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Release | Chiaki Uzurahashi
WONDER CREW president Tomonori Watanabe challenges to be the no.1 food company from Sapporo
Make people who interact with happy. WONDER CREW has various style restaurants with high aspirations and venture spirits. Tomonori Watanabe is the president who leads the firm. He pushes his will towards the food industry in Japan itself as well.
"Irasshaimase" as a "Thank you"Tomonori was working for a men's clothing sales company previously. When Tomonori read a book about a famous entrepreneur, he was really touched by the way of thinking and started to aspire to start a business himself.
He was grown up in an Okonomiyaki restaurant, which was run by his parents, and was helping the restaurants since he was a little child. When he was alone in the restaurant, the word "Irasshaimase" popped out from his mouth. ("Irasshaimase" is a Japanese word for welcoming someone into a restaurant) "I think I was purely happy when the customer came into our restaurant". When he think back about that, it was "Irasshaimase" as a "thank you" to the customer.
He experienced a part-time job at a restaurant when he was a student. He realizes that he could not say "Irasshaimase" with the emotion of gratitude. When he thought about starting a business, he chose the food industry because of his experience, he wanted to become a person to say good "Irasshaimase".
▲The president of WONDER CREW Tomonori Watanabe. Tomonori was born in Sapporo in 1982. He graduated Hokkaido Asai Gakuen University (Now Hokusho University). After graduated the school, he started working for a men's clothing sales company. After three and half years, he switched his career to the food industry. Opened "Veggy's House" in Chuo-ku Sapporo in 2011 when he was 27. Founded WONDER CREW in 2011.
Expanding restaurants ambitiously, using the strength of "design"Italian style bar using Hokkaido vegetables called "Veggy's House" was founded in 2010. Since then, he has expanded new restaurants which use Hokkaido ingredients in reasonable prices. Now WONDER CREW has 16 restaurants mainly in Sapporo with different brands, such as oyster bar "FISHMANS", "Hokkaido Italian Izakaya EZOBARU BANG!BANG!" and "Taishu Niku Sakaba Gatsuri".
▲EZO BARU BANG!BANG! provides Italian food with Hokkaido ingredients at a reasonable price. The picture is EZO BARU BANG!BANG Sapporo Odori. (Photo provided by WONDER CREW)
One of the strengths of WONDER CREW is that they do the creative works such as the web and interior design on their own. "To make the restaurant attractive and to be distinct comparing to other restaurants, design is necessary. The food industry will be a battle of creative so we will work on creative as if we are designing the eating situation itself."
The design production operated by WONDER CREW called "STUDIO WONDER" involves art director and graphic designer Sou Nomura, one of the directors of WONDER CREW mainly. The promotion tools of WONDER CREW's restaurant "Sanchi Chokuso Odoroki Kaisen Izakaya Uotto", which was one of the works of Nomura, won the second prize award in "Sapporo ADC Competition & Award 2015".
Make people who shine with happiness in the food industryThere are 40 members of WONDER CREW and the average age is 26. Including part-time workers, members are more than 150 people. Tomonori remains firm with the belief "people is everything". He says "company must be a place where people can grow up. Our company is young and there are many young people so I put emphasis on educating humanity".
To keep members' motivation high and make members who work together turn to the same good direction, Tomonori hosts "Nomiken" once in three months, closing all the restaurants. This is a training with different themes each time, and after the training, they eat and drink and interact with people including clients.
"My principle is to make 4 people happy: customers, employees, employees of clients, and residents of the area where our restaurants are".
To open a restaurant is to involve to town development. "I want to participate in revitalizing the town or the area through our restaurants and business".
▲"Lobster mayonnaise", one of the most popular dishes in EZO BARU BANG!BANG! (Photo provided by WONDER CREW)
Tomonori is very serious to change the food industry. To make the industry more attractive, and give more chances for new challenges to people in the industry.
"The job in the food industry is so much fun because it is a job to make customers happy. Speaking of management, the fun part is that the numbers change when location, business category, and customer needs matched. There are members who want to start a business in future. Life is one time so I want our members to challenge and I would love to back up them."
Tomonori also wants to establish a cooking school. "There are many people who get a job but give up when they are in training because they cannot do what they want. It is a pity. Can't we make young people become independent cooks earlier? I believe if I keep saying this, someone who will cooperate shows up so I will keep saying."
To his destinationTomonori started the franchise of EZO BAR BANG!BANG! last year, and two restaurants in Takamatsu (Kagawa prefecture) and Matsumoto (Nagano prefecture) opened outside of Hokkaido. There are more and more restaurants planned to open spreading all areas in Japan.
Tomonori has 3 goals to achieve by 2020; make the company listed, increase the number of restaurants to 70 including franchise, and make 10 entrepreneurs. "We will become a leading company in the food industry and become the no.1 food company in Japan". Tomonori puts passion to his words.
▲The picture taken with staffs at the 5th-anniversary party for Veggy's House. (Photo provided by WONDER CREW)
Tomonori, who keeps challenging, has a strong feeling;
"When I was a student, I could not do well in studying or working. A person like me can start a business and can enjoy working with people who are related and members in the same generation. I want young generations to feel something from me."
Tomonori keeps going with his staffs, seizes chances, and never stops to challenge.
Related Links- WONDER CREW
Text / Hokkaido Likers writer Chiaki Uzurahashi
Photo / Hokkaido Likers photo writer Minako Takada
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Grey Box, Six Foot and independent developer YAGER have launched Dreadnought, the class-based spaceship action game, into free-to-play open beta for all users on the PlayStation 4 computer entertainment system via the PlayStation®Store. The console-exclusive Hactar Hero Ship, a new ShareFactory theme, and two dynamic themes for the title are also available for free today through … Continue reading
Category Archives: Sony News
SUPERHOT and SUPERHOT VR Coming to PS4 Next Week
PlayStation fans are about to get SUPERHOT times two with next week's release of the original SUPERHOT for PS4 and SUPERHOT VR for PlayStation VR. PS+ members save an additional 20% when pre-ordering SUPERHOT or SUPERHOT VR, as well as scoring an exclusive PS4 theme. The FPS where time moves only when you move, SUPERHOT became an instant-hit upon release … Continue reading
'Fortnite' Gets Four PlayStation-Exclusive Heroes
At E3 this week, Epic Games announced that PlayStation 4 players who purchase one of the Founder's Packs for Fortnite will receive bonus hero cards. These exclusive hero cards are only available to PlayStation players, and sport a unique color scheme that players can use to show off their Sony pride. Fortnite will be available … Continue reading
PS4 Technical Beta Announced for 'GWENT'
CD PROJEKT RED, creators of The Witcher series of games, announce a weekend-long PlayStation 4 Technical Beta for GWENT: The Witcher Card Game. In GWENT: The Witcher Card Game, gamers clash with their friends in fast-paced duels that combine bluffing, on-the-fly decision making and careful deck construction. The game is played over a best-of-three series … Continue reading
'Invisible, Inc.' Console Edition PS4 Launch Details
Klei Entertainment, developers of Don't Starve and Mark of the Ninja, have announced that their spy game, Invisible Inc., is set to release on April 19 via PSN for the PS4. Priced at $19.99, Invisible Inc. Console Edition will be launching with the entire game that was available on PC as well as with the … Continue reading
'Beyond: Two Souls' Coming To PS4 Next Week, 'Heavy Rain' Next March
Quantic Dream co-CEO, Guillaume de Fondaumiere, has just announced via the Playstation Blog, that Beyond: Two Souls is coming to PS4 next week complete with "graphical improvements and fan-requested features." It'll be available to download on the Playstation Store for $29.99 on November 24th. The PS4 edition of the game promises a number of improvements including: … Continue reading
Film Noir Horror Game 'Dollhouse' Coming To PS4
Creazn Studio has announced that its film noir horror game Dollhouse will be coming to the PS4 as well as PC. Taking inspiration from the visuals and bleakness of 1940's film noir, Dollhouse tells the story of Marie, a detective trying to unravel her past. She slowly begins to discover that she might not be who … Continue reading
'Journey' Probably, Maybe Coming To PS4 on July 21
According to a listing on the Docs For Playstation site, Journey could be set to release on the PS4 on July 21. The site has that date listed for its release as well as game guides and manuals for the game. It also lists the name of the game as Journey Beta meaning the game … Continue reading
'Uncharted: The Nathan Drake Collection' Coming to PS4 This October
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'Until Dawn' Finally Gets a Release Date, New Trailer Too
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Two 'Batman: Arkham Knight' PS4 Bundles Announced
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'Tropico 5' Coming To PS4 in April
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$5 PSN Flash Sale Now Live, 20+ Titles Available
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subset: commoncrawl


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Field recording from a forest area at Røsnæs in Denmark. Recorded February 2008. Forest surrounding, birds and crows singing, a pigeon and a single woodpecker appears, wing flutters. Distant cars, a distant helicopter and general distant rumble.
Recorded with Zoom H2 build in microphones.
Good work !
Amazing recording, thanks!
Great stuff! Thanks loads :-)
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Do It Yourself Loan Modifications
- May 20, 2010
- Posted by Mcfarlin Law, in Loan Modification
Several Internet based companies have started to advertise "do it yourself" loan modification services. These companies promise to teach borrowers the basics of loan modifications while providing all of the necessary forms, documentation and information a borrower will need to successfully apply for a modification. Programs also promise to teach borrowers how to negotiate with their lenders.
If any borrowers are facing foreclosure and are considering purchasing a do it yourself loan modification program, don't.
To begin with, many of these programs don't even allow homeowners to see what they are buying until after a credit card has been processed. Many homeowners will be disappointed to find out that their "do it yourself" program is nothing more than an instruction manual full of information that could just as easily have been found on the web for free.
Homeowners must also consider the urgency of their situation. A foreclosure can be very time sensitive to many homeowners. What homeowner has the time or the patience to learn everything they need to know about a loan modification while their lenders are actively pursuing the foreclosure of the home.
Also, loan modification negotiation skills are not something that can be learned from a book or a class, and especially not over night. Negotiation skills are learned by sitting in on and conducting actual negotiation sessions. Many lenders can recognize an untrained negotiator from a mile away and are very skilled at breaking down the confidence of an untrained negotiator. Once the lender gains control over the negotiator, the negotiation is over.
Since foreclosure laws vary from state to state and from lender to lender, the information contained in do it yourself loan modification programs will not apply to every person the program is marketed to. Many homeowners will be very disappointed if they discover the tactics outlined in the do it yourself program don't apply to their specific state or lender.
To be blunt, these do it yourself loan modification services have "scam" written all over them. Borrowers often make uninformed decisions when faced with the stress of an impending foreclosure, but the stress should never lower a person's guard. Borrowers must be very cautious before approaching any company or service that offers a "quick fix", "guaranteed results", "overnight success", or the simplicity of a do it yourself program. A home is too large of a risk to take on an Internet purchased do it yourself program. Borrowers should use their money to secure the services of a trained attorney who can represent them every step of the way. Not only will this keep a homeowner from having to do all of the work themselves, but it will guarantee them that an individual with knowledge of mortgage laws and experience negotiating with lenders will be handling their case.
It is fine if a borrower wants to learn more about loan modification services for their own benefit, but their own home should not be used as the guinea pig to measure their skills. The lessons learned can be far more costly than the cost of the Internet purchased course.


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Honor the Heroes of the Game, Preserve Its History, Promote its Values & Celebrate Excellence Everywhere
Share your pictures, videos, and stories from your visit to the Pro Football Hall of Fame on our social media. We might even feature you in one of our promotions!
Wednesday, Aug 01 2012
Watch Curtis Martin's career highlights as presented by NFL Films.


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Learn Infusionsoft
Infusionsoft Training with expert Sharon Broughton
Learn from the "expert"! Sharon's training covers all aspects of Infusionsoft and delves deep into the areas that directly impact and support successful system use for business owners. This course is useful for both business owners or your Virtual Assistant support staff to learn the ins-and-outs of Infusionsoft.
What will we be covering?
- The differences between other systems and Infusionsoft so you can take advantage of features in Infusionsoft
- Setting up products and membership programs
- Merchant account and gateway setup
- Customizing the cart, order forms and web forms
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- Templates, broadcasting and sending ezines
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- Affiliate program set up
- Campaign builder (basic and advanced features)
- Plug ins, customizing, other programs that integrate with Infusionsoft (like CustomerHub and iMember360… and more)
For more information, just click here.


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Create a virtual photography, video, or art competition and use social media to ask your followers to vote for their favorite. Also, be sure to check out the World Oceans Day photo contest with the United Nations.
Sample Event: World Oceans Day Photography Competition
World Oceans Day is a great opportunity to gain support and raise funds for your conservation initiatives. Create an online fundraiser and spread the word on social media.
Looking to support World Oceans Day directly? Make a donation on the website or visit the Facebook page to make a donation or create a fundraiser. Thank you for your support!
Sample Event: Clean Ocean Access Fundraiser on World Oceans Day
Have a live stream for your followers on social media! Have a 360 video of the inside of an exhibit at an aquarium, zoo, or beach; record prominent speakers; or give an online tour of what you are doing for World Oceans Day. Whatever you choose, be sure to make it easily accessible, fun, and interesting!
Sample Event: "Souls of the Vermilion Sea" Online Film Screening and Facebook Live Filmmaker Q&A
Publish social media posts leading up to World Oceans Day. Whether it is daily or weekly, continue to remind people to celebrate World Oceans Day, get involved, and take action for the ocean. Download free World Oceans Day social media resources and graphics.


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The core of your being is illuminated by maintaining a youthful, breezy adaptability, as there's an uncanny ability to adjust to life's situations irrespective of what's thrown at you.
However, many times this is perceived as a lack of seriousness about life and that's far from the case. Inside, you struggle from bouts of inadequacy because you always think that you should be "better" in relation to anything you touch or any interaction you have. There's a tight grip on your heart and the playfulness serves as a way to ease that weight.
Taking responsibility for your emotional life is one of your greatest lessons to learn because at your highest potential you're designed to live a life of integrity where the public and private you is aligned. You don't have to fake it. The reality of your feelings is one of your greatest allies. So, curbing the tendency to bury significant emotions under a veneer of jokes only enables you to further evade the root of issues that need to be addressed.
Once that accountability is established, you'll feel more grounded and confident. In addition, you're able to more effectively deal with the feelings of others with an air of detachment which doesn't say " I don't care", but rather finds an objective space which will prevent you from being swayed into unhealthy decision-making.
With your Sun/Moon blend, the "youth" and the adult can work in unison. Understand that all work and no play indeed makes you dull, but also there are definitely matters in life which require the sober, ponderous and strategic approach that is naturally yours.
Balancing your sociability and solitude is also a key theme when it comes to your cosmic signature. You need time to process things and this needs to be accepted. You will need to find the sweet spot between focus and variety. You can't be everywhere at once and this can scatter your energies as well as your brain.
If you're able to successfully navigate this terrain, you will undoubtedly feel as if your ideas and your life in general has gained an "elastic pragmatism" where you can entertain possibilities, but have the common sense to know which ones are worth entertaining and which ones are merely flights of fancy.
At best, you possess a solid, hard-won emotional savvy along with a mental agility that can strip people and situations down to the bare bones. At worst you refuse to allow yourself the space needed to address your inner needs and overcompensate by convincing yourself and others that your life is nothing but fun and games.
Photo by Jhonis Martins from Pexels


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Returns and Refunds
If we have made an error with your order, or you have received your order in a damaged or faulty condition, you can return it to us within 7 days of the date you received the item, unopened with any seals intact and we will issue a full refund for the price you paid for the item. The refundable amount will include a deduction for any discounts you received on your original order, at a level proportional to the normal value of the returned item(s). Please note: We can only accept the return of opened items if they are faulty.
Instructions for returning items to us and our policy on the refund of delivery charges are carefully outlined in the following sections
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Your Statutory Rights (Applies to UK Customers Only)
Under UK law, you have the right to withdraw from your purchase of an item within seven working days of the item's delivery to you. We regret that we cannot accept cancellations of contracts for the purchase of products where the item has been opened and unsealed.
If you take delivery of a www.animalife.co.uk package and the contents have been damaged in transit, you can return the item(s) to us within 7 days of receipt for exchange.
How To Return An Item
Our returns policy means that if we have made an error with your order, or you have received your order in a damaged or faulty condition, you can return it to us within 7 days of the date you received the item, unopened with the seals intact and we will issue a full refund for the price you paid for the item. Please note: We can only accept the return of opened items if they are faulty.
Please follow the steps below to enable us to process your refund efficiently:
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This offer is available on products exclusivley in the "INTENSE" lifestage range and limted to the largest item in the range and purchased between 01/11/15 to 01/06/18.
If after using a loading serve to start and then used every day until you have used all the product, you are not happy with the results, we will refund you the purchased price excluding any delivery charges, while stocks last, limited to one purchase per customer per household. All you need to do is send the used and empty product back to us (tub and lid) by recorded delivery in a cardboard box, including your original receipt, original delivery note and enclose a letter (confirming your name, address, telephone and email address) with why you are not happy with the results to:
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text: The Starving Artists Show is a wonderful change of pace; a different type of show. Sell your odd pieces and clear your inventory at this end-of-season outdoor show. The fun event brings an eclectic crowd looking to buy unique art. This is a show where artists can discount, reduce, and generally sell off extra art inventory. Visitors come looking for bargains on everything from jewelry to ceramics to fine art. This unique art fair offers artists the opportunity to sell off their inventory at the end of the season; great art at great prices! Festival dates are September 24-25, 2016. Apply now before the January 25, 2016 deadline! Apply Now!


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It is recognized that cancer cells exhibit highly elevated glucose metabolism compared to non-tumor cells. We have applied in vivo optical imaging to study dynamic uptake of a near-infrared dye-labeled glucose analogue, 2-deoxyglucose (2-DG) by orthotopic glioma in a mouse model.
Methodology and Principal Findings
The orthotopic glioma model was established by surgically implanting U87-luc glioma cells into the right caudal nuclear area of nude mice. Intracranial tumor growth was monitored longitudinally by bioluminescence imaging and MRI. When tumor size reached >4 mm diameter, dynamic fluorescence imaging was performed after an injection of the NIR labeled 2-DG, IRDye800CW 2-DG. Real-time whole body images acquired immediately after i.v. infusion clearly visualized the near-infrared dye circulating into various internal organs sequentially. Dynamic fluorescence imaging revealed significantly higher signal intensity in the tumor side of the brain than the contralateral normal brain 24 h after injection (tumor/normal ratio, TNR = 2.8±0.7). Even stronger contrast was achieved by removing the scalp (TNR = 3.7±1.1) and skull (TNR = 4.2±1.1) of the mice. In contrast, a control dye, IRDye800CW carboxylate, showed little difference (1.1±0.2). Ex vivo fluorescence imaging performed on ultrathin cryosections (20 µm) of tumor bearing whole brain revealed distinct tumor margins. Microscopic imaging identified cytoplasmic locations of the 2-DG dye in tumor cells.
Citation: Zhou H, Luby-Phelps K, Mickey BE, Habib AA, Mason RP, Zhao D (2009) Dynamic Near-Infrared Optical Imaging of 2-Deoxyglucose Uptake by Intracranial Glioma of Athymic Mice. PLoS ONE 4(11): e8051. https://doi.org/10.1371/journal.pone.0008051
Editor: Pedro Antonio Valdes-Sosa, Cuban Neuroscience Center, Cuba
Received: August 28, 2009; Accepted: November 3, 2009; Published: November 30, 2009
Copyright: © 2009 Zhou et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported in part by the North and Central Texas Clinical and Translational Science Initiative (NCTCTSI) Pilot Grant Award as part of the NIH CTSA Grant UL1 RR024982 and DOD IDEA Award W81XWH-08-1-0583 (DZ). Imaging was fulfilled by the Southwestern Small Animal Imaging Research Program (U24 CA126608), the Caliper IVIS Spectrum under NIH 1S10RR024757-01 and the CRi Maestro was provided by the Joint Program in BME through a DOE grant #DE-FG02-05CH11280. MRI experiments were performed in the Advanced Imaging Research Center, an NIH BTRP # P41-RR02584 facility. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Glioblastoma multiform (GBM) is a lethal intracranial cancer, which exhibits a relentless malignant progression and is highly resistant to conventional multimodal therapies. GBM is characterized by the nature of extensive infiltration into surrounding normal brain tissue, which results in incomplete tumor resection and consequent recurrence [1], [2]. It is imperative to improve diagnostic imaging to evaluate intracranial tumor growth and therapeutic responses. Optical imaging has been rapidly adapted to cancer research. Recently, optical imaging using fluorescent dye labeled tumor-specific molecules has been successfully applied to imaging glioma in preclinical animal models based on overexpression of such markers in glioma [3]–[5]. In the clinic, several recent studies have demonstrated the ability of fluorescence imaging to facilitate radical resection of GBM during surgery [6]–[8]. Promising results reported by Stummer, et al. have shown that gross total resection of glioma guided by intraoperative fluorescence imaging is associated with improved prognosis of the patients with a median survival of 15–18 months, compared to 10–12 months after a subtotal resection or about 6 months after biopsy alone [8].
It is well recognized that cancer cells exhibit highly elevated glucose metabolism and up-regulated glucose transporters (GLUTs) compared to non-tumor cells. On this basis, 18FDG, the glucose analogue, has been used as the most common PET radiotracer to visualize clinical tumors and their metastases. However, a drawback of 18FDG PET for brain tumors is strong background signals of normal brain tissues, which often compromise the ability to diagnose the brain tumors. Moreover, PET has a low spatial resolution in spite of high sensitivity. Optical imaging by visualizing fluorescently labeled tumor cells has recently emerged as an attractive approach to facilitate identification of infiltrative tumors and sentinel lymph node metastases [9]–[14]. Alternative to radioactive deoxyglucose, fluorescent derivatives of 2-DG, e.g., 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose (2-NBDG) have shown a greater tendency to be delivered and trapped in tumor cells [15], [16]. However, the short wavelength of 2-NBDG (excitation, 475 nm; emission, 550 nm) limits its applications for in vivo imaging. Near infrared fluorescence has several advantages over the use of visible fluorophores including deeper penetration due to less tissue absorption and scattering of light, and minimal autofluorescence. IRDye800CW 2-DG (Li-Cor Bioscience), a NIR dye conjugated with 2-deoxyglucose (peak excitation 785 nm, emission 810 nm), has recently been developed and demonstrated as a tumor-targeting optical contrast agent in various tumors implanted subcutaneously in mice, which can be visualized in vivo by fluorescence imaging [17], [18]. Moreover, a recent study of pharmacokinetics has shown that there is essentially no retention of the dye in normal mouse brain 24 h after injection [19]. Thus, the 2-DG NIR dye may serve as an ideal contrast agent for optically imaging brain tumors.
In this study, we first applied bioluminescence imaging (BLI) and MRI to assess non-invasively intracranial tumor growth in an orthotopic glioma model in nude mice. We then exploited IRDye800CW 2-DG for dynamic in vivo imaging of these brain tumors. In vivo observations were validated by both ex vivo fluorescence imaging of cryosections of tumor bearing brain tissues and histological staining. Finally, fluorescence microscopic studies were performed to identify locations of the 2-DG dye in tumors and normal brain.
Immunohistochemical study showed extensive expression of luciferase in the U87-luc cells of intracranial tumor tissues (Fig. 1A). Longitudinal BLI studies revealed a weak signal initially 11 days after tumor implantation, which became stronger on follow-up to day 24 (Fig. 1B). The mean light intensity detected on day 24 for the group of tumor bearing mice was significantly greater than on day 11 (16×107 versus 11×106 photons/s; p<0.05). MRI studies based on T2-weighted and T1-weighted contrast enhanced images confirmed an intracranial tumor located in the right side of the brain of each animal (Fig. 1C). A significant correlation was found between BLI signal and actual tumor volume measured by MRI (r = 0.8, p<0.05; Fig. 1D). Tumor volume determined on the last MRI scan (one day before the in vivo fluorescence imaging) varied from 42 to 136 mm3 in the seven animals. There was no significant difference in tumor volume between the group receiving the 2-DG dye or the control dye when the in vivo fluorescence imaging was performed (p = 0.5).
A. Immunostaining with anti-luciferase antibody showed extensive expression of luciferase in tumor tissues. B. BLI showed increasing light intensity in the mouse brain over time. C. Transaxial MRI detected an intracranial tumor on consecutive slices (arrows) based on both T2- weighted and T1-weighted contrast enhanced MRI, which was found to grow in size on the follow-up study (84 mm3 on day 17, 136 mm3 on day 24). D. A good correlation between the BLI signal and actual intracranial tumor volume was observed (r = 0.8, p<0.05).
The real-time whole body NIR optical imaging, acquired immediately after i.v. injection of the 2-DG dye, IRDye 800CW 2-DG (15 nmol/mouse) or the control dye, IRDye 800CW carboxylate (15 nmol/mouse), visualized the first pass of the dye sequentially through various organs and tissues (Fig. 2). By narrowing the field of view (FOV), brain-focused in vivo imaging showed dynamic changes in fluorescence signal intensity on the tumor side of the brain (right) in comparison with the contralateral normal brain (left). Relatively higher signal intensity was observed for the 2-DG dye in both sides of the brain than the rest of the mouse body (Fig. 3). Time course curves showed that the signal intensity in both sides of the brain had a maximum value of 1.2×107±0.2×107 counts/s 10 min after injection, and gradually washed out over 4 h. There was no significant difference in signal intensity between the two sides of the brain during the first 4 h after injection (Figs. 3A and 4; p>0.3). Twenty four hours later, a dramatic drop in signal intensity was observed. However, significantly higher light signal was clearly seen in the tumor side of the brain than the contralateral normal side (1.8×105±0.2×105 versus 0.7×105±0.4×105; p<0.05; Fig. 4A). A mean tumor/normal ratio (TNR) for the group of 4 tumors was 2.8±0.7 (Fig. 4B). In contrast, the control dye showed little difference (0.54×105±0.1×104 versus 0.43×105±0.3×104; TNR = 1.1±0.3; Fig. 4B and C). To confirm that the signal originated from the brain tumor, we performed surgical procedures to reflect scalp and further remove skull to expose the brain. A brighter and more focused light signal emitted from the region of the tumor implantation was seen (Fig. 3A). Stronger contrast was achieved by reflecting the scalp (TNR = 3.7±1.1) and further removal of the skull of the mouse revealed a highest TNR of 4.2±1.1 (Fig. 4C). For the control dye, similar to the observation in the intact animals, no significant difference was found (TNR = 1.4±0.2 and 1.4±0.1, respectively; Fig. 4C). Comparison of in vivo BLI and 2-DG fluorescence imaging of intracranial tumors also showed a strong linear correlation albeit in a small number of animals (n = 4; r = 0.9; Fig. 4D). After sacrificing the mice, the whole brain was dissected and ultrathin sections (20 µm) of tumor bearing brain were imaged using the same fluorescence imaging system as used for in vivo. Most intriguingly, distinct tumor margins, even the tracks of migration of small number of tumor cells, were identified based on the fluorescence images, which correlated well with histological staining of the tumor (Fig. 5). In good agreement with in vivo imaging data, uptake of the 2-DG dye was found to be significantly higher in tumor than the contralateral normal brain (TNR = 3.7±1.1; p<0.01; Fig. 5I). Finally, fluorescence microscopy showed more uptake of the 2-DG dye in tumor regions, compared to the control dye (Fig. 6). Co-registration of the NIR signal with the DAPI stained nuclei showed a cytoplasmic localization of the 2-DG dye (Fig. 6G).
A series of whole body images was acquired before and after injection of near infrared dye labeled 2-DG, IRDye800CW 2-DG, via a mouse tail vein. Selected images captured the first pass perfusion of the dye into various internal organs: heart and lung area at 1.2 s, brain and upper limbs at 2.5 s, kidney at 4.5 s, etc. Principal Component Analysis of the kinetics was used to identify tissue regions based on DyCE software as shown in the color presentation (top left) and representative time courses (bottom right).
A. After i.v. injection of IRDye800CW 2-DG repeated in vivo fluorescence imaging was performed. During the first four hours, unmixed images showing stronger signal of the dye in the brain area, but there was no obvious contrast between the tumor side of the brain and the normal brain. However, 24 h later, the light signal remained only in the tumor side of the intact brain. Even better contrast was seen after reflection of the scalp or further removal of the skull. B. Normalized emission spectra showing the near infrared 2-DG dye with a peak emission wavelength at 810 nm (red), while the background signal (white) was at ∼770 nm. C. As a control of the 2-DG dye, IRDye800CW carboxyl was injected into a mouse bearing orthotopic glioma. No significant difference in light intensity between the two sides of the brain was observed at any time points.
Mean light intensity curves shown for the tumor side (solid square) and the contralateral normal brain (open circle) of intact mice. A. For the 2-DG dye (n = 4), the curves were essentially identical for the first 4 h post injection, but after 24 h, there was a significant difference (1.8×105±0.2×105 versus 0.7×105±0.4×105; p<0.05). B. In contrast, the control carboxylated dye showed no difference during the whole course (n = 3). C. Time course of tumor to normal tissue ratio (TNR) showing a significantly higher TNR in the 2-DG dye group (mean = 2.8±0.7; solid bar) versus the control group of the control dye (n = 3; 1.1±0.3; open bar) 24 h after injection (p<0.05). An even bigger TNR was obtained in the 2-DG group by removing the scalp (3.7±1.1) or skull (4.1±1.1). D. A strong linear correlation was observed between light intensity of in vivo BLI and 2-DG NIR imaging of intracranial tumors (r = 0.9).
Unstained coronal brain sections (20 µm) containing tumor tissues were obtained immediately after the 24 h in vivo image for ex vivo imaging. For the tumors with 2-DG dye, representative images for a larger (A) and a small (D) intracranial tumor showed predominant accumulation of the dye within the tumor mass. Tumor mass was clearly delineated from the surrounding normal tissues. Even a track of migrated tumor cells was depicted in each case (arrows in A and D), which correlated well with cresyl violet staining of the corresponding regions enlarged from B and E (arrows in C and F). Arrow heads refer to the dentate gyrus. The control dye showed no significant difference between the tumor regions and the normal brain (G and H). Tumor/normal ratio (TNR) of the 2-DG dye was significantly higher than that of the control dye (3.7±1.2 vs. 1.4±0.1; p<0.05; I).
The unstained frozen sections of tumor bearing brain tissues, also used in Fig. 5, were scanned under dark field (A and D). Near infrared signals (B and E) of the same fields detected with an infrared filter were overlaid on the dark field images (C and F). Significantly higher signal was seen in tumor tissues of the mouse injected with the 2-DG dye, compared to the one with control dye. Co-staining of nuclei of tumor cells with DAPI indicated cytoplasmic localization of the 2-DG dye (red; G).
We have demonstrated the feasibility of using the NIR dye labeled 2-deoxyglucose, IRDye800CW 2-DG, for in vivo fluorescence imaging of orthotopic glioma in a mouse model (Fig. 3). Ex vivo fluorescence imaging of tumor sections and microscopic imaging confirmed significantly higher accumulation of the 2-DG dye in intracranial tumors than in normal brain. Good correlations were found between each imaging modality in terms of in vivo evaluation of intracranial tumor burden (Figs. 1 and 4D).
Small animal imaging has been increasingly adapted for preclinical cancer research. In vivo imaging promises greater efficiency since each animal serves as its own control and multiple time points can be examined sequentially. In particular, multimodal imaging approaches provide comprehensive information about both tumor anatomy and pathophysiology and even molecular mechanisms [4], [20]–[23]. In this study, we combined optical imaging (bioluminescence and fluorescence imaging) with MRI to study longitudinal development of intracranial tumors and their uptake of a glucose analogue, 2-DG. There was good agreement between increased BLI signal intensity over time and enlarged tumor volume measured by MRI, as also reported by others previously for intracranial tumors in rodent models [4], [24]. Thus, the cheap, fast and high-throughput BLI seems to be just as effective in monitoring the deep-seated orthotopic tumor models as the expensive and time consuming MRI. However, BLI does not provide anatomic details due to the poorer spatial resolution and limited depth of light penetration. Recent progress in BLI tomography should facilitate three-dimensional analysis. In addition to providing detailed anatomic structure, MRI is also useful to obtain pathophysiological information, i.e., tumor vascular perfusion and permeability, oxygenation, and apoptosis or necrosis, though not reported here.
Fluorescence imaging can be used to evaluate small reporter molecule pharmacokinetics, avoiding the need for genetic modification of cells. In particular, near infrared fluorescence (NIR) imaging has found a greater potential for clinical application because of its long wavelength (650–900 nm), where light absorbance and scattering are significantly lower, and autofluorescence of normal tissues is also greatly reduced [25]. In view of this approach, various target-specific NIR conjugates have been reported for targeting tumor imaging, e.g., tumor integrin, αvβ3 [4], tumor growth factors or their receptors [18], [26]–[30], glycoprotein [31], or tumor specific protease [21]. Successful applications in various preclinical tumor models have been reported, though most of these studies were performed on surface tumors. In vivo NIR imaging of deep-seated orthotopic tumor models, in particular, intracranial tumors, remains challenging. By targeting the overexpressed αvβ3 in tumor, Hsu, et al. recently reported visualization of orthotopic glioma of mouse in vivo by NIR imaging via Cy5.5-RGD. Peak of tumor uptake was found 2 h post injection and TNR = 2.6 was achieved with a craniotomy [4]. More recent work with an orthotopic mouse brain tumor McCann, et al. successfully applied fluorescent molecular tomographic imaging to monitor protease activity in tumor by using protease activatable fluorescence, ProSense680 (peak light emission at 680 nm). By co-registering fluorescence images with MRI, localization of the active protease in tumor was determined [5].
Here, we applied the commercially available NIR labeled 2-deoxyglucose to imaging intracranial tumors based on the simple mechanism of differential levels of glucose metabolism between tumors and normal tissues. Tumor cells both require more energy for their higher proliferation rate and utilize the inefficient glycolytic pathway to produce energy. Thus, tumor cells need more glucose compared to normal cells. On this basis, 18FDG has been used as the most common PET radiotracer to visualize clinical tumors and their metastases. However, 18FDG PET imaging of brain tumor is often compromised by strong background signals of normal brain tissues. Due to the short half life of fluorine-18 (<2 h), PET imaging is normally performed within 1 h after injection of 18FDG. The lower contrast between tumor and normal brain may be partly attributed to this timing, at which a maximum ratio of uptake is not reached. Indeed, we found no significant difference in light signal in the tumor side of the brain versus the normal side of the brain during the first 4 hours post injection. The peak tumor to normal ratio was actually observed at 24 h, which is consistent with a previous study of subcutaneous U87 tumors using cy5.5 labeled D-glucosamine. However, a control dye, cy5.5-NHS used in that study also produced a high TNR [16]. Our study is also in a good agreement with another study that detected essentially no retention of IRDye800CW 2-DG in normal brain after 24 h assessed by ex vivo imaging [19]. In addition to tumor diagnostic imaging, pyropheophorbide labeled 2-deoxyglucosamine has shown a potential for photodynamic therapy on tumors [32], [33].
Our results of ex vivo fluorescence imaging showed that the 2-DG dye distributed well into the whole tumor despite some heterogeneity (Fig. 5). This observation may suggest delivery and distribution of the 2-DG dye to the tumor does not need the BBB disruption, which is the prerequisite condition for the dyes currently used for neurosurgery. Indeed, uncoupling of tumor vascular perfusion and permeability and uptake of FDG has been reported previously [35], [36]. However, it is still possible that the dye first leaks out through the disrupted BBB, from which it diffuses into the whole tumor. Further studies will be necessary using earlier stage of tumors, when the Gd contrast leakage is not obviously seen by T1-weighted contrast enhanced MRI, to prove this hypothesis. Furthermore, the usefulness of the 2-DG dye to stain the infiltrative tumor cells is limited by the tumor model used in this study. The U87 tumor has relatively sharp tumor-brain boundaries. GBM models showing more aggressiveness will be needed to test the ability to stain the finger-shaped infiltrative tumors. Instead of the established GBM cell lines, using surgically resected tumor tissues directly from GBM patients and passing them in animals will generate stable orthotopic GBM xenografts, which show oncogene patterns very similar to primary tumors of patients.
In summary, fluorescence imaging of deoxyglucose uptake in more clinically relevant orthotopic glioma models has not yet been reported. Our results may suggest an optimal time for imaging brain tumors based on the glucose analogues. The near-infrared dye labeled 2-DG may serve as a novel imaging probe to noninvasively monitor intracranial tumor burden in preclinical animal models. From a clinical perspective, development of NIR labeled tumor-specific molecules may have the potential to identify the infiltrating glioblastoma intra-operatively, and to improve the extent of resection of selected tumors.
Materials and Methods
Orthotopic Glioma Xenografts
All animal procedures were approved by the Institutional Animal Care and Use Committee of University of Texas Southwestern Medical Center. Human glioma U87 MG cells (ATCC, Manassas, VA, USA) were infected with a lentivirus containing a firefly luciferase reporter, and highly expressing stable clones were isolated. A ∼1 cm long incision of skin was made along the midline of the brain of an anesthetized nude mouse (BALB/c nu/nu; Harlan, Indianapolis, IN). Using a high-speed drill, a 1 mm burr hole of the skull was made in the right hemisphere, anterior to the coronal fissure. About 5×104 U87-luc cells in 3 µl mixture of PBS and Matrigel (25%, BD Biosciences, San Jose, CA) were injected directly into right caudal diencephalon, 1.5 mm beneath the dura mater using a 32G Hamilton syringe. Usage of a 32G fine needle minimizes tissue damage. The burr hole was filled with bone wax and the scalp was closed with sutures.
In Vivo BLI
BLI was initiated for monitoring intracranial tumor growth about 10 days after tumor implantation and repeated once a week using the IVIS Spectrum system (Caliper, Xenogen, Alameda, CA). The tumor bearing mice (n = 7) were anesthetized (isoflurane/O2 in an induction chamber; isoflurane from Baxter International Inc., Deerfield, IL) and a solution of D-luciferin (120 mg/kg in PBS in a total volume of 80 µl; Biosynthesis, Naperville, IL) was administered s.c. in the neck region. Anesthesia was maintained with isoflurane (2%) in oxygen (1 dm3/min). Five minutes after luciferin injection, an array of various exposure time (1, 5, 30, 60 s) was applied for image acquisition. Data were quantified with the Living Imaging software by using absolute photon counts (photons/s) in an ROI, manually drawn to outline the BLI signal of the brain.
In Vivo MRI
Once a BLI signal was observed, MRI was performed to assess tumor volume and growth by using a 4.7 T horizontal bore magnet with a Varian INOVA Unity system (Palo Alto, CA). Each mouse was maintained under general anesthesia (air and 2% isoflurane). A 27 G butterfly (Abbott Laboratories, Abbott Park, IL) was placed in a tail vein for contrast agent administration. Pertinent image slice positions were based on fast sagittal scout images. T1-weighted (TR = 250 ms; TE = 20 ms; slice thickness = 1.5 mm; FOV = 25×25; in plane resolution 195 µm) and corresponding T2-weighted (TR = 1900 ms; TE = 80 ms) spin-echo multislice coronal images were acquired. T1-weighted contrast enhanced images were acquired after i.v. bolus injection of the contrast agent Gd-DTPA-BMA (0.1 mmol/kg body weight; OmniscanTM, Amersham Health Inc., Princeton, NJ) through the tail vein catheter. We determined tumor volume on T2-weighted images by manually outlining the enhancing portion of the mass, excluding congested CSF signal in ventricles, on each image by using standard "browser" software provided with the Varian Inova imaging system. The area measurements were automatically calculated and multiplied by the MRI section thickness to calculate a per-section tumor volume. The total tumor volume was obtained by summing the volume calculations for all sections.
Near Infrared Fluorescence Imaging
In vivo real-time imaging of first pass perfusion.
When the intracranial tumors grew to larger than 4 mm in diameter based on T2-weighted MRI, in vivo fluorescence imaging was performed using a Maestro imaging system (CRI Inc. Woburn, MA). Each mouse was anesthetized by i.p. injection of a ketamine/xylazine cocktail (40 µl). A 27 G butterfly (Abbott Laboratories) was placed in a tail vein for administration of the 2-DG dye, IRDye800CW 2-DG (15 nmol/mouse in 150 µl saline, Li-Cor Biosciences, Lincoln, NB) or a control dye, IRDye800CW carboxylate (15 nmol/mouse in 150 µl saline, Li-Cor Biosciences). Based on the methodology developed by Hillman and Moore [37], an excitation filter (671–705 nm) was applied. A series of whole body images was acquired before and after a bolus injection of the 2-DG dye or the control dye. Typically, 5 frames/s for the first 20 s, 2 frames/s for the next 30 s and 1 frame/s for the rest of ∼2 min were acquired. Image processing and data analysis were performed using the DyCE software provided with the Maestro software 2.8.
In vivo dynamic imaging of 2-DG uptake by intracranial tumors.
The brain-focused fluorescence images were acquired before and 10 min, 30 min, 2 h, 4 h, and 24 h after injection of the 2-DG or the control dye. A set of filters specifically for NIR imaging (excitation, 671–705 nm; emission, 730–950 nm) was applied. At each time point, the mouse was anesthetized by i.p. injection of the ketamine/xylazine cocktail (40 µl). Immediately after the last image at 24 h, the scalp of the mouse was surgically reflected to expose the skull and a fluorescence image was acquired. Before sacrificing the mouse, the skull was removed to expose both sides of the brain tissues and a last in vivo image was obtained. The whole surgical procedure and imaging was completed within 15 min on the anesthetized animals and no obvious bleeding was observed.
Ex vivo fluorescence imaging on cryosections of tumor bearing brain.
Immediately after sacrificing the animals, the whole brains were dissected and embedded in O.C.T. and then transferred to −80°C freezer. On the second day, a series of coronal sections (20 µm) of the brain was cut. The frozen sections containing tumor tissues were identified by cresyl violet staining. The adjacent tissue sections were used for ex vivo fluorescence imaging and fluorescence microscopic study, as described in detail below. The ex vivo images were taken by using the same Maestro system as used for in vivo studies. Typically, an exposure time of 2.5 s was applied for image acquisition.
Analysis of Fluorescence Imaging
Fluorescence images were obtained using the Maestro system and processed with the Maestro software 2.8. The spectrum of background signal (peak emission ∼770 nm) was first obtained from a mouse before the IRDye 800cw dye injection, while the spectrum of IRDye 800cw dye (peak emission ∼810 nm) was detected from a solution of the dye in PBS (0.1 nmol/µl). The spectra were then imported and used to unmix the NIR dye signal from the background signal for both in vivo and ex vivo studies. The whole set of in vivo images of an individual mouse, obtained before and at various times after injection of the dye, was examined for quantification. A common ROI was based on the most obvious signal of the tumor, which was observed on the image of the skull-deprived mouse 24 h after injection in each of the 2-DG animals, was applied on both the tumor side and the contralateral normal brain of each image. A total of photon counts (counts/s) in the identical ROI was used for comparison of dynamic change in signal intensity.
Histology and Fluorescence Microscope
Cresyl violet staining was performed on the cryosections (20 µm). Tumor margins were determined under microscope and correlated with the NIR light signal acquired by ex vivo fluorescence imaging. The cryosections, unstained or counterstained by 4′,6-diamidino-2- phenylindole (DAPI), were used for fluorescence microscopy study. The NIR fluorescence signal was detected using a Zeiss AxioObserver (Carl Zeiss MicroImaging, Inc., Thornwood, NY) equipped with NIR filters. The NIR signals were recorded and overlaid on either a dark image or an image counterstained by DAPI of the same field. For luciferase staining, monoclonal mouse antiluciferase mAb (1∶150; AbD Serotec, Raleigh, NC) and cy3-conjugated goat anti-mouse secondary antibody (Jackson Immunoresearch Laboratories, West Grove, PA) were used.
We are grateful to Drs. Li Liu and Phillip Thorpe and Mr. Abhijit Bugde for technical and collegial support.
Author Contributions
Conceived and designed the experiments: BM AH RM DZ. Performed the experiments: HZ KLP DZ. Analyzed the data: HZ KLP AH DZ. Contributed reagents/materials/analysis tools: HZ KLP BM AH RM DZ. Wrote the paper: BM RM DZ.
- 1. Furnari FB, Fenton T, Bachoo RM, Mukasa A, Stommel JM, et al. (2007) Malignant astrocytic glioma: genetics, biology, and paths to treatment. Genes Dev 21: 2683–2710.
- 2. Wen PY, Kesari S (2008) Malignant gliomas in adults. N Engl J Med 359: 492–507.
- 3. Hsu AR, Hou LC, Veeravagu A, Greve JM, Vogel H, et al. (2006) In vivo near-infrared fluorescence imaging of integrin alphavbeta3 in an orthotopic glioblastoma model. Mol Imaging Biol 8: 315–323.
- 4. Hsu AR, Cai W, Veeravagu A, Mohamedali KA, Chen K, et al. (2007) Multimodality molecular imaging of glioblastoma growth inhibition with vasculature-targeting fusion toxin VEGF121/rGel. J Nucl Med 48: 445–454.
- 5. McCann CM, Waterman P, Figueiredo JL, Aikawa E, Weissleder R, et al. (2009) Combined magnetic resonance and fluorescence imaging of the living mouse brain reveals glioma response to chemotherapy. Neuroimage 45: 360–369.
- 6. Laws ER, Parney IF, Huang W, Anderson F, Morris AM, et al. (2003) Survival following surgery and prognostic factors for recently diagnosed malignant glioma: data from the Glioma Outcomes Project. J Neurosurg 99: 467–473.
- 7. Barker FG, 2nd, Chang SM (2006) Improving resection of malignant glioma. Lancet Oncol 7: 359–360.
- 8. Stummer W, Pichlmeier U, Meinel T, Wiestler OD, Zanella F, et al. (2006) Fluorescence-guided surgery with 5-aminolevulinic acid for resection of malignant glioma: a randomised controlled multicentre phase III trial. Lancet Oncol 7: 392–401.
- 9. Sevick-Muraca EM, Sharma R, Rasmussen JC, Marshall MV, Wendt JA, et al. (2008) Imaging of lymph flow in breast cancer patients after microdose administration of a near-infrared fluorophore: feasibility study. Radiology 246: 734–741.
- 10. Parungo CP, Ohnishi S, De Grand AM, Laurence RG, Soltesz EG, et al. (2004) In vivo optical imaging of pleural space drainage to lymph nodes of prognostic significance. Ann Surg Oncol 11: 1085–1092.
- 11. Burton JB, Johnson M, Sato M, Koh SB, Mulholland DJ, et al. (2008) Adenovirus-mediated gene expression imaging to directly detect sentinel lymph node metastasis of prostate cancer. Nat Med 14: 882–888.
- 12. Kaushal S, McElroy MK, Luiken GA, Talamini MA, Moossa AR, et al. (2008) Fluorophore-conjugated anti-CEA antibody for the intraoperative imaging of pancreatic and colorectal cancer. J Gastrointest Surg 12: 1938–1950.
- 13. Hayashi K, Jiang P, Yamauchi K, Yamamoto N, Tsuchiya H, et al. (2007) Real-time imaging of tumor-cell shedding and trafficking in lymphatic channels. Cancer Res 67: 8223–8228.
- 14. Hoffman RM (2005) The multiple uses of fluorescent proteins to visualize cancer in vivo. Nat Rev Cancer 5: 796–806.
- 15. O'Neil RG, Wu L, Mullani N (2005) Uptake of a fluorescent deoxyglucose analog (2-NBDG) in tumor cells. Mol Imaging Biol 7: 388–392.
- 16. Cheng Z, Levi J, Xiong Z, Gheysens O, Keren S, et al. (2006) Near-infrared fluorescent deoxyglucose analogue for tumor optical imaging in cell culture and living mice. Bioconjug Chem 17: 662–669.
- 17. Kovar JL, Simpson MA, Schutz-Geschwender A, Olive DM (2007) A systematic approach to the development of fluorescent contrast agents for optical imaging of mouse cancer models. Anal Biochem 367: 1–12.
- 18. Sampath L, Kwon S, Ke S, Wang W, Schiff R, et al. (2007) Dual-labeled trastuzumab-based imaging agent for the detection of human epidermal growth factor receptor 2 overexpression in breast cancer. J Nucl Med 48: 1501–1510.
- 19. Kovar JL, Volcheck W, Sevick-Muraca E, Simpson MA, Olive DM (2009) Characterization and performance of a near-infrared 2-deoxyglucose optical imaging agent for mouse cancer models. Anal Biochem 384: 254–262.
- 20. Zhao D, Richer E, Antich PP, Mason RP (2008) Antivascular effects of combretastatin A4 phosphate in breast cancer xenograft assessed using dynamic bioluminescence imaging and confirmed by MRI. FASEB J 22: 2245–2451.
- 21. Weissleder R, Pittet MJ (2008) Imaging in the era of molecular oncology. Nature 452: 580–589.
- 22. Serganova I, Blasberg RG (2006) Multi-modality molecular imaging of tumors. Hematol Oncol Clin North Am 20: 1215–1248.
- 23. Beattie BJ, Forster GJ, Govantes R, Le CH, Longo VA, et al. (2007) Multimodality registration without a dedicated multimodality scanner. Mol Imaging 6: 108–120.
- 24. Rehemtulla A, Stegman LD, Cardozo SJ, Gupta S, Hall DE, et al. (2000) Rapid and quantitative assessment of cancer treatment response using in vivo bioluminescence imaging. Neoplasia 2: 491–495.
- 25. Adams KE, Ke S, Kwon S, Liang F, Fan Z, et al. (2007) Comparison of visible and near-infrared wavelength-excitable fluorescent dyes for molecular imaging of cancer. J Biomed Opt 12: 024017.
- 26. Ke S, Wen X, Gurfinkel M, Charnsangavej C, Wallace S, et al. (2003) Near-infrared optical imaging of epidermal growth factor receptor in breast cancer xenografts. Cancer Res 63: 7870–7875.
- 27. Hama Y, Urano Y, Koyama Y, Choyke PL, Kobayashi H (2007) Activatable fluorescent molecular imaging of peritoneal metastases following pretargeting with a biotinylated monoclonal antibody. Cancer Res 67: 3809–3817.
- 28. Chang SK, Rizvi I, Solban N, Hasan T (2008) In vivo optical molecular imaging of vascular endothelial growth factor for monitoring cancer treatment. Clin Cancer Res 14: 4146–4153.
- 29. Diagaradjane P, Orenstein-Cardona JM, Colon-Casasnovas NE, Deorukhkar A, Shentu S, et al. (2008) Imaging epidermal growth factor receptor expression in vivo: pharmacokinetic and biodistribution characterization of a bioconjugated quantum dot nanoprobe. Clin Cancer Res 14: 731–741.
- 30. Runnels JM, Zamiri P, Spencer JA, Veilleux I, Wei X, et al. (2006) Imaging molecular expression on vascular endothelial cells by in vivo immunofluorescence microscopy. Mol Imaging 5: 31–40.
- 31. Zou P, Xu S, Wang A, Povoski S, Johnson M, et al. (2009) Near-Infrared Fluorescence Labeled Anti-TAG-72 Monoclonal Antibodies for Tumor Imaging in Colorectal Cancer Xenograft Mice. Mol Pharm 6: 428–440.
- 32. Zhang M, Zhang Z, Blessington D, Li H, Busch TM, et al. (2003) Pyropheophorbide 2-deoxyglucosamide: a new photosensitizer targeting glucose transporters. Bioconjug Chem 14: 709–714.
- 33. Zhang Z, Li H, Liu Q, Zhou L, Zhang M, et al. (2004) Metabolic imaging of tumors using intrinsic and extrinsic fluorescent markers. Biosens Bioelectron 20: 643–650.
- 34. Kremer P, Mahmoudreza F, Ding R, Pritsch M, Zoubaa S, et al. (2009) Intraoperative fluorescence staining of malignant brain tumors using 5-aminofluorescein-labeled albumin. Neurosurgery 64: 53–60; discussion 60-51.
- 35. Tseng J, Dunnwald LK, Schubert EK, Link JM, Minoshima S, et al. (2004) 18F-FDG kinetics in locally advanced breast cancer: correlation with tumor blood flow and changes in response to neoadjuvant chemotherapy. J Nucl Med 45: 1829–1837.
- 36. Galie M, Farace P, Nanni C, Spinelli A, Nicolato E, et al. (2007) Epithelial and mesenchymal tumor compartments exhibit in vivo complementary patterns of vascular perfusion and glucose metabolism. Neoplasia 9: 900–908.
- 37. Hillman EM, Moore A (2007) All-optical anatomical co-registration for molecular imaging of small animals using dynamic contrast. Nat Photonics 1: 526–530.


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Islands. The amount so reserved shall be allotted to eligible institutions and shall be available only for the purpose of providing workstudy assistance to such students." E L I M I N A T I O N OF AVERAGE H O U R S OF EMPLOYMENT L I M I T A T I O N NON-REGULAR E N R O L L M E N T PERIODS
Atrte. p. 1029.
g j ^, 136, Section 444 of the Higher Education Act of 1965 (as amended by this part^ is amended by adding at the end thereof the following new subsection: "(c) For purposes of paragraph (4) of subsection (a) of this section, in computing average hours of employment of a student over a semester or other term, there shall be excluded any period during which the student is on vacation and any period of non-regular enrollment. Employment under a work-study program during any such period of non-regular enrollment during which classes in which the student is enrolled are in session shall be only to the extent and in accordance with criteria established by or pursuant to regulations of the Commissioner." REVISION OF M A I N T E N A N C E OF E F FOR T REQUIREMENT
Post, p. 1033.
SEC. 137. Effective for fiscal years ending on or after June 30, 1970, section 444(a)(5) of the Higher Education Act of 1965 (as amended by this part) is amended to read as follows: " (5) provide that the institution will meet the requirements of section 464 of this Act (relating to maintenance of effort);" ADMINISTRATIVE
SEC. 138. Effective for jfiscal years ending on or after June 30, 1970, section 444(a)(2) of the Higher Education Act of 1965 (as amended b j this part) is amended by striking out all that follows "administrative expenses" and inserting in lieu thereof "in accordance with section 463 of this Act;". E L I G I B I L I T Y OF PROPRIETARY INSTITUTIONS
Ante, p. 1029. Post, p. 1032.
SEC. 139. Effective for fiscal years ending on or after June 30, 1970— (1) Section 443(b) of the Higher Education Act of 1965 (as amended by this part) is amended by striking out "or" before "an area vocational school", and by inserting before the period at the end thereof the following: ", or a proprietary institution of higher education (as defined in section 461(b) of this Act) ". (2) Section 444(a)(1) of such Act (as amended by this part) is amended by inserting after "work for the institution itself" the following: "(except m the case of a proprietary institution of higher education),". PART D—COOPERATIVE EDUCATION PROGRAMS GRANTS TO INSTITUTIONS OR HIGHER EDUCATION FOR PROGRAMS OF COOPERATIVE EDUCATION; GRANTS AND CONTRACTS FOR TRAINING AND RESEARCH IN COOPERATIVE EDUCATION
SEC. 141. Title IV of the Higher Education Act of 1966 is amended by redesignating part D as part F, by redesignating sections 461 79 Stat. 1251through 467 as sections 491 through 497, respectively, and by in425? 44^1^ 443',591! serting after part C the following new part:


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5 Tips to Create The Best Landing Page Design
B2B brands and organizations that employ effective landing pages can expect an increase in leads, a higher ROI and an improvement in their company's image. Landing pages need to employ effective design, and special care needs to be taken regarding how a landing page is presented to ensure your landing page converts. Here are five tips to ensure your landing page contributes to brand development:
1. CTA Design as Competitive Advantage
Subtle design elements can help a call-to-action (CTA) pop off the page. That's why you should consider using a strong font color, appropriate size text, and a minimal background to draw the user's eye. Small details make a difference in web design. One case study showed that by changing the CTA button from a dark blue to an eye-catching green helped increase conversions by 35 percent.
2. Draw A User Down The Funnel
Research has shown that most users view web pages in an "F" shape. That means readers start by looking at the left-hand corner and slowly make their way towards the middle section on the right side of the web page. Place key information in this "F" pattern to guide the visitor towards the final CTA on the right side of the page.
You should also think about implementing directional cues that reinforce your landing page's goals. These cues don't necessarily have to be an arrow pointing to the CTA; you can also have a person in a photograph looking in the CTA's direction, which is a subtle yet more powerful directional cue for visual branding. It's all about using design for a competitive advantage.
3. Reduce Distractions
While your website wants to provide easy navigation, your landing page wants to do the opposite. That's why you want to remove unnecessary navigation elements and links that might lead a visitor away from your page. Your goal is to keep the user there and ensure they perform the right actions.
Be sure that you also don't bog visitors down with difficult and lengthy fields. Just ask for their name and e-mail address. These fields should also be "above the fold," which means the visitor doesn't have to scroll down to sign up with your website.
4. Think About The Unique Needs Of Your Organization
It's important to think about your business and its unique branding when you design a page. For example, less exciting businesses can think about trying to generate excitement with landing page videos. One study found that landing page videos led to an 80 percent increase in conversions in certain cases.
Nonprofits may want to consider employing powerful imagery that highlights their cause. For example, a CTA imposed on the background of a clear-cut rainforest can trigger an emotional reaction that gets people on board with a nonprofit's mission.
5. Testing is Key
Often, it's worth experimenting with different landing page designs to overcome the challenges facing B2B brands. A/B testing can help you understand which ones convert at a higher rate.
For example, imagine you're a startup that doesn't have the brand appeal that larger companies have. In that case, you want to try arranging social proof like user testimonials, industry awards and media coverage at the top of the page instead of the bottom. This could immediately reassure new customers that you're a professional company that offers valuable products or services. However, only testing will help you understand whether this strategy actually pays dividends.
Ultimately, landing page design is both an art and a science. If you want a corporate rebrand or brand refresh that gets results, it's a good idea to work with a company like Novation Consulting. Our web and graphic design experts can help you produce landing pages that deliver real value and contribute to brand development.


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Home » Food Menu » Breakfast Menu December 2019
Date: 01 Sun 2019
Author: elaine
Breakfast Menu December 2019
5 April | 2019
Easter Afternoon Tea
Join us from the 15th – 22nd April...
3 February | 2019
Valentines High Tea*
Link to Menu:-Valentines 2019
I will certainly be going back to Cuthberts, it was a lovely relaxing cafe.
Gorgeous little tearoom! I almost don't want to tell you about it!
*Your name:
*Your E-mail:
*Your Message:
Contact Us
Copyright © 2018 Cuthbert's Bake House. All Rights Reserved.


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Bandana Scarf Skull€ 12,95 € 9,95
Bandana Scarf Sea Camouflage€ 12,95 € 9,95
Bandana Scarf Forest Camouflage€ 12,95 € 9,95
Black gloves with white skeleton print€ 14,95 € 9,95
- Lightweight balaclava
- Materials: 65% polyester and 35% cotton
- Close-fitting
- Cut-away at eye level
- One size fits most
- Skull print
- Suited for outdoor sports
- Ideal wear for snowboarders and bikers
- Wearable with or without helmet/ski specs
A balaclava is an ideal item to provide you with that extra little protection against weather and wind when you're driving down the road or gliding downhill. This balaclava is made of 65% polyester and 35% cotton and is close-fitting.
It's of course possible to combine this balaclava with a crash helmet and motorbike or ski specs. For outdoor sports a balaclava is an ideal solution which lies between no face protection at all and a full-fledged face mask.
Click on the photo to enlarge.
Item number: FM 5-006 (E5)


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"Whisk" Dad away this Father's Day! The last thing Dad needs is yet another tie! So this year give him something to wet his whistle!
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Save 25% off all single bottle craft and micro beers Saturday only! While supplies last.
Limit 12 per guest
750 ml. and smaller
No other discounts apply
No breaking up multi packs
Offer good Saturday, June 15th
"Whisk" Dad away this Father's Day! The last thing Dad needs is yet another tie! So this year give him something to wet his whistle! This whiskey cocktail will fire up the taste buds with some spicy ginger beer and fresh blackberries.
Purchasing whiskey for your father can be daunting, but never fear: We've made it simple and easy! The first step of the process is to determine what style of whiskey your dad already enjoys. If you purchase a style of whiskey you know he enjoys, but one notch better than he usually purchases, you'll be in good shape. If you don't know what whiskey your father likes, or if he's new to whiskey, then we've got you covered, too! Below are a few common styles of whiskey (with recommendations).
American Bourbon whiskey is smoother and sweeter than other whiskies. "Straight" bourbons cannot have added colors, flavors or other spirits, must be distilled from at least 51% corn grain and aged at least two years in charred, new oak barrels (plus a handful of other rules). Some whiskey (like Leopold Bros Small Batch Whiskey) cannot be called "straight bourbon" due to the minimal aging, but should not be discounted as excellent whiskey. The rules get a bit crazy, but don't let them get in your way of purchasing good whiskey. For seasoned whiskey drinkers, we recommend unique, small-batch or single-barrel whiskies like Rowan's Creek bourbon, Basil Hayden's, or Leopold Bros Small Batch Whiskey. For fathers new to whiskey, we recommend Buffalo Trace, Bulleit bourbon, or Knob Creek bourbon.
Rye whiskey is sharp and spicy, and has similar rules to straight bourbon, but must be distilled from at least 51% rye grain. Rye is the classic mixing whiskey for many cocktails, including the Manhattan. Rye was popular during prohibition, since Canadian rye distillers remained open while their US competitors shut down. For seasoned rye lovers, we recommend Masterson's rye, Whistlepig rye, or High West Rendezvous rye. For rye newbies, we recommend Redemption rye, Bulleit rye, or Templeton rye.
Single malt Scotch Whisky (the "e" was dropped in the Atlantic at some point) is made in Scotland with malted barley and often has a strong, smoky, peaty profile not for the faint of heart. These range from light and delicate like the Glenlivet 12-year-old, to rich and peaty like Lagavulin 16-year-old. The US, Japan, and other countries produce single malt whiskey, but it can't be called "Scotch" unless it is from Scotland and follows specific rules. For seasoned Scotch drinkers, we'd recommend getting something with more smoke and peaty character, most of which will say "Islay" on the bottle (like Laphroig, Lagavulin, Bowmore or Ardbeg). For dads just beginning to explore single malts, take a look at the wide variety of Glenlivet or Glenfiddich single malts.
The only downside to adding ice to a fine glass of bourbon is that normal cubed ice tends to melt quickly and dilute the bourbon's flavor. But at last we've found a solution to this problem: Tovolo Sphere Ice Molds, which create ice "cubes" in a perfect, spherical shape. Because they have a larger surface area than a cube, these ice spheres chill your drink more quickly, but are slow to melt, allowing you to enjoy your bourbon from start to finish.
We love Hak's Chipotle Bourbon and we think dad will too! It's got a robust taste of caramelized onion and garlic, a smack of bourbon, and smoked and roasted chilies that comfort the palate with a slow–release heat. And we're not the only ones who love this barbecue sauce! Chef Sharone Hakman wowed the judges on Fox's MasterChef reality cooking competition with his secret weapon: uniquely crafted barbecue sauces.
Handmade in store by our talented meat specialists, this pork tenderloin has a juicy, savory and uniquely delicious flavor profile. We use PrairieFresh 100% Natural pork tenderloin, seasoned with whiskey fennel and breaded with Italian parmesan panko. Cooked to perfection with our simple cooking instructions, this tenderloin is sure to please Pop!
This deliciously rich cheddar from Cabot Cooperative in Vermont is excellent with tart apples and it's perfect paired with Bulleit bourbon.
Steak: It's what's for Father's Day. This robust marinade is perfect to
accentuate the meaty flavor this big steak provides. Dig In!
Our wine tasting events are a fun way to spend a Friday night, 5 p.m. - 7 p.m. at participating Nugget Markets locations! Tastings are only $5/flight and include a voucher toward a future purchase from our wine department. Attendees must be at least 21 years of age.
Now in its 37th year, the 4th of July Criterium bike race comes to downtown Davis. We're a sponsor of this all-day community event, where kids to the pros come out to race. Hosted by the Davis Bike Club, proceeds from this event benefit the club and its Helmets for Kids outreach. www.davisbikeclub.org.
We love supporting our local Independence Day events and celebrations in our various local communities. This year, our sponsorships support the City of Davis (cityofdavis.org), Cameron Park CSD on June 29th (cameronpark.org), July 4th Run for the Woodland Schools Fdn (woodlandschools
foundation.org), City of Woodland (cityofwoodland.org) and Sacramento's Spirit of the Pocket Parade (valcomnews.com).
You are receiving this message because the email address [email address suppressed] was subscribed to our list.
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Copyright © 2013 Nugget Market, Inc.
168 Court Street, Woodland, CA 95695 | nuggetmarket.com
Prices valid June 12-18, 2013.


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The new Daddy D's serves the exact menu as at the original location — only Isaiah rules the smoker. He says his dad taught him everything he knows, right down to peeling the potatoes, and serving a consistent product is what it's all about.
"My dad trained me, and he wouldn't even let me touch the pit for the first three years," Isaiah tells Ravenous Traveler. "I never saw myself doing this growing up. I thought I'd go to the NFL. But I fell in love with it. I view it like football: It's not just what you do, it's how you do it."
Isaiah says Daddy D's make its own "family style" of barbecue, with influence from his father's home state of Louisiana. On the menu are chopped beef brisket, chicken, pork ribs, pulled pork, cole slaw, potato salad, and more.
|Isaiah and Karolyn Vercher [Photo: Daddy D's Southern-Style Barbecue]
The new Daddy D's opened at 11817 NE 117th Ave. in the last week of January. Like the original, it smokes its meats in a smoker beside the Shell station. Currently, there's some outdoor seating beneath the Shell station roof, and coming soon are picnic tables and a large tent for additional seats. It's open Tuesday through Saturday, from 11 a.m. to 7 p.m., or until the meat sells out (if it's later in the day, you can check availability by calling 360 356 8357).
And Isaiah had some other big news to share: The first full-fledged brick-and-mortar location of Daddy D's is slated to open behind the Oak Tree Casino in Woodland, Washington, right off I-5. Isaiah says his father will finally have the restaurant he's always wanted, and the menu will definitely expand. Daddy D's first brick and mortar aims to open late spring 2018, at 1243 N Goerig St. (formerly Premium Smoked Meats).
Here are some more things to think about:
"I love the physical form of making barbecue," says Isaiah. "It's a craft, and I love the everyday learning. I don't think anyone could ever say, 'I mastered barbecue.' And barbecue also brings people together. I've met some of the coolest people in the world over barbecue, and while I know a lot of good things attract a lot of good people, I gotta say barbecue really brings great people together."
|Some things to think about [Photo: Daddy D's Southern-Style Barbecue]
- Top Portland Barbecue Moves into the Moda Center [Eater PDX]


text: Home : Worship : Messages : Messages 2010 Planting Dreams in the Dark Today's Services were a unique offering from our Intern Minister, Sue. There is no traditional sermon available for streaming, however, you may purchase the recording of this Service from our Bookroom on Sundays. Also, don't forget to plant your wild flower seed paper. Attachment Posted on Sun, December 26, 2010 by Sue Magidson, Intern Minister


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Years ago, I worked with a woman who became extremely anxious when she needed to buy a gift for someone. At the time we worked together. Our office building was attached to a mall and during the holiday season we would go shopping during our lunch hour. One year she bought ten different presents for her husband. It wasn't that she was giving him ten gifts. She had planned on giving him one gift. One day she bought a gift, then worried he wouldn't like it. The next day we returned it and bought him a different gift. We returned that one and found something else. We did this ten times. I think we would have continued except she ran out of days and gave him the last thing she bought. Back then, this wasn't considered an anxiety disorder. It was considered high-strung, nervous or even neurotic.
Gift-giving anxiety, according to D.B.Wooten (2000), is a form of social anxiety. The amount of anxiety one feels at giving a gift is dependent on the need for approval and the fear of being seen or judged in a negative way. People with gift-giving anxiety might think:
- "What if my husband doesn't like his gift?"
- "What if my children think I was cheap?"
- "What if my children think I bought more for their brother or sister?"
- "What if my boss thinks my gift is inexpensive or he doesn't like the color?"
- "What if my sister already has this item?"
- "Will my gift to Dad be as good as my brother's gift to him?"
According to Wooten, the higher your need to impress, the more anxious you will feel when giving a gift to someone. Below are some ideas to help you overcome or manage your anxiety about giving gifts this holiday season.
Ask. If you are not sure what someone on your list would like, ask. After all, you are buying a gift because you want to make this person happy. You might be surprised at the response you receive, your friend, relative, co-worker might appreciate that you took the time to find out what they wanted instead of buying some generic gift.
Find creative solutions. Some people on your list may not need or want anything. They may appreciate, however, spending time with you, having you baby-sit or having their car washed. Gifts don't always need to be material items. They can be the gift of time. (Great for those that have a holiday budget.)
Buy gifts based on your budget. Sometimes we want to show-off and buy someone the most expensive gift. These aren't necessarily the best gifts. Gifts that are thoughtful and hold some meaning for the recipient are the best gifts. The amount of money you spend is not important.
Remember it really is the thought, not the gift, that counts. Think back to the gifts you received. Which ones do you remember? You probably remember the gifts your children gave you, a card that said, "You are the best." You probably remember those that were given with love. Use this as your guide. Give your gifts with love and they will also be appreciated.
Focus on the person rather than you. So many times we buy gifts that we want to give rather than gifts the recipient wants to receive. Think about the recipient's life. What would make their life easier, bring a smile to their face, bring joy to their day.
No matter what gift you give, if you give a gift from your heart, with love, it will be remembered cherished for years to come. Lou Tice of the Pacific Institute says it all, "Put aside the expectations and the stress, and remember that it is the time devoted, the intentions and the love involved that make any gift truly special. When all is said and done, it won't be the gift that is remembered. It will be those moments of true connection with loved ones that live in our hearts and minds forever."
Wooten, D.B. 2000. "Qualitative Steps toward an Expanded Model of Anxiety in
Gift-Giving", Journal of Consumer Research, Vol. 27, No. 1, pp. 84-95.


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|Applying for your loan is easy!
Unlike some lenders, we have NO application fee, and all referred customers receive a FREE consultation with multiple quotes from the lowest-rate lenders we work with! Please select one of the following options that is convenient for you.
1. Apply On-line: Complete the online Application Form now. This is recommended if you are ready to get pre-approved to buy a home, or to refinance one you already own. The process takes approximately 10 to 15 minutes. You will have the opportunity to save your work and return to it later. Please complete the form as best you can, and skip questions to which you don't know the answers. We can fill in any missing details later!
2. Apply over the Phone: 724-774-9464 Please leave a message if we are busy assisting other customers, and we will call you back as soon as possible to gather your information via phone.
3. Download the loan application, complete it as best you can, then mail or fax.
Helpful Tips – Document Checklist
The following Document Checklist will help you in filling out your loan application , and preparing for the pre-approval and loan process if approved for financing!
_____ Most recent 30 days' worth of paycheck stubs (for all jobs you have)
____ Social Security, Pension, Disability Income Award Letters (if applicable)
_____ Copy of your 2 most recent federal tax returns, including all schedules and accompanying W2's, 1099's and K1's. **If you have not filed your 2 most recent tax returns, please let us know immediately**
_____ Last 2 month's checking and savings bank statements as well as any other assets used for down payment or reserves. ALL pages of your statements are required, even those intentionally left blank.
_____ Copy of any large NON-PAYROLL deposit items on your bank statements. All deposits need to be sourced. We will help you with these once statements are reviewed.
_____ Copy of current mortgage statements on any current properties you own, as well as the associated property's tax bill and home insurance premium cost info (if applicable).
____ Proof your earnest money check /wire cleared (once it does clear your bank). A copy of the check or wire as well as an updated transaction history or bank statement showing the debit.
____ Home insurance agent contact info (needed right after inspections complete, usually end of week 2)
____ Copy of your driver license and social security card, and DD214 copy for Veterans.
Other important info:
Please do not open any new credit during the financing process, and please do NOT change employers. Please also know it is very normal for us to ask for other supporting information or documentation from you once the above list is reviewed.


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Anne M. Get all the facts for a more clear viewpoint with Viktor Mayer-Schönberger. Schuller – opportunities by word of mouth and referral marketing remain largely unused Munich, 2nd June 2009 companies ignore the possibilities of Word of mouth and referral marketing and thereby miss valuable opportunities in the new customer business. 40 percent of all companies are engaged at all not about referral marketing. 49 per cent do so at most places. Only 11 percent operate referral marketing as defined and systematic process. This is the result of a representative survey of 300 senior executives of the German economy in the framework of the excellence barometer 2009. Anne M. Schuller, an expert in loyalty marketing, referred to these numbers as sobering.
We have currently not only with an economic crisis, but primarily a crisis of confidence to do it. Therefore companies need to just now show that they belong to the good. It was most credible, if not from the company is claimed, but testified of their thoroughly satisfied customers. Because consumers less and less trust the advertising of the service providers on the market. Rather, they rely increasingly on the messages of their environment or follow the corresponding instructions in the Internet. Online recently various possibilities to improve the recommendation business opens the company, so the eight books and bestselling author continues. The new recommendation marketing so linked the offline to the online world. Recommended who is recommended to shows but only the University lecturer.
An internal base should be designed first. Based on the referral marketing is then in a four-step management process systematic control, design and measure. In her book, future trend referral marketing consultant shows Schuller practical and many examples, as is the fact that translate into. Such an approach can largely replace advertising budgets and firmly to support sales activities. As the recommended client acts as a free seller. More Info:


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|Course Title
|Certificate in Abnormal Psychology (CAP)
|For whom (target group)
|Abnormal Psychology is a complex, controversial and a very fascinating field of study in applied psychology. The modern psychological community has made tremendous strides in better understanding, diagnosing and treating abnormal disorders of the mind compared to centuries and even decades ago. While there is still much stigma attached to this branch of mental illness, it is less so than in the past. This course will introduce you to the many faces of abnormal psychology. It will discuss old and new ideas surrounding this branch of psychology and will offer general information about how abnormal and clinical psychologists and psychiatrists identify and treat these types of mental illnesses.
Whether you are considering an education or a career in abnormal psychology or if you are simply interested in learning more about this fascinating field of study, this course will introduce you to a better understanding of the intricacies of the human mind and the ever evolving study of Abnormal Psychology and Psychopathology.
|Entry Requirements
|University Graduates or Members of recognized Professional bodies, or School Teachers or Assistant Teachers, or Professional / Academic Diploma or Advanced / Higher (National) Diploma holders or Holders of a Certificate / Diploma in Education, Psychology or Counselling, and Age not less than 19 years, and Fluency of the English language is essential.
|Course Duration
|2.5 Months – part time study
|Contact Hours
|3 Hrs. per week
|Total No. of Classes
|10 class-contact sessions (10 Weeks duration)
|Total No. of Contact Hrs.
|30 Hrs. of Lectures and discussion time
|Class Times
|Fridays: 9.00 a.m. to 12.00 noon
|Course Syllabus Outline
Mode of Final Examination and Assessment
The award of the IMH Certificate in Abnormal Psychology (CAP) will be contingent upon passing a 2.5 hour internally set unseen Essay type final Examination paper to be answered on a closed book examination mode under supervision at the IMH premises.
|Regular Group Course Fees
|Rs. 31,000/- + Rs. 1,000/- for registration (Could be paid by cash, credit card or cheque in two equal installments of Rs. 16,000/-)
|Individual Course Fees
|Rs. 63,000/- + Rs. 1,000/- for registration (Could be paid by cash, credit card or cheque in two equal installments of Rs. 32,000/-)
|Self-Study Course Fees
|Rs. 19,000/- + Rs. 1,000/- for registration
|Next Course Start Date
|Group Class: Friday 16th March 2018 (A maximum of only 10 students will be enrolled for the group class)
Individual Class: On a mutually agreeable timetable schedule – date and time (to be decided jointly by the participant and the resource person).
|Resource Person
|Dr. Sir Romesh Jayasinghe, FRSPH(UK) FTBCCT(UK) MBCS(UK)
Psychotherapist, Certified Master Life Coach (USA)
Certified Clinical Hypnotherapist (ICBCH, USA)


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Ujoy brings you Swords of Immortals, a blend of mystical magical world and mortal combat. Get ready to witness some action that will thrill your all levels of consciousness. This game is providing you with the best fighting environment with the intense and interesting story line. A lot of dungeons and real time battles are here for you. Go for 1v1 battles, play across the servers and quench the thirst of combat. Moreover, you don't have to play alone, there are chances to play along with your partner.
This is a full time action opportunity. Don't lag behind on this one. Use your sword to fly around the beautiful scenes and vivid landscapes of the mystical world. This is going to be the best times where action combines with story. Moreover, stop by the dungeons to secure the wealth and move from place to place looking for adventure. Furthermore, play in multiple combat modes and show the world your skills. Huashan Dual, Team Dungeon, Guild War, Faction Battle Field and 1v1. All of these modes will give you an opportunity to show case your wits and strategy.
In addition to these features, this game has an interesting story line with some awesome graphics. The aura of this game and the graphics will leave you dumbfounded. The magical effects of sounds and the combined game effects will soak your wits into the gaming experience. This would be the best shot you could ever have in the world of martial arts. A shot not worthy to be missed. Do you think you have what it takes to be the best fighter of the mystical world? Moreover, upgrade your tools and dungeons as you escalate through the journey off myth and adventure. Make some alliances to have them by your side. Play the game of loyalty by sticking to your honor. These friends will stand by your side at the time of chaos.
This game is available for both the Android & iOS devices and can be downloaded from their respective stores. However, if you wish to play Swords of Immortals on PC, you have landed on the right page. Follow the steps below to Download Swords of Immortals for PC on Windows 10, Windows 7, Windows 8.1 / 8, Windows XP & MAC OS. So, don't wait and barge in the world of combat with all your strength.
Download Swords of Immortals For PC Windows & Mac
- At first, download and install BlueStacks App Player or Rooted Bluestacks completely. You can also have a look on the guide: How To Install Android Apps for PC via BlueStacks 2.
- Once downloaded and setup, open BlueStacks.
- Now in BlueStacks click on the Search Button in the very first row on BlueStacks home screen.
- Now in the search box type "Swords of Immortals" and get the manager in Google Play Search.
- Click the app and install it.
- Once installed, find Swords of Immortals in all apps in BlueStacks, click to open it.
- Use your mouse's right button/click to use this application.
- Follow on-screen instructions to learn about Swords of Immortals and play it properly
- That's all.
In addition, you can also try the new Remix OS 2.0 or AndyOS to run Swords of Immortals for PC. Android Emulators for PC guides have already been drafted for your ease so do check them out. Also, do not forget to leave your queries in the comments box. That's all!