E.F. interpreted results of experiments; L.B., R.G., J.R., and E.F. prepared figures; L.B., R.G., J.R., and E.F. drafted manuscript; L.B., Y.J., R.G., J.R., and E.F. edited and revised manuscript; L.B., J.-C.R., Y.J., R.G., J.R., and E.F. approved final version of manuscript; J.-C.R., R.G., J.R., and E.F. conception and design of research.
- Copyright © 2015 the American Physiological Society


text:
We recommend upgrading to the latest Internet Explorer, Google Chrome, or Firefox.
If you are using IE 9 or later, make sure you turn off "Compatibility View".
Handpicked brands for you
Black Gold: A Solo exhibition by Roshi K @roshi_k Hosted by Little Pink Monster Rogue Gallery •1913 E. 17th St• •Sat. Aug. 29th• •7-10 pm• With Edible and Audible Refreshments provided by: •DJ Irie Samurai •Rollin Smoke BBQ •Big Bend Brewing...


text:
30 Day Returns
Express Delivery Options Available
Free UK Delivery when spending £25+
Compston Corner, Cumbria, LA22 9DS
Words by tcsh
on 07/07/2011 15:13:13
Salomon have allowed us a sneaky peak at their new running pack that we'll be stocking from next Spring. The Salomon XA Skin Pro 16 + 4 takes inspiration from their current range of specialist running packs such as the XT Advanced Skin 5 and features a stable, limpet like fit, numerous bottle pockets and stowage options, and a tailor made hydration system is also included. The pack also has an expansion zip that expands its internal volume from 16 litres up to 20. This makes the pack great for endurance running events such as the Montane Lakeland 100, the Hardmoors 100 or the Ultra Marathon Du Mont Blanc.
The XA Skin Pro Pack has a really novel design, being somewhere between a conventional rucksack and a piece of clothing. The 'waistcoat' design offers superb comfort and amazing stability when running with it loaded. The front is secured by a innovative stretchy strap system that is secured across the chest and abdomen. The straps clips into a plastic rail on the left shoulder strap, doing away with regular clips to save weight. There are also twin bottle / energy gel pockets up front on the shoulder straps and stretchy wand pocket on the pack itself, giving you loads of storage options for your gear.
There is a small zipped compartment on the rear of the pack and the main compartment is expandable via a zipped bellows section. This allows you to tailor the capacity of the pack to the activity you're doing.
The Salomon XA Skin Pro 16 + 4 is a very cool little pack and is super stable when loaded, but it certainly won't suit everyone – this really is a piece specifically focused towards the endurance trail runner and adventure racer. That said, more and more people are turning to trail and endurance running as their sport of choice, and Salomon are very much at the forefront of that trend. We think they've produced an exceptional little pack here and think that it'll really appeal to Salomon's core running market.
Sign up to the climbers shop newsletter today!
30 Day Returns
Free UK Delivery when spending £25+
Sign-up below and you'll received sale previews, exclusive offers, first-looks at the latest products & Personalised recommendations
My message Close


text:
Top kickboxer Tiffany van Soest has sadly had to resort to selling her world championship Lion Fight belt in order to pay her bills.
In extremely sad circumstances, the Lion Fight featherweight champion has accused the promotion of not paying her for her fights. The result of this has left Tiffany Van Soest in debt accumulated from her preparations that went into winning the belt. Sadly, Tiffany Timebomb has announced that she may have to sell her belt just to get by.
Van Soest has been one of the most exciting female prospects in the world of combat sports in the last few years. Displaying incredible talent inside the ring, Van Soest has also shown her business savvy, signing with EPOK Agency to help nurture her career outside of the octagon.
At Lion Fight 22 Van Soest defeated Bernice Aldis for the featherweight title in spectacular style, splitting Aldis' face wide open with a vicious elbow.
The 26 year old Californian native exposed the promotion online via her own social media and naturally Lion Fight promotions has received heavy criticism.
With the vocal support Tiffany Timebomb has received online, we can hopefully expect Lion Fight promotions to right their wrongs and pay the talented fighter. A career in the world of MMA can be a lucrative one, but almost always a short one. Fighters have a limited amount of time to get in, earn some serious cash and leave in one piece. The perfect example of this would be Conor McGregor, who has been vocal about making as much money as possible before leaving the sport.
Lion Fight has yet to respond to the claims, and still champion Tiffany Timebomb proudly on their homepage as one of the promotions top stars.
Give us YOUR thoughts in the comments below!


text: Track Sample Source Code Submit a Sample Source Code Sample Source Code: SureDone C# Sample Code by SureDone Followers eCommerce, Inventory, Lists, Platform-as-a-Service The SureDone C# Sample Code by SureDone demonstrates authentication to send requests and responses via API. It provides endpoint in version 1 to interact with the interface. Summary Followers Comments Summary Followers Comments Followers (0) Track this Sample Source Code Sorry, No Followers


text: Hey guys, I have two new cuttle babies and 3 on the way (hopefully). I have one that was born on the 3rd so about 3 days old. I know they can go 2 days to 2 weeks before they eat thier first meal. I have some mysis on the way. But I tried an amphipod today and no such luck in the first 10 min. After that I stopped watching. Anyways, How long should I wait before I get worried that they are not eating and not going to make it? They seem pretty content staying in the algea. If in a week he hasnt eaten I know I will be freaking out so what should I do in that situation. Also do I just put the food in close to them? Cause I put them in and they( the food) run off. At this age will they chase them down? Thanks for all info in advance. I thought I knew a little about what Im doing, but now that its here I'm freaking out a little lol


text:
The First Men Trailer (2015)
01 January 2015 Drama, Comedy, Thriller 14 mins
An overextended teacher bumps into a troubled student while at the mall with her mom. An adaptation of Stacey Richter's Pushcart Prize winning short story.
Watch the official The First Men 2015 trailer in HD below or find more The First Men videos on Vidimovie.


text:
Visvesvaraya Technological University (VTU) on Tuesday, announced that the even semesters would commence from February 13th, 2017. The Vice Chancellor approved of the schedule yesterday.
In the academic calendar issued earlier last year, the even semesters were to start from 2nd February but the postponement in few exams led to this delay in next semester's classes.
The circular was issued yesterday and notified that the even semesters for B.Tech/ B.E./ B.Arch/ M.Tech/ M.Arch/ MBA/ MCA courses will kick-off from 13th February. However this excluded 4th semester MBA and M.Tech students.
The examinations for odd semesters of these courses were held during December 2016-January 2017. The exams including practical and theoretical, had begun on 1st December 2016 and ended in mid-January.
Coming to more important issues, that is, the declaration of results of odd semester exams. it's clear now that VTU results for the exams held during Dec 2016 to Jan 2017 will start rolling out soon now. A timeline indicating from next week could be termed as anything but obvious. As it is the case with VTU, the results declaration is usually a mess, we've seen that last semester and you students know that too.
Expectedly, the 7th semester results for B.E graduates can be out in few days. Even though the date for starting of next semester is out, it doesn't mean that results are sure shot going to be out before the said date. Sometimes, as we've witnessed in the past, students go on attending weeks of classes for the semester they don't even know they've qualified for or not.
Nonetheless, stay in touch with our blog/page here at ExamsWatch.com/tag/VTU for latest updates on VTU results. Also, follow us on Facebook here and here to make sure you get alerted first.
Note: Remember to search for ExamsWatch VTU when you go out searching for news on VTU results.


text:
Dear Hounds,
Wondering who among us is sorting out ways to enjoy eating out with friends while keeping to our shrunken budgets. In that vein, can you suggest your favorite BYOBs? Preferably with no corkage, but I know I'm pushing my luck. Some that I remember that I enjoyed were:
-"A" in upper Manhattan (no corkage)
-Ivo and Lulu (no corkage)