UTV INC ARCTIC CAT WILDCAT CLUTCH COOLING FAN KIT
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Clutch Cooling Fan Kit To Improve Clutch Air Flow
Cooler Belt Temps To Get Longer Life Out Of CVT Belt
Fits 2 and 4 Seat Arctic Cat Wildcat 1000 Models
Wired To Key On Ignition
Is your Wildcat’s belt getting too hot? Well ours is, so we have a clutch cooling fan kit to help ventilate the clutch system and keep it nice and cool for longer belt life. Comes with all parts needed to install for a keyed on/off fan. Will fit both the 2 and 4 seat models of the Arctic Cat Wildcat 1000 (Limited & X models).
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Total Hotel Score
Tiangeshan Road, Haitian Tourist Resort 276825, Rizhao
Wujiatai Village, Wolongshan Street, Shanhaitian Tourism & Vacation Area, Rizhao, Shandong 276500, Rizhao
Rizhao City No.33 Villa in Taigongdao Garden, Taigongdao 1st Rd, Donggang District 276826, Rizhao
Taigong one road 276800, Rizhao
* Distances are measured as a straight line, calculated from a starting point to a destination point
All bookings are confirmed instantly. No waiting, no email exchanges, no confusion.
Featuring diversified activities, Rizhao Haohao Hotel is situated along Bihai Road. Free WiFi and free parking service are available.
Port of Rizhiao is a 18-minute drive from Rizhao Haohao Hotel. Rizhao West Station is a 20-minute drive away. Guests can ask for surcharged shuttle service.
All rooms in Rizhao Haohao Hostel are equipped with a flat-screen TV. Guests can find free toiletries and slippers in their bathroom.
Rizhao Haohao Hotel can arrange diversified activities, including windsurfing, fishing and skiing . Free use of bicycles is available. The reception will offer tour guidance and free luggage storage.
Guests are required to show a valid government-issued ID card or passport upon check-in.
Toilet, Bath or Shower, Slippers, Shower, Free toiletries
Private entrance, Tile/Marble floor, Desk, Carpeted, Cleaning products, Heating, Dressing Room, Air conditioning, Wardrobe/Closet, Fan, Clothes rack
Flat-screen TV, Computer, TV
City view, Mountain view, Sea view
This twin room has a flat-screen TV and a balcony.
Slippers, Bath or Shower, Shower, Free toiletries, Toilet
Tile/Marble floor, Private entrance, Air conditioning, Cleaning products, Carpeted, Clothes rack, Desk, Wardrobe/Closet, Fan, Dressing Room, Heating
TV, Computer, Flat-screen TV
City view, Sea view, Mountain view
This double room features a balcony and a flat-screen TV.
After booking, all of the hotel’s details, including telephone and email address, are provided in the online booking confirmation, confirmation email.
Fishing, Windsurfing, Skiing
Shuttle service, Bikes available (free), Shuttle service (additional charge)
Luggage storage, Tour desk, Private check-in/check-out, Express check-in/check-out
Shared lounge/TV area
Daily maid service
Heating, Air conditioning
Free WiFi! WiFi is available in the hotel rooms and is free of charge.
Free parking! Free public parking is possible at a location nearby (reservation is not needed).
From 07:00 Until 23:00 hours
Until 12:00 hours
All children are welcome.
There is no capacity for extra beds in the room.
Free! Pets are allowed on request. No extra charges.
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domain: travel


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The sterling silver cuff bracelet has an oval bezel mounting onto which you can set an 19mm x 27mm oval glittering and colourful cabochon to artfully embellish the bracelet design. It is a popular choice for many jewelers, jewelry designers and artists to add beautiful and a true statement piece to their designs.
The whole top has an elegant curve adjustable to fit your wrist and bring sophistication to any look during the day or evening.
The bracelet isrhodium plated to ensure a lustrous, highly durable, tarnish resistant surface.
This bracelet has in inside diameter of 2.2″ (51mm) and an 19mm x 27mm oval bezel mounting.

domain: artistic


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Gift set – Non-alcoholic
Surprise with a personalised beer gift set with your own photo and text as a gift!
The non-alcoholic gift set is filled with 5 different kinds of non-alcoholic beer. Design your own print with your photo and text here! With a personalised design your beer gift set will be even more fun to enjoy with your friends and family, for a birthday or to bring out a toast to a special occasion!
Contents: Bavaria (300ml), Bavaria Radler (300ml), Amstel Radler (300ml), Palm green (250ml), Jupiler (250ml).

domain: hobby


Makers of applications that locate drunk-driving checkpoints are misunderstood, defenders said Wednesday, a day after four U.S. senators called for smartphone makers to pull applications from their services.
The applications do more than identify drunk-driving checkpoints set up by police, and the DUI (driving under the influence) checkpoint functionality actually aids police, said Joe Scott, CEO and founder of PhantomAlert, one of the companies targeted by the senators.
“They’re misjudging us,” Scott said Wednesday. “It’s a safety tool. It’s approved by a lot of police departments. How is that we’re being sanctioned? It just doesn’t make sense.”
When users of PhantomAlert report DUI checkpoints, it often appears to users that there are more checkpoints than actually exist, Scott said. The result is the app “deters people from drinking and driving,” he added. “We’re like a force multiplier for them.”
The Association for Competitive Technology (ACT), a Washington, D.C., trade group, also questioned the request from Senators Harry Reid of Nevada, Charles Schumer of New York, Frank Lautenberg of New Jersey and Tom Udall of New Mexico. The four Democrats sent a letter to smartphone software vendors Apple, Google, and Research In Motion Tuesday, asking them to stop selling DUI checkpoint apps.
The letter did’’t name the DUI checkpoint software, but Alert was one of several companies targeted, a Senate spokeswoman said. Similar software includes Cobra’s iRadar, Trapster, and Fuzz Alert.
On Wednesday, the senators reported that RIM would remove the DUI checkpoint apps. “Drunk drivers will soon have one less tool to evade law enforcement and endanger our friends and families,” the senators said in a statement. “We appreciate RIM’s immediate reply and urge the other smartphone makers to quickly follow suit.”
But the apps in question contain publicly available information provided by police departments as well as reports from drivers, said Morgan Reed, ACT’s executive director. The social-networking and law enforcement information makes the apps very popular, he said.
“While I applaud the senators for seeking to curb drunk driving, their criticism of online travel apps misses the point,” he said. “Law enforcement authorities have embraced these services, expressing their strong approval for products that reduce speeding and improve traffic safety.”
The apps provide drivers warnings about other potential roadway problems, Reed said. “Any one of the programs’ users can submit a warning about a traffic obstruction as simply as e-mailing a friend or posting a message on their Facebook profile,” he added. “The suggestion that the government should compel Apple, RIM, or other mobile application stores to block programs that simply allow users to report information based on location is misguided at best. Having the government act as arbiter of which products should be sold in stores is a slippery slope that few would welcome.”
Asked if the senators’ letter amounted to the government compelling action of the smartphone makers, ACT spokesman Jonathan Godfrey suggested it did. “When the Senate majority leader [Reid] and his senate colleagues send a letter urging you to do something, implicit is that they can have hearings and examine legislative fixes if you don’t address their concerns,” he said.
When police departments set up DUI checkpoints, they often advertise their plans, added Scott. “All we’re doing is we’re taking information and pushing it toward the drivers,” he said. “A lot of police departments are saying that part of the campaign is awareness. If PhantomAlert gets the word out … that’s going to deter a lot of people from drinking and driving.”
PhantomAlert, available since 2008, points out speed traps, red-light cameras, high-accident intersections, speed bumps and other traffic hazards.


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Boots responds to criticism on MUR targets
Record learning outcomes
Pharmacists working in Boots UK have been â€œremindedâ€ of company guidance around Medicine Use Reviews, following claims in The Guardian this week that the company is driving staff to achieve MUR targets largely for financial gain.
â€œOur professional standard is clear: services must be for the benefit of patients not the attainment of numerical targets and are fundamental in helping customers make the most of their medicines in managing their conditions effectively. We have today reminded our pharmacists of our existing guidance (previously issued in 2014) as we fully support their professional judgment to assess the appropriateness of a clinical service and these services,â€ said Boots UK, in a statement.
â€œOffering the best care to patients is at the heart of everything we do and this includes offering pharmacy services that are relevant to the changing needs of patients and healthcare systems. Our pharmacists are empowered to use their professional judgment to assess the appropriateness of a clinical service, and we make it clear to our colleagues that these services should not be undertaken inappropriately.â€
The company defended the role of community pharmacy in offering services such as MURs and itâ€™s own approach.
â€œProviding accessible healthcare through community pharmacies has always been integral to Boots mission. We believe community pharmacy today plays a vital role in supporting the UK health system and we actively support NHS initiatives such as medicines usage reviews.
The statement also said:
â€œWe donâ€™t recognise the claims in todayâ€™s press, which are not representative of the 60,000 colleagues who work for Boots UK and our commitment to developing healthcare further. The health and wellbeing of all our colleagues is, and always has been, a priority for the business.â€â€¨
â€œOver the last ten years, our group has continued to significantly re-invest profit right across Boots UK. We are absolutely clear that the drive for strong financial performance has never been to the detriment of our constant priority on pharmacy, and delivering the best healthcare services in the communities we serve.â€
Boots dispenses more than 220 million prescriptions every year in the UK, said the company. â€œWe strongly believe that community pharmacy today, more than ever, plays a vital role in supporting the UK health system.â€

domain: health


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My husband and I were impressed with the food and service here. The decor is lovely and it feels very cozy inside.
The elk loin and peppered steak were prepared very well. The Sundance salad had the perfect amount of dressing, and the beet starter was cool and refreshing.
The berries and custard were a perfect dessert, the beignets were a little dense for my taste.
Great choice for a special night out.
Own or manage this property? Claim your listing for free to respond to reviews, update your profile and much more.

domain: food


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Discontent is simmering among even voters of the extreme right for Kateel's alleged non-performance during the last two terms.
However, there is no palpable excitement either in the opposite Congress camp which has fielded a young candidate in Mithun Rai, belonging to the same Bunt community as Nalins and who toes a soft Hindutva line in his campaign.
The reason is simple.
Exemplifying the voter mood was the unprecedented massive response to Modi during his campaign here Saturday that saw the entire 15-km stretch from the Mangaluru airport to the public meeting venue overflowing with delirious supporters.
Taking note of people perched on treetops to listen to him, Modi was even worried about their safety.
"I will be hurt if anything happens to you," a concerned Modi told them.
While on his return to the airport too, thousands of people lined up the streets, forcing slow movement of the convoy. Modi, on his part, did not disappoint them as he stood in his vehicle and greeted them.
In the constituency sharply divided on communal lines, Congress had been able to keep it as their fortress till 1991.
But ever since the emergence of BJP as a national player and "protector of Hindutva", it managed to snatch the seat from Congress in 1991 and since then never had to look back.
We all know that Kateel has done very little for the constituency during the last two terms. But Hindus will forget their disenchantment with him as they want Modi back as Prime Minister, who they think, is the man to protect the nation, its traditions and ideals, said K Ramesh, an advocate from Sullia.
BJP has succeeded in projecting national security as the key issue in the elections in Dakshina Kannada.
A good number of people, especially from the business community, are enamoured by the Prime Minister's leadership image, the trump card of BJP.
The common people and the lower middle class, though hit hard by demonetisation and GST, are also often swayed by communal emotions stirred up by the strong right wing outfits here.
Besides the RSS, right wing outfits like Hindu Jagarana Vedike and Sri Ram Sene have a strong presence here.
The activists of such groups often resort to moral policing and self-styled fight for cow protection, fanning communal tensions.
The RSS leaders, who had earlier dropped hints that the incompetent Kateel was no longer acceptable to them, changed their stance once it was clear he was being re-nominated.
Sometimes, people have to tolerate even incapable people, keeping in mind the interests of the nation, a senior leader said.
For the Congress, which has gambled on the young Mithun Rai, an upset win is the only thing to hope for.
Various local issues, including incomplete flyovers, excessive collection from toll gates and the merger of Vijaya Bank, which originated in the district, with Bank of Baroda that directly affect the people, are being taken up in their campaign.
Congress leaders in the district, many of whom were aspirants for the seat, are exhibiting a rare unity in their campaign for Rai as he has been handpicked by AICC president Rahul Gandhi to give representation to youth in the state list.
Rai is seen in his campaign wearing saffron shawls and chanting Hanuman chalisa at meetings.
Justifying Rais overtures to the majority community saying he has been chanting prayers right from his childhood, Congress leaders feel that with a boy-next-door image, he is likely to get a chunk of votes from the youth.
Rai has another advantage with open support of the Left and farmers groups.
Ravi Kiran Punacha, state secretary of the Karnataka Rajya Raitha Sangha, who also offered support to Mithun Rai, charged that the plight of farmers and common man will be more miserable if Modi comes back to power.
With only three days left for polling, political watchers are waiting with fingers crossed on the outcome.
BJP is expecting a smooth sail on the Modi factor while the Congress hoping for a miracle.
(This story has not been edited by Business Standard staff and is auto-generated from a syndicated feed.)

domain: sociology


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Poetry Ireland connects poetry and people. In October 2019 Poetry Ireland launched their inaugural Poet in Residence with poet Catherine Ann Cullen, the residency was completed in October 2021. As part of her residency, Catherine Ann ran numerous projects, both online and offline, to engage people and communities in poetry. Piquant worked with Poetry Ireland and Catherine Ann to produce a video that captured the legacy of the Poet in Residence. Letting Poetry In was the section winner in the Human Spirit category at the 2022 International Docs Without Boarders Film Festival. The film was also selected for screening at the 2023 Berlin Short Film Festival.
What we did:
- Video Production
- Concept Development
- Script Writing
- Camera Operation
- Video Editing
- Motion Graphics Design
- Voice Over Recording
CONCEPT DEVELOPMENT — Letting Poetry In is a video that celebrates the legacy of Poetry Ireland’s Poet in Residence Catherine Ann Cullen. The video combines interviews, poetry and documentary footage to showcase the work and impact of the residence period. It was clear that this story needed time and space to breath; as such we produced a 5 minute lyrical documentary that creates space for the poetry, and showcases the transformational impact of the residency on the people who engaged in the projects. A carefully crafted soundscape including the diegetic sounds of the city brings a deeper connection to the poetry, the people and their stories.
SCRIPT WRITING — Creating the script of this video, we knew that it was a demanding story to tell. Catherine Ann Cullen touched the lives of so many different groups and individuals and they needed a place in the video. Scripting thought provoking questions gave the video participants the opportunity to share their experiences and ideas. The structure of this video intertwines poems read to-camera throughout the narrative to encourage the viewer to relax and enjoy the craftsmanship of poetry.
VIDEO PRODUCTION — The Piquant Team travelled to Poetry Ireland HQ in Dublin’s north inner city to conduct interviews with Catherine Ann Cullen and others to give a full perspective of the impact she left. These interviews act as the foundations of the video with documentary footage captured at the Poetry Ireland heritage building and Dublin street scenes to bring a deeper impact to the story we are telling. The amazing talent of the poets was captured by our team as we recorded an A cappella reading of their poems. This brings poignancy to the video footage captured and allows the viewer to feel the power to the words in motion.
MOTION GRAPHICS — Due to the Covid 19 pandemic which erupted in the middle of the residency Catherine Ann moved a lot of her projects online. We wanted to capture how people engaged with the projects on social media platforms and so used motion graphics to visually showcase the poetry created in reaction to Catherine Ann’s prompts. Piquant’s video production and motion design teams worked collaboratively to produce the on-brand motion graphics that work to build the narrative, and communicate the impact of the poetry projects using social media during Covid lockdowns.


Opinion: Can George Bush recover his authority in time for history's verdict?
Friday, 17 March 2006
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BUSH, George W.
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To welcome someone into your home is to show them what you think’s important. Remember the first friend-date you had as an adult? For mine, I was staging my coffee table with smart books and a candle (I should’ve dusted the dust jackets first for maximum credibility).
Now, you know you’re my friend if you walk into the space now occupied by Lego, library books, and eleven hundred little scraps of paper Louisa tells me are “bookmarks.” (I love you and I can’t keep up with my many, many children, is what my living room says.)
But to welcome someone to your hometown is to show them something deeper, something maybe mitochondrial. Ah yes, I’ve said when I see friends’ hometowns. I can see this place in you.
So, that’s happening next week at a book event in the place where I graduated, got married, and flee to when I need my mom.

domain: literature


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Upromise is a free rebating program that helps families save money for college. Upromise members receive rebates from participating companies ranging from 1/3% to 10%. The rebates can be deposited directly into section 529 plans set up for the member's designated children.
Participating companies include more than 15,000 grocery and drug stores, 8,500 retail stores, 7,000 restaurants, 100 online stores, and 50,000 realtors. Major companies participating in Upromise include AT&T, America Online, American AAdvantage, Avis, Borders Books & Music, CVS, Century 21, Circuit City, Citi, Coca-Cola, Coldwell Banker, Exxon/Mobil, General Motors, Giant Eagle, McDonald's, Sharper Image, Staples, Starwood Hotels and Toys R Us.
If you register your credit cards, you may find that many of the retailers you already patronize participate in Upromise. The Upromise network is very large.
Other ways to earn rebates include online shopping through the Upromise web site, the Bank of America Upromise credit card (gives you an additional 1% rebate on eligible purchases) and special Upromise programs. The Upromise credit card does not charge an annual fee. (However, if you use this credit card with Quicken, it only downloads transactions once a month, unlike the daily downloads available with other credit cards. If you want more frequent downloads they charge a monthly fee.)
You can automatically have your Upromise balance invested in a section 529 plan. You can also transfer it as a gift to another Upromise member, donate it to the Upromise Education Foundation, or redeem it as cash. Your friends and relatives can become Upromise members even if they don't have children, and designate your children's section 529 plans to receive their Upromise rebates.
Upromise agreed to be acquired by Sallie Mae in June 2006. Starting in June 2007, Upromise reward balances may be applied quarterly to reduce the loan balance on loans serviced by Sallie Mae.
|Home | Loans | Scholarships | Savings | Military Aid | Other Types of Aid | Financial Aid Applications
Answering Your Questions | Calculators | Beyond Financial Aid | Site Map | About FinAid®
|Copyright © 2016 by FinAid Page, LLC. All rights reserved.
Mark Kantrowitz, Founder

domain: economics


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Taking its name (and theme tune by Rocky Roberts and Luis Bacalov) from Sergio Corbucci’s influential 1966 spaghetti western Django, Tarantino’s new film sees this most reflexive of American auteurs turn his hand to the Western.
Django (Foxx) is a slave freed from leg irons by Dr King Schultz (Waltz) a German dentist turned bounty hunter. Schultz is seeking some outlaws but has no idea what they look like, but has heard of a slave who could identify the men. Which leads him to track down the slave traders who recently bought the unfortunate Django. Django has been separated from his wife after they tried to flee a plantation, and Schultz makes him a deal, partner with him in the bounty hunting business and the romantic German will help him track down his wife once spring comes and the snows thaw. Of course, it soon emerges that out of irons Django is a natural with a different type of metal, lead.
As the reference to the Corbucci’s film indicates the primary source of cinematic inspiration for Django Unchained is not the classic westerns of John Ford, Howard Hawkes or even the revisionism of Sam Peckinpah, it is the European emulations and mutations of this most American of genres. Italian westerns had a complex relationship to American genre, and to the history and symbolism of America itself. While American westerns up to the 1960s could still celebrate the notion of manifest destiny with a straight face the more cynical and left leaning Italian writers and directors brought eyes altogether more jaundiced to the format. The West portrayed in the spaghetti westerns was violent, unjust and founded on motivations of greed and revenge. Heroes became gave way to anti-heroes.
Tarantino’s film is everything we have come to expect from him. It is sprawling, episodic and somewhat baggy in its story and structure. This is perfectly in keeping with the western genre and in fact the plot is uncharacteristically linear for Tarantino, sticking to one story and one central character and proceeding from A to B (with a few flashbacks to fill in the backstory). There is plenty of talk and humour (often carrying a whiff of the gallows) and the film dishes out short, sharp shocks of violence. But there is something here that Tarantino has arguably not had in one of his films since 1997’s Jackie Brown: soul. Just as many of the European westerns Tarantino references tried to skewer American history, Tarantino has in his sights a dirty great elephant squatting in the room. That would be the shame of slavery. The film very quickly and brutally dispenses with the traditional romanticised Johnny Reb western bullshit that glorifies the south, something that even featured in that most libertarian of 70s westerns, Eastwood’s The Outlaw Josey Wales. Tarantino holds the glory of the west and the spread of white culture at the expense of other races in contempt. The world of Django Unchained is one of plantations, slave owners, idiot clansmen, oppression and brutality. This is a Tarantino film to be sure, but one that is about something.
However, don’t get the wrong idea; this isn’t Lincoln, this is an exploitation movie through and through. Tarantino paints the screen red with sudden outburst of cartoonish violence. The tone is ironic and comedic (the film was nominated in the drama section of the Golden Globes, despite being funnier than all of the films in the musical or comedy category). But at unexpected and shocking moments the film will drop the jocularity and ironic distance for sudden moments of horrible racism and violence of a less Tex Avery nature. Two sequences in particular stood out for me as genuinely horrible, a brutal "Mandingo" fight in which two slaves are forced to fight to the death like dogs for the amusement of their white "masters", and the terrible fate of a runaway slave later in the film. Depressingly, Tarantino has again been dragged into the ever recurring debate about screen violence inspiring real violence in the wake of recent tragedies. But in these scenes he shows again how deft a director he is in his use of suggestion. It’s not what you see, it’s what you think you see.
Performances and casting approach perfection in this picture. The leads are terrific – Foxx is stoic and simmers with righteous rage, while Waltz is superb as a cultured killer of warmth and humanity. The relationship of Django and Schultz make this also a great buddy film. In the film’s second half the pair track Django’s wife (Washington) to the plantation of Calvin Candie, a vile slave owner played with disturbing relish by DiCaprio. DiCaprio is despicable, but perhaps the most evil character in the film is his black butler Stephen, played by Jackson in truly intimidating form. Stephen is the ultimate Uncle Tom, almost more devoted to the principles of slavery than Candie is. It is a chilling performance.
Technically the film looks great as shot by veteran cinematographer Robert Richardson. Tarantino die hards will feel a pang of pain that this is the first of his films not to be edited by the late Sally Menke, but Fred Raskin (who was an assistant editor on Kill Bill) does a great job. The soundtrack is another romp through the director’s record collection and mixes scores from spaghetti westerns, soul music, country and hip hop. Some have complained about this, but it is no more anachronistic that the music used by Corbucci or Leone on their westerns, and it works goddamit.
This is a blazing, full bore return to form from Tarantino. The director has, in my opinion, drifted somewhat since Kill Bill: Part 1. I didn’t much care for Kill Bill: Part 2, disliked slasher/car chase hybrid Death Proof immensely, and whilst Inglourious Basterds contains some of the best work Tarantino has ever done, it was also undisciplined and subject to frequent longueurs. Time and further viewings are needed to form a full opinion, but my gut feeling is that Django Unchained may be as good as Pulp Fiction. Whatever, this savage, brutal, hilarious film is almost indecently entertaining.
EXTRAS ★★ For such a magnificent film, the bonus material is rather sparse: the featurette Reimagining The Spaghetti Western: The Horses & Stunts of Django Unchained (13:51); the featurette The Costume Designs of Sharen Davis (12:07); the featurette Remembering J Michael Riva: The Production Design of Django Unchained (12:54); the Tarantino XX Blu-ray Collection Promo (1:25), which is a trailer for a 10-disc box set; and the Django Unchained Soundtrack Promo (00:22), which is an ad for the soundtrack. No audio commentary, no deleted scenes. Pretty poor, really.


AMO Supports Traineeship Program to Prepare the Next Generation of Manufacturing Experts
Today, the Office of Energy Efficiency and Renewable Energy’s (EERE) Advanced Manufacturing Office (AMO) announced $2.5 million in funding to support university-led traineeship programs that address workforce training needs in the early-stage technology area of advanced materials and process technologies in energy-related manufacturing. EERE will competitively select U.S.-based institutions of higher education to implement masters-level training programs designed to train a new generation of advanced manufacturing engineers to fill workforce needs across industry, national labs, and universities.
These traineeship programs will focus on advancing critical STEM disciplines and competencies specifically relevant to the AMO mission where other U.S. government or academic workforce development programs either do not exist or where DOE-relevant, early-stage technology areas are not being leveraged to support the DOE mission.
With new knowledge and skills in advanced materials and process technologies for manufacturing, the students in this traineeship program will have the potential to advance new technology research and development leading to meaningful impacts across the energy and manufacturing sectors.
This new generation of advanced manufacturing technical experts will need to be trained if the private sector is going to translate early-stage research and development research to U.S. leadership in energy and advanced manufacturing. This investment in the future workforce, informed by the most relevant early-stage research and development, will enable the growth of the advanced manufacturing field and bolster U.S.-based manufacturing and innovation.
EERE will competitively select one or more U.S. institutions of higher education to implement masters-level training programs. Selected institutions will receive up to $2.5 million to establish programs in which students gain hands-on, industry-related experience as well as cross-cutting technical skills development of importance to advanced manufacturing in areas including data analysis, information systems, modeling and simulation, and process design. Concept paper applications are due August 8, 2017.
View the full solicitation and submission instructions.
EERE's Advanced Manufacturing Office supports early-stage applied research and development of new materials, information, and processes that improve American manufacturing’s energy efficiency, as well as platform technologies for manufacturing energy-related products. This technology innovation in advanced manufacturing is a foundation for economic growth and jobs in the U.S.


The Government must set out a clear vision for what will replace the Green Deal following an announcement that the scheme has effectively been scrapped, according to the Federation of Master Builders (FMB).
Brian Berry, Chief Executive of the FMB, said: “The Government’s announcement that it will provide no further funding to the Green Deal Finance Company and will also stop any future funding of the Green Deal Home Improvement Fund is the final nail in the Green Deal’s coffin. The Green Deal was the greatest flop of the last Parliament – it failed spectacularly in its mission to incentivise millions of house holders to improve the energy efficiency of their homes. However, the Government would have been wise to reform, rebrand and relaunch the Green Deal rather than scrap it altogether.
“What’s clear is that the need to improve the energy efficiency of our properties is an increasingly pressing priority but the Government is showing very little leadership or ambition. The goal of insulating a million more homes over the next five years is a meagre target when you consider that around five million homes were provided with energy efficiency improvements through various schemes over the past five years.
“The Government and industry should not be defeated by the poor results driven through the Green Deal and instead we must learn the lessons and move forward. There are lots of measures which could kick start activity – the Government should provide home owners with zero interest loans to make their properties energy efficient. This move could generate more tax and has worked in Germany.”


text:
Tuesday, September 5, 2006
News staff writer
August 21, 2006
Highway 35 between White River Sno-Park and Cooper Spur Road will close at noon Tuesday, Aug. 22 for up to 48 hours to allow for burn-out operations close to the highway and accommodate low-level air support in the continuing effort to control the Bluegrass fire on Mount Hood.
During times when the highway is open the following restrictions continue until further notice: Highway 35 is closed from 7 a.m. to 9 a.m. The closure will allow the Forest Service to move equipment and personnel. Between 9 a.m. to 9 p.m. pilot cars and follow vehicles will control both directions of traffic. From 9 p.m. until 7 a.m. no vehicles will be allowed to stop or park at any trailheads between Cooper Spur Road and the junction of Forest Road 48.
During closure periods, motorists should use Interstate 84, U.S. Highway 26 or U.S. Highway 97 (commercial trucks should use 97) as alternate routes.
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Lawnmower torches Arbor Vitae on Portland Drive
The riding lawn mower driven by Norma Cannon overheated and made contact with dry arbor vitae owned by Lee and Norma Curtis, sending more than a dozen of the tightly-packed trees up in flames. The mower, visible at far right, was totaled. No one was injured; neighbors first kept the fire at bay with garden hoses and Westside and Hood River Fire Departments responded and doused the fire before it reached any structures. Westside Fire chief Jim Trammell, in blue shirt, directs firefighters. The video was taken by Capt. Dave Smith of Hood River Fire Department. Enlarge

domain: news


I thought I would take a moment to respond to Monique Beyerle’s distorted reporting (letter) on trapping in New Mexico in the Feb. 24 edition of the Los Alamos Daily Post.
I would encourage everyone to get online and read the New Mexico Dept. of Game and Fish’s five pages of regulations pertaining to trapping of wildlife in NM that is included in the current years hunting rules and info. Trapping is highly regulated – much like hunting or fishing. I certainly support the notion that everyone is entitled to their opinion – but stating that trapping endangers citizens and their children is a HUGE distortion of trapping fur bearers.
The traps used will not maim or cause irreparable harm (amputation?) to hikers or those simply enjoying public lands in NM. Clear distortions and accusations like this are simply propaganda intended to mislead people in supporting a ban on trapping on public lands in NM. This is just another attack on the rights of men and women who hunt, fish and trap on public lands throughout the State.
Wildlife policy should be administered by professionals and not politicized by conservation groups who seek to limit use of public lands to activities that they deem “appropriate.” The rights of hunters and trappers to use public lands have come under increasing pressure from “Anti” groups for decades. The anti-trapping movement is just one more ploy to decrease the access to public lands and individual rights which are guaranteed to all citizens.
I would encourage readers to do independent research on the issue and they will find that trapping does not adversely affect wildlife populations. In reality, some trapping of predators may allow deer, elk and bighorn sheep to flourish or re-establish populations in new areas. Readers should also keep in mind that the resurgence of CONSERVATION of wildlife in North America was facilitated by HUNTERS and the fees they paid to engage in time honored pursuits. Check it out … read about the Dingell-Johnson and Pittman-Robertson Acts and their impact on wildlife in the U.S.
In the meantime, let’s leave wildlife management to the professionals who have been trained to manage our collective resources. Leaving wildlife management to the ballot-box and funded advertisements to politicize the issues is a clear disservice to our environment and individual rights.


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Is the OG 'Fortnite' Map Coming Back? Here's What We Know So Far
Many longtime fans have wondered if the OG 'Fortnite' map would ever be coming back to the battle royale game. As it turns out, it might. Here's what we know.
What was just rumors in 2021 is shaping up as a possible reality now: OG Fortnite may finally return due to the Creative 2.0 update that just released. With Creative 2.0, teams of creators could try their hand at recreating the Chapter 1 Fortnite map with the new tools available.
It wouldn't be the first time third-party creators made the original Fortnite map; however, now it's possible through official means on the Unreal Engine Editor and shareable between the community. Here's a breakdown of everything we know so far about the OG Fortnite map coming back.
Is OG 'Fortnite' coming back? Yes, but it's still debatable how it'll hold up.
One of the best creator teams, Atlas, revealed they were working on making the OG Fortnite map shortly after the Creative 2.0 announcement from Epic Games.
The project doesn't have an official release date yet, but the small teaser posted on Twitter from Atlas says the fan-created map is "coming soon." The team also created a separate Twitter account for the OG map to share progress and announcements for the project.
As expected, other creators willing to recreate the original with its iconic points of interest and lootable weapons will join the fray, so it's unclear which imitation will stand out most. "2000 other people will do this, but who will do it the best," said one Twitter user under the Atlas announcement post.
Whichever recreation holds up over time will likely be the defacto version that everyone plays, but as things stand right now, the Atlas project seems like a clear winner already.
The Atlas creator team posts an image of the OG 'Fortnite' map on Twitter.
The creator team dropped an early bird's eye view of the original map on March 17 via Twitter to show their progress. Locations such as Tilted Towers, Wailing Woods, and Retail Row appeared in their original spots all those years ago.
Since then, the Atlas page has posted a small survey on whether players would like a classic Loot Pool or a mixed version and promotional marketing for the upcoming map. Details on the release date are still under wraps, but players could expect a release soon since the creator team got hands-on time with Creative 2.0 earlier than others.
You can follow their Discord page to stay in the loop on new information about the project.

domain: game


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Katherine Langford Nude Pics and Videos
32 Super Sensual and Sexy Pictures of Katherine Langford
Katherine Langford Hot Sexy Bikini Pictures Are A Charm For Her Fans
She had attended different acting classes and workshops accordingly she juggled three part-time jobs.
Or should we say cums back? Katherine appeared in the play named Hotel Sorrento sponsored by her school.

domain: entertainment


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An overwhelming majority of justices of the Supreme Court (SC) are expected to concur with the draft ponencia of Associate Justice Lucas Bersamin that declares the Disbursement Acceleration Program (DAP) as unconstitutional.
The Manila Times’ source in the tribunal said Bersamin’s 95-page ponencia raised four points on why DAP was illegal.
“Ang ganda ng pagkakasulat ni Luke [Bersamin’s nickname] mahirap kontrahin. Eh magaling naman talaga si Luke. Kaya sure na unconstitutional ang DAP [The draft decision is well written, it would be hard to question it. Bersamin is really good. It’s sure that DAP will be ruled as unconstitutional],” the source said.
The source added that Bersamin’s ponencia declares DBM Circular 541 unconstitutional and orders the issuance of a “prohibitive injunction” against the circular from the Department of Budget and Management.
The circular, issued by Budget Secretary Florencio “Butch” Abad on July 18, 2012, allowed the DBM to withdraw and pool the unobligated allotments of agencies “with low levels of obligations for continuing and current allotments.” These funds were then used for other priority programs and projects.
Several groups and individuals, including the Integrated Bar of the Philippines; Philippine Constitutional Association; Volunteers Against Crime and Corruption; Carol Araullo; Pork Barrel System Petitioners led by Greco Belgica, Reuben Abante and Jose Gonzales; former Iloilo Rep. Augusto Syjuco; and Citizens Crime Watch as represented by lawyers Jose Malvar Villegas Jr. and Manuelito Luna, filed petitions questioning the legality of the DAP at the Supreme Court.
The respondents are President Benigno Aquino 3rd and Abad.
Malacañang cited poverty alleviation as a major reason behind its creation of the program.
The SC en banc is expected to vote on Tuesday, July 1, on whether DAP is constitutional.
In his ponencia, Bersamin raised four grounds on why DAP is unconstitutional—cross border tossing of funds, use of funds for unlisted projects, implementing new projects even if concerned agencies had no savings to fund such projects and using DAP funds for projects that did not undergo bidding.
The magistrate said the SC “ought to halt the cross border tossing of funds,” referring to the transfer of government funds from one branch to another. The justice, in particular, pointed out the illegal transfer of funds by the executive department to the Commission on Audit and the Commission on Elections and the legislative branch.
This “cross-border” scheme was first employed by Malacanang at the height of the impeachment trial of former Chief Justice Renato Corona. The Times source said most of the justices are of the opinion that the scheme violates the doctrine of separation of powers.
Although the Constitution allows the President to use savings under his “fiscal autonomy”, the magistrates are of the opinion that he cannot transfer savings to other branches of the government.
The SC magistrates also pointed out that projects that are not listed under the General Appropriations Act (GAA) must be declared illegal and unconstitutional. They said the GAA has the force and effect of a law since it is passed by Congress and signed by the President of the Philippines. As such, the SC justices see that a prohibitive injunction must be issued in funding projects that are unlisted under the GAA.
Some justices also believe that it is unconstitutional to fund projects if there are no savings to begin with for these projects to get started, as well as bankrolling with DAP funds projects that are awarded despite the absence of public bidding.
It was Sen. Jose “Jinggoy” Estrada who exposed DAP when, in a privileged speech, he claimed that the senators were bribed by as much as P50 million each to convict Corona. He said the money given to the senators came from DAP.


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The mining wastes have different physical and chemical properties, which bring on various potential environmental influence.Different types of mining wastes are generated from according to the type of deposit and the technics used for mining and for ore processing or quarrying process. Mining wastes is the coarse or fine residues generated from the quarrying and processing of crushed-rock Aggregates. Sometimes, with proper disposal, these waste are saleable products, they have economic value. These recycled wastes could be used for site restoration, landscaping, construction etc. here are some common types of mining wastes.
Waste rock is unused extraction products generally stored for long period in a landfill site nearby the mining center, for saving transport costs. Generally, opencast pits and quarries generate much more mining waste than an underground mine. The main type of waste rock is generated by surface (or barren rock) stripping to expose the shallow ore. these barren rocks are generated by the passages (shafts, crosscuts). There are huge volume of waste rock are generated from mining and quarrying activities everyday.
Tailings (processing waste)
At a mine, an ore mill normally abuts on the extraction centre to produce the first marketable products (metallic concentrates, sorted ore, and ingots). The technological processes are very different according to the ore properties and various technics (flotation, leaching, and biotechnology). These units produce various types of waste, which can include:
- aqueous solutions from cyaniding,
- slurries of finely ground particles that have undergone one or more types of physical or chemical treatment, and which frequently contain one or more industrial additives that have participated in the conversion process (xanthates, miscellaneous salts, starch, etc.). These tailings are normally dumped in a sort of lagoon or settling basin within an embankment at the exit of the mill;
- in some case, atmospheric releases from sulphide roasting. Emissions to air would result in “environmental impact”.
Mill waste is generally referred to as tailings, or releases or effluents. It is generated by the various mining processes to meet final application demand. Waste is generated at all levels of the recovery process to upgrade the minerals, within the same process chain, and is considered as ultimate or stripped of useful elements.
Intermediate storage of products, ore stockpiles are not waste and are normally temporary dumps of lean ore at the mine site, depending on the cut-off content, which may vary with time. This type of mine project management is included in the overall mining plan. This management requires maintenance on the mine site for a period sometimes longer than a decade. Selectivity materials correspond to ores of lower grade than the limit assay. These ores have a content which, while lower than the limit content when stored, can be handled later without loss in the processing plant when it is not be saturated, in which case it is treated as lean ore.
In nature, the ores of metals such as iron, copper, lead, aluminium, and other metals are found in impure states, often oxidized and mixed in with silicates of other metals. During smelting, when the ore is exposed to high temperatures, these impurities are separated from the molten metal and can be removed. The collection of compounds that is removed is the slag. Slags are produced in many metallurgical operations throughout the world and are considered a by-product of the production of iron and steel.

domain: chemistry


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Michael McKean is an American actor, and director who is best known for My Little Pony: A New Generation (2021), Playing God (2021), This Is Spinal Tap (1984), A Mighty Wind (2003), and Better Call Saul (2015). Born on October 17, 1947, Michael hails from New York City, United States. As of 2021, Michael McKean‘s age is 74 years. Check below for more deets about Michael McKean. This page will put a light upon the Michael McKean bio, wiki, age, birthday, family details, affairs, controversies, height, weight, caste, rumors, lesser-known facts, and more.
|Stage Name||Michael McKean|
|Real Name||Michael McKean|
|Birthday||October 17, 1947|
|Age||74 years (As of 2021)|
|Birthplace||New York City, United States|
|Hometown||New York City, New York, United States|
Education & Qualification
Age and Birthday
Michael McKean was born on October 17, 1947 in New York City, United States. Accordingly, he celebrates his birthday on October 17 of every year. You can wish him a happy birthday on October 17 of every year.
Michael McKean‘ father’s name is and his mother’s name is .
- Michael McKean was born in New York City, United States, as Michael John McKean.
- He is an actor, and director, best known for My Little Pony: A New Generation (2021), Playing God (2021), This Is Spinal Tap (1984), A Mighty Wind (2003), and Better Call Saul (2015).
- Michael has played the role Richard of ‘Pink Skies Ahead’, Roy of ‘Helpsters’, and Jack of ‘Grace and Frankie’.
- He has nominated for several awards, including Academy Awards, USA for Best Music, Original Song in A Mighty Wind (2003), and Academy of Science Fiction, Fantasy & Horror Films, USA for Best Supporting Actor on Television in Better Call Saul (2015).
- Michael has been married to Annette O’Toole, on 20 March 1999.
- He married to Susan Russell, on 10 October 1970, and they got separated in 1993, they have two children’s.
- Likewise, he could be found on Instagram under the username @mjmckean.
Google Search Trends
Michael McKean‘s search trend from the last 12 months (The below graph report is directly fetched from the ‘Google Trends’):
- Does Michael McKean know cooking? – N/A.
- Does Michael McKean smoke? – N/A.
- Does Michael McKean drink alcohol? – N/A.
- Does Michael McKean go to the gym? – N/A.
- Michael McKean’ hobbies are reading, photography, learning, traveling, internet surfing.
- Michael McKean’ father name is .
- Michael McKean’ mother name is .
- Michael McKean’ hometown is New York City, United States.
- Michael McKean’ birthday date is October 17, 1947.
- As of 2021, Michael McKean’ age is 74 years.
- Most searched terms about Michael McKean on Google and Bing are Michael McKean marriage, Michael McKean age, Michael McKean wiki, Michael McKean photos, Michael McKean lover, Michael McKean instagram, Michael McKean facebook, Michael McKean family, Michael McKean salary, Michael McKean height, Michael McKean bio, Michael McKean income, Michael McKean house, Michael McKean latest news, Michael McKean tiktok, Michael McKean musicaly.
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@pete-s said in Raspberry Pi 4 as IT Workstation:
But that means that you can run the lighter 32bit OS version that is faster AND it is already overclocked and includes a massive passive heatsink.
But the web browser plus whatever else is running in addition to the OS will still eat up what's left of the RAM pretty easily I would think. My phone uses more than 4g of ram easily.
Not really. It's actually hard to use up the 4GB RAM on the RPI4 as a normal user. Remember that everything running on it is lean, like LXDE for example.
I'd say that if you're happy with the RPI4 performance as a desktop then there's a 95% chance that 4GB RAM will be enough.
If you're a power user then RPI4 is too slow and you'll probably looking at something with at least 16GB RAM anyway.
This is why I like the RP400. It's the CPU, more than the RAM, that is the bottleneck for use and the RP400 is like 15% faster.


21 Nov ATV & UTV ESCAPE Magazine Nov/Dec 2018 Issue LIVE!
ATV & UTV ESCAPE Magazine November/December 2018 issue now LIVE!
The November/December 2018 issue of ATV & UTV ESCAPE Magazine is now live! Check it out on the side for a host of great articles from the off-road world, including:
- 2019 Can-Am Maverick Sport review
- 2019 Yamaha YXZ1000R SS review – part 1
- Guide to preparing for High Altitude Riding
- Buyer’s Guide: Holiday goodies!
- Is Honda coming out with a Sport SXS?!
- First Impression of the Nikola Electric UTV
- Behind The Helmet with Rick Sosebee!
- and of course the ever popular reader’s rides 🙂
All of this and more can be found in this bi-monthly magazine. We hope you enjoy the work from all of us at TGH and the ATV ESCAPE networks. It’s one of our favorite magazines ever!
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GungHo Online Entertainment, Inc. develops and operates " Ninjara, " a competitive Ninjagam action game for Nintendo Switch.
The game, which has surpassed 7 million downloads worldwide, is a light and dynamic action game that is free to play and easy to play.
On June 25, 2021 (Friday), "Ninjara" celebrated its first anniversary! Congratulations!
In the "Ninjara 1st Anniversary Commemorative Broadcast" broadcasted to commemorate the first anniversary, various new information was announced under the title of "Ninjara 1st Anniversary Project"! According to the preliminary announcement, "We are planning to announce information on a major collaboration to commemorate the first anniversary." ...... finally revealed the details!
The collaboration between "Ninjara" and the popular anime "Demon Slayer" will take place this summer!
Ninjara" x "Demon Slayer" Collaboration Information Summary!
As part of the "Ninjara 1st Anniversary Project", an in-game collaboration with the "Demon Slayer" anime has been announced!
The collaboration will take place in the month of lacquer ......, which means that it will start in July!
Demon Slayer" collaboration period
July 20, 2021 (Tue) 11:00 - August 31, 2021 (Tue) 10:59
Take on the form of Sumijiro and travel around the metropolis!
This collaboration will feature a number of avatar items based on characters from "Demon Slayer"!
Some of the character coordinates were previewed on the "Ninjara 1st Anniversary Broadcast"!
Character coords based on the motifs of "Tanjiro Kamado", "Priestess Kamado", "Anzujiro," "Zenitsu Wagatsuma" and other Onigakurei members will appear in the game! Character coordinates will also be available!
In addition, items based on the motif of the biggest nemesis "Onimai Tsuji Musho" will also appear. ......
Will players themselves be able to dress up as Sumijiro and his friends and engage in fierce battles in the big city? We look forward to more information!
The "Tsuji" in Onimai Tsuji is correctly spelled with a single dot.
The teaser movie of the "Demon Slayer" collaboration is now available on YouTube!
Tanjiro Kamado and other "Demon Slayer" characters are running through a big city in the "Ninjara" world.
The unique cartoon-like 3D model of the Ninjara design has a great impact! Demon Slayer" has collaborated with a variety of content, but this design is quite fresh, don't you think?
Ninjara TV #9 1st Anniversary Special
In addition, "Ninjara TV #9 1st Anniversary Special" will be broadcast live on YouTube Live and Twitter today, Friday, June 25, starting at 7:00 p.m.!
In addition to the information announced in the "1st Anniversary Broadcast," more campaign information for Japan will be announced!
Be sure to check it out!


The winner in the Respect category is Todd Fling. To the s, the feeding granularity of large jaw crusher that can break materials tons per hour has got nearly mm. Underground station - An enlargement of an entry, drift, or level at a shaft at which cages stop to receive and discharge cars, personnel, and material.
Measured coal resources — Coal for which estimates of the rank, quality, and quantity have been computed from sample analyses and measurements from closely spaced and geologically well-known sample sites, such as outcrops, trenches, mine workings, and drill holes.
Outside covered braai area leading from the dining room and leads out to the garden. Development mining - Work undertaken to open up coal reserves as distinguished from the work of actual coal extraction. Descriptive of certain rocks and minerals. Timber - A collective term for underground wooden supports.
Its thickness may vary from a few to several thousand feet.
The boats, 20 meters long and 30 tons each, were loaded into the aircraft using ramp and rails as well as a crane with ton lifting capacity.
Ranks of coal — The classification of coal by degree of hardness, moisture and heat content. It is similar to coal ash. All stoves complete and ready to use.
Drainage - The process of removing surplus ground or surface water either by artificial means or by gravity flow. The Group business units have carried Brow - A low place in the roof of a mine, giving insufficient headroom.
Much of the chlorine for bleach, cleansers, and disinfectants is produced using asbestos products. White damp - Carbon monoxide, CO.
Also, a tunnel driven from one seam to another through or across the intervening measures; sometimes called "crosscut tunnel", or "breakthrough". The former is also called swing jaw crusher, while the latter does arc swing, and at the same time it also make top and bottom motion, so it is also called jaw crusher with complicated swing.
This became the seventh success of the Group in the most prestigious Russian airline competition. Dust consisting of particles of coal that can pass through a No. Used for detonating explosives.
Roll - 1 A high place in the bottom or a low place in the top of a mine passage, 2 a local thickening of roof or floor strata, causing thinning of a coal seam.
It is the third aircraft of the modification financed by Volga-Dnepr Group. Usually has a high Btu value and may be referred to as "soft coal. Coal Gasification — The conversion of coal into a gaseous fuel.History of crushers, crushing machine and crushing equipment.
Jaw crusher, roller crusher, hammer mill all is a type of crusher. The Development Of Jaw Crusher Machine Crushers Jaw crusher in the mining industry use is very extensive, then jaw type crushing machine's history is Jaw Crusher, jaw crushers Through independent research and development, our jaw crushers of big and mid-bid model are placed in leading class in china.
Mineral Resources in United Arab Emirates. TheUnited Arab Emirates is abundant with various types of mineral swisseurasier.com is the eighth largest producer of crude oil in the world accounting for % of the total crude oil production.
In machinery terms, the Cracker was a large jaw crusher that broke down large pieces of rock to smaller more manageable pieces of rock. In early less developed mines this job was done manually (also with explosives) This job title would refer to someone who oversaw the. A. Abutment - In coal mining, (1) the weight of the rocks above a narrow roadway is transferred to the solid coal along the sides, which act as abutments of the arch of strata spanning the roadway; and (2) the weight of the rocks over a longwall face is transferred to the front abutment, that is, the solid coal ahead of the face and the back abutment, that is, the settled packs behind the face.
Coal miningWorld Coal Association. In this area you can find information on how the coal industry invests in the protection of precious resourcesCoal & water Coal miningcoal mining today.Download


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Forbo/Marmoleum Neutral pH Cleaner
A low toxicity, low VOC, readily biodegradable, concentrated floor cleaner that is specially designed for everyday floor cleaning and may be used with all Forbo flooring products to remove routine soils. It performs excellent for use in mop and bucket cleaning or automatic floor scrubbers. Neutral pH will not harm floor finishes.
DIRECTIONS: Dilute 1/2 – 1 ounce of product per gallon of unheated tap water. Fill mop bucket or auto scrubber with the solution. Mop and/or scrub the floor with the cleaning solution. Pick up soil and excess solution with clean a mop. Rinse mops frequently and change solution as needed. The floor should be rinsed with clean cool water and allowed to dry thoroughly.
Approximate Spread Rate: 32 Gallons of cleaner, 128 Gallons of Cleaner
Applicator: Microfiber Applicator or Finish Mop
Country of Manufacture: CN


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Looking for a New Doctor? Nearly Half of People Have the Same Goal, Survey Suggests
The results found patients value knowledgeable, empathetic physicians. If you’re seeking a new one, follow these expert tips.
We all know the difference a good doctor can make, but finding that person isn’t always easy.
A new survey from Everyday Health and Castle Connolly suggests many patients are willing to make the effort. Nearly half said that they’re considering a switch this year. Twenty percent of people said looking for a new doctor is something that’s always on their to-do list or something they consider every year, while another 25 percent of respondents said it’s now on their mind more than ever. The survey involved close to 1,300 consumers and 287 healthcare professionals.
When choosing a new doctor, patients said they most valued the ability to listen, followed by their experience and accomplishments.
Living through the COVID-19 pandemic may have more people focused on their physical and mental health, and motivated to make a change if they don’t have a strong relationship with their doctor, says Arefa Cassoobhoy, MD, MPH, who specializes in internal medicine.
“We all know someone who had COVID-19, or someone who had a medical issue and because of the pandemic they had to use telemedicine, or someone who went to the hospital and had a rough experience because it was so crowded,” Dr. Cassoobhoy says. “All of these health issues are so much more real and in our face right now, and that may make us more motivated to find physicians who meet our needs."
What Patients Value Most in a Doctor
What patients want more than anything is for doctors to make space for their questions and listen to their concerns, the survey suggests. Sixty-one percent of respondents listed this as one of their top two priorities in searching for new physicians.
Doctors’ credentials mattered but not quite as much. Forty-eight percent of participants ranked this in their top two priorities. Another 45 percent prioritize compassion, while 23 percent cited access to online scheduling, telemedicine, and the time it takes to get an appointment.
Younger patients put more stock in finding doctors who had respect and sensitivity for their race, ethnicity, gender identity, and socioeconomic status. This was a top priority for 1 in 4 participants 18 to 24 years old, but ranked 23 percent for patients overall.
How Patients Are Seeking Out New Care
When asked to rank the best resources for finding new doctors, 42 percent of all survey respondents said they would rely most on recommendations from doctors they already receive care from. Another 29 percent of people said their top source of recommendations would be friends, family, and coworkers. And about 1 in 5 people said they would be most likely to scour websites with physician ratings and reviews when seeking out a new doctor.
“Asking a doctor you like for a referral is smart because they may steer you to a clinician with a similar approach to patient care,” Cassoobhoy says. “Turning to friends and family is also a big help because they’ll tell you practical things like how easy it is to find parking or how long they keep you waiting or whether the nurses and staff are approachable."
Why Patients Are Looking for a New Doctor Now
Some people may also feel like it’s time for a change because they feel like they’re not as healthy now as they were before the COVID-19 pandemic, Cassoobhoy says. Many of us are experiencing unexpected and unwelcome life changes, she explains, and these stressors can manifest in subtle ways, such as with changes in eating and sleeping habits, or with more obvious effects, like worsening depression or anxiety. In the survey, about 54 percent of doctors reported that their patients’ health was worse now, while only 29 percent of patients said this was true of their personal health.
Similarly, 87 percent of physicians saw deterioration in both physical and mental health for their patients, compared with 63 percent of patients.
Doctors may see things differently because they’re thinking about changes that don’t necessarily cause symptoms, Cassoobhoy says. They might, for example, say patients aren’t as healthy because they gained 5 to 10 pounds or experienced small increases in blood pressure or blood sugar.
“Patients think they’re still healthy because they don’t notice changes, but doctors know the risk of diabetes and heart disease and stroke have increased even though the patient standing in front of them says they feel fine and don’t feel any different from a year ago,” Cassoobhoy says.
What to Do if You’re Looking to Change Your Doctor
There are several things you can do as you look for a new doctor to help ensure that the change results in somebody who meets your needs.
Cassoobhoy says a well-trained doctor will listen to you and ask thoughtful questions as you tell your story. When they share their thoughts with you, you should be able to ask questions in return, she adds. Last, the office visit should feel collaborative and respectful, finishing with a logical, clear plan of what the doctor and his or her staff will do next, and what you need to do.
Also, the U.S. Department of Health and Human Services (HHS) advises people to do what a lot of survey respondents are already doing: Ask your doctor and trusted friends and family members for recommendations.
Once you have somebody in mind who seems like a good fit, check with your insurance company to make sure the provider is covered by your health plan, HHS advises. Double check this by also calling the doctor’s office to confirm this on their end before your first appointment.
While you’re on the phone confirming the doctor accepts your insurance, there are some other things you should ask, according to HHS. Find out what hospital the doctor uses, who will see you if your doctor isn’t there, whether the doctor treats your specific medical conditions, and whether the doctor speaks your preferred language. You may also want to ask about the cancellation policy, how long it takes to get appointments, how long checkups typically last, and whether the doctor can run lab work or X-rays in the office, HHS suggests.
During that first visit, think about how the doctor and office staff make you feel, HHS advises. If the doctor doesn’t make you feel comfortable, show respect for what you have to say, know your medical history, and spend enough time with you then it may be worth continuing your search.
“There are plenty of wonderful doctors out there,” Cassoobhoy says. “It may seem like a daunting task, but it’s well worth the effort to look for a doctor and office staff that value communication and kindness along with great medical care.”
The Everyday Health–Castle Connolly Physician-Consumer Parallel Pandemic POV Survey was conducted in November and December 2021 across three audiences: a research panel of 1,007 consumers; 289 Everyday Health visitors, and 287 Castle Connolly healthcare professionals.

domain: medical


Army Of Lovers
Army of Lovers is a Swedish dance music group which formed in 1987, and has had a number of hits in Europe throughout the 1990s with songs such as "Crucified", which was number one on the Eurochart for eight consecutive weeks. It remains their best known song internationally. The name of the band alludes to a documentary Armee der Liebenden oder Revolte der Perversen of the German gay rights activist Rosa von Praunheim, the title of which in turn alludes to the Theban Band. Army of Lovers reformed with Alexander Bard, Camilla Henemark and Jean-Pierre Barda in late 2012 to enter the Melodifestivalen 2013 with the song "Rockin' the Ride" in hopes of representing Sweden in the Eurovision Song Contest 2013 in Malmö. They failed to progress to the final and a few days later issued a press release to announce that Henemark will be replaced by former member Dominika Peczynski for the remainder of their on-going reunion.


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Pork chops are browned and simmered to tenderness with honey and fresh apricots for a lovely spring and early summer treat.
Posted: May 24, 2011
| Cooks I Like
Posted: Apr. 27, 2013
Posted: Aug. 10, 2011
A new recipe revealed each day until Christmas. Check it out!
Get time-saving recipes to save your busy life.
Get the season’s best recipes for holiday feasting.
Read our allrecipes.com blog


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A number of cancers are characterized by overexpression of the epidermal growth factor receptor (EGFR), a membrane protein which mediates cell growth, proliferation, and differentiation in multiple tissues. Many epithelial tumors have been found to express high numbers of EGF receptors,126.96.36.199.6.–7 and the receptor levels are associated with poor clinical prognosis in cancers of the bladder,4 lung,5 and breast.4,6 Antibodies for EGFR tagged with fluorescent probes have been used as contrast agents to image EGFR overexpression.8 The EGFR-induced shift in the plasmon resonance has also been used as an image contrast method.9 Both of these methods provide at most a 10-fold intensity contrast factor.
To overcome the limited contrast, we explore the use of surface-enhanced Raman spectroscopy (SERS). By exciting a sample with laser light, energy is transferred to (Stokes process) or lost (anti-Stokes process) by the medium in an inelastic scattering process referred to as Raman spectroscopy. The scattered light is a fingerprint of the set of atomic vibrations, which is unique to the arrangement and types of atoms under study. In general, 1 in photons is scattered inelastically in this manner. By tuning the incident laser frequency to the plasma frequency of the nanoparticle assembly, an overall enhancement of 7 to 8 orders of magnitude of the scattered signal is possible;10 this is referred to as the surface-enhanced Raman scattering effect. Over the past few years, there has been a flurry of activity in the application of Raman spectroscopy to problems in biology. Sha et al.11 have reviewed the field of the application of SERS to oncology. The idea of using SERS to image cancerous tissue was exploited by Kah et al.12 A self-assembled SERS-active, gold nanoparticle (AuNP) monolayer film tagged with anti-EGFR was used to distinguish cancerous and normal cells in saliva for early diagnosis of oral cancer. Qian et al.13 have also employed the SERS to target EGFR overexpressing cells in vivo using mouse xenograft tumor models. Approximately 1.4 to reporter molecules (e.g., malachite green) are adsorbed on each 60-nm gold particle and stabilized with thiol-polyethyleneglycol (thiol-PEG). Their SERS particles exhibit a signal that is 2 orders of magnitude larger than quantum dots, highlighting the power of the technique. Keren et al.14 have demonstrated the multimodal SERS detection using antibody-tagged Oxonica nanoparticles. Kim et al.15 developed the antibody-conjugated, silver nanoparticles-embedded silica spheres to target HER2 and CD10 on cellular membrane constructs. None of these reports have mapped the distribution and fate of the nanoparticles within a single cell. Our study is an extension of an earlier contribution16 and aims to determine the microscopic distribution of antibody-tagged AuNPs in order to map the cellular distribution of EGFR in both normal and cancerous cell cultures.
Materials and Methods
Gold Nanoparticle Synthesis
The AuNPs used in the experiment were synthesized using the well-known method of Turkevich et al.17 and Frens.18 A 0.01% by weight gold chlorate solution was heated with stirring to boiling in an extremely clean round bottomed flask fitted with a refluxer in a fume hood. The flask was heated in a bath of silicon oil to ensure even heat distribution. A precise volume of 1% by weight sodium citrate was added to reduce the gold salt with continued boiling. The amount of sodium citrate added to the stirring gold solution ultimately determined the final size of the AuNPs, with smaller volumes of citrate producing larger particles.
Gold Nanoparticle Characterization
Three techniques were used to characterize the AuNPs and their derivatives: UV-visible spectroscopy (UV-Vis), transmission electron microscopy (TEM), and dynamic light scattering (DLS). From UV-Vis spectroscopy, the approximate size, plasmon absorbance wavelength, and concentration of the nanoparticles were obtained. The UV-Vis analysis was performed using a UV-1700 (Shimadzu) or ND-1000 (Nanodrop) spectrophotometer. The concentration of nanoparticles can be calculated using the absorbance maximum:
Antibody Conjugation to Gold Nanoparticles
To test the effectiveness of anti-EGFR antibodies, a series of experiments were run. Antibodies specific for human EGFR, produced in mice (Sigma-Aldrich, E2156, Monoclonal anti-EGFR-human, clone 225, purified mouse immunoglobin, Immunogen—partially purified EGF receptors from human A431 cells), IgG polyclonal nonspecific antibodies (Sigma Aldrich, I8765, Mouse IgG-technical grade from normal mouse serum), and antibody free solutions were incubated with nanoparticles. AuNPs of approximately were diluted in a ratio with 50 mM HEPES and then centrifuged at 3000 rcf for 1 h at 4°C to concentrate. This gives an approximate volume of 4 mL and concentration of . A solution of antibodies at a final concentration of was prepared in 25 mM HEPES. This solution was added to the nanoparticle solution. An equal volume of PEG solution was added dropwise. The solution was incubated overnight at 4°C or until use. Unattached antibodies were removed by centrifuging at 3000 rcf for 1 h at 4°C just prior to use. The solution was resuspended in 50 mM HEPES, giving an approximate volume of 1 mL. This solution was then characterized with UV-Vis. A shift of 6 to 10 nm indicates that the antibodies are attached. The approximate concentrations of the components in 1 mL of the prepared solution were: nanoparticles—, antibodies—, and PEG—. The antibody-free, or bare nanoparticle, solutions were subjected to the same procedure to account for any losses.
Two types of cells were used: EGFR overexpressing human epidermoid carcinoma A431 cell line from ATCC and normal human bronchial epithelial (NHBE) cells from Lonza. The antibody-conjugated AuNPs () were then attached to the cells (), leading to an approximate ratio of . In a biosafety cabinet, the prepared antibody solution was pipetted into a dish of cells where the cells had been grown on a coverslip. The cells were counted to give no more than 0.5 million for an experiment. The solution was incubated with the cells for 30 min and washed with sterile 50 mM HEPES. The coverslip was flipped into a clean glass bottomed Petri dish. A polymer spacer was pressed against the coverslip to secure it. The dish was filled with HEPES buffer (pH 7.4) and loaded into a custom-made Raman sample holder.
Raman spectra were obtained with a LabRam confocal micro-Raman system (Jobin Yvon Inc., Edison, New Jersey), which consists of a grating spectrograph coupled to an Olympus BX40 microscope with a motorized stage. A 632.8-nm He-Ne laser excited the Raman spectra by focusing through a objective with a coverslip correction collar. The incident power at the sample is . The Raman scattered light was collected in a confocal arrangement by the same objective and spectrally dispersed using a Horiba Jobin Yvon Raman Spectrometer onto a thermoelectrically cooled CCD detector. The system also has a video system for white light image capture, a computer to collect and process data, and a motorized stage for translation along , , and .
The system was calibrated with the silicon line at (at least 2000 and up to of signal was achieved). A grating of , a confocal pinhole of , and an input slit width of were used. Spectra were collected with a 1 s integration time, averaged over two scans. A step size of or less was utilized, typically , to map the spectral features of a cell. In this confocal arrangement, the depth resolution is . The intensity distribution in a narrow spectral range was used to produce a Raman map of the cell of interest.
Results and Discussion
Nanoparticle Preparation and Characterization
To produce gold colloidal solutions, 1 mL of 1% by weight sodium citrate was added dropwise to a boiling solution of 100 mL 0.01% (by weight) gold salt at 100°C. The reaction is complete when the solution turns a reddish color (in 5 to 10 min). The gold colloidal solution was characterized by UV-Vis spectroscopy, TEM, and DLS. An analysis of the TEM images in Fig. 1(a) reveals particle diameters of approximately , while the DLS histogram [Fig. 1(b)] implies an average particle size of approximately 34 nm. An increase is expected since DLS measures the hydrodynamic radius including the citrate capping molecule. UV-Vis absorption [Fig. 1(c)] exhibits a maximum absorption at 532 nm for the bare nanoparticles (blue curve) and 536 nm for AuNPs tagged with anti-EGFR (pink curve). The latter curve is obtained after eliminating unattached anti-EGFR by centrifugation at 3000 rcf. The 4-nm red-shift between the maxima of the absorption spectra is consistent with the attachment of anti-EGFR to the AuNPs, which is ultimately due to the increase in the index of refraction of the medium surrounding the AuNPs.
Anti-EGFR-Tagged Gold Nanoparticles
The anti-EGFR-tagged AuNPs were aggregated by adding 0.01 M PBS (pH 7.4) to the gold colloidal solution. Figure 2 is the Raman spectra of the point indicated in the optical image of an aggregate (inset). The data were highly reproducible if the laser power, under the objective, is kept below . The spectral features are specific to the attached antibody. Huang et al.19 reported the SERS spectra of anti-EGFR. The peaks reported by them are comparable with those found here (Table 1). The absence of an amide I band near is striking, as this feature is normally prominent in the nonSERS Raman spectra of proteins. This band requires the presence of the entire backbone in the enhancement area in a parallel orientation. Its absence implies that only the side chain groups were enhanced.
Peak positions, relative strength, and tentative assignments for the SERS spectra of anti-EGFR. Peak positions obtained from Lorentzian fits (very strong—vs, strong—s, moderate—m, weak—w, and very weak—vw).
|Raman shift (cm−1)||Relative peak intensity||Assignment|
|282±1||vw||}S-Au plus torsional|
|367±3||vw||O-Au plus torsional|
|383.6±0.3||m||}N-Au plus torsional|
|432±1||vw||Ring bending/torsional mode|
|449.9±0.9||vw||Ring torsion of phenyl20|
|467±1||vw||Valine or glutamic acid21|
|475.0±0.9||vw||Valine or glutamic acid21|
|506.6±0.2||m||}S-S stretching vibrations in cysteine residues19,20|
|551.5±0.5||w||Tryptophan, histidine, or arginine21|
|603.8±0.2||m||C─S stretch glycine, glutamate, or phenylalanine21|
|639±1||vw||C─S stretch and C─C twist of tyrosine20|
|703±1||w||C─C twist of tyrosine or phenylalanine20|
|716.1±0.2||m||Deformation vibrations of COO− (Ref. 19)|
|737.4±0.6||w||C─S stretch20 or deformation vibrations of COO− (Ref. 19)|
|812.6±0.5||vw||Tyrosine ring breathing vibrations (exposed)22|
|856.9±0.9||w||Tyrosine ring breathing vibrations (buried)19,22|
|870.2±0.6||vw||C─C stretch in proteins20|
|896.7±0.5||w||C─C stretch in glycine19|
|947±4||vw||C─C stretch proline and valine20|
|1007.1±0.2||vs||Ring C─C stretch in phenylalanine or tryptophan19|
|1077.9±0.3||w||Tryptophan or C─N stretching mode of proteins20|
|1129±0.2||vs||Symmetric C─C vibrations19|
|1141.2±0.5||m||Symmetric C─C vibrations19|
|1240.2±0.4||s||Tryptophan ring vibrations19|
|1269.1±0.2||vs||C─H bending mode|
|1317.1±0.4||w||Carbonyl symmetric stretch|
|1379.6±0.1||vs||Carbonyl symmetric stretch|
|1422.0±0.5||w||COO− stretch in IgG aspartic or glutamic acid or C═C stretch20|
|1490.5±0.6||m||Tryptophan, tyrosine, ring stretch/breathing23|
|1552±3||vw||C─C stretch of tryptophan23|
|1584.5±0.3||w||Asp, glu, C═O stretch23|
|1619.6±0.2||w||C═C stretch of tryptophan or phenylalanine20|
|1749.2±0.4||w||C═O stretch of aspartic acid23|
|2878±1||w||CH2 asymmetric stretch of proteins20|
|2934.3±0.1||vs||C─H vibrations in proteins; Chain end CH3 symmetric band|
Proteins are expected23 to bind to the nanoparticles through those amino acids which form complexes with gold; among them sulfur containing cysteine, nitrogen containing histidine and tryptophan, and the basic amino acids lysine and arginine, at high pH above their isoelectric point (pI). The acids, aspartic and glutamic acids, would also be expected to adsorb when deprotonated, as would tyrosine through the oxygen atom side group following the loss of hydrogen. At the pH used in these experiments, 7.4, the deprotonated acids would be expected to bind to the nanoparticles, but not the bases. The results outlined in Table 1 confirm this hypothesis. The tentative peak assignments are taken from a variety of sources. Cysteine, histidine, tryptophan, tyrosine, aspartate, and glutamate peaks dominate the spectra below , while the high-frequency peaks are likely due to C─H and vibrations of these amino acids. In addition, among these amino acids, few characteristic peaks appear below . The appearance of low-frequency features arises from the binding of sulfur, nitrogen, and oxygen to the AuNP; for which the magnitude of charge transfer decreases in the same order, sulfur-nitrogen-gold. This has the effect of reducing the contribution of charge-transfer effects to the resonance enhancement in SERS, reducing the strength of these modes, in the order of 302, 397, and .
Six modes exhibit very strong enhancement, corresponding to vibrational frequencies of (phenylalanine or tryptophan), (symmetric C─C stretch), (C─H bending modes), (carbonyl symmetric stretch), and (C─H stretch).
Raman Spectra of Cancerous and Normal Cells
A 0.5 mL of A431 and NHBE cells was pipetted into glass bottom Petri dishes and left to attach overnight (for A431 cells) or for 2 days (for NHBE cells) in an incubator (5% , 37°C). 0.5 mL of the anti-EGFR-tagged AuNPs, at a concentration of , was added to the cells 30 min prior to the Raman experiments.
Raman spectra were collected by a point mapping scheme, with a step size of along and . At each point, spectra are collected for 1 s, averaged over two scans. Figure 3 shows the Raman spectra following excitation at four locations of the cell, indicated in Fig. 4(a). All peaks in the spectra are due to SERS. The simple Raman spectrum without SERS is basically the same as the background noise (spectra 4). It is clear that the SERS spectra collected at different locations vary. Figure 4(b) is a SERS image obtained by mapping the intensity of the peak, while Fig. 4(c) is a SERS image obtained by mapping the intensity of the peak.
The SERS images exhibit a number of interesting features [Figs. 4(b) and 4(c)]. First, there is no enhancement in the nucleus (Nu). The signal from within the nucleus shows a featureless background. Second, SERS spectra are observed in the areas near the surface and within the cell membrane. This observation can be reconciled with the fact that the EGFR is recycled by the cell through the process of endocytosis. Furthermore, electron micrograph images presented in supplemental Fig. 5 of Ref. 13 reveal AuNPs within endosomes of cells tagged with AuNPs. Moreover, the Raman spectra are different in the four spots indicated in Fig. 3. This implies that the environment in the immediate vicinity of the AuNPs differs. Within endosomes, the EGFR is broken down into protein fragments which would produce significantly different spectra. When the image of any vibrational mode (e.g., 1450 or ) in a given Raman spectrum is mapped, one observes little change in the Raman image (i.e., its spatial intensity distribution). While we have presented only two examples, this observation appears to be generally true. Selection of any other mode in a given spectra gives rise to a similar image, implying that the Raman image is consistent with the distribution of nanoparticle aggregates.
Table 2 lists the observed Raman shifts for each of points 1 to 3 in Fig. 3. Most of the peaks are common to anti-EGFR, as listed in Table 1. There are additional peaks which can be mostly attributed to the appearance of tyrosine and tryptophan residues, and C─C and C═O stretches of amino acids. New peaks appear at points internal to the cell, where the lysozyme would begin breakdown of the protein, freeing those that are already bound to the nanoparticle to make an even stronger connection and undergo reorientation. The conclusion can be drawn that there is a rearrangement of the amino acids upon endocytosis to make tryptophan and tyrosine residues, which both contain ring systems, more prominent in the spectra.
Peak positions, relative strength—ST, and tentative assignments for the SERS spectra in the vicinity of nanoparticles, located within or on the surface of A431 cells. Peak positions obtained from Lorentzian fits (very strong—vs, strong—s, moderate—m, weak—w, and very weak—vw). Trp—tryptophan, Tyr—tyrosine, Phe—phenylalanine, Arg—arginine, and Pro—proline.
|Point 1 spectra peaks (cm−1)||ST||Point 2 spectra peaks (cm−1)||ST||Point 3 spectra peaks (cm−1)||ST||Assignment|
|859.0±0.3||w||Phe21 or Tyr22|
|884.86±0.07||vs||Trp, ring breathing mode, or CH2 proteins20|
|920.8±0.2||m||C─C stretch of proring20|
|981.3±0.1||s||Trp, Tyr, ring deformation|
|1000.6±0.6||w||Phe, ring stretch20|
|1050.1±0.1||m||1044.6±0.3||m||Trp, ring stretch23|
|1133.9±0.5||w||C─C skeletal stretch transconformation20|
|1218.1±0.2||s||1215.0±0.5||w||Trp, Tyr ring deformation23|
|1273||w||1273||w||1269.2±0.3||m||Tyr, ring deformation|
|1330.2±0.4||w||Trp, Tyr ring stretch23|
|1356.6±0.1||s||Trp, ring stretch23|
|1384.3±0.8||w||Tyr, ring stretch|
|1458.8±0.5||m||1449.26±0.06||vs||CH2 deformation/C═C bending mode of phe20|
|1552.9±0.3||s||1545.3±0.6||w||1554.6±0.9||w||C═C Trp stretch20|
|1563.7±0.6||w||Trp, Tyr, ring stretch23|
|1582.05±0.09||vs||Asp, Glu C═O|
|1600.27±0.08||vs||1601±2||w||Trp, Tyr ring stretch23|
|1622.6±0.3||w||C═C stretch Trp, Tyr|
One can estimate the enhancement by considering the Raman signal generated by a bulk sample of anti-EGFR and the reduction in the number of anti-EGFR molecules contained within an AuNP aggregate. The numerical value of the enhancement factor is determined by comparing the Raman intensity per molecule in the presence or absence of nanoparticles with all other conditions being constant (e.g., laser power). Its value is given by , where is the number of analyte molecules in the illuminated volume in the presence of nanoparticles and is the number of molecules in the illuminated volume in the absence of nanoparticles. We estimate that the enhancement ranges from to .
Figure 4(d) shows the optical image of an NHBE cell. Raman images of the [Fig. 4(e)] and [Fig. 4(f)] peaks show little or no enhancement over the entire cell. The only exception is a region in the lower right of the Raman images, which is outside the cell [Figs. 4(e) and 4(f)].
The optical image [Fig. 4(d)] clearly shows an aggregate of AuNPs, adhering to the Petri dish, that was not removed by the washing process. The spectra (Fig. 5) are very similar to the SERS anti-EGFR spectra of Fig. 2. The similarity of the spectra indicates that the nanoparticles retain their anti-EGFR coating. Other than this artifact, the Raman images and spectra are featureless and weak.
Figure 6 is a plot of the Raman spectra from the cells in Figs. 7(a) and 7(d). These spectra are completely featureless and are comparable with the background intensity of the cell nucleus spectrum (Fig. 3, point 4). Figure 7 shows the control experiments with A431 cells incubated with bare AuNPs [Fig. 7(a)] or nonspecific polyclonal antibody-tagged AuNPs [Fig. 7(d)].
Figure 7(b) shows the SERS intensity map at of the cells incubated with the bare AuNPs, while Fig. 7(c) is the intensity map at . The maps are very similar. The SERS intensity map at for the nonspecific antibody-tagged AuNPs is shown in Fig. 7(e) and the map is shown in Fig. 7(f). These maps are also similar, however, the spectrum at seems to have slightly higher background intensity. The reduced intensity of the normal cell spectra [Figs. 6, 7(b)–7(c), and 7(e)–7(f)] as compared with the intensities obtained from the anti-EGFR functionalized A431 cancer cells [Figs. 3 and 4(b)–4(c)] corresponds well to expectations.
We have targeted EGFR in EGFR-overexpressing cells of the A431 human epidermoid carcinoma cell line (one million per milliliter) and in NHBE cells by binding anti-EGFR-tagged 30-nm AuNPs. We observed dramatic SERS effects in the A431 cells. As expected, control experiments with NHBE cells and with bare or nonspecific antibody-tagged AuNPs bound to A431 cells produced insignificant SERS enhancement. The spectra are associated with residues which have functional groups which associate with gold, including the sulfur containing cysteine and nitrogen containing histidine and tryptophan. Aspartic and glutamic acids would also be expected to adsorb when deprotonated, as would tyrosine through the oxygen atom side group following the loss of hydrogen. Evidence for rearrangement of these amino acids upon uptake into lysosome and endosomes is evident when comparing the SERS spectra of anti-EGFR-coated nanoparticles with spectra from cells. Tyrosine and tryptophan residues dominate.
Raman signal enhancements are observed for A431 cells and reflected the AuNP distribution. AuNPs are located on the surface as well as on the interior of the cell, while being excluded from the nucleus. The EGFR is recycled by the cell through the process of endocytosis and is not found in the nucleus. Moreover, EGFR is known to distribute nonuniformly on the cell surface. Thus, the SERS map is consistent with the expected distribution of EGFR. The SERS enhancement factor estimated from our data is between and . Studies are underway to optimize the AuNP size and laser excitation wavelength. The NHBE cells do not show signal enhancements. A contrast factor of at least is estimated, which means that a SERS image should have orders of magnitude more contrast than a typical fluorescence image or one generated by white light scattering from nanoparticles. A SERS approach to imaging EGFR distribution is, therefore, very promising.
We are indebted to the Natural Science and Engineering Research Council of Canada, the Canada Foundation for Innovation, Canadian Institute for Health Research, National Cancer Institute of Canada, and the Canadian Dermatology Foundation for financial support. We are also indebted to Drs. B. Frisken and N. Forde, both of Simon Fraser University (SFU)—Physics, for making their lab available during the sabbatical of KCH. In particular, KCH wishes to acknowledge the help of Dr. Frisken and her students (David Lee and Rasoul Narimani) for helping with the Dynamic Light Scattering experiment and for providing office space during his sabbatical at SFU. KCH and LL would like to thank Dr. Forde, her students, and postdocs, especially Astrid van der Horst, Suzana Kovacic, and Laleh Samii, for their willingness to help with experiments and equipment (including UV-Vis spectrometer). KCH is grateful also to XKC for his tremendous support during his 6-month sabbatical term at SFU.
Leanne J. Lucas graduated from Dalhousie University with a Bachelor of Science, Honors Co-op in chemistry (2010), and a Master of Science in physics and atmospheric science (2013). Her research interests include Raman spectroscopy, proteomics, nonconventional cancer diagnosis and treatment methods, and renewable energy. She was the first African Nova Scotian to receive a graduate degree from the Department of Physics and Atmospheric Science (2013). She currently works at Maxxam Analytics in Bedford, Nova Scotia.
Xiaoke K. (Michael) Chen is a senior lecturer at Simon Fraser University. He received his PhD degree from the City University of New York in 1991. He is the author of 42 journal articles. His current research interests are Raman spectroscopy and physics education.
Aaron J. Smith is a scientist at Tesla Motors. He received his PhD degree in physics from Dalhousie University in 2012. His current focus is on advance lithium-ion battery testing and analysis.
Mladen Korbelik is a distinguished scientist at the British Columbia Cancer Agency (Vancouver, British Columbia) and professor at the Pathology and Laboratory Medicine Department, University of British Columbia. He has authored around 150 journal papers or book chapters. His research interests include the use of photodynamic therapy for treatment of solid malignant tumors and the associated inflammatory/immune host response, vaccines, and other forms of cancer immunotherapy.
Haishan Zeng is a distinguished scientist at the British Columbia Cancer Agency and a professor of dermatology, pathology, and physics at the University of British Columbia. His research has been focused on developing optical imaging and spectroscopy techniques for improving early cancer detection. He has published over 123 refereed journal papers, 1 book, 8 book chapters, and has 20 granted patents.
Patrick W. K. Lee is a professor and Cameron Chair in Basic Cancer Research at Dalhousie University. He received his PhD degree in biochemistry from the University of Alberta in 1978. He is the author of more than 100 journal articles. His current research focuses on oncolytic viruses, oncogenes, and tumor suppressors.
Kevin Cecil Hewitt is an associate professor of physics at Dalhousie University. He received his BSc degree in physics and biology from the University of Toronto and a PhD degree in physics from Simon Fraser University, in 2000. He is the author of more than 30 journal articles and has written 1 book chapter. His current research interests include Raman spectroscopy, holography, and simulated Raman spectroscopy.

domain: biology