text:
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Hey PEOPLE!10:07 PM
Well, this is Alexa Sleadd, thought I'd report on today's hapnins...
Grandma and I drove all over Ashland this morning taking pictures, and now we are in Willows, CA (just south of Redding) at a hotel.
Now, at this hotel, I dove into the pool and was delighted at how warm the water was. I swam for about 8 minutes, until this guy comes walkin' out. His face was beet red, he was sporting a grungy tank-top, shorts, bit of a beer belly and long scraggly hair. I didn't seriously look at him for more than a third of a second, it was all I needed to get creeped out. I nonchalantly got out of the pool and sauntered back to my hotel room. My heart was pounding. Now I feel dumb for over-reacting, and I can't blame the fellow for closely resembling a creep, but better safe than sorry, eh? I kinda wish my older bro were here. Ah well, must fend for myself.
I've taken lots of pictures today.
You can't see it well, but there's a glass window showing the engine underneath. Amazing!

subset: commoncrawl


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Unformatted text preview: No, his eyes dragged themselves open,
from darkness to dim. It was his body that reacted, shrugging upward and throwing out an arm to grip the air.
The voice calmed him now. "Sorry it's taken so long. I think people have been watching me. And the man with
the identity card took longer than I thought, but—" There was a pause. "It's yours now. Not great quality, but
hopefully good enough to get you there if it comes to that." He crouched down and waved a hand at the
suitcase. In his other hand, he held something heavy and flat. "Come on—off." Max obeyed, standing and
scratching. He could feel the tightening of his bones. "The card is in this." It was a book. "You should put the
map in here, too, and the directions. And there's a key—taped to the inside cover." He clicked open the case as
quietly as he could and planted the book like a bomb. "I'll be back in a few days."
He left a small bag filled with bread, fat, and three small carrots. Next to it was a bottle of water. There was no
apology. "It's the best I could do."
Door open, door shut.
Alone again.
What came to him immediately then was the sound.
Everything was so desperately noisy in the dark when he was alone. Each time he moved, there was the sound
of a crease. He felt like a man in a paper suit.
The food.
Max divided the bread into three parts and set two aside. The one in his hand he immersed himself in, chewing
and gulping, forcing it down the dry corridor of his throat. The fat was cold and hard, scaling its way down,
occasionally holding on. Big swallows tore them away and sent them below.
Then the carrots.
Again, he set two aside and devoured the third. The noise was astounding. Surely, the Führer himself could
hear the sound of the orange crush in his mouth. It broke his teeth with every bite. When he drank, he was quite
positive that he was swallowing them. Next time, he advised himself, drink first.
Later, to his relief, when the echoes left him and he found the courage to check with his fingers, ea...
View Full Document
This note was uploaded on 01/17/2014 for the course ENG 99 taught by Professor Michal during the Winter '13 term at CSU Sacramento.
- Winter '13
- Michal
Click to edit the document details


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Honors College Research Scholarships
The Honors College may provide up to $1,000 to offset costs associated with a student's research or creative project. This is a competitive process; awards will be determined by the quality of submissions and the availability of funds. The deadline for applications is November 17, 2017.
Honors College Research Scholarship Guidelines and Application
- Applicants must be currently enrolled in the Honors College.
- The proposal must be approved by the student's faculty research advisor before submission.
- Applicants must submit the scholarship application and budget form and any other supporting documents electronically.
- Applicants must provide a justification for each item in the proposed budget. Only items listed approved as allowable costs can be purchased. Indicate if any budget items will be supported from another source (e.g., your college, department, faculty grant, or personal funds).
- Recipients must submit a progress report, approved by the faculty mentor, to the ARCH office at the end of the fall semester.
- At the completion of the project, scholarship recipients must submit a detailed two‐page summary, approved by the faculty research mentor, describing the results/outcomes of the project. The report must be turned in to the ARCH office for final approval.
- Projects involving human subjects, vertebrate animals, or recombinant DNA require prior approval by the FIU Institutional Review Board.
- Students are required to present their work at the Undergraduate Research Conference at FIU (CURFIU) in the spring.
View the Allowable Costs information before filling out the application.
Complete the ARCH Research Scholarship application form here.
Complete the ARCH Creative Project Scholarship application form here.
Research Conference Scholarships
The Honors College provides up to $500 to offset costs associated with an external research conference. This is a competitive process with the granting of awards determined by the quality of submissions and the availability of funds.
Complete the Honors College Research Conference Scholarship application form here.
External Sources of Research Funding
The following information regarding funding is from the Web-GURU Guide for Undergraduate Research, a comprehensive resource for student researchers. If you interested in ARCH, or are currently active in the program, it's highly recommended that you visit the Web-GURU website and explore everything they have to offer.
There are four basic types of funding sources:
- Government
- Private industry
- Foundations
- Professional organizations
Many government agencies provide funding to individuals and teams of researchers to work on specific types of research projects. Examples of government agencies that provide financial support include:
- The National Science Foundation (NSF) (science, engineering, and education);
- The National Institutes of Health (NIH) (health related projects in science and engineering);
- The Department of Energy (DOE) (energy-related research); and
- The National Aerospace Agency (NASA) (research related to aerospace science and engineering)
Many private companies provide grants and/or materials and supplies in support of research. Examples of companies that provide targeted support include:
Privately funded foundations are another good source of funding for research. Examples of foundations that provide financial support to researchers include:
- Howard Hughes Medical Institute (HHMI)
- Research Corporation (RC); and
- The Camille and Henry Dreyfus Foundation
Professional organizations represent still another possible source of funding for your research project. Examples include:
There are a number of useful tools that you can use to identify possible funding sources. For example:
- The American Association for the Advancement of Science's (AAAS) Grants Net
- The U.S. Department of Health and Human Services' Grants; and
- The U.S. Department of Education's e-Grants
However, the majority of these provide support to scientists and engineers with advanced degrees. That said, there are a number of associations and agencies that provide support for undergraduate research. Web-GURU has compiled Program Listings that will assist you in identifying possible funding sources specifically targeted to undergraduate researchers.
For more information:
Associate Dean Juan Carlos Espinosa
Honors College
(305) 348-4100


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TEHRAN, Iran (AP) Iran's foreign minister on Tuesday denied his country has slowed its nuclear program, contradicting remarks by the head of the U.N. nuclear agency that Iran appeared to have paused uranium enrichment.
Foreign Minister Manouchehr Mottaki said uranium enrichment was continuing unabated.
"Iran's legitimate activities with the aim of producing fuel ... are continuing their natural trend," Mottaki said at a news conference. "There has been no change in that course."
Iran's Atomic Energy Organization also said Tuesday there had been no change in the enrichment schedule.
The head of the International Atomic Energy Agency, Mohamed ElBaradei, said Monday that Iran appeared to have at least temporarily paused on the development of its uranium enrichment program. Enriched uranium is used to fuel nuclear power stations and, taken to a higher degree, can be used in the manufacture of nuclear bombs.
The IAEA has installed cameras at the enrichment center in Natanz.
A pause could be an attempt to ease Iran's conflict with the Security Council, which is considering harsher sanctions on the Islamic Republic.
But underlining that any pause was informal and temporary, ElBaradei, in separate comments, repeated the main finding of his recent report on Iran, saying that "Iran has not suspended its enrichment-related activities" as demanded by the Security Council.
And he also said his agency cannot be sure if Iran's nuclear activities are peaceful or a cover for a weapons program until the country cooperates with his experts.
China urged Iran on Tuesday to fully open its nuclear program to U.N. inspections, as a meeting of the International Atomic Energy Agency moved toward slashing technical aid programs for the country because of its defiance of the U.N. Security Council.
"We welcome, support and call upon Iran to step up cooperation with the International Atomic Energy Agency," Chinese Foreign Minister Li Zhaoxing told reporters in Beijing, picking up on a theme raised by ElBaradei at the Vienna meeting.
Unless Iran takes "the long overdue decision" to cooperate with the IAEA, it "will have no option but to reserve its judgment about Iran's nuclear program," he said.
Iran, which says its nuclear program is intended only to create fuel for power plants, has enriched small quantities of uranium to the low level suitable for nuclear fuel generation. The U.S. and its allies fear that Iran could build nuclear weapons with larger amounts of more highly enriched uranium.
Hard-line Iranian President Mahmoud Ahmadinejad had been expected to announce last month that Iran had started installing 3,000 uranium-enriching centrifuges at a facility in the desert outside the central city of Natanz, where it has about 500 centrifuges above and below ground. But the announcement never materialized, an apparent step back that ElBaradei appeared to confirm Monday.
Up for review as early as Tuesday at the Vienna meeting of the IAEA's 35 board nations will be a Feb. 22 report from ElBaradei finding that Iran has set up hundreds of centrifuges as part of its continuing enrichment efforts.
The board was expected to approve last month's decision by ElBaradei to suspend nearly half the technical aid his agency provides to Iran. Only North Korea and Saddam Hussein's Iraq have faced such punishment in the past.
The European Union said it backed the suspensions, saying it "supports the (IAEA) ... views" on the 23 projects that could be suspended — 18 contained in the most recent recommendations and five frozen earlier.
The board also will be reviewing North Korea's apparent willingness to ultimately dismantle its nuclear arms-making capabilities.
The U.N. Security Council's permanent members and Germany are currently discussing strengthening sanctions imposed on Iran in December for its refusal to halt enrichment.
Meanwhile, the official Islamic Republic News Agency reported on Tuesday that Iran's military tested anti-aircraft defenses at another nuclear facility, near the central Iranian city of Isfahan. The report did not elaborate and officials at the Isfahan Governor Office declined comment.
The report said the tests lasted three hours. Isfahan residents were notified in advance so the tests would not be mistaken for an airstrike against the plant, nine miles outside Isfahan.
Copyright 2007 The Associated Press. All rights reserved. This material may not be published, broadcast, rewritten or redistributed.
Conversation guidelines: USA TODAY welcomes your thoughts, stories and information related to this article. Please stay on topic and be respectful of others. Keep the conversation appropriate for interested readers across the map.


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Hey all
I got to thinking here at work while at lunch counting down to my 5 days off that starts in just over 2 hours how everyone around here celebrates around this time of year. We have the Secret Santa thing going on (and I am in on that), but that is based on the Pagen Christian thing, so I am wondering what do you do for the holidays? Do you rock Kwanzaa? Or maybe Chanukah is your thing!? (7 days of gifts!)
Plus, since I do not celebrate Kwanzaa or Chanukah, would anyone mind explaining the customs and what not. I am some people have no clue about either.
If I have missed any holidays that you may celebrate please fill us in!
Oh, and keep the comments civil, I would hate to start blocking people because they are being assholes.


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Celine Dion Sheet Music - My Heart Will Go On on Chromatik/
Composer James Horner persuaded a lot of people to make this famous vocal version of "My Heart Will Go On" to happen. Ignoring Titanic director James Cameron's wishes to keep the song an instrumental, film composer approached Celine Dion to record her vocals on it... which she initially refused. After some convincing and some extra electronic textures, Dion agreed and recorded one of her most classic songs in a single take. For a singer to bust out such a legendary vocal delivery after one take (which she had rarely done in the past), requires serious talent and practice. Take a cue from Celine and practice your heart out on this operatic classic on Chromatik.


text:
Product code: HD-317
Product type: Made
Energized by: Dr. Puneet Chawla
Marketed by: Live Vaastu Private Limited Faridabad 121001 Haryana.
Net Weight: 30 GMS
Check delivery and payment options at your location
Expect delivery in 3 - 7 days
Auspicious Swastik sticker with Swastik symbols can be easily pasted on walls and doors. These auspicious symbols are an invitation to the divine energies to bless that area with auspiciousness. It symbolize blessings of Lord Ganesha and Goddess Lakshmi. It is stuck on entrance of prayer room or home or business premises. It stands for good luck, wealth and fame. Swastika Sticker is a lucky charm, good for wealth and prosperity
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Weight: 30 GMS
Width: 10 CM
Height:10 CM
Please Note: All products sold on Live Vaastu are energized by Dr. Puneet Chawla


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Heshana Khagan
|This article needs additional citations for verification. (January 2008) (Learn how and when to remove this template message)
Heshana Khagan (r. 610–617, died 619), was the second Khagan and third ruler of the Western Turkic Khaganate. He was the son of Niri Qaghan. About 605 rebels drove him from Dzungaria to the Ili River area. He as defeated by Sheguy and fled east to Karakoja (Gaochang) and a year later to Chang'an.[1] By 619, he was no longer in control of his khaganate and was at Chang'an, the capital of China's Tang Dynasty, when Emperor Gaozu of Tang, giving in to Eastern Turk pressure, had him delivered to Eastern Turk emissaries to be executed.
Names:Chinese: 曷娑那可汗, Modern Chinese: (Pinyin): hésuōnà kěhàn, (Wade-Giles): ho-so-na k'o-han, Middle Chinese (Guangyun) [ɣɑt.sɑ˥˩nɑ˩ kʰɑ˥ɣɑn˩˥] or 曷薩那可汗/曷萨那可汗, hésànà kěhàn, ho-sa-na k'o-han; at one point known as Chuluo Khan (處羅可汗/处罗可汗) and (泥厥處羅可汗/泥厥处罗可汗), personal name Ashina Daman (阿史那達漫/阿史那达漫, āshǐnà dámàn, a-shih-na ta-man.
- ^ Karakoja, Niri and 605 are from Christoph Baumer, History of Central Asia, volume two, 2014, (index). The rest is probably from Lev Gumilyov.
Heshana Khagan
|Preceded by
Tardush Qaghan
Niri Qaghan
|Khagan of the Western Turkic Khaganate
|Succeeded by
|This article related to Central Asian history is a stub. You can help Wikipedia by expanding it.


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Chennai | Coimbatore | Delhi | Mumbai
Om Sai International is a reputed Manufacturer, Exporter and Supplier of Black Pepper Seeds . Known as the 'king of spices', Black Pepper Seeds are one of the most exotic Indian spices. We offer different varieties of Black Pepper Seeds such as malabar garbled, panniyur-1, karimunda, kuthiravally, arakkulam munda, balankotta, kalluvally, etc. We make use of a combination of natural advantages and organic techniques t
Om Sai International
353, Market Yard, Gultekdi, Maharashtra , India
We are pepper black pepper: features: 1) color: black, green, red, yellow . 2) type: pimento (hot and sweet). 3) size: large, medium and small. Packaging: 10kgscarton, 15kgsbag wrapped with papper in cartons payment term: 100% irrevocable, at sight lc. Or 100% advance t.t.
Sun And Sons Sea Air Land Impex
No. 134, 2nd Floor, Dudhane Building, Nda Road, Warje, Maharashtra - 411058, India
We are offering black pepper seeds. Our company is one of the leading exporters and suppliers of black pepper seeds that are characterized by rich flavor and sharp & penetrating smell. Hailing from tamil nadu state, we have been providing whole black pepper seeds, commonly known as kali mirch
M/s. Gulmohar International Inc
Ground Floor 678, Ghorpade Peth, Khadakmal Aali, Opp. Saint Teresa Church, Maharashtra - 411042, India
Based in India, we endeavor to export and supply the purest and freshest quality of White Pepper Seeds for our esteemed clientele across the globe. Stringent quality controls are exercised by our dexterous team while sourcing, packaging and distribution of White Pepper Seeds, to adhere with international food and safety standards. Priced reasonably, we are well-equipped to timely tackle retail or bulk orders of White
Ht- Drk International / Habibee Traders
A3 Mahesh Paradise, Nagras Road, Aundh, Maharashtra - 411007, India
Based in Pune, Maharashtra we are a successful manufacturer, exporter, and supplier of Organic Black Pepper Seeds . Pepper is probably one of the most commonly and widely used seasonings in the world. These are grown in neat and hygienic conditions using organic fertilizers. The seeds we offer are thoroughly processed and dried to achieve their deep black color. These small seeds can be used as they are or grou
The Organica
A4, 9, Meera Nagar, Lane-7, Koregaon Park, Maharashtra - 411001, India
Keeping the stern check on packaging procedures, we ensure that the products are packed using the finest packaging material, followed by a stringent check of the whole batch by the professionals. We are a quality-centralized firm and make use of only high quality material for manufacturing the offered lot. Also, we ensure that the material is sourced from the reliable places.
Avanish Expro Enterprises
Silver Crescent A402, Old Mundhwa Road, Kharadi, Maharashtra - 411014, India
Post your Buy Requirement
Deals in Turmeric Seeds, Corriander Seeds, Black Pepper Seeds
Kp, Maharashtra - 411001, India


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So it happened! He asked on New Years right after our kiss. It was so beautiful!
He didn't even read about the different ways I would propose to myself and somehow it just worked out. The universe has a funny way of working out doesn't it?
The night as a whole was pretty awesome (aside from Chad's friend going all drunken asshole on me – but I'm choosing to focus on the positive. Ain't nobody got time for meanness) and Chad was the sweetest. When he asked me, two things popped in to my head.
- We "officially" get to start planning our life together
- I get to have the same name as Chad and our kids
- Okay, well 3 things – I have to make it Facebook official – so all of my friends who aren't in Wisconsin will know what the deal is.
How Did He Propose?
Right after our New Year's kiss, he pulled me to the side and asked me to marry him and spend forever with him. I, of course, being all girly begin to cry and say yes. We kiss, he puts it on my hand and then I go screaming to Chad's friends although the whole place seemed to have heard since I don't know how to be quiet.
I talked to everyone and their mom that night, letting them know I was engaged and just being my bubbly outgoing self. I talked to one couple who was super cool and just real chill. They told me their story and it was the sweetest. I talked to a bunch more people and eventually I lose track and the night gets blurry (everyone kept buying us congratulatory shots). Somehow I barely remember getting back to our hotel room with sloppy joes, which were absolutely delicious – I think.
You know, my favorite part of the proposal was that he asked my dad. That was the most important part to me. My parents and I don't have the same beliefs as far as their religion goes, but the love I have for them and the respect that Chad shows my parents means a lot to me and to know that he did that, really meant a lot to me.
Wedding Planning Ideas
It is so funny, I started reading a book called The Desire Map and my opinion about what my wedding should look like has changed drastically. If you would have asked me a few days ago, I would have said, I want my dream wedding at the Monona Terrace but I realize that it isn't my true wedding want. I don't want some super classy, high-priced, have to invite people that I don't even want there sort of wedding.
I want to have a celebration of the life we have made together so far and the new life going forward that we will be creating together with our kids. I think we focus too much on the wedding and less on what happens after a wedding. And so I want to celebrate us but without the 15 – 20k price tag that comes with it.
Maybe a weekend in Vegas with only a few people that we want there and then just renting a hall out here when we get back to celebrate with the people who didn't come or who we didn't invite to the wedding. I don't know yet. I just got engaged and Chad and I have to work out more of our beliefs and morals together first before we can really set anything in stone.
I just found out one of his old best friends and their wife separated and I don't want to be that stat, I want to make sure that we aren't that couple, that we communicate (even when we don't want to) and that we have the same values as we move forward.
Ok, I was going to try to find a nice way to end this, but I'm hungry and ready for a tasty protein shake.
Oh! Come follow us on Facebook, you will get to stay in the loop on all wedding stuff and revelations or holy shits (about pricing of course) as we go :0).


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Carter, Betty (10 items)
Welcome back! Whilst you were away we added the functionality to view auctions and bid in pounds.
Carter, Betty: Information on the Catawiki site about Betty Carter 1955-1969 guest: Ray Charles , 1995.
At its heart, Catawiki is an online catalogue with images of, and information on, collectibles. Our catalogue is compiled by users in a similar way to Wikipedia.


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6 cars 12 seats Playground Kiddy Rides Motorless Slide Dragon Train Ride
Introduction of amusement ride roller coaster
Kids amusement park direct manufacture slide dragon is a kind of hot amusement equipment in the park,it's very popular with children and some young people.The product is suitable for playgrounds, squares, parks, kindergartens,shopping malls, restaurants, communities,residential quar-ters and other types indoor and outdoor venues.
Parameter of amusement ride roller coaster
|kids roller coaster kit slide dragon coaster rides
|Total height
|cotton cloth and plastic film
|1 set
Pack & loading amusement rides in our plant
1) Seaworthy package cotton fabric & plastic film wooden case
2) We can also pack rides according to your requirements
1.Guarantee of ride is 12 months, during this period the unartificial faults of all components is free to change.
2.More than 10 professional technicists and special hotline help installation.
3.Profound marketing strategy help clients make profit.
4.Technician install in clients site.
5.Technician training for operation and maintainance.
6.The regular survey for rides analysis&suggestion on operation.
Company information:
Henan Weiyuan Machinery CO.,LTD can manufacture amusement rides for customers, like the mini roller coaster, the carousel horse, the disco tagada rides, pirate ship rides, flying tagada, coffee cup rides and others, high quality products and service for customers, looking forward to make long term cooperation with customers.
1.if i want to make the some of the designs myself, is that ok?
yes, for the details, customers can send your requirements to us, then we will make it for you, the colors and light can be customized according to needs.
2.what about your payment term and delivery time for the mini roller coaster?
our payment term is T/T, L/C for this rides, and delivery time will be 20 workdays after get the deposit.
If you are interested in our mini roller coaster, welcome to send inquiry or leave message to us, will reply you as soos as possible.
3) We'll load the rides in reasonable and saving-space way
6 cars 12 seats Playground Kiddy Rides Motorless Slide Dragon Train Ride Images


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|Home | About | Journals | Submit | Contact Us | Français
The Gtr1 protein of Saccharomyces cerevisiae is a member of the RagA subfamily of the Ras-like small GTPase superfamily. Gtr1 has been implicated in various cellular processes. Particularly, the Switch regions in the GTPase domain of Gtr1 are essential for TORC1 activation and amino acid signaling. Therefore, knowledge about the biochemical activity of Gtr1 is required to understand its mode of action and regulation.
By employing tryptophan fluorescence analysis and radioactive GTPase assays, we demonstrate that Gtr1 can adopt two distinct GDP- and GTP-bound conformations, and that it hydrolyses GTP much slower than Ras proteins. Using cysteine mutagenesis of Arginine-37 and Valine-67, residues at the Switch I and II regions, respectively, we show altered GTPase activity and associated conformational changes as compared to the wild type protein and the cysteine-less mutant.
The extremely low intrinsic GTPase activity of Gtr1 implies requirement for interaction with activating proteins to support its physiological function. These findings as well as the altered properties obtained by mutagenesis in the Switch regions provide insights into the function of Gtr1 and its homologues in yeast and mammals.
The small GTPases of the Ras superfamily have been implicated in diverse cellular functions, including regulation of ion channel activity, cytoskeleton reorganization, nucleocytoplasmic transport, vesicular trafficking, cell proliferation and differentiation [1]. All Ras-GTPases share conserved amino acid sequences, the G1 to G5 motifs, which regulate the GDP/GTP exchange and GTP hydrolysis, and thus trigger multiple intracellular signaling cascades [2]. Switch I and Switch II are flexible regions around G2 and G3, respectively, which change conformation upon GTP/GDP exchange, thus facilitating interactions with its effectors, GTPase-activating proteins (GAPs) and guanine nucleotide exchange factors (GEFs) [3,4]. Ras-GTPases generally exhibit slow dissociation of GDP for GTP and low intrinsic GTP hydrolysis rate, which are strictly enhanced by GEFs and GAPs, respectively [5].
The Gtr1 protein of Saccharomyces cerevisiae (S. cerevisiae) is a multifunctional GTP-binding protein, involved in phosphate acquisition through modulation of Pho84 transport activity [6], ribosome biogenesis [7] and epigenetic control of gene expression [8]. It has also been shown that the Gtr1 protein is a subunit of the EGO/GSE complex, which is indispensible for intracellular sorting of the amino acid permease Gap1 [9], Gtr1 has been implicated in regulation of the TOR signaling cascade in response to amino acids [10]. Most recently, it has been proposed that the Switch regions are essential for activation of the TOR1 complex (TORC1) [11]. Despite an otherwise low sequence similarity, the G motifs in Gtr1 display high conservation with other Ras GTPases, and are located within the N-terminal GTPase domain of the protein [12]. The C-terminal domain of the protein has been implicated in self-interaction [13] and protein-protein interactions [11]. In contrast to Ras homologues, the Gtr1 protein lacks lipid modification motifs in its C-terminal region, and the G4 motif (H/NKXD) contains a histidine instead of an asparagine residue [6].
Gtr1 belongs to the distantly related RagA family, which displays a low sequence similarity with the Ras, Rab, Ran, Arf and Rho proteins [7,13]. The Gtr1 protein displays 52% and 47% sequence identify with the mammalian Rag GTPases RagA and RagB, respectively [7]. Notably, recent studies have shown that the Gtr1 and RagA proteins share a similar mechanism of amino acid-mediated TOR activation, indicating that these proteins are functionally conserved in eukaryotes [14]. S. cerevisiae also contains Gtr2, which corresponds to RagC/RagD in humans. Like the mammalian Rag GTPases, GTP-bound Gtr1 and GDP-bound Gtr2 form a stable heterodimeric complex in vitro[13]. The GTP-bound form but not the GDP-bound Gtr1 interacts with itself, whereas Gtr2 can interact with itself only in the presence of GTP-bound Gtr1 [13]. Most recently, the 3D-structure of the Gtr1-Gtr2 complex in the GMP-PNP bound form was resolved at 2.8Å resolution [11]. The structure has brought insights into the location of G domains and Switch regions. Based on this, it was proposed that upon nucleotide exchange the Switch regions change conformation, allowing for interaction with Raptor and activation of TORC1 [11]. In the same report, it was shown that the C-terminal domains of the two proteins mediate heterodimeric complex formation that is indispensible for activation of TOR signaling pathways.
Previous studies have localized the Gtr1 protein both to the nucleus and to the cytoplasm [15], where it interacts in a GTP-dependent manner with diverse cytoplasmic and nuclear proteins, such as Ego1, Yrb2, Nop8 and Ego3 [16-19]. Based on competition studies, it was proposed that the Gtr1 protein displays higher affinity for GTP than for GDP [13]. The GTP-bound form of Gtr1 interacted with the Rpc-19 subunit of RNA polymerases I and III in yeast two-hybrid assays, indicating a role in the assembly of RNA polymerase [20]. Recently, the Vam6 protein, known as a GEF for Ypt7, was shown to assist the exchange of GDP/GTP on the Gtr1 protein in vitro[14]. Moreover, Binda et al. [21] have reported that Vam6 activates the Gtr1 subunit of the EGO complex during TORC1 function.
Gtr1 harbors all the necessary structural elements for functioning as a GTPase. Previous attempts to estimate the GTPase activity of Gtr1 [13], RagA [7,] and RagC proteins [16] resulted in either none or too low activity to be measured. Here we quantify the intrinsic GTP hydrolytic activity of purified recombinant Gtr1 protein and several variants using a radioactive GTPase assay, and in addition study their tryptophan fluorescence emission properties.
S. cerevisiae full-length Gtr1 proteins (wild-type, cysteine-less (C-less), Arg37Cys and Val67Cys) were expressed in Escherichia coli and purified in a single step by affinity chromatography essentially as described [12]. The complete procedure is provided in Additional file 1. The protein expression levels and purity of the preparations were assayed as described [12]. The oligomeric state of Gtr1 was assayed by native gel electrophoresis as described [22]
The GTPase assay was carried out using purified recombinant protein at a final concentration of 0.15mgml-1 (4μM), 185 nM GTP and 15 nM [α-32P] GTP (3000Ci/mmol, 1Ci=3.7x1010 Bq; PerkinElmer, Boston, USA) in a 50μl containing assay buffer (25mM HEPES/NaOH, pH 7.4, 5mM KCl, 5mM MgCl2 and 100mM sucrose) as described in Ref. [23].
The enzymatic reaction was carried out at 37°C for the indicated time periods (0, 15, 30, 60 and 120min), and terminated by the addition of an equal volume of 4M formic acid. A volume of 2.5μl (62.5 nCi) quenched reaction mixture was spotted onto a poly (ethyleneimine)-cellulose plate (Merck, Germany) and the nucleotides were separated by thin-layer chromatography using 0.75M KH2PO4 (pH 3.65) as elution buffer. The radioactive nucleotides were detected by phosphorimaging (BAS-1500, Fuji, Japan). The radioactive GDP spots were quantified using MultiGauge software (Fuji, Japan). The data are given as means±SD and were obtained from 2–3 independent experiments. Initial velocities for GTP hydrolysis were measured using the same assay conditions at varying [α-32P] GTP concentrations (0.1 to 1.2μM) for 60min. The data were plotted as a function of GTP concentration, and fitted to the Michaelis-Menten equation. A Lineweaver-Burk plot of the data was used to determine the values for the Michaelis-Menten constant (Km) and the maximum velocity (Vmax) parameters. The catalytic constant kcat was calculated as the Vmax/Et ratio, where Et is the enzyme concentration.
Tryptophan (Trp) fluorescence spectra of purified recombinant Gtr1 proteins were recorded using a Fluoromax-3 spectrofluorometer (Horiba Jobin Yvon, Japan) in the presence of guanine nucleotides. The interaction of the protein with guanosine 5'-O-(3-thiotriphosphate) (GTPγS) or GDP was measured using 0.019mgml-1 (0.50μM) protein and 25μM nucleotides in the GTPase assay buffer. The reaction mixture was incubated for 120min at 37°C prior to fluorescence measurements [24]. The intrinsic protein fluorescence was excited at 297nm, and emission spectra were recorded in the range 297 to 400nm at 25°C. The spectra of free nucleotides in assay buffer were subtracted from the collected sample dataset.
Sequence alignments of Gtr1 from S. cerevisiae with other yeast and mammalian members of RagA family and with human H-Ras were obtained using ClustalW (Figure(Figure1).1). The location of G motifs, Switch regions and residues mutated in this work are highlighted in the alignment. The position of the nucleotide-binding domain of Gtr1 (residues 1–185), of the Switch I and II regions, of the mutated Arg37 and Val67 residues, and the tryptophan residues, Trp60, Trp167 and Trp175, are indicated in the X-ray crystal structure of Gtr1 (Figure(Figure22).
The GDP/GTP exchange and intrinsic GTP hydrolytic activity are the most important functions for all regulatory GTPases. In order to characterize the properties of Gtr1, an N-terminal His6-tagged version of the full length Gtr1 protein was stably over-expressed using a bacterial production system and purified by a single step affinity chromatography [12]. Based on SDS-PAGE analysis, the purity of the recombinant protein was estimated to 95% and the Mr to 36kDa. The identity was confirmed by Western blot analysis using an anti-His antibody (Figure(Figure3).3). The oligomeric state of the recombinant protein is monomeric since the recombinant protein migrates at 36kDa in native gel electrophoresis (Additional file 2: Figure S1).
Trp fluorescence has been widely used as a tool to study structural changes upon nucleotide binding to Ras GTPases [23,24] The Gtr1 protein harbors three Trp residues, namely Trp60, Trp167 and Trp175 (Figures1 and and2).2). Conformational alterations upon nucleotide binding to Gtr1 could result in changes in the local environment of these Trp residues in the protein structure. FigureFigure44 shows the fluorescence emission spectra of purified Gtr1 wild-type protein in the presence of GDP or GTPγS. When loading the Gtr1 wild-type protein with nucleotides, we observed a blue shift of 2nm in the tryptophan fluorescence emission between the GTPγS-bound form (329nm) and the GDP-bound form (327nm) (Table(Table1).1). This indicates a more hydrophobic localization of the tryptophan residues upon GDP binding, and thus a more compact overall conformation. Furthermore, the data presented in FigureFigure44 indicate that the recombinant Gtr1 wild-type protein has retained the nucleotide binding functionality. Taken together, our results indicate that Gtr1, like other GTPases, can adopt two distinct conformations, corresponding to the GTP-bound and GDP-bound state, which can be monitored by tryptophan fluorescence. This tool has been used to study several mutants in the work, as described below.
Previous attempts to measure the GTPase activity of Gtr1 and RagA proteins have failed [7,13], and thus the proteins were considered to lack GTPase activity in the absence of activators. A weak GTPase activity has been indicated for RagC [16]. Here we measure the intrinsic GTPase activity of Gtr1 using [α-32P] GTP and purified wild-type Gtr1 in assay conditions similar to those previously described [23]. As shown in FigureFigure5A,5A, as result of the hydrolytic activity, GDP was produced in amounts increasing with the incubation time in samples containing purified protein, whereas GDP production was absent in control samples devoid of protein. The chromatography plates were overloaded with radioactive sample, and therefore, the corresponding decrease in the GTP amount resulting from its hydrolysis could not be detected. The ratio between the amount of GDP and the total amount of GDP+GTP reached a maximum of 30% at the incubation time of 120min, in line with the previous report for RagC 16].
Next, we determined the enzymatic kinetic parameters of the protein under similar assay conditions as described in Methods. Values of 1.0μM and 0.1nmol GDP mg protein-1min-1 were obtained for the Km and Vmax parameters by Michaelis-Menten analyses (Figure(Figure5B5B and insert). The obtained values were used to calculate the kcat of the protein, 0.004min-1. For reference, the kcat values determined for three representative GTPases, namely Gial, Ras and EF-TU are 3, 0.3 and 0.003min-1, respectively [25]. The finding of an extremely low GTPase activity implies a strict regulation, for example interaction with other proteins, such as Gtr2. In addition, Vam6 has been proposed as a GEF for Gtr1 [14,21]. It will be important to understand the way this key GTPase is regulated by activating proteins to control various processes.
We have previously used EPR spectroscopy analysis on site-directed mutants of Gtr1 and showed that only the N-terminal part of the protein harbors the structural elements involved in GTP binding [12]. In the same study, we examined the conformational changes upon GTP binding of Gtr1 in the regions around residues Arg37 and Val67. These residues are located at the border of Switch I and II region, respectively (Figures1 and and2).2). Sequence analysis indicated that the Arg37 residue is highly conserved within the RagA family, but is not present in other Ras GTPases (Figure(Figure1).1). On the other hand, the Val67 residue is not conserved in either Rag or other Ras proteins. Residues Arg37 and Val67 were, however, chosen for mutagenesis since the activities of the two Switch regions could be monitored without completely abolishing GTP/GDP-binding. We have tested the functional importance of these residues in Gtr1 by measuring the GTPase activity of Arg37Cys and Val67Cys mutants constructed in a Cys-less background (Cys replaced by Ser). In addition to offer the ability to modify the cysteine thiol-group with spectroscopic probes, the relatively low bulkiness of the cysteine side chain as well as its chemical ambivalence makes the cysteine residue a suitable replacement to study the dynamics of the Switch regions. As shown in FigureFigure1,1, the amino acid sequence of the Gtr1 protein contains five native cysteine residues, of which Cys62 and 180 reside in the N-terminal GTPase domain, whereas Cys217, 257 and 284 are located in the C-terminal domain. The creation of a functional Gtr1 Cys-less, in which serine residues replaced all five native cysteine residues, has been previously described [12]. All three mutant proteins (Cys-less, Arg37Cys and Val67Cys) were purified under similar conditions as the wild type (WT) Gtr1 protein. Western blot analysis shows similar levels of the expressed proteins (Figure(Figure6A).6A). We first analyzed the GTPase activity of the Cys-less mutant. The amount of GDP produced by the Cys-less mutant was found to be slightly higher than that in the WT at 30min, followed by a gradual decrease to WT levels during prolonged incubation with [α-32P]GTP (Figure(Figure6B).6B). This result indicates that cysteine residues may not be essential for the GTPase activity, which is consistent with previous reports on other Ras GTPases [26,27]. The Arg37Cys mutant exhibits a diminished GTP hydrolytic activity (60% of WT levels) throughout the incubation period (Figure(Figure6B6B and C). In contrast, the Val67Cys mutant displayed about 25% higher GTPase activity than the WT protein. These results indicate the importance of Arg37 and Val67 for the intrinsic GTPase activity of Gtr1, in the absence of interacting proteins such as GEF and Gtr2.
Next, we investigated if the effect of mutations on the GTPase activity could be explained by the dynamics in the protein structure. For this purpose, all mutant Gtr1 proteins were analyzed for tryptophan fluorescence in a similar manner as performed for the WT protein. As observed in the WT, the emission maxima in all mutant proteins, displayed a blue shift between GDP and GTPγS forms, (Figure(Figure77 and TableTable1),1), indicating that environmental changes for the Trp residues contribute to the observed blue shift. The blue shift from the GDP to the GTPγS states in Arg37Cys was the smallest (1nm) whereas in Val67C is the largest (5nm). Notably, a blue shift was also visible in both the GDP and GTPγS-bound states of all mutants relative to the WT, indicating a more hydrophilic environment for the Trp residues in the mutants. Again, Arg37Cys and Val67Cys distinguished themselves from the Cys-less mutant since they display the largest blue shifts relative to the WT in both nucleotide bound states. A third interesting difference was the higher fluorescence intensity observed for both the GTPγS and GDP forms in the Cys-less mutant as compared to all other samples, and might be explained by changes in the degree to which Trp fluorescence is quenched by neighboring residues in this mutant. The observed difference cannot be attributed to possible nucleotide-induced aggregation of the protein as similar levels of the protein were indicated in all samples by native PAGE analysis (Additional file 2: Figure S1). Moreover, this difference may have no relevance for the GTPase activity in the Cys-less mutant, which displayed WT levels. Taken together, the fluorescence data indicate that distinct conformational changes occur in the Arg37Cys and Val67Cys mutants that could impact on their performance during the GTPase assays (Figure(Figure66).
Recently, the crystal structure of the Gtr1-Gtr2 complex, with the two proteins bound to GMPPNP, was solved [11]. This structural model served as a template to design and analyze the role of RagA in activating TORC1. Mutational analysis of several residues indicated that the surface charge potential is of great importance for RagA to interact with Raptor, and thus being able to activate TORC1 [11]. Of the two adjacent arginines close to the Switch I region and which are conserved amongst the Gtr1 and RagA region (Arg36, Arg37), we have mutated Arg37 to a cysteine and have observed a change in hydrolytic activity. The removal of one of the two Arg residues might alter the surface charge to a degree where a minimal conformational alteration could influence the nucleotide interaction. Whether this charge alteration effectively results in a conformational change that impairs nucleotide binding is currently not known. Our results indicate that a replacement of Arg37 with a cysteine leads to a decreased hydrolytic activity, supporting the importance of this residue.
Here we have employed a combination of structural, cysteine mutagenesis, radioactive GTPase assays and intrinsic tryptophan fluorescence approaches to study the biochemical activity of Gtr1 from S. cerevisiae. The data obtained reveal a very low intrinsic GTPase activity of the Gtr1 protein as compared to Ras GTPases, implying requirement for activating proteins, as previously reported [14,21]. This activity was found altered in Arg37Cys and also Val67Cys mutants of the Switch regions and associated with conformational changes, which are distinct from those in the WT and cysteine-less mutant. Despite the fact that both mutated residues are not fully conserved amongst Gtr1 homologues, alterations made in those positions have an influence on the GTPase activity and fluorescence. This indicates that both residues are of importance for the functionality of Gtr1. Our findings provide insights into the function of Gtr1 and its homologues in yeast and mammals.
GAP: GTPase-activating protein; GEF: guanine nucleotide exchange factor; GTPγS: guanosine 5'-O-(3-thiotriphosphate); TORC1: TOR complex 1; Trp: tryptophan; WT: wild type.
PS participated in experimental design and conducted the bulk of experimental work from protein overexpression to data analysis, and article writing. CS participated in the design and analysis of the GTPase activity data and in writing of the article. JOL generated the cysteine mutants and participated in the discussion of the fluorescence data. DS produced the structural figure and analyzed the fluorescence data. LRP participated in data processing and figure preparations. MA participated in protein expression and purification. BLP participated in the design, coordinated the experiments and writing of the article. All authors have read and approved of the final manuscript.
Supplemental methods - Expression in Escherichia coliand purification of Gtr1 protein [12].
Figure S1 - Assessment of the oligomeric state of recombinant Gtr1 protein by native gel electrophoresis. Figure S2 - Stability of the nucleotide-bound Gtr1 Cys-less protein assayed by native gel electrophoresis and Western blotting.
We are grateful to Prof. Peter Brodelius for providing us with the pET28a+ expression plasmid. This work was supported by grants from the Swedish Research Council to B.L.P. (621-2003-3558 and 621-2007-6144), C.S. (621-2007-5440 and 621-2010-463) and J.O.L. (522-2008-3724).
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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Tummy Tuck (Abdominoplasty) in Fort Lauderdale with Dr. John Michael Thomassen
A tummy tuck (abdominoplasty) is a surgical procedure that is designed to improve the appearance of the abdomen. Dr. John Michael Thomassen frequently performs tummy tucks for residents in and around Fort Lauderdale, Fla. During an abdominoplasty, Dr. Thomassen tightens the muscles of the abdominal wall, he also removes excess fat and unnecessary skin from the abdomen. Sometimes, during a tummy tuck, Dr. Thomassen also performs liposuction.
Reasons the Abdomen Becomes Less Tone
Various factors contribute to the loss of tone in the abdomen. These factors include the combined effects of gravity, aging, weight fluctuations, pregnancy/childbirth and a loss of skin elasticity.
What an Abdominoplasty in Fort Lauderdale Can Accomplish
When Dr. Thomassen performs a tummy tuck, he contours his patient's body to create a firm and flat abdomen.
Custom-Designed Treatment Plans to Meet Each Patient's Specific Needs
Each patient is unique, therefore, Dr. Thomassen takes the time to create each of his patients a treatment plan that is designed to meet his or her specific needs.
Who Should Consider a Tummy Tuck with Dr. Thomassen in Fort Lauderdale?
The majority of individuals who seek a tummy tuck in Fort Lauderdale are women who have had several pregnancies resulting in excess fat and skin on their abdomens. Women who intend to have more children should wait to have a tummy tuck because pregnancy can negatively affect the results of their abdominoplasty.
View Before & After Photos
Other individuals who choose to have a tummy tuck include:
- Men and women who have a protruding abdomen with excess fat and/or skin that is resistant to diet, and exercise.
- Healthy individuals who are within 25 pounds of their recommended body weight and who have realistic expectations as to what a tummy tuck can accomplish.
- Individuals who have lost a substantial amount of weight and have excess skin; however, an individual who plans to lose more weight should postpone a tummy tuck until his or her weight loss goal is reached.
An Initial Consultation for a Tummy Tuck at Thomassen Plastic Surgery in Fort Lauderdale
Patients should be candid during their initial consultation: All of the questions that Dr. Thomassen and his staff ask are intended to prevent complications and ensure the safety of the patient.
During an initial consultation, Dr. Thomassen will want to know about the patient's medical history, including:
- The medications, herbal supplements and vitamins he or she takes
- Prior treatments and surgical procedures
- Allergies
Dr. Thomassen will ask whether the patient smokes, drinks alcohol or uses recreational drugs as well as the patient's motivation for seeking an abdominoplasty. The use of tobacco, alcohol and recreational drugs can have a profound impact on the body's ability to heal itself. Individuals who smoke need to quit at least a month before and avoid smoking one month after their tummy tuck procedure. Smoking and second-hand smoke cause a decrease in blood flow, which can result in infection, skin loss, poor wound healing and increased scarring.
After Dr. Thomassen reviews the patient's medical history, he will want to examine his or her abdomen. Dr. Thomassen may choose to take pictures of the patient's abdomen to use for reference as he creates the patient's surgical treatment plan. In addition, these photos may be referred to during the abdominoplasty itself. For comparison purposes, Dr. Thomassen may also take photos of the patient's abdomen several months after his or her tummy tuck procedure.
A Tummy Tuck with Liposuction
Many of Dr. Thomassen's patients request a tummy tuck in conjunction with liposuction. The best candidates for this combination procedure are individuals who have excess fat and loose skin on their abdomens.
Preparing for an Abdominoplasty in Fort Lauderdale
Prior to a tummy tuck, Dr. Thomassen will most likely request that the patient visit his or her physician to have a thorough physical exam and routine blood work. Patients who are 40 years of age or older may need to have an electrocardiogram (EKG/ECG) prior to their tummy tuck procedure.
Certain medications need to be avoided for at least two weeks before surgery and one week after their surgical procedure. These medications include nonsteroidal anti-inflammatory drugs (NSAIDs) like aspirin and ibuprofen. NSAIDs are known to cause bruising and abnormal bleeding.
The Day of the Abdominoplasty
Patients need to fast for at least eight hours before their tummy tuck with Dr. Thomassen in Fort Lauderdale.
Since an abdominoplasty is an outpatient procedure, patients need to bring a licensed driver with them on the day of their procedure. A tummy tuck can take two or three hours to complete.
1. Shaving and Marking the Abdomen
The patient's abdomen is shaved and marked prior to surgery. These markings are used as a guide during the surgical procedure.
2. Anesthesia
The majority of Dr. Thomassen's patients receive general anesthesia: All anesthesia is administered by a licensed anesthesiologist.
3. Making the Incision
Once the patient is anesthetized, Dr. Thomassen will create an incision that extends across the lower abdomen (directly above the pubic area) from hip to hip.
If an individual is also having liposuction, the liposuction procedure is performed first. Additional incisions will need to be made. These small incisions are necessary as they are used to insert the cannula (the thin tube used to remove excess fat).
4. The Fat and Skin Are Lifted
The skin and fat are gently lifted off of the abdominal wall's muscles.
5. Tightening of the Abdominal Muscles
If necessary, Dr. Thomassen uses sutures to tighten the abdominal muscles.
6. Excess Fat and Skin Are Removed
After tightening the muscles, Dr. Thomassen removes any excess fat and skin.
7. Dr. Thomassen Redrapes the Skin
After removing the excess skin and fat, the skin is redraped over the abdominal muscles. To hasten the healing process, Dr. Thomassen may place a drain beneath the incision. A drain is a thin, rubber tube that assists in draining fluid that accumulates beneath the incision.
8. Closing the Incisions
To minimize scarring, Dr. Thomassen carefully brings the skin together and closes it with sutures.
9. Dressing is Applied and an Abdominal Binder is Placed
A sterile dressing is applied over the incisions. Then, a compression garment is placed to help support the abdominal wall as it heals. Compression garments and abdominal binders also decrease post-operative swelling, which reduces the incidence of bruising.
10. Recovery Room
Patients will remain in the recovery room for approximately an hour following their abdominoplasty.
11. Results
Patients will notice an improvement in the contour of their abdomen immediately: As swelling subsides, the contour of the abdomen will continue to improve.
Recovering from an Abdominoplasty in Fort Lauderdale
Patients will wear the compression garment or abdominal binder for approximately three weeks after their surgery.
Drainage Tube Removal
The drainage tubes that Dr. Thomassen placed will be removed at his office in Fort Lauderdale several days after the procedure.
At-Home Recovery
Most patients need to spend several days in bed directly following their tummy tuck: Patients should expect to feel some discomfort and pain. Dr. Thomassen will provide patients with medications and tips for reducing their pain level.
Returning to Work
While each patient is unique, the majority of patients will require 14 days off of work to recover.
Dr. John Michael Thomassen is a plastic and reconstructive surgeon in Fort Lauderdale who frequently performs tummy tuck procedures for individuals who want to improve the way their abdomens look. If you are interested in learning more about abdominoplasty and liposuction, or any of the other procedures that Dr. Thomassen performs, please contactThomassen Plastic Surgery today.
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Fort Lauderdale Tummy Tuck Surgery (Abdominoplasty)
Desire for an abdominoplasty to improve the appearance of the abdominal wall is a common concern. Anomalies of the abdominal wall can result from many varied experiences. Weight fluctuations throughout life can lead to excess skin that will not go away with diet or exercise. Pregnancy can lead to not only excess skin but also a weakness of the abdominal wall that results in a protruding abdomen, also recalcitrant to exercise. Prior abdominal surgery can result in scars and contour irregularities that can only be corrected with surgery. Endogenous and exogenous hormones and medications can also have an effect on our body contours that may be permanent and only corrected with body contouring surgery. Finally our own genetics and age related changes can result in excess skin and fat that requires more than diet and exercise to correct.
A history of any medical conditions, particularly heart, lung, liver, or rheumatologic conditions need to be discussed in the initial evaluation. A history of smoking is a significant risk factor for wound healing. Current recommendations are that patient stop smoking for a minimum of 4-8 weeks prior and after surgery. The use of nicotine patches is not an adequate measure in the perioperative period.
The physical exam focuses on evaluation of the entire abdomen, umbilicus, mons pubis, flanks and back rolls. Any prior surgeries and the resulting scars need to be considered when planning an abdominoplasty procedure. If an incision appears to interrupt the blood supply to the abdominal wall, then a variation of the surgery needs to be performed to ensure proper wound healing. The use of liposuction with an abdominoplasty helps address fullness noted in the mons pubis, hips and flanks, and dorsal back rolls.
The skin quality needs to be assessed as part of the exam. The presence of striae indicate a loss of elasticity of the skin and may affect the way the surgery is performed. Excess skin is the main indication for an abdominoplasty procedure and the amount of excess needs to be qualified to choose the right procedure for the patient. Patients who have abundance of adipose tissue but do not require skin or muscle treatment, i.e., have a good skin tone and reasonable potential for the skin to contract after removal of substantial volume, may be better candidates for liposuction alone as opposed to an abdominoplasty procedure.
The quality of the musculofascial layer is also important to evaluate for any weaknesses. More commonly, patients exhibit a weakness of this fascia termed rectus diastasis. This leads to a bulging abdomen that can only be fixed by plicating the fascia and restoring the rectus muscles to their original position. More severe defects of the fascia consist of hernias, particularly umbilical hernias and ventral hernias. These may need to be ruled out and if present, will be corrected at the time of the abdominoplasty.
The innervation of the abdominal wall is provided by lateral branches of intercostal nerves. These branches remain intact after the procedure so numbness in the anterior abdominal wall after surgery will improve and sensation should return to normal. Other nerves in the abdominal wall like the lateral femoral cutaneous nerve and the ilioinguinal and iliohypogastric nerves are susceptible to injury during closure so particular attention needs to be directed at these.
The presence of intraperitoneal fat or fat inside the abdominal cavity deep to the fascia cannot be treated with an abdominoplasty procedure. This fat is only reducible through diet and exercise. If fullness due to intraperitoneal fat is excessive, this may preclude undergoing a successful abdominoplasty.
Preoperative preparation
Once the decision to pursue an abdominoplasty is made, blood work is obtained and possibly a medical clearance from your primary care doctor is obtained to ensure you are at optimal health for the surgery. In some patients the use of anticoagulants is recommended due to their having a high risk of blood clots develop in the lower extremities after the surgery.
Day of Surgery
The day of surgery, markings are made on the patient's body with the patient's choice of swimwear or underwear. Incisions are placed immediately above the pubic region with a horizontal lateral extension, so that it would be hidden beneath current attire.
The surgery is typically performed under general anesthesia with a board certified anesthesiologist. When you wake up from the procedure, you will have an abdominal binder with dressings and should be comfortable. You may be admitted overnight or go home after the surgery. The pain medicine prescribed to you by Dr. Thomassen will keep you comfortable during the early postoperative period.
This procedure is indicated when there is minimal extra fat and skin in the lower abdomen only, and only fascial weakness in the lower abdomen below the umbilicus. It removes an elliptical area of skin in the lower abdomen that may be affected by prior scars or striae and does not address the upper abdomen. The incision line is not as long as a standard abdominoplasty incision. Liposuction is added to improve the overall result.
The umbilicus is not relocated with this procedure. The fascia below the umbilicus is plicated so any bulging in the lower abdomen is corrected. In some cases the umbilicus can be "floated" by cutting the stalk and reattaching it lower on the abdominal wall. Indications for this are limited.
This procedure is not indicated for most patients since most patients would benefit for more skin removal and plication of the entire length of the abdominal wall fascia.
Full Abdominoplasty
First described by Pitanguy and Callia in 1967, this is the procedure most commonly performed in patients desiring improvement of their abdomen and torso. It involves a low anterior curvilinear incision extending from one anterior superior iliac spine (ASIS) to the other, umbilical translocation and full fascial plication. The umbilical stalk is kept attached and the umbilicus is brought out through a new incision in the abdominal wall skin. Sometimes, plication of the external oblique aponeurosis is added for improved contouring of the waist.
Liposuction has been proven safe to undergo along with this extensive dissection and is typically added to improve the overall contour of the result. It can reduce the thickness of the upper abdominal area. Liposuction and wedge reduction (both vertical and horizontal) are the most common techniques used to treat the mons pubis and excess volume in this area.
Sometimes an abdominoplasty can be combined with a hysterectomy or other gynecological procedures for patients interested in avoiding two recovery periods. Studies have proven the safety of these procedures.
Fleur de Lis Abdominoplasty
In this variation, the horizontal elliptical excision of lower abdominal wall skin is performed as well as a vertical elliptical excision in the midline. This results in a vertical scar that extends from the xyphoid to the pubis, and then a second horizontal scar extending from each hip. This procedure is indicated in patients who have lost significant weight and have excessive skin in the midline. It aids in contouring the waist for a more feminine outline; however the additional vertical scar needs to be considered carefully.
Because conventional abdominoplasty procedures involve significant undermining, there is increased potential for complications, such as skin necrosis, infection, wound dehiscence, seroma, and hematoma. In addition, plastic surgery procedures are associated with a high incidence of adverse effects when performed in patients who smoke. These complications are related to the release of nicotine and carbon monoxide during smoking, which decrease blood flow and oxygen delivery and induces thrombogenesis. One recent study of 132 patients who underwent abdominoplasty reported a complication rate of 48% in smokers.
Patients should be advised to discontinue smoking four to eight weeks before the surgical procedure and for an additional four weeks after surgery. Serum levels of nicotine have been shown to return to normal when patients abstain from smoking for eight weeks. In addition, studies of patients who underwent flap reconstruction showed that complications were decreased significantly when smoking was discontinued a minimum of four weeks before surgery.
Some patients are at high risk for developing a clot in the lower extremity deep vein system during or after the surgery. The use of graded compression stockings worn preoperatively and 7 days postoperatively, intermittent pneumatic compression devices placed before surgery and until discharge, strict intraoperative and perioperative warming, and ambulation within the first hour of the operation are important to reduce this risk. High-risk patients are treated with an anticoagulant after surgery to help reduce this risk.
Seroma is a fluid collection that can occur under the skin flap. Postoperative seroma in abdominoplasty is the most common complication in this type of surgical procedure and is avoided with the use of drains, quilting sutures, and compression garments.


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25thframe.co.uk > Action | Sci-Fi | Thriller > Death Race
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UK release date: 26th September 2008
Highest chart position: 4
Weeks on box office: 3
Total UK gross £1,920,818 and the 2092th top grossing film in the UK
UK BBFC Certificate: 15Genres:


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We're beyond blessed to be part of Byron Spirit Festival 2017 with 2 offerings at this magical events alongside some truly magical, mystic and talented musicians, artists, teachers and facilitators. Yoga Rhythms line up is RIA & FRANKO HEKE with more TBA. "Honouring spiritual practices of past, present and future that support us to embody our roles as Custodians for our Earth, our Body and…Find out more »


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Camden , SC health care quality comes down to access, affordability and outcomes. Kershaw County ranks 14 for health outcomes 13 for health factors, and 21 for healthy behaviors in SC.
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Dennis Hissink : August 5th 2003 - 23:56 CET
Leica announces stylish Leica D-LUX
The LEICA D-LUX is a new digital compact camera from Leica Camera AG, Solms with a high-performance LEICA DC VARIO-ELMARIT f/2.8-4.9/5.8-17.4 mm ASPH. triple zoom lens. "The LEICA D-LUX has been given a timeless design which will survive short-lived fashion trends. It also offers many possibilities of individualization," says Roland Wolff, Product Manager of the Compact Cameras business unit at Leica Camera AG. The small and elegant LEICA D-LUX has a clear array of controls and is easy to use. The design of the LEICA D-LUX is reminiscent of the use of classic forms for Leica's rangefinder cameras. It is a modular composition of basic geometric figures such as semi-circles and rectangles.
The result is a timelessly elegant shape that fits comfortably in the photographer's hand. "Like all our compact cameras, the LEICA D-LUX was designed as a longlife product. The three-year warranty, which is unusually long for a compact camera, underlines the company's policy of value retention. Leica's typically long product life cycles ensure that the camera will not be replaced by a successor model after only a few months," explains Roland Wolff.
On top of this, owners are given an optional guarantee in the form of the free "Leica Passport". In the first year of ownership, the LEICA D-LUX is ensured against all damage, even if caused through the fault of the user, e.g. by knocks or water. The normal warranty conditions then apply in the second and third year.
The LEICA D-LUX has an aluminum body. Great attention has been paid to the smallest of details: for example, the main controls, the lids of the battery compartment and the compartment for the various connections are made of metal and not imitation-metal plastic.
In the growing digital camera market, the LEICA D-LUX takes account of the increasing desire of quality-aware consumers to stand out from the crowd by using individual products with their own personality. The LEICA D-LUX will be produced in comparatively small quantities of a few thousand a month. Its unique nature is also reflected in its valuable accessories - the high-quality leather case is offered in five different colors and models. Another individualizing feature is Leica's unique offer to engrave a name in the camera body on request (as an alternative to the "Leica passport").
The LEICA D-LUX sports a 1/2.5" CCD image sensor with 3.2 million effective pixels and a triple zoom lens LEICA DC VARIO-ELMARIT f/:2.8-4.9/5.8-17.4 mm ASPH. which was designed at Leica's Optics Center in Solms. "The Leica brand stands for top optical quality and is inextricably linked with photographers who capture the decisive moment in a highly emotive way," says Roland Wolff. The LEICA DC VARIO-ELMARIT f/2.8-4.9/5.8-17.4 mm ASPH. corresponds to a 35 to 105 mm lens for 35 mm photography and consists of seven elements in six groups, including three aspherical elements. The optics were individually matched to the 3 megapixel image sensor. Their interaction with the electronics and software of the LEICA D-LUX results in true-to-life color rendering, excellent definition and superlative brilliance.
The minimal delay of the shutter release, fast data storage and signal processing of the LEICA D-LUX are genuine advantages in practice, making the camera ready for virtually non-stop operation. Series can be taken at a rate of four frames per second. A series can consist of up to five pictures in high JPEG quality or eight pictures at the maximum compression setting. In addition, video films can be recorded with sound - the only time limit being the size of the memory card (360 seconds for the 64 MB card, for example). The LEICA D-LUX offers a 114,000-pixel 1.5" monitor for viewing the picture during and after the exposure. Using the second battery, the camera is ready for more than two hours of action in active mode.
The LEICA D-LUX comes with two rechargeable lithium ion batteries and a generous 64 MB SD card. The memory card stores 45 to 553 images, depending on the resolution and compression setting. The delivery also includes a high-quality leather case for transporting the camera, a matching carrying strap and a second case for keeping the extra battery and another SD card, an external power/charger unit and extensive ADCSee 5 PowerPack (PC) software.
The LEICA D-LUX will be available at Leica stockists' from May 2003.
LEICA DIGITAL-MODUL-R in preparation for 2004
The new Casio Exilim EWC-1 waterproof case
New MINOX 4 Megapixel camera DC 4011
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Ps. this week also marked the first week since Hadley's been born that I've managed to get a post up every day! ICYMI: Cyber Monday Sales // Thanksgiving Recap // Hadley 5 Months // Gift Guide: Beauty Lover
Ps. this week also marked the first week since Hadley's been born that I've managed to get a post up every day! ICYMI: Cyber Monday Sales // Thanksgiving Recap // Hadley 5 Months // Gift Guide: Beauty Lover
20 comments
One of the good things about Jesse's job starting later in the morning than his previous job is that he comes home at a later time, which means most things I order online have usually already arrived at the house before he is home.ReplyDelete
That ornament is so cute! I bought a snowman ornament this year for my older sister and her family that is holding snowflakes with each of their names in a snowflake. I ordered it and received it weeks ago and of course 2 days ago she tells me that she is pregnant (which I'm overjoyed about). Just means they'll be getting a new ornament for Christmas next year with the new baby's name included. And Rory will have a cousin that will only be about 6 months younger than her. So I'm excited about that.
Someone had a very good mail week! Love the caption - dog not for sale LOL!! Happy Friday girl! xo, Biana -BlovedBostonReplyDelete
Love the new mirror that you guys got--and that is a great price!ReplyDelete
My packages have started rolling in, too. Hell, one of them came the day after I ordered it and I was like "what magical elves work for Under Armour?" hahah Hoping for the next few to arrive today because on of them has MG's birthday gifts in it, and its his birthday today...oops! haha
Have a great weekend!
I can't believe I haven't picked up that mirror yet, it's gorgeous and such a good price! My Black Friday and Cyber Monday packages have started rolling in too - Nathan about dies every day! Have a great weekend, love!ReplyDelete
As Mark would say "blogger Mail day" in a weird sing song voice ;-) You definitely got some great buys!ReplyDelete
Love the ornament! I've been looking at them on amazon and can't decide between a snowman or a snowflake! Have a great weekend!ReplyDelete
Girl I have honestly had busy mail weeks where I ran home on my lunch break to grab packages. haha it's so sad! Or I deliver things to my parents house and pick them up when I get my kids ;-) Yay for updating your mantle mirror! Hope you have a great weekend with your Christmas party!ReplyDelete
Ahh - first Christmas ornaments. so special to have - you'll keep it forever.ReplyDelete
I was spoiled by having H home most of last week too, the struggle is real this week! In love with that ornament! I've been a bad mom, and need to research those before it's too late! Happy Friday gurlie <3ReplyDelete
Green Fashionista
Woohoo for posting every day! This week has totally felt like the longest ever! Loving that ornament girl!ReplyDelete
That mirror! I'm telling myself I need for somewhere in our house!ReplyDelete
I love collecting Baby's 1st Christmas ornaments. I really like the spode ones, got a picture one and plan to do a footprint one for Fletcher. Brady's judgy puppy eyes in the picture are so funny. Remind him of all the great deals you found before he judges your stack of boxes hahaha :)ReplyDelete
That ornament is SO cute! So great for Hadley's first Christmas! :)ReplyDelete
That ornament is so cute! We need to get a first Christmas ornament still. I looked a few places and didn't see one that stood out. Love that sweater!! And that pie sounds soooooo good right now!! Yum! Hope you enjoy your first Christmas party of the season, have a great weekend!ReplyDelete
I love that mirror! I'm looking for a gold mirror for baby girl's nursery, I never think about looking at Target!ReplyDelete
I did notice your daily posts last week - get it, girl! I love when all the packages start arriving because it's basically like early Christmas. Hope you had a nice weekend!ReplyDelete
Love that mirror! And how sweet is that ornament?! Hope you had a great weekend!ReplyDelete
Southern Style
Kudos to you for keeping up with the blog posts! I don't know how you do it! lol! That chocolate pie looks yummy! Love your new mantle mirror!ReplyDelete
Great job getting all those posts up this week! You go momma! I don't think I've done that yet either haha. We've been looking at one of the Lenox ones for T's first ornament too... There's a Rudolph one I think I'm going to buy :)ReplyDelete
oh wow, that mirror is gorgeous! i want it. and how cute is the ornament? love it. what a sweet first ornament :)ReplyDelete


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So Titanic 3-D is coming to town. Yay - pause for golf clap - personally I'm more excited for BATTLESHIP. Don't fault me for it, Hunger Games definitely ain't coming. Something about post-apocalyptic authoritarian dictatorships just doesn't go over here. Ah, well.
Fifteen years ago, Titanic made a big impression in China back when there were even fewer foreign movies screening (horrors! I can't imagine). At the time, Jack and Rose's steamy car scene ran in its entirety, nudie painting and all. I mean, after all, it was pg-13 and everything, right?
We wont be so lucky the first time around. The nude scene has been cut, says Ministry of Tofu. And some people are pretty steamed about it. This satirical phony official government explanation for striking the scene made me laugh out loud [translation also from MoT]:
"The State Administration of Radio, Film and Broadcast finally gave an explanation of its decision to remove the nude scene it OK'd 15 years ago: 'In light of the specialness of 3D movies, we are concerned that viewers may extend their hands for a touch during the scene and hit heads of viewers sitting in their front, which may result in disputes. In consideration for building a society with spiritual civilization, we decide to remove the scene.'"


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Tuesday, January 19, 2010
The Lovely Bones: The Movie
Warning, mild spoilers for the book.
I was hesitant to watch this film as The Lovely Bones is one of my favorite books. But color me impressed I loved it. It was good, honest, adaption with great special effects.
I am glad Peter Jackson chose relative unknowns for the roles of Susie, Ruth, Lindsay, and Ray. It made the film feel more realistic than it would've been had some teen idol had been cast. I wasn't too impressed with casting Rachel Weisz and Mark Wahlberg as Susie's parents, they acted well they just didn't seem to fit in.
The actress who really stole the movie was Susan Sarandon, as Susie's boozing, fun-loving grandmother. Her role was expanded from what was in the book and simply made the movie. Stanley Tucci did well as Susie's killer, but without the back story provided in the book, he became just another psycho.
Peter Jackson stays mostly true to the book. However, in order to condense it into a movie certain plot points were dropped such as Susie's mother's affair and her friendship with Ray's mother. The characters of Ray and Ruth were diminished as well as Lindsay's relationship with her boyfriend, Samuel. This is kind of a bummer considering the large impact these characters had in the novel.
Even the changes that Peter Jackson did for the story such as Holly's true identity and the expanded character of Susie's grandmother worked for the film. Rarely do changes in the plot do that and it only enhances the charm of the film.
If you liked the book, you'll like the movie. Peter Jackson did a phenomenal job bringing the story to life.
Posted by Jennifer Turner at 5:39 PM
Labels: Based On The Novel
1 comments:
- Alison said...
I LOVED this book. I'm pretty upset that my local movie theater isn't playing the movie!
Hearing that a book version lover was impressed with the movie makes me even more anxious for when it's released on DVD!
- January 19, 2010 at 9:13 PM
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Doctors Lounge - Pharmacy Answers
"The information provided on www.doctorslounge.com is designed to support, not replace, the relationship that exists between a patient/site visitor and his/her physician."
Forum Name: Pharmacy & Drug Topics
Question: Any safe anti-inflammatories/pain relievers for asthmatics?
|dacubster - Mon Dec 10, 2007 11:55 pm
Share |
I am a 24 year old female with a history of severe asthma, bronchitis, hyperthyroidism, IST, and depression.
I recently reinjured my shoulder (from a preexisiting injury) and the doctor suspects a labral tear. I am currently waiting on physical therapy and then possibly surgery.
However, since the reinjury occured I have been unable to straighten my arm and have been in significant pain. My primary doctor and orthopedic surgeon have tried numerous drugs which have either not worked or I have had bad reactions to.
I have been on many doses of steriods to no avail. I have been on the anti-inflammatory Etolodac, which caused a severe asthma attack. I have also been on PROPOXYPHENE which I also did not have a good reaction with. They will not try most pain relievers because I have had severe breathing problems in the past with even small doses of Vicodin and Morphine. I have tried over the counter relievers (tylenol, etc), but they do little to help.
The problem is that the doctor does not want to try anything (especially anti-inflammatories) else because they are worried about setting off another asthma/breathing problem. This has left me in severe discomfort and stiffness. Are there medicines that I could safely take without having an affect on my breathing?
|Dr. Chan Lowe - Thu Dec 13, 2007 10:54 pm
Share |
Hi Dacubster,
This is difficult. If you are having worsened asthma symptoms with some non-steroidal anti-inflammatories the risk of having problems with others is high. For pain, I would recommend talking with your doctor about other pain medicines that are not in this category of meds.
You may also want to consider seeing a pulmonologist to help get your asthma under better control so that you may be able to use the non-steroidals.
Best wishes.
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First thing you do when you start with Android Development is learning how to debug.
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Dive in android source code and see for yourself how things work.
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Go to github, go to code.google and scan through good open source projects.
You will find tons of gold in them. Implementation, best practices, smart hacks.
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There are tons of blog on android which are regularly updated.
Follow devs on google plus, great place to stay updated.
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Latest posts by sunny (see all)
- Kindergarten Kids Word Games - March 8, 2016
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- How to Hide WhatsApp Photos and Videos from Android Gallery - April 7, 2014


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The Walt Disney Co. topped fiscal third-quarter earnings expectations by hauling in 77 cents per share, a 15 percent improvement over last year and 4 cents better than analysts had predicted.
Revenue also topped expectations, coming in at $10.7 billion, up 7 percent. Net income was $1.5 billion, up 8 percent.
Chief executive Robert Iger credited the company's Media Networks, especially ESPN, along with Parks and Resorts and Consumer Products for the strong results, though not Studio Entertainment, which was responsible for the bulk of a $34 million impairment charge.
Of Disney's five segments, only Studio Entertainment posted lower revenue this quarter compared with the same quarter last year. Revenue fell 1 percent to $1.62 billion, and operating income at the studio plunged 60 percent to $49 million.
Cars 2 and Thor had tough comparisons with last year's Toy Story 3 an Iron Man 2, but Pirates of the Caribbean: On Stranger Tides offset much of the damage.
Disney shares were rising 2 percent in after-hours trading.
Disney shares, which had been up 9 percent on the year as recently as last month, have taken a nosedive along with the rest of the market. In August alone, the stock has plunged 10 percent. On Tuesday, though, the shares gained 5 percent to $34.70 on a strong market rebound, though the stock remains down 8 percent on the year.


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