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Who're we? We are a New York Tristate Space primarily based Skilled Automotive detailing service. we offer an reasonably priced value that provides professional, high quality auto look providers to auto fanatic and on a regular basis hardworking people. We use only the highest quality merchandise and the latest innovations to ensure that your vehicle gets the correct care and protection it deserves. We're enthusiastic about automobiles and won't decide on anything less than perfection. NB: If one disc is like this and the opposite one on the other side seems OK you might have wheel cylinder component failure and the automotive should not be driven besides fastidiously to the repairer. Volvo will present its dedication to station wagons, whose sales are being hit by the global growth in SUVs, when it unveils its new V90 later this month. Step 15. Wash and dry your microfiber towels SEPARATELY from any other laundry. Use scorching water, detergent, but no cloth softeners as it would damage the microfibers. Dry them on low heat so you will not melt the fibers. Retailer them for subsequent time you might want to wash your car.
We're excited to reach this agreement with IMSA to campaign the RC F GT3 at such a excessive level. Our hope is to build a robust basis for the new F Efficiency Racing group during 2016 season, seizing the opportunity to develop the workforce and the race automobile in anticipation of the multi-automobile effort throughout the 2017 season," said Jeff Bracken, Group Vice President and Basic Supervisor, Lexus division.
If in case you have accomplished a Basis/HND course and wish to study this diploma with us in 2017, you may be able to begin in Yr two/three (level 5/6). Please be aware that the modules for a 12 months two/three start shall be completely different to the ones listed above. You have to to obtain the 2016/17 programme for extra information on which modules you'll be finding out.
German designs began accepting acceptance afterwards the Eighties, notable afterwards the buildup of Audi. Volkswagen, which was abased on Marcello Gandini and Giorgetto Giugiaro and Karmann, afterwards shaped the abreast architectonics accent forth with Audi. BMW's attack into sports auto apparent a brand new pattern in auto architectonics because it alleged for a sporty-wanting accustomed auto with Giovanni Michelotti, afterwards added by Ercole Spada applicable into the Eighties, and Klaus Luthe till mid-Nineties. The American constructed-in artist Chris Bangle murderer by BMW in late-Nineties to re-define the forged and he acclimated new particular person columnist know-how for admixture curves abacus controversial administration components in his designs.
Experience : Being a UPESite is without doubt one of the most fascinating moment in my life. 12 months by Yr i learnt many new things, rectified my mistakes, briefly I am leaning to grow to be a matured particular person. Engineering is quite challenging however I enjoyed alot and being a Energy System Engineer I'm now extra technically lively in at this time's world in energy sector which is the spine of any nation. UPES offered me a platform to specific myself on a challenging surroundings through which by means of numerous ups and downs I am completely satisfied to say that i am on the right track.


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City Life Church, Cambridge started 24 years ago with a vision, passion and boldness to do something risky. It's been a blast, and like everything worthwhile, it's been difficult as well as joyous. Still, we see God has brought our time to an end. As of June 10th CLC Cambridge will disperse, sending our final number out to change the world--the mission God gave us right from the very start. Thank you to everyone who's joined the amazing adventure! There is so much to be grateful for. We still identify as family and we remain friends as we enjoy finding out what God has for us all next. --Kevin, Tom, Lyn & Tim
2018 Season of Prayer
Praying is really important for CLC. We were born from prayer & we've enjoyed many milestones along our 20 years history–talking & listening to God along the way.
I remember when we were considering renting Portugal Place — the student house that embodied our unconventional approach to blessing students while blurring boundaries and experimenting with words like community and authenticity. This great blessing was a decision which was preceded by an answered prayer–this one was for the down-payment (as our requests to God can be practical and so can His answers!)
A few years before that CLC was bursting with vision to reach students and see Jesus do something new in their world. So what did we do? We dreamed and we talked and we acted. But also we prayed.
The heartfelt and persistent prayer of a righteous man (believer) can accomplish much [when put into action and made effective by God—it is dynamic and can have tremendous power]. James 5:16b
We prayed specifically, effectively, passionately, asking Him to send us three key students as we didn't know anyone inside the illustrious walls and gates of our city's fine university. And, He did! Not right away, but thanks to His righteousness and love, He heard our trusting requests and answered with a 'Yes!'.
More recently, only last year, I've been asking us to join me praying for visitors and guests and new folks to join CLC. My specific prayer has been:
- add our next youngster
- add or next family
- add our next worker who we can release to take us forward reaching out
Well, have you met our young guests in December?! We've had grandchildren and a grandparent–what fun! Come on, let's keep asking, seeking and knocking!
Our season of Prayer starts this month and it's a special chance for us to meet Him together, hear Him for our church and our communities.
We know prayer is about so much more than just asking. While we talk and listen to Him, He changes us, blesses us, grows us more and more into the people that He always intended us to be! Sometimes it's great to be with Him, to enjoy Him with our agenda's set aside. That's what we'll be doing Be Still and Know that He is God at our Sunday evening, Jan 7th, gathering at the Cades. You are welcomed to come around 8pm and we'd love you to join your prayers together.
love & blessings,

subset: commoncrawl


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A Novel Radio Frequency Coil Design for Breast Cancer Screening in a Magnetic Resonance Imaging System.
Faculty Advisor or Committee Member
Reinhold Ludwig, Advisor
Faculty Advisor or Committee Member
John M. Sullivan, Jr., Committee Member
Faculty Advisor or Committee Member
William R. Michalson, Committee Member
Faculty Advisor or Committee Member
Fred J. Looft, Department Head
Magnetic Resonance Imaging (MRI) is a widely used soft tissue imaging technique that has gained considerable success because of its sensitivity to several tissue parameters. However, commercially available whole-body imaging systems with large encircling radio frequency (RF) and gradient coils are less efficient when the goal is to obtain detailed, high-resolution images with high specificity and sensitivity from localized regions of the body such as the female breast. This research addresses these problems by proposing a new design in RF coil development for breast cancer screening in a conventional 1.5T MRI system. The new design provides two resonant receiving modes that operate in a quadrature configuration, and a region of interest (ROI) that closely conforms to the shape of the female breast. We adopted an optimum design strategy that combined the analytic Biot-Savart intergral equation with the Method of Moment formulation in the development of electromagnetic models and simulation tools. These models were used to analyze the magnetic field distribution and the spatial field coverage, as well as the magnetic field uniformity in the ROI. Results from our analysis were employed in the construction of a highly scalable prototype. The validation of our design strategy is confirmed by comparisons with the commercial Ansoft HFSS v8.5 finite element package.
Worcester Polytechnic Institute
Degree Name
Electrical & Computer Engineering
Project Type
Date Accepted
Copyright Statement
All authors have granted to WPI a nonexclusive royalty-free license to distribute copies of the work. Copyright is held by the author or authors, with all rights reserved, unless otherwise noted. If you have any questions, please contact firstname.lastname@example.com.
Repository Citation
Obi, Aghogho A., "A Novel Radio Frequency Coil Design for Breast Cancer Screening in a Magnetic Resonance Imaging System." (2004). Masters Theses (All Theses, All Years). 91.
Magnetic Resonance Imaging, Radio Frequency Coil, Breast Cancer, Imaging Systems, Magnetic resonance imaging in medicine, Breast, Cancer, Diagnosis, Radiofrequency spectroscopy


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Application Travel Agency LB V1.5 is an integrated environment of a Travel Agency Management . The user application is able to manage accounts of:
- clients,
- partners,
- airline and shipping companies,
- tours and hotels.
Detail the features of the application are presented below:
Customers stored their personal data , their bookings airline and ferry tickets , as well as tours and hotel reservations.
At the same time in the payments tab displayed , debts and their sets, while there is also the possibility of customer analytic movement printing .
Day Fund
At day fund printed all credits and debits in the office space that the user specified. So it may not be only day fund , but the week , month , etc. The application supports only costs to partners . There is possibility of recording and other expenses , such as rent, staff payments , electricity, telephone , etc. , in addition to the implementation of the module business costs , with additional costs.
Office Moves
On office moves are shown in detail all the dummy sets and can filter for controlling certain period.
Partners Moves
In partner moves recorded data of the partner and office payments to him. Are aggregated data obtained automatically from the moment of reservation the customer tab in this partner.
There is also the possibility of applying a time filter as well as detailed prints.
Air-Sea Lines Moves
In this tab recorded data of air and ferry companies that works the tourist office and are aggregated data , so you can control the traffic that makes office with that company .
This page gives more opportunity for the Office to better negotiate with the representative of a large company.
There is the potential to filter , and detailed print .
Debts customers and partners
These two tabs appear total customer debts and debts to partners of the tourist office .
It is possible to print data in any case including contact numbers .
Create / Change Tour – Employees
In this region out tours provided by the tourist office , as well as employees who work in it .


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Professor Geoffrey Beattie BSc PhD CPsychol CSci FBPsS FRSM FRSA is a prize-winning academic with a First Class Honours degree in psychology from the University of Birmingham and a Ph.D. in psychology from the University of Cambridge. He was awarded the Spearman Medal by the British Psychological Society for 'published psychological research of outstanding merit' and the internationally acclaimed Mouton d'Or for his work in semiotics. He is both a Chartered Psychologist and a Chartered Scientist. He is also a Fellow of the British Psychological Society, a Fellow of the Royal Society of Medicine, a Fellow of the Royal Society of Arts and an ex-President of the Psychology Section of the British Association for the Advancement of Science (B.A.).
He is the author of 24 books published by Granta, Victor Gollancz, Chatto & Windus, Penguin/Mainstream, Weidenfeld & Nicolson, Orion, Headline, Routledge etc. His books have been translated into six languages including Italian, Chinese and Taiwanese. He has published over a hundred academic articles in a range of journals which include Nature, Nature Climate Change and Semiotica.
His book 'The Psychology of Language and Communication' with Andy Ellis was published in the Psychology Press/Routledge Classic Editions series for 'timeless classics' in 2017. His new book 'The Conflicted Mind: And Why Psychology Has Failed To Deal With It' will be published later in the year by Routledge. He is currently working on a novel for Gibson Square set in Sheffield during the Thatcher era.
He was the resident on-screen psychologist for eleven series of Big Brother on Channel 4 from 2000-2010, and has presented a number of television series, including 'Life's Too Short' (BBC1), 'Family SOS' (BBC1), 'The Farm of Fussy Eaters' (UKTV) and 'Dump Your Mates in Four Days' (Channel 4).
He has appeared as an expert analyst and commentator on BBC News, Euronews (see below), Russia Today, ITV News (including his own strand 'The Body Politic' on News at Ten on the run up to a General Election), CNN, Sky News, BBC News 24, BBC World Service, BBC Sport, News Asia, GMTV, Channel 4 News, Channel 5 News, BBC Breakfast, Good Morning America, Tonight with Trevor McDonald etc. and been interviewed on numerous radio programmes, including the Today Programme, Midweek, Woman's Hour, Science Now, All in the Mind, Word of Mouth, the BBC World Service 'World of Books' programme etc.
He has also appeared on numerous documentaries for the BBC, ITV, Channel 4, Channel 5 (image below from a documentary on Michael Jackson) and Sky television.
In addition, he has carried out media work on behalf of a range of organizations, including ITV, Universal Pictures, Department for Work and Pensions, NHS, Nivea, Barclays, Virgin Broadband, Royal Mail, Disney, Learndirect, Expedia, Teletext, UKTV, Zurich Financial Services, Esure, BT Broadband, Virgin Trains, Shell, Colgate, Kleenex, the Foreign and Commonwealth Office etc.
He has given numerous keynote addresses to both academic and non-academic audiences including Unilever, P&G, ITV, HSBC, Tesco, Marketing Forum, PricewaterhouseCoopers, Annual Conference of High and Supreme Court Judges in Ireland, Marketing Society, Equality Challenge Unit, Malaysian Olympic Association, British Council, Samaritans and the Central Office of Information, and written extensively for The Guardian, The Observer, The Observer Magazine, The Independent, The Independent on Sunday and The New Statesman, as well as contributing to Granta magazine, over a number of years.
He presented the Star Lecture at the University of Manchester in 2011 and he will present a lecture on 'Visible Thought: How spontaneous hand movements connect to thinking' at the Museum of London on behalf of Gresham College (founded 1597) in April 2018.


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Harrington Chats Vision With Local NAACP
Instead, she is seeking to create a culture and environment where causes such as drug or mental health problems are addressed in the community so the crimes don't happen again.
"My vision, and it is a big vision, is for structural change and to reallocate resources from a system of prosecuting people, processing through the courts, and incarcerating people to a system that is more of a social safety net, holistic health-based system," Harrington told the local branch of the NAACP Wednesday evening.
"But that is big. I can't do that by myself."
The Berkshire County branch of the NAACP had already brought criminal justice to the top of its priorities two years ago. Now with a new district attorney, the organization invited Harrington to a meeting to talk about the system moving forward. It is just the latest event the NAACP has put on to better connect with officials in the community.
Harrington outlined the vision she had campaigned on, one of changing to a system that focuses more on diverting people from jail and finding alternative methods to rectify behavior than chalking up conviction numbers. She hopes to move the low-level crimes into diversion programs and focus prosecutorial resources and efforts on violent criminals.
"I do not believe the House of Correction should be the drug treatment provider, or any kind of treatment provider, of choice. I think that jail is appropriate for people who are dangerous but I don't think people should go to jail to get drug treatment or to get therapy," Harrington said, "My goal is to divert people into programs that part of the community."
The most logistical issue facing such efforts is a lack of resources. While there are professional providers of such service, the Berkshires doesn't have enough for the demand.
That is a problem that is out of Harrington's hands for the most part. But she told the crowd at the Berkshire Athenaeum that she has been setting the framework by meeting with local organizations doing that work and state and federal lawmakers on getting grant funding to expand.
"I'm working for more resources but that is a big challenge," Harrington said.
While resources may be needed for better access to programs, the district attorney is currently creating the pathways to get there that hadn't existed before. One of the first actions teed up is the development of a juvenile diversion program.
Helen Moon, a city councilor and Harrington's campaign manager, has taken a newly created job in the office as director of special projects. Moon's first task is to build that juvenile diversion program.
Moon told the audience that she currently developing a model, particularly looking at the Middlesex model, as one that will treat juveniles as juveniles. She said she is also looking to develop task forces consisting of prosecutors from the office, school administrations, police, and state Department of Children and Families in each of the three areas of the county -- North, Central, and South -- to focus on at-risk youth.
"We want to make sure [we find] people who believe in this model of treating juveniles like juveniles and the role it can play in the DA's office. We are going to invite people to create that task force," Moon said.
Harrington hopes to create similar programs for those who struggle with addiction or mental health as well. But, she admits she has a bridge to cross with police officers.
"They feel like they do their job and arrest people that are committing crimes, that are not getting along in the community, and bring them to court and they are just let go. They call it 'catch and release.' They are very frustrated," Harrington said.
"My pitch to them is that these people don't belong in jail. We have to come up with the programming to address whatever the underlying issues are."
She said she has brought in training programs for the assistant district attorneys who serve as the "front line" of the office. They will be reflective of the ideals of the office.
"I hold everybody in our office to a high standard," Harrington said.
Internally, she is also tightening up policies. Auditor Suzanne Bump presented a series of recommendations to improve operations that Harrington said she is implementing. Harrington also created a director of operations position, being filled by Dina Guiel, who worked on the Harrington campaign as well, to tighten up the organization's policies and procedures. One such task is to tighten up how drug forfeiture money is handled so it goes back to the office in a timely manner, which was cited by Bump.
"I created a new position, chief of operations, who is going to help make sure that kind of things doesn't slip through the cracks," Harrington said.
The district attorney added that she is also limiting what the office goes after for forfeiture.
She is working with the attorneys and with defense attorneys to stray a bit from cash bail, which she says hurts poor people. She is looking to be more creative with conditions of release to ensure that the person returns to court to answer to charges and doesn't get jailed just because of financially unreasonable demands.
While there are many ways Harrington is looking to keep people from jail, there are some crimes she won't tolerate. She said she will "aggressively" prosecute cases of domestic violence and pledged to do more on that subject.
"I've heard from so many women who have tried so hard to get justice, tried to get restraining orders, tried to get help from law enforcement, and it hasn't been there. We have a disconnect," Harrington said.
"The police officers I am working with, I know want to help and want to do the right thing. We need to figure out where that disconnect is and to make sure that people who are victims are heard and we are protecting them."
She promised to review 15 years worth of unindicted cases of sexual assault and contact the victims and see if they want to move forward and to bring charges in cases where it is warranted.
"I have heard from victims who felt that their cases were not prosecuted and they should be. My intention is to review those cases, to contact the victims and determine what cases have probably cause and what victims want to seek justice," Harrington said.
When asked about marijuana, Harrington said that is trickier. She said the office is developing a policy around marijuana cases now.
"We are working on developing our marijuana policy. It is my belief that people who are involved in retail sales of marijuana on the black market certainly do not belong behind bars. But I am a mom. I am concerned about protecting young people from marijuana. Marijuana is devastating on developing brains," Harrington said.
She added, "we have had some really violent crimes in our community that came about starting as marijuana deals. It is not without its danger. It is a complicated issue. I don't want people in jail because of marijuana but we are also working on how we can protect the public."
Harrington spoke and answered questions with members of the NAACP for more than an hour Wednesday night.
Local NAACP President Dennis Powell thanked Harrington for having the conversation and said it is important that the NAACP has a continual dialogue with elected officials moving forward.
"It is really important in really changing the narrative at the state house as well," Powell said.
Tags: criminal justice, district attorney, harrington, naacp,
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text:
Separate names with a comma.
Discussion in 'Challenge Pad' started by londoncuppa, Jul 21, 2017.
do we need to upload each card , each day? single or on the template?
Here's the link to the thread, Cath. It has all of the information, along with the downloads. If you have any questions after you read it, just let me know in that thread or send me an IM. We're having the kick off chat tomorrow. Thanks for much for your interest. I hope you'll play along with us!


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This program will set up your camps and events. An event can be a lesson, clinic or any activity where you need a schedule. You can also use the Daily Sheet function to create handouts, lesson plans, flyers, etc. You can then either print or email them as .pdf files. Here are some samples:
Camp Schedule
Participant Handout
Lesson Plan
Event Flyer


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Peter C. Beirne
Peter Beirne helps victims of asbestos exposure obtain justice from the corporations responsible for causing them to suffer life-threatening harm. He is an experienced advocate in the courtroom, adept at puncturing the claims and deflating the arguments those defendant corporations make in their bids to escape liability.
Mr. Beirne joined Weitz & Luxenberg in 2015 and works with the Asbestos Litigation team in our Los Angeles office. He began the practice of law in 2004.
Mr. Beirne first worked as an associate attorney in a Los Angeles law firm that focused its practice on mass tort litigation. He subsequently joined another Southern California firm that handled, among other things, personal injury cases.
Later, Mr. Beirne opened his own law office. His practice areas included personal injury, civil rights, criminal defense, employment law, entertainment law, intellectual property, business litigation and probate. In 2012, he closed his solo practice to join another law firm where he began concentrating on representing clients injured by asbestos.
Prior to gaining admission to the California bar, Mr. Beirne studied law at Notre Dame Law School. While attending Notre Dame, Mr. Beirne served as treasurer, and then president, of the school's Irish Law Society. During his final year at Notre Dame, he also held the post of the law school's Honor Code prosecutor.
Mr. Beirne was notably active in moot court: he served on the school's Moot Court board, was a member of the 7th Circuit Appellate team, and participated on the Barristers National Mock Trial team. He found his moot court experiences so rewarding that he later accepted an invitation to act as mock-trial coach for students at South Pasadena, California, High School from 2006-07.
Year Joined Firm
- 2015
Areas of Practice
- Mesothelioma and Asbestos
Bar Admissions
- California, 2004
- U.S. District Court Central District of California, 2004
- U.S. Court of Appeals 9th Circuit, 2004
- Notre Dame Law School, Notre Dame, Indiana
- J.D. - June, 2003
- Honors: Secretary, Irish Law Society, 2001-2002
- Honors: President, Irish Law Society, 2002-2003
- Honors: Notre Dame Barristers Trial Team, 2001-2002
- Honors: Member, Notre Dame Moot Court, 2001-2003
- Honors: Member, Notre Dame 7th Circuit Appellate Team, 2003
- Honors: Notre Dame Student Bar Association Honor Code Prosecutor, 2002-2003
- College of Charleston, Charleston, SC, USA
- Major: Theater
- Employment Law and Legal Issues regarding Business Operations and Employee Relations, California TCM University, 2010
Honors and Awards
- Los Angeles Rising Star Attorney, Super Lawyer Magazine, 2005
Professional Associations and Memberships
- Pasadena Bar Association, Member, 2006 - Present
- Los Angeles County Bar Association, Member, 2004 - Present
- State Bar of California, Member, 2005 - 2006
- American Bar Association, Member, 2002 - Present
- California Employment Lawyers Association, Member, 2004 - Present
- Irish Law Society, Secretary, 2001 - 2002
- Irish Law Society, president, 2002 - 2003
- California Bar Association
Past Employment Positions
- Matthews & Rager, Associate Attorney, 2005 - 2006
- Girardi & Keese, Associate Attorney, 2004 - 2005
Ancillary Businesses
- Entertainment Contract Preparation
- English


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Principal's Message
Tēnā koutou i runga i ngā moemoeā me ngā tumanako o te tau hōu. Ka tangihia, ka mihia, ka poroporoakitia ki ngā mate huhua kua wheturangitia. Haere atu rā, moe mai, okioki e. Kāti ngā pitopito kōrero mo rātou mā, kei te mihi, kei te mihi, kei te mihi.
We sincerely hope that all of our whānau have had a great break with friends and family, and we welcome everyone back to a very positive year at Allandale School.
We've had a wonderful start to the new year with lots of happy children re-uniting with their friends and getting to know their teachers. It was great to see so many new faces at our pōwhiri this morning! We've had a large number of new enrolments to start the school year and our roll is currently just over 400 students.
Kia tau te rangimārie,
Drew Manning (Principal)
Allandale Primary School
Allandale Primary School
167 King Street,Whakatane, 3120
Ph: (07) 308 7645
Cell: 021 025 01768
Fax: (07) 307 0868
Email: email@example.com


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Included in your booking
Don't see an amenity you're looking for? Ask the host, Michelle
Don't see an amenity you're looking for? Ask the host, Michelle
Location Description: Multiple sections of the venue are available for rent. Outdoor area is perfect for any open-air networking events. Access to multimedia LED screens make it perfect for any music video, film/photo shoots or as the backdrop to a panel series. Office spaces and conference also available for corporate meetings or company functions. Exterior has a huge parking lot area organizers can rent out for additional fees. Street metered parking also available. Amenities include: WiFi, indoor conference room, projector, whiteboard, Parking Lot and Street Parking, Outdoor, kitchenette, Handicap Accessible, Pet Friendly. Additional Services: We can provide tech engineers and custom motion graphic design services for additional fees. Mandatory one tech engineer to operate LED Glass walls. Tables, chairs and other furniture rentals available for additional fees. Space is located in a gated, safe office building complex in the heart of financial district in Downtown Los Angeles. Due to the shared nature of our building, at least one staff person must be on site during your event at the cost of $30 hour until load out. Amenities WiFi, Chairs, Multimedia video walls, Tables, Picnic Benches, Projector, Stage Lighting, AV Equipment, Outdoor Area, Wheelchair Accessible, Restrooms, Street level access ADD ON AMENITIES FEES: Coffee & Tea: From $6-10/person Meals & Snacks: From $10-45/person Security: From $30-55/hour Production Assistant: From $20-50/hour Tech Engineer: From $40/hour Motion Graphic Designer: From $40/hour
We follow Peerspace's cancellation, Overtime and Insurance policies.
All ages
Guests may cancel their Booking until 7 days before the event start time and will receive a full refund (including all Fees) of their Booking Price. Guests may cancel their Booking between 7 days and 24 hours before the event start time and receive a 50% refund (excluding Fees) of their Booking Price. Booking cancellations submitted less than 24 hours before the Event start time are not refundable. Learn more
It was such a great experience shooting here. Michelle was absolutely awesome. She was with us the whole time, and always on the requests that we have. They offered their LED screen for our shots, and even helped us to edit the videos we wanted to play. We will definitely recommend this location to other projects. Thank you!


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Health Insurance FAQ'S
Health Insurance FAQ
I lost my job and my health insurance, what options do I have?
In most states you have the option to apply for COBRA through your previous insurance company.
Often we are able to find an individual or family health insurance plan at rates that are better than COBRA. Be sure to ask
your agent about your options.
My COBRA runs out at the end of this month, is there anything I can do?
Many people are eligible for individual or family plans at prices that are better than COBRA.
We have "Guaranteed Issue" plans for individuals who have completed 18 months of coverage under Cobra
I own my own business but don't have any employees. What kind of health insurance is available?
A business owner can choose from individual, family, or group
health plans
"Business Group of One" is particularly appealing to individuals who have a medical history that wouldn't qualify for
individual insurance.
Do you pull a
credit report
when looking for insurance quotes?
For health insurance, insurance companies and agents do not generally pull
credit reports
while providing quotes insurance.
What other benefits do your policies offer?
As an independent agency, we don't work with just one company. Each company provides a myriad of benefits and options. If you
are having trouble navigating these waters, please take time to talk with one of our agents.
Many of our companies offer additional benefits to their policies. Including but not limited to: Supplemental Death Benefits,
ID Theft Protection, and single deductibles.
Does it cost more to work with a broker?
In most states, brokers are not allowed to charge any additional fees. This means that the rate the insurance company filed
with the state is the price you pay.
How much coverage do I need?
How much could you afford to pay out of pocket if:
You went to the doctor?
You had to purchase medicine?
You had to stay a night in the hospital?
You broke a bone?
You found out you had a serious illness?
These are all things to consider when purchasing your health insurance. Every policy provides different levels of protection
for different events. If you have any questions, please consult with one of our licensed agents.
Loveland Office: 970-222-7202
Designed and created by Angela DiMichele
Leivestad Agency
Denver Office: 303-500-3386
Connect to us:


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Friday 26th October – 18:00 / Verdensteatret
Konrad "Conny" Plank is undoubtedly one of the most innovative sound designers of his time. The recordings realized in his sound studio near Cologne between the 60s and 80s, revolutionized the music world. He was the pioneer of "Krautrock" and paved the way for the revolutions of electronic and pop music respectively. Bands and artists who were fathered by the sound wizard (mostly before they became world famous) all agree that their careers would have been different without the talents of Conny Plank. Kraftwerk, Neu!, Can, Whodini, Brian Eno, DAF, Gianna Nannini, Ultravox and the Eurythmics are just some to be named.
Conny Plank died much too early aged 47. For his wife, the actress Christa Fast, and his 13 year old son Stephan, he left behind a sound studio known near to everywhere. 25 years later his son embarks on a journey to find his father, his importance and his impact. Because all these musicians know Stephan from a young age from the studio, this film brings an unusually intimate and emotional insight to the artists, their work with Conny Plank and all that defines music production.
The documentary is the search for the person and producer Conny Plank, a journey through music history, a son getting to know his father and a soundtrack that has influenced millions of music fans over three decades.
Directed by Reto Caduff and Stephan Plank, CONNY PLANK: THE POTENTIAL OF NOISE is presented in collaboration with c/o Pop in Cologne, as part of We Are Europe.


text: Building a Feadship is seen as the pinnacle of super yacht ownership. Every Feadship is entirely custom designed and built by hand to suit the wishes of the owner, without any limitation. We created a platform that would capture this story from every angle: from the design process to the finished product and humbling passion and craftsmanship of the workers.


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John Scalzi, a science fiction writer whose blog is one of my favorites, wrote a couple weeks ago about the issue of piracy in the book industry. Summing up his position, Scalzi thinks that worrying about piracy is a stupidity.
He sees two categories of book pirates:
First, the idiots, who won't buy the books (in electronic or physical form) whether they have the money or not. They are simply not interested in paying for the goods.
Second, the occasional pirates, who download books because they don't have the money to pay for the books they want to read. The reasons are many: they are cash-strapped college students, the books are not physically available where they live and it would be too expensive to import, and so on. Those persons will likely start paying for books when money starts flowing towards them, especially books from writers they learned to appreciate when they couldn't still pay for their books.
I agree with Scalzi. I know people who fit both categories and they are easily distinguished. As an avid reader, I once belonged into the second category, although I now rarely download a book I don't own physically (except, of course, in the case of e-books). When I do, it's often because I need to find something quickly in the book or solve some problem and it's easier to "pirate" a electronic version temporarily than manually search five hundred pages for a specific sentence.
Also, when I belonged into the second category for the main reason Scalzi cites (wanted to read the books, couldn't pay for them — especially where newer books where concerned and the only other available choice was spending half my weekly wages in a single three-hundred-page book), I kept a list of the books I downloaded, and have bought most of them by now. The ones I haven't bought yet are likely to be published in Brazil soon, and I'd rather buy them here to minimize costs (I'm still not rich, you know).
Anyway, what I find most interesting in the whole discussion is how some authors who otherwise share the same views on the genre take directly opposing views on the issue of piracy. Some believe is the rise of e-books as the main form of publishing is quite inevitable. Others, like Harlan Ellison, are prepared to fight tooth, nail and processes at the mere mention of their books in any kind of digital format (as if paper would somehow preclude their works from being pirated).
Which is a pity. I would gladly surrender my money to some writers were they willing to understand this new market — which, in my opinion, has little to do with the issue of intellectual property. The Long Tail is a reality in the Internet, and, as such, is beyond immediate frontiers. Those writers — Harlan Ellison, for example, is quite technophobic — think it's possible to prevent their works from being distributed in P2P networks. More than that, they ignore the fact that many of their readers would pay for electronic versions of their works if only the price were realistic, adjusted to the realities of the medium (easy replication, limited availability, etc.) and market demands, as Fictionwise has shown.
Scalzi also talks about his own experience (shared by many people I know) of searching for ways to monetarily compensate writers for reading their books by indirect means — buying them as used books or lending them from libraries, for example, especially when the books were out of print. In my experience, this kind of behavior is mostly limited to readers of science fiction, fantasy, and horror, but serves to show that the undiscriminated bashing of those "pirates" is really excessive.
Obviously, there are other pertinent issues that I'm not considering here, that people like Cory Doctorow and others far more intelligent than me are talking about, and this entry is not the place to write about them. But I'm happy to see writers openly declaring their disdain for the stupidity demonstrated by members of the industry and other writers who can read the writing in the wall.


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Translational control mechanisms in cutaneous malignant melanoma: the role of eIF2α
- Immacolata Maida†1,
- Paola Zanna†1,
- Stefania Guida2Email author,
- Anna Ferretta1,
- Tiziana Cocco1,
- Luigi Leonardo Palese1,
- Paola Londei3,
- Dario Benelli3,
- Amalia Azzariti4,
- Stefania Tommasi4,
- Michele Guida4,
- Giovanni Pellacani2 and
- Gabriella Guida1Email author
© The Author(s) 2019
- Received: 21 September 2018
- Accepted: 2 January 2019
- Published: 11 January 2019
Melanoma cells develop adaptive responses in order to cope with particular conditions of tumor microenvironment, characterized by stress conditions and deregulated proliferation. Recently, the interplay between the stress response and the gene expression programs leading to metastatic spread has been reported.
We evaluated levels and localization of eIF2α/peIF2α in V600BRAF and wtBRAF metastatic melanoma cell lines by means of western blot and confocal microscopy analyses. Furthermore, we performed a sequence analyses and structure and dynamics studies of eIF2α protein to reveal the role of eIF2α and its correlations in different pathways involved in the invasive phase of melanoma.
We found peIF2α both in cytoplasm and nucleus. Nuclear localization was more represented in V600BRAF melanoma cell lines. Our studies on eIF2α protein sequence indicated the presence of a predicted bipartite NLS as well as a nuclear export signal NES and an S1 domain, typical of RNA interacting proteins. Furthermore, we found high levels of transcription factor EB (TFEB), a component of the MiT/TFE family, and low β-catenin levels in V600BRAF cells.
Based on our results, we suggest that peIF2α nuclear localization can be crucial in ER stress response and in driving the metastatic spread of melanoma, through lysosomal signaling and Wnt/β-catenin pathway. In conclusion, this is the first evidence of nuclear localization of peIF2α, representing a possible target for future therapeutic approaches for metastatic melanoma.
- Metastatic melanoma
- eIF2α
- MiT family
- Nuclear peIF2α
- Structural analysis
Melanoma is one of the skin tumors with increasing incidence in the general population [1]. It is a particularly severe form of skin cancer due to its ability to metastasize, even several years after the removal of the primary melanoma, and to its resistance to medical therapies in case of metastasis. One of the peculiar characteristics of melanoma is the involvement of several molecular pathways in its development and progression. This feature is associated to diverse escape mechanisms to target therapies (i.e. anti-BRAF) for the metastatic disease.
The most recent knowledge of melanoma biology focuses on the role of endoplasmic reticulum (ER) stress, autophagy and translational reprogramming. In brief, tumor microenvironment conditions, characterized by stress stimuli and a push to fuel continue proliferation, induce the development of cellular adaptive responses. In particular, these stress signals induce the activation of the unfolded protein response (UPR) [2–4], and translational block, in order to avoid the accumulation of proteins entering the ER. The translational block is achieved through the phosphorylation of the alpha subunit of eukaryotic translation initiation factor 2 (eIF2α) which is one of the key regulator of the Integrated Stress Response (ISR). ISR is a common adaptive pathway, activated in response to diverse stress stimuli, in order to restore cellular homeostasis [5]. The central core of ISR response is eIF2, a heterotrimeric protein constituted by α, β and γ subunits, an essential factor involved in the initiation phase of eukaryotic translation. eIF2 is a G-protein that, in its active GTP-bound form, interacts with Met-tRNAi promoting its binding to the 40S ribosomal subunit and determining the correct localization on the start codon. The codon-anticodon interaction triggers GTP hydrolysis, whereby an inactive eIF2-GDP complex is released. eIF2 reactivation is catalyzed by the recycling factor eIF2B that allows GDP/GTP exchange. Indeed, the recycling of the factor can be inhibited by the phosphorylation of its alpha subunit at Ser 51. In its phosphorylated form, eIF2 behaves as a competitive inhibitor of eIF2B, preventing GDP/GTP exchange and thereby reactivation of eIF2. eIF2 phosphorylation is triggered by various stress conditions, such as viral infection or nutrient deprivation, which lead to the activation of specific kinases (PKR, PERK, GCN2, HRI). The phosphorylation of eIF2α determines an increase in the expression of the transcription factor ATF4, a key component of the ISR, in order to promote cellular recovery. ATF4 is a transcription factor of the ATF/CREB family that regulates the expression of genes involved in oxidative stress, amino acid synthesis, differentiation, metastasis and angiogenesis and it is induced by stress signals including anoxia/hypoxia, ER stress, amino acid deprivation, and oxidative stress [6].
There are evidences that eIF2α expression levels and phosphorylation status increase in tumours compared with normal tissue [7–11]. In melanoma, it has been previously demonstrated that in V600BRAF metastatic melanoma cell lines eIF2α is more phosphorylated, respect to wtBRAF melanoma cells [12].
A recent study [13] shows the important interplay between the ISR and the gene expression programs leading to metastatic spread, regulated by MITF [13]. MITF is a transcription factor that drives the expression of genes involved in cell differentiation, proliferation and survival. It plays an important role in melanocyte development by regulating the expression of genes implicated in melanin production and its levels have been associated to melanocytes transformation to melanoma [14, 15]. One of the components of the MITF/TFE family of transcription factor [16] is TFEB, well-known for its key role in autophagy activation and recently proposed as a master regulator of ISR [5].
Several pathways involved in cellular proliferation, translational reprogramming, together with molecular adhesion variations (i.e. β-catenin), seem to be the key events that drive gene expression to an invasive phase of melanoma [12, 13]. Because of its pivotal role in translation initiation, eIF2α is the main target of translational regulation mechanisms. Thus, the aim of the current work is to perform a detailed evaluation of eIF2α/peIF2α levels and location. In addition, we estimated the correlation of eIF2α with different pathways involved in the invasive phase of melanoma in an in vitro melanoma model.
Materials and methods
Cutaneous melanoma cell lines
Hmel1 and M3 were metastatic melanoma cell lines extracted in our laboratory and proceeding from patients coming from the south of Italy. These cell lines have been described in Zanna et al. [17], and Ferretta et al. [12]. Hmel1 and M3 represent the metastasis harboring V600BRAF mutation. HBL is a metastatic melanoma cell line, kindly provided by Ghanem Ghanem. It was used as control cell line because it is wt for BRAF. In addition, hmel9 cells, non-metastatic V600BRAF-mutated, previously described [12, 17] were used.
Western blots and cell fractioning
Cells were lysed as described in Herraiz et al. [18]. Bicinchoninic acid reagent, from Sigma Aldrich (St. Louis, MO), was used to quantify proteins. Electrophoresis and Western blots were performed as described in Ferretta et al. [12]. 30–50 µg of the cell lysate were loaded on a SDS-PAGE (10% or 13% for TFEB) and transferred to a nitrocellulose membrane (GE Healthcare, Little Chalfont, UK) for Western Blot analysis. The polyclonal antibodies, anti-eIF2α and anti-peIF2α Ser 51 were purchased from Cell Signaling Technology (Danvers, MA); the polyclonal anti βcatenin antibodies and the secondary antibody-HRP conjugated were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). The mouse anti Flag M2 monoclonal antibody was from Sigma. TFEB antibodies and the anti-mouse HRP secondary antibody were from Millipore (Burlington, Massachusetts, US). Protein loading was assessed reprobing the blots with anti-βtubulin antibodies (Sigma Aldrich, St. Louis, US). All the antibodies were used according to the manufacturer's instruction. Chemioluminescence was detected using the Super signal western Pico Pierce or the chemiluminescent Pierce ECL Western Blotting Substrate (Thermo Scientific, IL). The densitometric profiles were performed using a Image lab 5.2 imaging software (Bio-Rad Laboratories) or Bio-rad GS 800, and analyzed by the Quantity One Software, all purchased by Bio-Rad Laboratories (Hercules, CA).
The M3 cell fractioning was performed as previously described in Zanna et al. [19], obtaining nuclear and cytoplasmic fractions used for western blot analyses. The purity of the nuclear and cytosolic fractions was verified by the presence of βtubulin immunoreactive band after immunoblotting with anti-βtubulin antibody).
Confocal microscopy
Cells grown on coverslips and washed with PBS, fixed with 4% paraformaldehide, and blocked with 20 mM glycine. Cells were permeabilized with 0.5% Igepal CA-640 in PBS. Cells were stained with anti-eIF2a and the anti-peIF2α diluted in PBS containing 1% BSA. Goat anti-rabbit Alexa 488 conjugated antibody Molecular Probes (Invitrogen, CA) in PBS/1%BSA was used. Incubations were performed at 4 °C. Samples were examined with a Nikon confocal microscope (Nikon Corporation, Tokyo, Japan).
eIF2α cloning in a pCMV-flag vector
eIF2α molecular cloning was performed on pCVM-flag (Sigma Aldrich, St. Louis, MO). eIF2α coding sequence was obtained by M3 cDNA by PCR using a proofreading polymerase (Kapa Hifi, Roche, Basilea Switzerland) and the primers eIF2αHINDIIFw (5′-CGCAAGCTTCCGGGTCTAAGTTGTAGA-3′) and eIF2αXbaIRv (5′-CGCTCTAGACACAAAGTTAATCTTCAGC-3′), purchased from Sigma. The PCR product and pCMVflag were digested for HindIII and XbaI restriction enzymes (Promega, Wisconsin, US), loaded on a 1% preparative agarose gel and eluted using the extraction kit purchased by RBC Biosciences (New Taipei, Taiwan). The ligation of the pCMV vector with the 947 bp PCR eIF2α was performed o.n. at 4 °C using a T4 DNA ligase (Promega) according to the manufacturer's instructions. Ligations were used to transform Top10 chemically competent E. coli (Thermo Fisher Scientific, Waltham, Massachusetts, US), the colonies were selected and minipreps were screened by digestion with HindIII and XbaI. The pCMVflag and pCMVflag-eIF2α were used to transfect 293T cells using lipofectamine 2000 transfection reagent (Invitrogen, Meridian Rd, Rockford, IL), according to manufacturer's instructions.
eIF2α structural analysis
The solution NMR structure of the alpha subunit of human eIF2 1Q8 K [20] was obtained from the RCSB PDB [21] available at http://www.rcsb.org. From each of the reported models in the 1Q8 K entry a separate pdb file was obtained by Tcl scripting in VMD [22]. These pdb structures were processed as described in [23, 24]. Briefly, structures in the data set were aligned to a reference by a rotation and translation matrix, by a Tcl script in VMD. The Cartesian coordinates of alpha-carbon atoms of the rotated structures were extracted and arranged in a matrix form in which each row represents a structure, and principal component analysis (PCA) was performed on the normalized data [25–28]. To visualize the obtained principal displacement modes encoded in the whole 1Q8K PDB entry, a nmd file was constructed [23, 24], which was analyzed by the VMD plug-in NMWiz [29].
The prediction of nuclear export signals (NESs) in the sequence of the alpha subunit of human eIF2 has been performed at the NetNES 1.1 Server [30]. Protein–protein interactions were obtained from the BioGridrepository [31, 32].
The prediction of eIF2α nuclear localization signals (NLS) were performed using the program cNLS Mapper (available at http://nls-mapper.iab.keio.ac.jp/cgi-bin/NLS_Mapper_form.cgi) using a cutoff 4. An additional analysis of the eIF2α functional domains has been performed using the SMART program (http://smart.embl-heidelberg.de/).
peIF2α localization in melanoma cell lines
As reported in Ferretta et al. [12] the non-metastatic V600BRAF hmel9 cell line, showed a lower level of peIF2α, as compared to metastatic melanoma cells [12]. Therefore, we extended the analysis of peIF2α cytoplasmic and nuclear localization to hmel9 cell line, finding lower nuclear level of peIF2α in non-metastatic V600BRAF hmel9 cells, as compared to metastatic V600BRAF M3 cells (see Additional file 1: Fig. S1).
eIF2α sequence analysis and eIF2α-flag expression and localization in 293T transfected cells
A further evidence of eIF2α nuclear localization was achieved generating a construct in which eIF2α was expressed in the pCMV-flag vector under the control of the cytomegalovirus promoter and transfected in 293T cells. We analyzed the total extracts of the 293T transfected with the pCMVflag-eIF2α expression vector in comparison with 293T cells transfected with the empty pCMV-flag vector. The exogenous eIF2α protein was expressed and recognized by western blotting using anti flag M2 antibody (Fig. 2b). To detect if eIF2α was transported from the cytoplasm (site of translation) into the nucleus, we isolated transfected 293T cytoplasmic and nuclear fractions (Fig. 2c). We performed a western blot using the anti-eIF2α antibody and we detected eIF2α-flag either in the nucleus and in the cytoplasmic fraction of the 293T transfected cells. The eIF2α-flag is expressed and recognized by the anti-eIF2α antibody (total form) like a slightly higher molecular weight protein, due to the presence of the flag, with respect to the endogenous eIF2α as shown in M3 cell line used as control (Fig. 2c). The fractions purity was determined using βtubulin as cytoplasmic marker (Fig. 2c).
eIF2α structure and dynamics
TFEB and βcatenin expression analysis
We have already reported lower levels of βcatenin in various V600BRAF melanoma cell lines as compared to HBL wtBRAF cells [19]. In order to estimate β-catenin levels also in M3 cell lines, western blotting analyses were performed. In Fig. 4b we show a 2-times lower levels of βcatenin in V600BRAF hmel1 and M3 cell lines as compared to HBL cells.
In this study, we analyzed phosphorylation, localization and conformational variations of the eIF2α subunit in metastatic melanoma cell lines with different BRAF mutational status, evaluating potential correlation with other pathways involved in the invasive phase of melanoma.
We have previously shown that peIF2α level is higher in cell lines proceeding from skin metastases (hmel1 and M3) than in cell lines from non-metastatic skin lesions (hmel9 and hmel11), harboring the V600BRAF mutation [12]. As a matter of fact, increased levels of peIF2α have been related to a worst melanoma prognosis [12] and to the invasive phase of melanoma [13].
The most striking result of the current work is the finding, in metastatic cell lines, of a nuclear localization of peIF2α, as compared to non-metastatic cells (Additional file 1: Fig. S1).
Since eIF2α is well characterized as a translation initiation factor, one would expect to find it in the cytoplasm and not in the nucleus. Nevertheless, another translation initiation factor, eIF4E, has already been reported to have a nuclear localization [8, 38–40]. During conditions of low nutrient availability, eIF4E can be sequestered by eIF4E-binding proteins (4E-BP), thus limiting the assembly of the eIF4F complex [41]. The nuclear import of eIF4E is mediated by the eIF4E-transporter, binding both eIF4E and the nuclear import receptors, importin α/β [38, 42]. In the nucleus, eIF4E is known to promote the nuclear export of a set of mRNAs [42–45].
In this study, the nuclear localization of eIF2α is corroborated by the finding that the protein sequence contains signals for both import in, and export from, the nucleus. Furthermore, structural studies indicate that these signals are exposed and available in the phosphorylated form of the protein (Fig. 3 and Additional file 2: Model 1, Additional file 3: Model 2), justifying the observation that the nucleus is especially enriched in peIF2α. Indeed, recent data suggest that the two domains of the eIF2α may move toward or away from each other, switching from a closed to an open conformation. However, the closed conformation is disrupted by phosphorylation of S51 [34], keeping the protein in a conformational "opened" status that exposes the nuclear signals necessary for the entrance and the exit from the nucleus. In addition, our results suggest that the transport of this factor out of the nucleus could be due to the interaction with NES sequence of the same factor with XPO1, as recently reported experimentally by Kirly [35], a well-known nuclear exporter of cellular proteins and RNAs.
The significance of peIF2α in the nucleus is still unclear. However, it is known that eIF2α can be phosphorylated by several kinases, activated in order to counteract adverse cellular conditions in tumors. Accordingly, tumor cells develop adaptive responses in order to cope with particular conditions of the tumor microenvironment, characterized by stress stimuli and a push to fuel continue proliferation. These stress signals induce the activation of the unfolded protein response (UPR) [2–4], which enables the accumulation of misfolded or unfolded proteins in the ER. Responses include an enhanced protein refolding, degradation of misfolded proteins, and translational block, in order to avoid the accumulation of proteins entering the ER. Therefore, ER stress, leading to eIF2α phosphorylation, inhibits eIF2B, which blocks the assembly of the ribosome translation initiation complex, thus reducing the rate of translation initiation [13, 46]. We have already demonstrated that eIF2α phosphorylation and ERK phosphorylation, due to constitutive activation of BRAF signaling in V600BRAF-mutated cell lines, are correlated. Furthermore, we also found different MITF levels in melanoma cell lines harboring BRAF mutations, as compared to wtBRAF. Accordingly, lower MITF levels in V600BRAF cells [12] (Fig. 3a) can be explained by the recently described transcriptional/translational coupled feedback loop in which MITF increases ATF4 transcription, and peIF2α promotes its translation. After that, MITF transcription and translation are blocked, respectively, by ATF4 and peIF2α-mediated inhibition of eIF2B [13].
On the other hand, ATF4 upregulation is also induced by TFEB, a component of the MITF/TFE family of transcription factors [16]. TFEB has a pivotal role in ER stress response and autophagy induction [47]. The high TFEB expression in V600BRAF metastatic melanoma cell lines found in the present work can be correlated with the high LC3II/LC3I ratio reported in Ferretta et al. [12], in the same cell lines, supporting the role of TFEB in autophagy.
Since MITF/TFE factors can regulate lysosomal signaling, including Wnt/β-catenin [36], we analyzed β-catenin levels. We found low β-catenin levels in V600BRAF cell lines, which can be correlated to the low MITF levels found in the same cell lines, as reported in Ferretta et al. [12]. Literature reveals that signaling through β-catenin enhances MITF expression. On the other hand, MITF, depending on its protein levels, can interact with β-catenin (alone or in a complex with Lef-1) to activate downstream targets [48]. The presence of low levels of β-Catenin, associated with low MITF levels in V600BRAF melanoma cell lines, could be linked to autophagy activation in these cell lines [49], as proved by the high TFEB levels revealed in this study. Interestingly, I.J. Davis group reported that TFEB can rescue MITF knockdown in cell sarcomas [50]. According with this study, our results indicate that MiT family members can compensate each other for oncogenic response in V600BRAF cells.
Translational reprogramming has been shown to drive gene expression programs leading to metastatic spread [13]. Therefore, eIF2α could affect the transcription of specific genes, and/or mediate the transport of specific mRNAs to the cytoplasm. In addition, eIF2α possesses a putative RNA-binding domain, and it has been suggested to interact with RNA polymerase [51]. Further studies will be performed to evaluate the nuclear eIF2α activity and to improve the emerging knowledge relating translational reprogramming to the invasive phase of melanoma, in order to contribute to the development of new target treatments for metastatic melanoma.
This is the first report of the nuclear localization of peIF2α in metastatic melanoma cell lines.
Our structural studies of eIF2α protein sequence show the presence of a predicted bipartited NLS as well as NES and an S1 domain. These findings suggest its entry and exit in/from the nucleus and its role as a modulating factor of oncogenic pathways, especially implicated in establishing and maintaining a metastatic phenotype.
Taken together, our results underpin the key role of peIF2α nuclear localization in ER stress response and in the invasive phase of melanoma.
Authors' contributions
The idea of this work IM, PZ, GG. PZ, AF, DB performed the experiments while LP performed the bioinformatic analysis. SG, TC, PL, AA, ST, MG, GP analyzed and interpreted, based on different expertise, the results. IM, PZ, SG, TC, LP and GG wrote the manuscript, that was finally approved by all authors. All authors read and approved the final manuscript.
We would like to thank Prof Paola Londei for hosting Dr Paola Zanna in her laboratory in Rome (Sapienza University) and Mary V. Pragnell for language editing.
Competing interests
The authors declare that they have no competing interests.
Availability of data and materials
The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.
Consent for publication
Not applicable.
Ethics approval and consent to participate
Procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2000.
This work was supported by grants from the University of Bari.
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Authors' Affiliations
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Heartworn Highways, James Szalapski, USA, 1976, 92 min., English
Released only briefly on VHS and passed around or talked up via word of mouth, the 70s cult classic Heartworn Highways intimately explores the roots of outlaw country music, offering a rare insight into the lives of the musicians who set the bar for those who came after.
Final Cut is a documenta 14 film program that takes place at Filmladen Kassel, a small arthouse cinema, during the final week of the exhibition. Daily from September 11–17, beginning at 8:30 pm, it presents a selection of riveting yet seldom-seen films compiled by Artistic Director Adam Szymczyk.
If not stated otherwise, all screenings show original versions with English subtitles.
Entrance fee: 7.50 € / reduced 7 €. Tickets for the screenings can be purchased from September 7 or on the day of the screenings, daily from 5 pm at Filmladen Kassel. Reservations are not possible.


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Reading level: grade 4. Interest level: ages 9-12. With black humour and touching moments, young Anna recounts the tale of the summer she finally got 2 pet hamsters, and her ensuing investigation into how the hamsters were massacred in the night. (Great Critter Capers series)
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For those who aren't quite up to date with the history of Apple, Steve Jobs left the company in 1985 and for a decade, went through quite difficult times. Doug Menuez had the chance to take some rarely seen photographs of Steve Jobs during that period and shares them here:
What is striking about those photographs is the way they tell a story—different when compared to the one about Jobs we all know. It is one of those scenarios where the picture speaks a thousand words.
In the first image Menuez shares, even though it is mostly obstructed by out of focus elements, you can see just enough of the facial expression and that deep glare in his eyes that puts all of the distractions in perspective and simply makes the image complete.
Doug Menuez had the exact intention, as he says:
"He kinda went into his own world briefly. And that is when I took the picture, when I saw him almost meditating."
This next image is again all about the eyes, but this time it is the hands that tell the story, obstructing a whole lot but leaving just enough room for the eyes to pierce through the fingers. It's excellent visionary portraiture.
The next few images are messy if you don't know what you are looking at, but once you realize what's going, on everything falls into place.
In this final image, it's obvious that Steve Jobs is ecstatic, but that wouldn't be so emphasized if there weren't the shared emotions of the other people in the frame to multiply the effect. Yes, they are out of focus but it is enough to feel the joy that they all share. It seems as if he is on the top of the world, and in that moment he probably felt that way, as well.
All of these photos are classic black and white photographs. There is no color to guide your eye. Here it is all about the framing, context, emotion, and the moment. The framing might not be perfect, nor the light. But everything else is executed in a maestral manner that it makes these images iconic and storytelling.
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Fighting six captains in a row (and decorating your wall with their heads) confirmed
Blitz Royale Arena Progress 04
by Adam
Slowly developing this Blitz Royale Arena into something neat. The latest additions even provide a hint to our plans for Hex Gambit's single player mode. When we shift our focus to single player full time, we'll tell you all about it!
Check back weekly for more progress on our upcoming turn-based strategy game, Hex Gambit. Buy a copy today to save 10% on our $14.99 launch price, then give us some feedback to help shape the final product! If you haven't already, be sure to sign up for our mailing list to get an email reminder for the official release on PC later this year.
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@Harti: I think you should know that all the colorful banners and lights that are happening to my maps stem from your feedback a while ago about everything being very gray and stony.
Continue the discussion talk.onemanleft.com


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In year 2000, after Kiehl's was acquired by L'Oreal, Kiehl's has swiftly expanded its brand presence into 37 countries across five continents. Its omnipresence proves that Kiehl's is truly an international brand.
Although internationalization provides brand opportunities to cut marketing cost and capture more customers, some challenges are unavoidable when brands go global. Since the brand entered Asia market, Kiehl's has been enjoying double digits growth. However, concerns about whether Asian consumers are able to appreciate its brand culture and to what extent that Kiehl's should customize in order to better meet Asian customers' needs, have existed all the time.
Overall, our group feels that Kiehl's should maintain a high level of standardization when expanding into different countries. As Kiehl's has been positioned as a luxury yet cult brand – which is highly accepted and valued by its customers – too much customization will eventually diminish its cultivated cult image. Kiehl's iconic symbols, such as medicine bottles-looking packaging, Mr. Bones and "lab ambassadors" enable the brand to stand out from the clutter. While these symbols may appear irrelevant to the masses, they certainly CAN and WILL attract those who are drawn to quirkiness.
Why Brand Extension?
Strong brand names deliver high sales and profits. As an established cult brand, Kiehl's can leverage on its brand strength to launch new products/services in a new category: there is lesser need to communicate awareness and imagery, since consumer knowledge is prevalent.
Kiehl's can harness on its strong "anchor" core product – its expertise in pharmaceutical manufacturing and chemistry – to launch other brand extensions. There also needs to be a broader market definition – that Kiehl's is not just a skincare brand, but a lifestyle and wellness brand which offers beauty and spa services.
However, we acknowledge that Kiehl's might face the risk of brand dilution, especially since Kiehl's portrays a cult brand image. Hence, the following recommendations for brand/line extension have to serve the primary purpose of adding customer value – both functional and emotional benefits (badge/status value).
Starting out as a pharmacy dishing out its very own formulated skincare, Kiehl's has been dedicated to health and wellness of its customers' skin for over 160 years. Not surprisingly, Kiehl's decided to extend into the health & wellness category by opening a spa in New York which uses none other than their amazing range of apothecary-inspired products!
The spa, named Spa 1851, was opened in July this year and features three full treatment rooms, a separate men's grooming station and interiors that boast vintage reclaimed ceilings, antique and oak fixtures. Although Spa 1851 has yet to release official sales figures, our group feel that it is a promising initiative and Kiehl's should introduce such an outlet to Singapore as well. Apart from facials, Kiehl's could also offer body treatments and massages.
We propose targeting Kiehl's existing customers as they tend to be loyal to the Kiehl's brand and would be more willing to give the spa a shot. In addition, the spa may help the brand acquire new customers as it gives non-users a chance to try Kiehl's products by going for a one-time facial before deciding if they do want to purchase Kiehl's products. In this way, non-users who have not tried Kiehl's products due to various reasons (e.g. they are too intimidated by the store's appearance or are too shy to ask for samples) will have the opportunity to try the brand's products without feeling any embarrassment or discomfort.
We propose that Kiehl's continue to focus on skin customization for customers. As such, Kiehl's Skin Treatment Spa will offer a complimentary skin consultation with a Kiehl's staff assistant before choosing the most suitable treatment from a plethora of treatment types (e.g. moisturizing, skin repair, anti-ageing facial treatments etc.)
All Kiehl's staff will also be decked in lab coats, and the storefront of the spa will look similar to Kiehl's current stores to provide a consistent brand image.
Natural Wellness Café
Our group also recommends opening a natural wellness café in the same vicinity of our Kiehl's Skin Treatment Spa. The café will not solely serve to fulfil the basic needs of our customers but as the name suggest, our wellness café also seeks to bring out the wellness in people through a careful selection of light and healthy food such as fruits and salads and beverages such as fruit juices and tea.
Kiehl's natural wellness café will specialize in tea for its healthy benefits. Teas with its antioxidant properties have been known to lower cholesterol levels, stimulate the immune system, delay aging and fight high blood pressure, etc. There's an old adage "An ounce of prevention is worth a pound of cure". In our café, we want to offer well-being tea that is light, medicinal and yet flavorful to promote the overall well-being and vitality of our customers, thereby completing their experience with us.
Kiehl's first started out as a pharmacy producing skin care products using the most unique natural ingredients. As a new brand extension to Kiehl's, Kiehl's should take advantage of their pharmaceutical background and launch a new range of medical cosmetics!
We aim to target women with this new "medical cosmetics range" as studies have shown that women tend to be more self-conscious and are more proactive in seeking solutions to resolve their skin problems. However, we noted that most females ignore their skin problems and apply more make-up to hide their skin imperfections instead.
Therefore, Kiehl's medical cosmetics provide the perfect solution – make-up which heals the user's skin as well as covers up all imperfections, thus giving users all the confidence they need!
To promote the environmental friendliness of Kiehl's products, we suggest installing a "refill station" at the proposed "Natural Wellness Cafe." Currently, Kiehl's has a few refillable products, although they are not promoted extensively. Since hygiene is a concern to many, we recommend accepting the empty bottles from the customers and retain them for cleaning at Kiehl's refill stations. The customer who brought the empty bottle will then be given another recycled bottle (which has been cleaned) at discounted price. This exchange program encourages users to be more environmentally-friendly as well as enhances Kiehl's mission to be an eco-friendly brand.
Currently Kiehl's has products lines for skincare, body, men and hair. However, Kiehl's has not (or rarely) introduced cosmetic-related accessories. Since Kiehl's is a 'cult' brand, it could leverage on its strong customers' loyalty and on L'Oreal's expertise to introduce several skincare-related accessories such as hair brush, eye mask, bath gloves, skin sponge and moisturizing socks, etc.
While our group has come up with several product extension for Kiehl's, we certainly do not recommend Kiehl's to implement all of them at once. Instead, we propose that Kiehl's approach each product launch with care and in the following sequence:
Since the first (and only) Spa 1851 has just started operations in New York early this year, we reckon that it would take some time for the Kiehl's management to evaluate the spa's popularity and sales estimates before deciding whether or not to introduce it to consumers overseas as well. Thus, we propose that Spa 1851 to be launched at a later time.
Similarly, for Kiehl's Wellness Cafe, we suggest that Kiehl's launches it last as it will be an entirely new establishment and Kiehl's needs time to research on the various F&B items to feature in its cafe. Furthermore, Kiehl's has to factor in start-up costs and source for suitable vendors/partners before embracing this new business.
Overall, our group feels that as a cult brand, Kiehl's has come a long way and it definitely has the potential to grow further. Consumers are constantly inundated with various product advertising and may inevitably suffer "ad fatigue". However, with Kiehl's quirkiness and the brand's trusted reputation, we believe that Kiehl's will definitely stand out among its competitors! Moving forward, we hope to see Kiehl's adopt more exciting product extensions and we are definitely eager to see how our proposed brand strategies will impact the brand in future!


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This goes beyond appalling, cruel and hypocritical: viciously and cruelly beheading a innocent creature while making the argument to protect life?
Sorry, he is a total and unredeemable asshole.
Debate continues to rage between pro-life and pro-choice activists about when life begins and must be protected.
One topic that hasn't been considered? The similarities between abortion and chicken slaughter. Well, now, thanks to one Missouri legislator, that is up for discussion.
In what was no doubt the most bizarre moment of last week, anti-abortion GOP state legislator Mike Moon prepared for the new "nothing but abortion" special session of the Missouri legislature by killing a chicken — and posting the video on Youtube.
"When the governor called for the second special session this year, I was right in the middle of my summer job.
But like any good career politician, when I get the call (breaks to behead chicken), I'm going back to work," Rep. Moon said on his video, according to the Missouri Times.
"God gave us man dominion over life. He allows us to raise animals properly and care for them and then process them for food so we can sustain life. And that's what I'm doing here with this chicken. So we've been called back to this special session for the primary purpose of supporting life, protecting the unborn specifically. I think we need to get to the heart of the matter here (pulls out chicken's heart). So today, I'm filing a bill that will lead to the stopping of abortion in the state of Missouri and I hope you'll support it."
Moon later told reporters that he wasn't really trying to compare butchering a chicken to abortion, despite the video's message.


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Here are the stories we'll be following the week of July 2, 2018:
Swiss involvement in an Israeli project to control rats using barn owls has become much more than just scientific cooperation. By expanding to neighbouring Jordan and the West Bank researchers hope the enigmatic birds will also help to build dialogue despite political tensions in the region.
Blockchain startups are denied basic banking services in Switzerland due to banks' fears of potential fraud and money laundering. Can setting up minimum standards for such startup companies encourage banks to deal with cryptocurrency firms?
We visit what is arguably the most environment-friendly student apartment in the world. Made from recycled materials "mined" from urban environments, the flat itself is designed to be dismantled in five years.
As the new president of the Parliamentary Assembly of the Council of Europe, Swiss Senator Liliane Maury Pasquier has her work cut out. She talks to swissinfo.ch about the difficult financial situation of the 47-member body and the impact of the "caviar gate" scandal that involved influence buying by Azerbaijan.
Ever wondered why dialects are flourishing in the German-speaking part of Switzerland but are restricted to a few isolated pockets in the French-speaking part? We try to make sense of the rise and decline of Swiss dialects.
In case you missed it:


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- This event has passed.
Greenovation: Urban Leadership on Climate Change
November 18, 2016 @ 12:00 pm - 1:30 pmFree
Joan Fitzgerald is a Professor of Urban and Public Policy at Northeastern University. She focuses on urban climate governance and the connections between urban sustainability and economic development and innovation. Her third book, Emerald Cities: Urban Sustainability and Economic Development (Oxford Univ. Press), examines how cities are creating economic development opportunities in several green sectors and discusses the state and national policy needed to support these efforts. Emerald Cities builds on her 2002 book, Economic Revitalization: Strategies and Cases for City and Suburb (Sage), which identifies strategies for incorporating sustainability and social justice goals into urban economic development planning. In 2012 she published a three-volume anthology, Cities and Sustainability. Fitzgerald has published in academic journals such as Environment and Planning C: Government and Policy, Local Environment: The International Journal of Justice and Sustainability, Economic Development Quarterly, Urban Affairs Quarterly, Urban Affairs, International Journal of Urban and Regional Research and the political quarterly, The American Prospect. She also blogs on the London School of Economics United States Policy and Politics site. Her academic and consulting work has been supported by the Funders Network for Smart Growth and Sustainable Communities, John D. and Catherine T. MacArthur, Annie E. Casey, Rockefeller Brothers, Rockefeller, Surdna, Century, and Robert Wood Johnson Foundations and the Urban Sustainability Directors' Network. She has also conducted research for the U.S. Department of Labor, the Massachusetts Technology Collaborative, the Boston Housing Authority and other government agencies. She is currently working on her next book, Greenovation, which examines how cities advance green technologies. In addition, she is examining governance of green stormwater infrastructure. She teaches Cities, Sustainability and Climate Change and Urban Revitalization. Before coming to Northeastern University, Joan taught urban planning and policy at the New School University, the University of Illinois at Chicago, and Ohio State University.


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Joining the Institute 10 June 1971, Richard Noble (recently retired HM, Inspector of Mines & Quarries) has been a committed and long serving member of the IQ. He has made huge contributions through his enjoyable and well-attended guest speaking slots at technical evenings and other IQ events.
Well regarded for channeling his wealth of knowledge and support along with guidance and advice from the HSE, he has additionally been hugely influential in the conception and success of the Yorkshire and Derbyshire Health & Safety days.
In recognition of his exceptional dedication and work, Richard became the 27th person in the IQ's 99-year history to receive the special accolade of an Honorary Fellowship, which was formally presented at the Yorkshire Branch technical evening in April.