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Notfromearth Italy
Notfromearth is an electronic music and visual project by Carlo Alfano. Notfromearth is a passion for sound research with
the strong desire to push cinematic sound beyond corners.
Carlo Alfano is a video-maker, musician, designer, who is interested in doing artistic research without giving a precise definition of its creative product. ... more
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Russian bank 'peeks' at privatization in Croatia
Russian investment bank VTB Capital plans in next years on Balkan invest few hundreds of million euro, said its leader at the opening of regional office in Sofija, pointing out among interesting opportunities privatization program in Croatia.
Investment branch of second in size Russian state loan giver should step up on theplace of investors from European union which are drawn from region due to loan and economy problems in parent country. "Actual economy climate in middle and eastern Europe and decreased presence of investors from European union in region offer to VTB great opportunity for spreading on local markets", said head director Atanas Bostandijev. "I'll be happy if in next years we invest few hundreds of million euro in different projects in Bulgaria, Romania, Serbia and Greece directly as a bank or through the support to our clients", added Bostandijev. Among "good opportunities" he pointed out energy, transport, telecommunication and agricultural sectors in region. "We will research privatization programs in Bulgaria, Serbia and Romania", said Bostandijev in interview for Bloomberg. "We are interested in planned sale of branches of Romanian railway for transport of cargo.Serbiaplans to sell the banks, which are also interesting. The sane goes for similar programs inSloveniaandCroatia", said Bostandijev.
For all questions for travelling to Russia, getting visa for Russia you can contact Atlantis Travel. We know ALL about Russia.
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Isabella De Santos – Taste Her Rainbow
Released: July 29, 2014
Today, we bring you another Money Talks original that involves a Sex Swing, a pole, and a dildo. Use your imagination and then see who Havoc was able to convince a sexy girl to take the challenge and pull off in this hot and daring stunt. After that, we caught up with Mirko and Ava, who were doing a little shopping for some fresh tits n ass. They both agreed on the lovely Isabella. Don't miss the hotties in this episode!


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Cancer Research UK is proud of its popular graduate scheme, which it has been running for 10 years. Its role in encouraging university leavers to join the charity sector has been recognised by inclusion in The Times league table of the top 100 graduate employers.
Garcia Williamson, the charity's head of learning and development, says the scheme is an excellent succession pipeline for the organisation. "It enables us to attract great talent and invest early in the development of graduates' careers," she says.
Since 2005, more than three-quarters (41 out of 53) of those finishing the two-year scheme have gone on to get permanent jobs in the charity. Many have progressed through it and now hold senior positions; one of them is a director.
Graduate schemes are less common in the charity sector than in the private sector. The Wellcome Trust and the learning centre provider IntoUniversity both run schemes similar to that of CRUK, and CharityWorks helps graduates find roles in other charities.
In 2014 CRUK received 3,000 applications for eight £24,000-a-year graduate training posts in the areas of fundraising and marketing, scientific strategy and funding, policy, information and communications, and finance and technology. Williamson says the charity looks for evidence of ability outside academic achievements, personal drive and a willingness to think commercially.
She advises people who are interested in applying for graduate traineeships to research them thoroughly. "Think about the charity's cause and whether you feel connected to it, because this shines through at interview," she says.
Anne Coleman
She was a communications graduate and is now executive project manager in the policy and information team
I felt strongly about CRUK's cause and the scheme appealed to me because I was unsure what area of comms I wanted to work in. This scheme provided the opportunity to explore different avenues on the four placements. There is additional training and development, and you're encouraged to take on challenging projects – so I felt I'd have the opportunity to learn and progress.
I found the courses, such as project management and influencing and persuading, had practical implications that have proved particularly helpful in my job. It was also good to get together in my graduate peer group to talk through challenging situations. I found advice from the other graduates and their experiences helped me to develop my skills.
I'd definitely recommend this route if you're looking for a challenge, opportunities to try things outside your comfort zone and the chance to develop your skills in a supportive and motivating workplace. But it's not the easy route - you need to be prepared to work hard.
It's important to be articulate about why you want a place on the scheme, what you can bring to the role and what you hope to gain from the experience. Think about what separates you from the crowd. Throughout the interview process I had to talk about times when I'd demonstrated competencies such as problem-solving, leadership and building relationships. I made sure I could cite examples from my academic, professional and personal life that reflected my abilities. It's also good to let your passion for the cause and enthusiasm for the role shine through.
Emily Stewart
She is in her second year of the graduate scheme, currently in the innovation team
I feel incredibly privileged to be here. Not only do I get to work in a sector and for a cause that I feel passionate about, but the development opportunities and training are excellent. I applied for the scheme because I wanted to develop commercial awareness and transferable skills, and gain an understanding of how an organisation of this size works, while contributing to a cause I care about.
I studied philosophy and psychology, so my first role, as business improvement manager for the treasury department, was a baptism of fire. But learning about finance was just the kind of new experience I wanted to gain, and I've always felt supported. Getting onto a graduate scheme is great when you're not 100 per cent certain where your skills might be suitable.
I'd advise anyone interested in a similar scheme to consider their true motivations and be ready for a challenge. If you can demonstrate your enthusiasm for the cause and are open to learning, then you'll already be in with a good chance. Schemes such as this one push you in every way because you're constantly proving your contribution and value to new teams in a short space of time. But that's a great lesson in itself.
If you're still studying, then make the most of the various opportunities to develop while at university. Then you'll have something to share and to talk about when it comes to applying for graduate schemes.


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Theology of St. Cyril of Alexandria.
London ; New York : T & T Clark, 2003
|Named Person:
|Cyril, Saint Patriarch of Alexandria; Cyril, Saint Patriarch of Alexandria; Cyril, Saint Patriarch of Alexandria; Cyrillus, Alexandrinus.
|Material Type:
|Document, Internet resource
|Document Type:
|Internet Resource, Computer File
|All Authors / Contributors:
Thomas G Weinandy; Daniel A Keating
|9780567603647 0567603644 1283201046 9781283201049
|OCLC Number:
|Reproduction Notes:
|Electronic reproduction. [S.l.] : HathiTrust Digital Library, 2010. MiAaHDL
|1 online resource (xv, 269 pages)
|Master and use copy. Digital master created according to Benchmark for Faithful Digital Reproductions of Monographs and Serials, Version 1. Digital Library Federation, December 2002.
|Cyril of Alexandria as interpreter of the Old Testament / Robert Louis Wilken --
Cyril and the mystery of the incarnation / Thomas G. Weinandy --
Theotokos : Mary and the pattern of fall and redemption in the theology of Cyril of Alexandria / Frances Young --
The mystery of the trinity according to Cyril of Alexandria : the deployment of the triad and its recapitulation into the unity of divinity / Marie-Odile Boulnois --
The fullness of the saving God : Cyril of Alexandria on the Holy Spirit / Brian E. Daley --
Divinization in Cyril : the appropriation of divine life / Daniel A. Keating --
Incorruption, anti-Origenism, and incarnation : eschatology in the thought of Cyril of Alexandria / John J. O'Keefe --
Cyril of Alexandria : bishop and pastor / John A. McGuckin --
'Apostolic man' and 'luminary of the church' : the enduring influence of Cyril of Alexandria.
|edited by Thomas G. Weinandy and Daniel A. Keating.
Publisher Synopsis
..."Here are welcome and expert introductions.... Debate about Cyril's undoubtedly powerful Christology will continue, with this collection an important widely-based contribution."--Sanford LakoffRead more...
Similar Items
Related Subjects:(9)
- Cyril, -- Saint, Patriarch of Alexandria, -- approximately 370-444.
- Theology, Doctrinal -- History -- Early church, ca. 30-600.
- Cyril, -- Saint, Patriarch of Alexandria, -- approximately 375-444.
- RELIGION -- Christian Church -- History.
- RELIGION -- Christianity -- History.
- Cyril, -- Saint, Patriarch of Alexandria, -- approximately 370-444
- Theology, Doctrinal -- Early church.
- Theologie.
- Cyrillus -- Alexandrinus.


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Top Ten Sexiest Fragrances for Valentine s Day If you are going out on the town this Valentine s Day or staying in for the evening the right perfume can definitely help set the mood for romance. With so many different fragrances to choose from, it can be difficult to find just the right perfume for this occasion. Here are some top picks for the sexiest perfumes to wear (or give) on Valentine s Day. Modern Classics Some scents have stood the test of time as sexy perfumes for Valentine s Day. These classic scents have been around for decades, but have a timeless feel. Several have been updated to keep them feeling ugg pink fresh and modern. (1)Lancme Tresor has been around for years and is still going strong. This perfume can be worn to the office and then out to dinner; it is sensual yet subtle, very feminine and considered a true classic. Nothing says Valentine s Day like (2)Chanel No.5, a timeless fragrance that truly evokes love and romance. Also try Chanel No. 5 Eau Premiere, an update of the classic scent that is a bit lighter than the original (yet still quite alluring.) (3)Guerlain Shalimar is a classic oriental blend that has been available for decades; also try Guerlain Shalimar Black Mystery (with notes of amber and vanilla anything with vanilla is a very sexy scent!) (4)L Air du Temps is the ultimate refined fragrance, a timeless classic by Nina Ricci. This scent proves that a lady can be very very sexy on Valentine s Day (think Grace Kelly.) Sexy and Provocative (5)Poison by Christian Dior is so sensual and seductive that it is best worn with a confident attitude. Poison might be considered a bit retro, but that s not a bad thing, as it has stood the test of time. It s a very heady perfume, rich, full bodied and unabashedly sexy for Valentine s Day. (6)Falling in Love by Philosophy is the most provocative scent in the Philosophy fragrance line. Their other scents are about smelling clean, fresh or feminine; this fragrance is all about sensuality and romance. Falling in Love is so sexy it can set the stage for a special Valentine s Day that s very romantic and memorable. (7)DKNY Delicious Night is the new fragrance by Donna Karan that lives up to its name. The scent is an intoxicating blend that smells like blackberries. Also by Donna Karan: Black Cashmere and Cashmere Mist where can i buy ugg boots in store are both all out sexy, seductive perfumes that should set the tone for a romantic Valentine s Day. Ladylike and Romantic Some fragrances are so sentimental and feminine, they are perfect for a romantic occasion like Valentine s Day. (8)Vera Wang Princess is a delightful floral blend with notes of vanilla and pink guava (it also comes in a beautiful, lilac colored, heart shaped bottle fit for a princess.) The beauty of the packaging, and the loveliness of the scent, also makes it an ideal Valentine s Day gift. (9)Nina Ricci Nina is a beautiful fragrance that also comes in a lovely bottle that will make the wearer (or the recipient) of this perfume feel like a fairytale princess. It has been around forever, and can evoke memories of the high school prom! However, it is a soft, sexy scent that smells a bit like baby powder. There is something inviting, approachable and comforting about this scent, which seems to have cult following. (A tip: layer Love s Baby Soft with L Air du Temps. Something about the combination of the two is soft, feminine and very sexy.) Many people find Bath Body Works Warm Vanilla Sugar a very sexy fragrance. This scent also seems to have a cult following. The perfume is delicious with vanilla notes, coconut and sandalwood. Bath Body Works sells this fragrance in ugg boots canada sale body lotion and there is a whole product line of Warm Vanilla Sugar, often sold in cute little gift sets (this is a big seller during the holidays, and would be perfect for Valentine s Day too!) An evening in fancy dress at a romantic restaurant might call for Poison, or Chanel No.5, or Falling in Love by Philosophy; however, for an evening at home with food, romance, wine, (or champagne!) and candles, then Warm Vanilla Sugar and Love s Baby Soft, or one of the floral romantic scents by Vera Wang or Estee Lauder, might be the perfect choice for Valentine s Day. These ten sexy perfumes are great to wear or give on Valentine s Day. The right fragrance can definitely set the mood for a very romantic evening!.
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Gassed Louisiana sinkhole family human rights plea exposes coverup
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Someone really needs to help these people!
Anonymous Coward 17338098
But isn't the mother responsible for the welfare of her own small children?
Anonymous Coward 13793225
You know most societies families neighbors help each other out. No one can do everything completely on their own.
What is wrong with you?
How dare you judge someone you don't even know.
Did you raise yourself and never need assistance or did you have a community helping you along, even invisibley it happens every day and it gives delight to all concerned.
Anonymous Coward 16213681
Chill out with your histrionics there . . .
If you actually took time to read my first post, you would see that I acknowledge that the gov SHOULD be evacuating everyone with a certain radius of the sinkhole.
But this girl is the mother of two very small children and it is HER responsibility to insure their safety.
She and her mother acknowledge that there were problems 9 years ago . . . and yet she brings two babies into that environment.
Now if she or someone who knows her wants to contradict that legitimately, they can do so. Otherwise, my interpretation of this is valid.
Hell yeah I can judge . . . because every irresponsible, reckless person in this country is part of the whole of this society and when they screw up, a little bit of society is screwed up.
And I don't know where people like you get the idea that it's perfectly acceptable for people to AUTOMATICALLY depend on their family and neighbors . . . perhaps if people were stronger and more determined to be self-sufficient, this society wouldn't be comprised of so many apathetic jackasses just wanting a handout and EXPECTING others to do their work and get them out of messes when THEY are perfectly capable of doing so or could have not been in the mess in the first place if they'd used better judgment.
There ARE those who work hard, act responsibly and are decent, genuinely good people who may need a helping hand in one way or another and THEY are the ones we should be willing to help.
And may I add this - WHERE is the father (or fathers) in this story?
Anonymous Coward 13793225
~~~~This~~~~ With a cherry on top, Bouya!
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DUMBO, Eberhard Faber Among Industrial Sites on LPC Agenda This Week
October 29th, 2007
Brooklyn's Industrial Waterfront is the focus of a major designation day at the Landmarks Preservation Commission next week. On Tuesday, October 30, the Commission will hold a public hearing on the proposed DUMBO Historic District and vote on the designation of the Eberhard Pencil Factory Historic District. Both sites were highlighted in the MAS's successful nomination of the Brooklyn industrial waterfront heritage to the National Trust for Historic Preservation's 11 Most Endangered Places for 2007. The Commission's action on DUMBO and Eberhard Faber follows the designation of the Domino Sugar Factory in September. DUMBO is significant for its association with the rise of the city of Brooklyn as a major American industrial center and is unique to New York City for its nineteenth and early twentieth century industrial buildings, Belgian block streets, and its location on the East River by the imposing anchorage of the Manhattan Bridge. Continue Reading>>


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Home > Groceries > Drinks > Tea, coffee & hot drinks > Green & white tea > Teapigs Green Tea With Mint (15 bags)
Teapigs Green Tea With Mint (15 bags)
Dietary Summary:
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Marrakesh Mint
In Stock
£3.95 Standard UK Delivery / Free Over £50 / Next Day Delivery Available
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Product description
A delicate green tea known as "precious eyebrows" blended with peppermint leaves for that traditional Moroccan experience.
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Joseph Orton
Joseph Orton (22-11-1994 / England)
Poems by Joseph Orton : 19 / 18
Chains - Poem by Joseph Orton
The chains that I thought bound us.
Are just in my head.
You see no chains.
You see nothing to bind us.
As much as it hurts to admit it.
The past can't be re written.
And thing won't go back to before.
I can't change fate.
And my fate is at a crossroads.
The littlest of things effect me in big ways.
And I'm quite ashamed of this flaw.
But I'm not ashamed of how little time is.
Paths can recross.
But these chains that I believed in.
Can never rebind.
Because they were never bound to begin with.
Hope? A myth.
Listen to this poem:
Poems by Joseph Orton : 19 / 18
Comments about Chains by Joseph Orton
Read this poem in other languages
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Poem Submitted: Saturday, May 4, 2013
Poem Edited: Wednesday, September 18, 2013
Joseph Orton's Other Poems
Famous Poems
Phenomenal Woman
Maya Angelou
Still I Rise
Maya Angelou
The Road Not Taken
Robert Frost
If You Forget Me
Pablo Neruda
Langston Hughes
Annabel Lee
Edgar Allan Poe
Caged Bird
Maya Angelou
Rudyard Kipling
Stopping By Woods On A Snowy Evening
Robert Frost
A Dream Within A Dream
Edgar Allan Poe


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MNRE to get energy efficient headquarters
The Atal Akshay Urja Bhawan, to be built at the central government office complex area in New Delhi, will house a pavilion showcasing renewable energy systems and devices
New Delhi: The new and renewable energy ministry is set to have modern headquarters named after former Prime Minister Atal Behari Vajpayee which will generate clean energy almost equal to its annual power consumption, the ministry said.
The Atal Akshay Urja Bhawan, to be built at the central government office complex area in the capital, will house a pavilion showcasing renewable energy systems and devices, the ministry said quoting Piyush Goyal, minister for power, coal, new and renewable energy and mines.
"Energy conservation and renewable energy are areas where India is soon going to be a global leader. This government is committed to strive for this mission and light up peoples' lives in an environmental friendly and sustainable manner," the statement said quoting Goyal.
The headquarters will be a symbol of the clean energy revolution in the country, said Upendra Tripathy, secretary in the ministry. India has set a target of having 175 gigawatt (GW) of renewable energy capacity by 2022, up from about 7 GW at present. A report from Bridge to India, a clean energy consultancy, released on Wednesday said that the country has added 513 MW of rooftop solar capacity over the past 12 months, growing at 113 % over the previous 12 months, taking the total installed capacity of such systems to 1 GW.


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What is the average cost for hypopigmentation treatments post laser tattoo removal?
Doctor Answers 4
There is no "Cure" for Hypopigmentation
Thanks for your question. You did not mention what laser was used in this treatment. Q-Switch or nanosecond lasers have a more inherent risk of complication over picosecond lasers.
Picosecond lasers fire 1000 times faster, using a pressure wave rather than heat to shear the ink molecules apart. After using the PicoSure for almost 2 years, we have only seen pinpoint hypopigmentation in a very small number of patients usually of darker skin tones. We cannot say it is impossible to get burned with a Picosecond laser, but it is highly unlikely. The 755 nm works well on almost all ink colors.
The next thing you need is patience. We tell every Pico tattoo removal patient to expect at least 6 treatments and don't be surprised if it is 10-12 or longer with thick tribal tattoos. We see some patients twice a year and their tattoos seem to clear up to 6 months. After using the PicoSure for 2 years, we have found it is best to wait 2-6 months between treatments. There is a risk of complications with any laser treatment. Minimize your risk by treating your tattoo as few times as possible. There is a limit to how small the laser can break up the ink. It's up to you and your body to take the ink away. We encourage our patients to drink plenty of water, sweat it out and eat a healthy diet between treatments.
We recommend finding an experienced physician in your area. Treatment prices vary by market. You are trying to reverse permanent ink. Good Luck and have patience!
I am sorry to hear that. There is no way to cure hypopigmentation. Instead skin matching ink is tattooed on pigmented areas, or you can try fraxel laser with latisse to re-pigment the area, but it does not work with all patients. I would suggest you to switch laser being used or change provider to avoid further pigmentation.
Learn more: http://www.myethosspa.com/services/tattoo-removal/
Hypopigmentation after Laser Tattoo Removal
This can be tattooed your normal skin colored or fraxel laser can be used with latisse to re-pigment the area. It will take a series of treatments both to remove the rest of your tattoo and re-pigment. Costs will vary, but see an expert. Best, Dr. Emer.
Learn more: https://www.youtube.com/channel/UCa6C2fuS-AgOnvBjr6_VSQg
You might also like...
Hypopigmentation from tattoo removal
There are no routine treatments for this problem. It is an extremely hard problem to fix. Sometimes it will improve itself over several months. Some people are doing PRP for this and others are using Latisse for this. I don't know success rates for this.
These answers are for educational purposes and should not be relied upon as a substitute for medical advice you may receive from your physician. If you have a medical emergency, please call 911. These answers do not constitute or initiate a patient/doctor relationship.


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Robert Schaffer has worked with leaders for 50 years but the consultant says in the Harvard Business Review that despite a dizzying rate of economic, social and environmental changes, the mistakes they make still fall into four classic behavioural traps
Senior managers love to set bold goals, but generally they fail to set proper expectations. It sounds grand to say "We're going to reduce the use of cash by 40 per cent next year" or "We are going to shift focus from mid-market customers to the upper end during the next two years." But usually those ambitions hit the ditch, because the leader hasn't spelled out a credible plan for achieving the goal or specified who exactly - one person - is accountable, both important parts of setting proper expectations.
Sometimes leaders describe what must be done but then sabotage themselves and the effort with some weasel words that let people off the hook. Take the understanding leader who says, "I know you have lost some people, Stan, but you have to really give it a go - we really need to increase sales in your territory."
With operating managers so preoccupied with the performance of their own units, they often elude responsibility for organization-wide performance. Instead, they delegate the responsibility for that upward, back to the senior manager.
Mr. Schaffer cites as an example the CEO of a large IT-based company who determined that demographic and technological trends would gradually render many of its business lines obsolete. He tried to draft some of the smartest people in some key units to help him develop new strategies. But their bosses objected, claiming that while they understood the dangers of obsolescence they had critical problems those individuals had to deal with immediately. The CEO acquiesced, and allowed them to delegate the responsibility back to him.
Staff experts who advise leaders, and consultants brought in to help with change, are great at conceiving new production systems, organizational structures, and marketing plans. But they usually don't assume responsibility for outcomes. This collusion - leaving nobody responsible - comes because the experts are uncertain they can work with the client company to produce results and the leader wants to provide some distance from the effort anyway. If it works, the leader will be happy to be hailed as a hero, but if it fails it's nice to say "even the experts couldn't solve this one."
When senior managers challenge their employees to improve sales or reduce costs or make other improvements, they are often told, "Yes, but first we must …" Maybe that first item on the agenda is the need for staff training or a market study, but today's management culture accepts that the first step to improving performance is finding new programs to achieve those gains.
That's because everyone wants to believe they are doing their utmost under the current operation. But Mr. Schaffer offers examples where big gains were made within existing systems thanks to focus and effort.
MARKETING: Building sales with small companies
Big businesses that serve other big businesses often fumble when they try to turn their attention to attracting smaller business as customers. MarketingSherpa editor Sean Donahue recently offered lessons taken from Rogers Communications, which was trying to develop business with companies that had 20 employees or fewer:
Create dedicated small-business sales and marketing teams that can target these small operations properly, because in many ways they act like consumers but want to be treated like a business. Rogers also created a dedicated call centre for small business support, rather than relying on the regular consumer support line.
Develop products specifically aimed for small business, rather than adding a few bells and whistles to consumer products or stripping features and functionality from products for larger enterprises. In 2009, for example, Rogers launched the first land-line phone system for small business.
Partner with other large companies trying to reach into this marketplace, to make the effort more affordable and effective. Rogers helped create an alliance called The Committed To Growing Small Business Coalition with companies such as HP and Staples.
Get involved with groups such as local chambers of commerce, universities, and business incubators that support small business. Rogers also started its own venture-capital fund for high-tech startups.
Buy from small businesses; they love to get their foot in the door of large companies but often are stymied by procurement policies. As Rogers has been finding, however, it can be a significant challenge to change those policies.
MANAGEMENT: Why comparing workers with their peers can backfire
It is usually assumed that employees who are benchmarked against each other work harder. But research by Wharton Management Professor Iwan Barankay, reported in Knowledge@Wharton, suggests this is flawed thinking.
He advertised jobs on the Mechanical Turk website that involved analyzing images. In the experiment's first stage, he posted two identical jobs, but one offered feedback on the worker's accuracy at the end of the assignment while the other didn't. He thought the first would be more popular, given conventional management wisdom that people want to know how they rate. But the job without the feedback attracted more workers -more than three times as many (254 to 76).
In the second stage, he randomly divided workers into a control group receiving no ranking; in another group, people were asked if they wanted to do more work and also received feedback with a ranking from previous jobs. More people in the group that didn't receive feedback returned for more work (66 per cent to 42 per cent), while those who received feedback were 22-per-cent less productive in their work.
Act and react
Reactive tasks are those driven by others while proactive tasks are driven by your goals. Entrepreneur Scott Scheper suggests that you should balance your time so 20 per cent is spent on reactive tasks and 80 per cent on proactive. howtogetfocused.com
Filtering e-mail
Kevin Rose, founder of Digg, advises you to create a VIP filter in your e-mail program that captures messages from your boss, investors, close friends and siphons them into a separate folder that you check first every morning. Kevinrose.com
Google mapping
Most companies pay considerable attention to how their website scores on Google searches. But you also need to make sure that you are among the top choices when somebody uses Google Maps to look for your type of business in a specific area. Monday Morning Memo
Reality check
When assessing mediocre or lousy employee you have, ask five simple questions: Who hired them? Who trained them? Who coaches them? Who supervises their work? Who keeps them around? Donald Cooper Corporation Business Management Newsletter
To-do on the Web
Thingler is a free service that allows you to establish a to-do list on a website that can be shared with collaborators on a project. The list is on a unique page, and anyone on the team can add or delete items that the others can see when they refresh the site. Lifehacker.comReport Typo/Error
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Susan Linn has been serving nonprofit organizations for more than 25 years. Susan's early career days as a direct service worker for struggling children and families continue to influence her leadership of programs and services today. Her leadership roles include two decades in upper management, fundraising leadership, and chief executive roles for schools, municipalities, higher education, and nonprofits. A graduate of Cornell University and San Diego State University, she has served on several Boards. Susan's spouse has epilepsy, and they have been involved with epilepsy-related causes in a number of capacities for many years. When not at work, Susan values her time with her children, 2 dogs, and 4 cats, especially time in the outdoors. Susan has lived in Providence and Bristol, RI as well as Fall River, Watertown, Woburn, and Shirley MA and Nashua, NH: she and her family frequent Maine for surfing in the summer!
Dr. Elaine Kiriakopoulos is a neurologist. A graduate of McMaster Medical School and the University of Western Ontario in London, Ontario, Canada. Dr. Kiriakopoulos completed her neurology residency and her Epilepsy/EEG fellowship at the Beth Israel Deaconess Medical Center in Boston. Her clinical and research career were based in the Department of Neurology at the Beth Israel Deaconess Medical Center and Harvard University. Dr. Kiriakopoulos's research interests have focused on advancing clinical imaging and EEG recording techniques to improve diagnosis, treatment, and patient outcomes. She has worked in the areas of localization in epilepsy and her work includes pre-operative brain mapping with functional magnetic resonance imaging and examining the use of high density electroencephalography in the ICU setting for early seizure detection and treatment. She has presented her research at the national and international level throughout her career. Dr. Kiriakopoulos serves as a member of the Editorial Board of the Epilepsy Foundation of America.
Dr. Kiriakopoulos has been a longstanding advocate of empowering patients with epilepsy through knowledge and accessibility to a comprehensive epilepsy care network of physicians, nurses, social workers and psychologists. Having joined the EFNE team, she hopes to continue to provide integral outreach to the epilepsy community of New England with valuable information, resources, and the vital support required to improve the lives of persons with epilepsy and their families. Dr. Kiriakopoulos resides in Winchester, Massachusetts with her husband and two children, and their mischievous West Highland terrier "Pearl". Her free time is spent traveling with family and in the garden testing the limits of her green thumb!
Early in her career, Judy developed a passion for philanthropy working as a staff assistant for the Founder of Cambridge College. She served the college for almost two decades. Her greatest achievement at Cambridge was to lead its first ever Capital Campaign, having raised nearly $40M, the first endowment. Over the course of her career she has continued to work for non-profit organizations in order to serve the greater good. In her free-time Judy likes to garden and to spend time with her teenage daughter and her 2-year old rescue dog. Like any native New Englander, she cherishes the warm seasons and loves the Atlantic Ocean. Judy holds an M.Ed. from Cambridge College.
Susan has an Associate degree in Sociology from Bunker Hill Community College and attends Southern New Hampshire University where she will earn her degree in Business Administration with a concentration in Organizational Leadership in 2017. She became a volunteer mentor for Epilepsy Foundation of America in 2000. Since then, she has provided more than 400 presentations on epilepsy and related topics to a wide variety of audiences, having worked for EFNE since 2001. Susan's involvement with EFNE began in 1996 as a volunteer and as the parent of a thirteen-year-old daughter with epilepsy. She and her daughter as well as their vast network of family and friends still participate as volunteers in dozens of EFNE walks, camps, and other programs every year.
Franky Descoteaux is a serial entrepreneur, who has dedicated her career to solving core challenges through the lens of innovation. In an effort to change the perception of Lowell, MA, she launched three retail businesses in downtown, which contributed to the City's revitalization. She sat on the Lowell City Council where she focused on building bridges on issues ranging from public safety to union rights. She also fought to ensure that entrepreneurs in Gateway Cities have access to the same education, resources and mentoring as their counterparts in Boston as Executive Director of a local non-profit, EforAll.
Franky holds a B.S. in Management of Information Systems from UNLV and an M.S. in Innovation & Technical Entrepreneurship from UMass Lowell. She in a Trustee for the Greater Lowell Health Alliance and Innovation Academy Charter School and teaches Entrepreneurship and Small Business Management at Middlesex Community College.
Franky is a Female Founder Fellow at Founder's Institute. She won the prestigious Entrepreneur of the Year award from Enterprise Bank as part of their Celebration of Excellence and is a recipient of the PepsiCo Diversity Award. Franky Descoteaux lives in Lowell, Massachusetts with her two talented children, two energetic dogs, and gifted artist/musician husband.
Bill has worked in the field of disability issues for over 45 years, starting his career as a special education teacher right after attending Syracuse University. In 1980, he became a founding member, and first Executive Director, of what was then called the Epilepsy Association of Greater Boston and is now EFNE. In 2005 Bill went to work for the national Epilepsy Foundation as Director of State Government Relations, until retiring in 2013. Retirement didn't last long! In 2014 he "came full circle" and again is with our local Epilepsy Foundation affiliate serving as Director of Advocacy and Public Policy for EFNE. He returns to the National office each Spring to conduct their "Teens Speak Up" program; bringing teens from across the country who live with epilepsy to learn about advocating for epilepsy and to visit with their legislators on Capitol Hill. Now living in Newport, RI, Bill can often be found on the highway traveling to Boston, Providence, Concord or Augusta to promote public policies beneficial to people with epilepsy and their families.
Jenna Whitney has a background in development and event planning, and worked previously at The University of Texas at Austin and Tufts University. She attended Simmons College, and holds a BA in Communications. In 2009, she volunteered with C.A.R.E. in Cambridge, MA and from 2010 – 2016, she was a volunteer with Austin Pets Alive! in Austin, TX. In 2014, Austin Pets Alive! named her a Platinum Volunteer for her consistent service to Neonatal Kitten Program. Jenna lives in Cambridge, MA.
Prior to coming on board with the Foundation, Francesco acted as the Business Manager for his parent's small business in Revere as well as running his own business for 10 years. He graduated from Suffolk University with a degree in Business. His personal experience with epilepsy comes from family members on his and his wife's side who have been diagnosed with epilepsy. Outside of the Foundation, he is a member of the National Soccer Coaches Association of America as well as the Fraternal Order of Freemasons.
Rachel Buckley started her professional career in October 2016 with Epilepsy Foundation New England. Prior to working at EFNE, Rachel took part in an internship with the Walt Disney World Company in Orlando, Florida. She graduated from Westfield State University with a Bachelor's degree in Psychology and a minor in Business. Rachel lives in Melrose, Massachusetts.
Sarah Robinson has worked for the past five years as a consultant with small nonprofits. Prior to starting her consulting practice, she worked as a Major Gifts Officer for a national health care non-profit, Executive Director for a statewide network, and as an Office Manager for an emerging community organization. Sarah holds an MS in Nonprofit Management from Antioch New England. Sarah and her husband live in Portland with their two very active children. In her free time, she gardens, snowboards, and volunteers for the PTO and Girl Scouts.
Zach attended University of Massachusetts Dartmouth, where he studied Crime & Justice Studies and Political Science. Zach has been an avid volunteer for the Coalition for Social Justice, has participated in two cross-country bike rides to raise money for cancer causes, and has been a community organizer and activist. Zach's personal experience with friends and loved ones with epilepsy and his desire to contribute to the nonprofit sector has led him to EFNE and the Matty Project.


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Description: In recent decades, the lives of people in their late teens and twenties have changed so dramatically that a new stage of life has developed. In his provocative work, Jeffrey Jensen Arnett has identified the period of emerging adulthood as distinct from both the adolescence that precedes it and the young adulthood that comes in its wake. Arnett's new paradigm has received a surge of scholarly attention due to his book that launched the field, Emerging Adulthood.On the 10th Anniversary of the publication of his groundbreaking work, the second edition of Emerging Adulthood fully updates and expands Arnett's findings and includes brand new chapters on media use, social class issues, and the distinctive problems of this life stage. In spite of the challenges they face, Arnett explains that emerging adults are particularly skilled at maintaining contradictory emotions--they are confident while being wary, and optimistic in the face of large degrees of uncertainty. Merging stories from the lives of emerging adults themselves with decades of research, Arnett covers a wide range of topics, including love and sex, relationships with parents, experiences at college and work, and views of what it means to be an adult. He also refutes many of the negative stereotypes about emerging adults today, finding that they are not "lazy" but remarkably hard-working in most cases, and not "selfish" but rather concerned with making a contribution to improving the world. As the nature of American youth and the meaning of adulthood further evolve, Emerging Adulthood will continue to be essential reading for understanding the face of modern America.
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List price: $31.95Edition: 2ndCopyright year: 2014Publisher: Oxford University Press, IncorporatedPublication date: 9/25/2014Binding: PaperbackPages: 416Size: 6.00" wide x 9.00" long x 1.00" tall Weight: 1.430Language: English
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What is the case that establishes who is an occupier?
What did Lord Denning establish in Wheat v E Lacon & Co Ltd?
What is the definition of a visitor and what is the section?
What was established in the case of The Calgarth 1927?
Can a visitors permission be revoked?
What is section 2 (1) of the Occupiers Liability Act 1957?
What is Section 2 (2) of the Occupiers Liability Act 1957?
What was established in Cunningham v Reading football Club 1992?
What case established that the occupier can be liable for the dangerous state of their premises?
What was established and what happened in Ferguson v Welsh 1987?
What case established that can occupier can be liable for injury caused to a visitor where the occupiers premises themselves are in a dangerous state as a result of activities that have and are happening on the occupiers premises?
What is the standard of care test?
What is the question that needs to be asked regarding standard of care?
What was established in the case of Simms v Leigh Rugby Football Club 1969?
What case established that account must be taken of the usual factors that impact up on the standard of care that D is expected to reach?
What section regarding visitors states the standard of care of a child?
What was established in Jolley v Sutton London Borough Council 2000?
what case established that the council should have taken into account the reasonable child?
What case other than Jolley established that the account of a reasonable child was not taken into account?
What section relates to the standard of care skilled visitors should apply?
What case demonstrates the legal principle regarding standard of care of skilled visitors?
What section regards warnings to visitors?
what are the two cases regarding warnings to visitors?
What section regards faulty work and visitors?
what case regards faulty work and visitors?
What section and what is the defence and case regarding defences for claims from visitors?
what sections define when a duty is owed to a trespasser?
what was established in Tomlinson v Congleton Borough Council 2004?
what case established the rules regarding duty of care of a trespasser?
What is the case regarding that the trespasser can be a visitor but then a trespasser may exceeding the scope of their permission?
What are the rules regarding the standard of care towards a trespasser?
What section regards the standard of care for both trespasser and visitor?
What section and case regards the defence for occupier against the trespasser?
what was established by the judge in Geary v JD Weatherspoon PCC 2011?
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\n\n\n\nRecent Hot Clicks\n02-04-08: Ovechkin's dream girlfriends\n02-01-08: Sexiest Female Athlete Commercials\n01-31-08: Personalities that ruin your Super Bowl party\n01-30-08: Casting "Super Bowl: The Movie"\n01-29-08: Watch Marisa Miller box\nBy Jimmy Traina \n\n\n\n\n\n\n\n\nThe Links\nBefore They Were Models\n\nIrina :: Pamela Hanson/SI\nSI.com's 2008 Swimsuit section launches next Tuesday, but thought we'd get the action going now. Some of our top models recently discussed what they did before hitting the pages of Sports Illustrated. For example, Irina (above) talks about her true love -- ventriloquism.\nThe 2008 NBA Oscars\nLe Basketbawl combines NBA midseason awards with the upcoming Academy Awards for this amusing column. \nFrom The Home Office...\n\nDavid Letterman offered up the Top 10 New England Patriots Excuses on his show last night. Even better was Dave taking some shots at Bill Belichick before going into the Top 10 list.\nSuper Stalker \nMister Irrelevant's/Sporting News' Chris Mottram has put together a must-see gallery of the stalking he did in Arizona last week. Keep in mind: nothing was Photoshopped.\nEli Vs. Elisha\n\nElisha Cuthbert :: Peter Kramer/Getty Images\nSuper Bowl hero Eli Manning and The Girl Next Door star Elisha Cuthbert have more in common than just a similar first name. Rumors and Rants pits these two against each other in a tale of the tape. (Warning: Link contains strong language.)\nGiant Bummer\nThe Sports Hernia has an inside look at what happened when Tiki Barber watched the Super Bowl, while LOL Jocks gives us the scoop on a conversation between Tiki and a former teammate.\nSumming Up The Weekend\n\nRyan Parker has made two new videos in honor of Super Bowl XLII. First up is a New York Giants victory song. And then we have a diddy about the newest case of Spygate.\nAn Amazing Journey\n\nYesterday we posted a video of an amazing Steve Perry-sound-alike singing Don't Stop Believin'. Well, several of you wrote in to tell us that the singer's name is Arnel Pineda and that Journey actually hired the guy a couple of months ago as its new lead singer. Bobby Dozier and Gabriel ofSanta Barbara, Calif., sent us this CNN report about Arnel getting the job. Dan in Salt Lake City sent us this story, while Jon in Rochester, Minn., sent us this one. Lastly, someone who signed their e-mail "Arnel Pineda DOES sing for Journey now!" sent us this link to Journey's Web site that details the whole situation. We thank all of you who wrote in and we promise to do a better job of keeping up with our Journey news. We'll wrap things up by giving you this link -- Arnel belting out Faithfully.\nToday in Campus Clicks \n\nWhich schools are endorsing which candidates? ... Offensive lineman gets Punk'd ... Iowa State student steals text book ... Video: The Best of Bobby Knight... more.\nGet More Links On Facebook\nFor exclusive links and photos that don't make this page, sign up for our Facebook group and get them out there.\n\n\n\nVideos of the Day\nBelichick Honored For His Work In Film\n\nJames Lipton interviews Bill Belichick. (Warning: Link contains strong language.)

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An untargeted multi-technique metabolomics approach to studying intracellular metabolites of HepG2 cells exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin
The study focused on the analysis of intracellular metabolites using NMR, LC-MS and GC-MS, with emphasis on the reproducibility and repeatability of the data. State of the art pre-processing and alignment tools and multivariate statistics were used to detect significantly altered levels of metabolites after exposing HepG2 cells to TCDD. Several metabolites identified using databases, literature and LC-nanomate-Orbitrap analysis were affected by the treatment. The observed changes in metabolite levels are discussed in relation to the reported effects of TCDD.
Untargeted profiling of the polar and apolar metabolites of
Metabolomics has been defined as the quantitative measurement of the multi-parametric metabolic response of living systems to patho-physiological stimuli or genetic modification
Recently, there has been an exponential growth in the number of published papers concerning metabolomics of a wide variety of systems
Owing to ethical concerns (animal welfare) and cost efficiency, there is a need to develop alternatives to conventional toxicity testing incorporating animals
The goals of the present work were two-fold. First, to develop an untargeted
TCDD is an agonist of AhR, a cytosolic ligand-activated transcription factor. Upon activation, AhR dimerizes with ARNT to form a heterodimer that binds to DNA sequences called xenobiotic response elements (XREs). Through such binding, AhR up-regulates the expression of several downstream genes including those encoding xenobiotic metabolizing enzymes such as Phase I (e.g. cytochrome P450 monooxygenases) and Phase II (e.g. glutathione S-transferases, sulfotransferases) biotransformation enzymes
In this study, the human hepatoma cell line HepG2 was chosen for experiments concerning
For the untargeted metabolomics approach used in this study, multiple complementary analytical techniques were applied to polar and apolar cell lysate fractions, i.e. NMR and GCMS to the apolar fraction, and NMR and LCMS to the polar fraction. Data analysis was performed using state-of-the-art software
Particular emphasis was placed on the repeatability of experiments, reproducibility of metabolic changes, normalization and validation of the results by literature comparison.
The advantages and limitations of omics techniques applied to
Cell extracts containing intracellular metabolites were separated into polar and apolar fractions and subjected to 1D 1H-NMR and LC-MS or 1D 1H-NMR and GC-MS, respectively. The majority of NMR and MS data obtained were complementary in terms of their information content. Raw data were pre-processed and aligned using specific software enabling statistical analysis. Subsequently, metabolites with significantly altered levels were identified using databases, literature and LC-nanomate-Orbitrap analysis. In this study, a particular focus concerned the repeatability of experiments with HepG2 cells obtained from different passages, and reproducibility of biological replicates from the same passage, and normalization of experimental data prior to comparison. Results were compared with data from previous studies concerning the effects of TCDD in non-metabolomics studies.
The first major problem addressed was the normalization of chemical profiles. In this study, typical changes in metabolite concentrations were between 20 and 100%. Compounds such as TCDD can influence overall cell metabolism and cause a decrease in the rate of cell proliferation. Therefore, the net effect of exposure can be an overall decrease of metabolites because there is less cell material. If this is not taken into account it will dominate the metabolomics results and metabolic effects may go undetected. Furthermore, in order to identify statistically significant changes in metabolite concentrations in the order of 20 to 100%, it was essential to have multiple replicates.
Preliminary studies in our laboratory have shown that the age of cell cultures (expressed as passage number) could influence the magnitude of the effect of the exposure on the cell metabolome. To address this issue, HepG2 cells obtained from different passage numbers were exposed to medium containing 10 nM TCDD or the vehicle, DMSO, for a period of 48 h.
The apolar fraction
NMR analyses of the apolar fraction were performed using the same deuterated chloroform. Therefore, the residual proton signal of chloroform was used as an initial absolute internal standard. This allowed the amount of lipids in solution to be estimated by calculating the ratio between the sum of the intensities of all spectral signals (TS (total signals) = signals 1-22 in Figure
Figure 1
Total 1H NMR spectral signals of the apolar fraction of HepG2 cells exposed to TCDD (above) or the vehicle control DMSO (below)
Total 1H NMR spectral signals of the apolar fraction of HepG2 cells exposed to TCDD (above) or the vehicle control DMSO (below). For both treatments, four spectra were overlaid. The numbers of the signals correspond to those indicated in Table 1.
Figure 2
Ratios between the intensity of some NMR spectral signals
Ratios between the intensity of some NMR spectral signals. A: Ratio between the intensity of the total NMR spectral signals (TS) and the intensity of the residual CHCl3 signal. B: Ratio between the intensity of the phospholipid signal in the NMR spectra (PL) and the intensity of the residual CHCl3 signal.
TS predominantly represented membrane constituents and triglycerides. Membranes are thought to be relatively constant in terms of composition. Since membrane constituents such as phospholipids (PL) are easily detected in NMR-spectra, it was assumed that generic phospholipid signals (such as signals 18 and 20 in Figure
The ratio between the intensity of the PL signal (signal 20 in Figure
When scaling on PL signals (Figure
Representative 1H NMR spectra replicates of apolar extracts of HepG2 cells scaled on the PL signal are overlaid in Figure
Table 1
Assignment of the 1H NMR spectral signals of the apolar fraction.
Peak number
Chemical shift (ppm)
Assignment a
Total cholesterol C18H3
Total cholesterol C26H3/C27H3
Acyl groups CH3 (all except n-3)
Total cholesterol C21H3
n-3 acyl groups CH3
Free cholesterol C19H3
Esterified cholesterol C19H3
Multiple cholesterol protons
Acyl groups -(CH2)n-
Acyl groups -OCO-CH2-CH2- and H2O
Multiple cholesterol protons
Acyl groups -CH2-CH=CH-
Acyl groups -OCO-CH2-
Acyl groups =HC-CH2-CH=
Choline N(CH3)3
Glycerophospholipids N-CH2
Glycerol backbone -CH2OCOR
Phosphatidylcholine PO-CH2
Esterified cholesterol C3H
Glycerophospholipid backbone >CHOCOR
Glycerol backbone >CHOCOR
Acyl groups -CH=CH-
Numbers correspond to those in Figure 1.
aProtons assignment based on
NMR data were pre-processed and aligned using a program developed in-house
Figure 3
PCA plots (after ANOVA p < 0.01) of the apolar fraction of HepG2 analyzed by 1H NMR and pre-processed and aligned using a program developed in-house.
PCA plots (after ANOVA p < 0.01) of the apolar fraction of HepG2 analyzed by 1H NMR and pre-processed and aligned using a program developed in-house. A: Spheres with the same colour are technical replicates of the same sample per passage number. DMSO30: green; TCDD30: red; DMSO7: lila; TCDD7: yellow; DMSO11b: light blue; TCDD11b: dark blue; DMSO17: orange; TCDD17: purple; DMSO11b: light green; TCDD11b: white. B: Samples re-grouped with regard to treatment (irrespective of passage number). DMSO: green; TCDD: red.
An alternative method of mining these normalized data sets is to perform a PCA after consecutively doing a 2Log transformation, a regrouping towards treatment (TCDD) and control (DMSO), and an ANOVA (p < 0.01) (Figure
The NMR data suggested that fatty acid composition changed as a result of TCDD exposure. Therefore, a GC-MS analysis of the fatty acid composition was performed on the apolar fraction of the cells. Consistent with the 1H NMR results, there was a difference in the apolar fractions of cells exposed to TCDD and DMSO with regard to the lipid content. Among the methyl esters of fatty acids that were identified, the arachidonic (AA, C20:4, n-6) and docosahexaenoic (DHA, C22:6, n-3) PUFAs had the same profile within the samples. The NMR data demonstrated that long chain PUFA content correlates highly with PL. Therefore, samples were normalized on the basis of the sum of the AA and DHA signals. This normalized dataset was subjected to MVA (2Log transformation, ANOVA with p < 0.01, followed by a PCA; see Figure
Figure 4
PCA plots (after ANOVA p < 0.01) after normalization of the apolar fraction of HepG2 cells analyzed by GC-MS
PCA plots (after ANOVA p < 0.01) after normalization of the apolar fraction of HepG2 cells analyzed by GC-MS. A: Spheres with the same colour are technical replicates of the same sample per passage number. For denotation of the colours, see legend Figure 3. B: Samples re-grouped with regard to treatment (irrespective of passage number). DMSO: green; TCDD: red.
In order to extract the most significant differences between the exposures, the samples were re-grouped with regard to the treatments and re-analyzed (PCA, after an ANOVA with p < 0.01; see Figure
Additional file 1
PDF file containing Figures S1, S2, S3, S4 and S5 and Tables S1, S2, S3, S4, S5 and S6, mentioned in the text.
Click here for file
The selection of peaks underlying the largest differences (and additionally surviving a Bonferroni correction) between the treatments were identified. Table
Table 2
Apolar metabolites significantly affected by TCDD (p < 0.01) and average fold change ratio between treatments (TCDD vs. DMSO).
C16:1 (n-6)
C16:1 (n-9)
C18:1 (n-9)
Total triglycerides
Total cholesterol ester
*Underscore letters indicate isomers (unidentified).
Individual passage number results are in Additional File
The polar fraction
The 1H NMR spectra of polar fractions were normalized using the PL/CHCl3 ratio described above (obtained from the corresponding apolar fractions) and presented in Figure
Figure 5
1H NMR spectra overlaid (4-fold) of the polar fraction of HepG2 cells exposed to DMSO (below) and TCDD (above)
1H NMR spectra overlaid (4-fold) of the polar fraction of HepG2 cells exposed to DMSO (below) and TCDD (above). The following metabolites are identified by using standards, from the literature and/or databases and the numbers of the signals correspond to those indicated in Table 3. 1&2: leucine and isoleucine; 3: valine: 4: threonine: 5: lactale; 6: alanine; 7: putrescine; 8: acetate; 9: N-acetyl-aspartate; 10: glutamate; 11: glutamine; 12: reduced glutathione; 13: oxidized glutathione; 14: acetone; 15: citric acid; 16: creatine and/or phosphocreatine; 17: choline derivatives; 18: taurine; 19: glycine; 20: serine; 21: nucleotides derived from uridine (UMP, UDP, UTP); 22: nucleotides derived from adenosine (AMP, ADP, ATP); 23: NAD+ and/or NADH; 24: tyrosine; 25: formate.
Figure 6
PCA plots (after ANOVA p < 0.01) after normalization of the polar fraction of HepG2 cells analyzed by 1H NMR and pre-processed and aligned using a program developed in-house
PCA plots (after ANOVA p < 0.01) after normalization of the polar fraction of HepG2 cells analyzed by 1H NMR and pre-processed and aligned using a program developed in-house. A: Spheres with the same colour are technical replicates of the same sample per passage number. For denotation of the colours, see legend to Figure 3. B: Samples re-grouped with regard to treatment (irrespective of passage number). DMSO: green; TCDD: red..
Table 3
Assignment of 1H NMR spectral signals of the polar fraction.
Peak number
Chemical shift (ppm) and multiplicity b,c
Assignment a
0.96 (t)
0.96, 1.05 (t)
1.01, 1.04 (d)
1.32 (d)
1.33 (d)
1.48 (d)
2.00 (s), 2.48 (dd)
2.08 (m), 2.34 (m)
2.12 (m), 2,44 (m), 3.76 (m)
2.18 (m), 2,57 (m), 2.98 (dd), 3.31 (dd), 3.78 (m)
Oxidized glutathione
2.18 (m), 2,57 (m), 2.98 (dd), 3.78 (m), 4.58 (dd)
Reduced glutathione
2.80 (dd)
2.55 (d), 2.67 (d)
3.04 (s), 3.93 (d)
3.23 (s)
Choline derivatives
3.26 (t), 3.41 (t)
3.56 (s)
3.83 (dd), 3.98 (dd)
6.10 (d), 6.14 (d), 8.18-8.22 (ov), 8.52 (s), 8.58 (s)
Nucleotides derived from adenosine AXP (AMP, ADP and/or ATP)
5.95-5.99 (ov), 7.94-7.97 (ov), 8.09 (d)
Nucleotides derived from uridine UXP (UMP, UDP and/or UTP)
6.01-6.20 (ov), 8.12-8.20 (ov), 8.79-8.88 (ov), 9.08-9.36 (ov)
Nicotinamide adenine dinucleotides (NAD, NADH, NADP and/or NADPH)
6.99 (m), 7.20 (m)
8.46 (s)
Numbers correspond with those in Figure 5.
aProtons assignment based on commercial standards, Human Metabolome Database
bs: singlet; d: doublet; dd: doublet/doublet; m: multiplet; ov: different overlapping signals
cOnly observed signals are shown
The normalized NMR data indicated that the levels of several polar intracellular metabolites decreased due to exposure to TCDD (Table
Table 4
Polar metabolites significantly affected by TCDD exposure.
Change fold ( 1 H NMR p < 0.01)
Change fold (LC-MS p < 0.01)
Leucine, Isoleucine
Pantothenic acid
Nucleotides (AXP and UMP)
Oxidized glutathione
Reduced glutathione
The fold changes are given as the ratio of the average content in TCDD and in DMSO. Individual passage number results are in Additional File
*Tentatively identified
The same extracts were analyzed by LC-MS in order to obtain more detailed information concerning the differences in polar metabolite levels due to treatment. The LC-MS data were pre-processed and aligned using MetAlign, the program developed in-house
Figure 7
PCA plots (after ANOVA p < 0.01) after normalization of the polar fraction of HepG2 cells analyzed by LC-ToF-MS and pre-processed and aligned using MetAlign (www.metalign.nl)
PCA plots (after ANOVA p < 0.01) after normalization of the polar fraction of HepG2 cells analyzed by LC-ToF-MS and pre-processed and aligned using MetAlign (www.metalign.nl). A: Spheres with the same colour are technical replicates of the same sample per passage number. For denotation of the colours, see legend Figure 3. B: Samples re-grouped with regard to treatment (irrespective of passage number). DMSO: green; TCDD: red.
Table 5
Masses and fragments of the masses of polar metabolites identified by means of LC-nanomate-Orbitrap.
Parent mass(M+1) 1
Deviation (mDa) 1
Fragments 1
Acetyl spermidine
Citric acid (NH4+)
Pantothenic acid
Glutathione reduced
Glutathione oxidized
1Experimental value obtained in our measurements
2Tentatively assigned formula(e) and/or identification
There were significant increases in citric acid (average ratio TCDD: DMSO = 1.7) and in GSSG (average ratio TCDD: DMSO = 1.3) and GSH (average ratio TCDD: DMSO = 1.2) contents in TCDD treated samples, in general agreement with the 1H NMR data. However, the GSH/GSSG ratio as determined from LC-MS data was reduced, in contrast to the NMR data. Generally, NMR data are more quantitative and reliable; furthermore, the LC-MS data were acquired at a later date than the NMR data. In a follow-up NMR study on the stability of the sample, it was observed that the intensity of the GSH signal decreased over time during storage whereas the intensity of the GSSG signal increased (see Figure S1 in Additional File
This reaction is known to be dependent on pH (i.e. more prominent at higher pH). Therefore storage conditions such as pH are important. NMR samples were measured directly after adding phosphate buffer to the freshly made extracts. In this study the UPLC-TOF/MS samples were stored at -20°C without the phosphate buffer and therefore under more acidic conditions (pH ca. 5 as a result of the extraction conditions). The maximum storage time before UPLC-TOF/MS measurement was 4 months. Under these storage conditions still some conversion to of GSH to GSSG was present as deduced from our experiments.
The main aim of this study was to demonstrate the feasibility of using metabolomics on intracellular compounds of
To substantiate the usefulness of the untargeted metabolomics methodology, the observed changes in metabolite levels were interpreted in the light of published data concerning the effect of TCDD. Ultimately, this was the validation of the procedures and protocols developed in this study. In general, a decrease or increase of an intracellular metabolite concentration is not necessarily an indicator of flux. Actual metabolite levels depend on how biochemical pathways/processes are regulated, how metabolites in pathways interact and how fast and in which directions reactions occur (kinetics). Cells tend to balance their metabolism after perturbation in order to maintain homeostasis. Therefore, it is likely that only changes in the main pools of substrates in the cell are measured when it is exposed to a toxicant. Inhibition/activation of a pathway may lead to increased or decreased levels of relevant pools of compounds. However, it is often difficult to determine whether inhibition or activation is relevant by observing metabolite concentrations at one time point. As discussed in more detail below, TCDD is known to have an effect on the proliferative capacity, metabolism and antioxidant status of the cell.
Effect on cell proliferation
Membrane constituents such as PL (Figure
Spermidine and N1-acetylspermidine were also found in lower levels after TCDD treatment. Polyamines have important functions in cell proliferation, differentiation and apoptosis, and they are essential for hepatic growth and regeneration
ODC is induced during conditions where there is enhanced gene activation and tissue growth. It has been reported that ODC induction is inhibited by TCDD
Therefore, it is possible that induction of polyamine synthesis in cultured HepG2 cells was inhibited by TCDD, resulting in the low content of spermidine.
Effect on metabolism
Changes in metabolic profiles due to TCDD treatment of HepG2 cells indicates major changes in the general metabolism of these cells, which involves fatty acids, amino acids and nucleotides. This is reminiscent of wasting syndrome
The decreases in triglycerides, cholesterol ester and unsaturated fatty acids observed in the 1H NMR analysis of the apolar fraction, together with the decreases in several fatty acids observed in the GC-MS data, indicate that under the conditions used in this study, TCDD affected lipid metabolism.
Using GC-MS analysis it is possible to determine the nature of the fatty acids that are most affected by exposure to TCDD (Table
Pantothenic acid and AMP are metabolites found in lower concentrations after exposure to TCDD (Table
Furthermore, decreases in the content of propionyl- and butyryl-carnitines were observed when cells were exposed to TCDD. These short chain acyl-carnitines are metabolic products of the reaction of acyl CoA and carnitine, which is mediated by transferases in the mitochondria
The level of citrate increased, whereas the level of lactate decreased, after exposure. Citrate is a feedback inhibitor for glycolysis at the level of phosphofructokinase. Lactate is a product of the conversion of pyruvate by lactate dehydrogenase. These two metabolite changes can indicate that glycolytic activity is lower because of an influx of acetyl-CoA, for instance due to fatty acid beta-oxidation. This is in agreement with the breakdown of triglycerides and degradation of fatty acids, and changed activity in mitochondria (see above).
Together, these metabolic effects are reminiscent of a wasting syndrome in which fatty acid metabolism and energy metabolism (glycolysis, TCA cycle) are affected, and the amino acid and nucleic acid pools are reduced. These latter effects could contribute to the observed decrease in cell proliferation. However, it is not clear from the data whether beta-oxidation increased or if fatty acids were transported out of the cell. In adipose tissue, TCDD mobilizes fatty acids towards the plasma
Effect on antioxidant status
NMR and LC-MS spectra demonstrated an increase in reduced (GSH) and oxidized (GSSG) glutathione content in samples treated with TCDD (Table
GSH acts as a nucleophilic "scavenger" of numerous compounds and their metabolites via enzymatic and chemical mechanisms (converting electrophilic centers to thioether bonds) and as a substrate in the glutathione peroxidase (GPx)-mediated reduction of lipid hydroperoxides and of H2O2 to water
In relation to the synthesis of GSH, Boverhof and co-authors
Other effects associated with TCDD treatment
Creatine is decreased in cells as a result of TCDD treatment (Table
The content of taurine, a non-protein sulfur-containing β-amino acid, is increased after the exposure of cells to TCDD. Taurine plays an important role in several biological processes including anti-oxidation, detoxification, membrane stabilization and maintenance of osmolarity
An increased signal (average ratio TCDD: DMSO = 1.5) tentatively assigned to uridine diphosphate-N-acetylgalactosamine (UDP-NAcGal) and/or uridine diphosphate-N-acetylglucosamine (UDP-NacGlu) was observed after TCDD exposure (Table
UDP-NAcGal and UDP-NacGlu are N-acetylhexosamines involved in several biosynthetic reactions including O-glycosylation of proteins on serine and threonine residues. O-linked glycosylation involves the transfer of the N-acetylhexosamine group from the UDP-acetylhexosamine, which is catalyzed by glycosyltransferases
Lower activity of these enzymes could theoretically produce an increase in the levels of UDP-N-acetylhexosamines. In agreement with this, decreased expression of GALNT1, the gene encoding the UDP-N-acetylgalactosamine transferase, was observed by Kim and co-authors
Changes in the intracellular levels of UDP-N-acetylhexosamines, as observed in this study after exposure to TCDD, may indicate that protein O-glycosylation is affected. Altered expression levels of glycosylated protein have been described after TCDD exposure
We have no explanation for the decrease in concentration of N-acetyl aspartate. This metabolite is normally expressed at high concentrations in the brain and has been implicated as an osmolyte
The present study demonstrates that untargeted profiling of the polar and apolar extracts of
This study is an example of how complementary analytical techniques (NMR, GCMS, LCMS) can be used on the same samples and provide complementary data. The untargeted nature of the experiments aids in finding differences in profiles, which are then subjected to identification approaches. As discussed, normalization and sufficient biological replicates (and repeated experiments) are essential for reliability (in terms of statistical analysis) in detecting relatively small differences in analyte concentrations.
Maintaining homeostasis could be the driving force in cells responsible for keeping concentration differences small. The effects are predominantly centered on storage pools of metabolites.
Minimal essential medium (MEM glutamax), non-essential amino acids, sodium pyruvate, penicillin/streptomycin and fetal bovine serum were purchased from Gibco BRL (Breda, The Netherlands). Ammonium acetate (NH4Ac), sodium hydroxide (NaOH), sodium chloride (NaCl), dipotassium hydrogenphosphate (K2HPO4), monopotassium hydrogenphosphate (KH2PO4), boron trifluoride (BF3), EDTA and deuterated chloroform (CDCl3), deuterium oxide (D2O) and deuterated methanol (CD3OD) were obtained from Merck (Darmstadt, Germany); methanol (MeOH) and iso-octane from Biosolve (Valkenswaard, The Netherlands); DMSO and TMSP from Sigma-Aldrich (St. Louis, MO, USA) and TCDD (CAS no. 1746-01-6) from Cerilliant (Round Rock, TX, USA). All chemicals and solvents were purchased in the highest purity available. Ultra-pure water was obtained using the PureLab equipment from Rossmark (Ede, The Netherlands).
Cell culture treatment and disruption
HepG2 cells were obtained from the ATCC (Rockville, MD) and cultured in MEM glutamax supplemented with 1% non-essential amino acids, 1% sodium-pyruvate, 1% penicillin/streptomycin and 10% fetal bovine serum (FBS). The cells were incubated at 37°C with 5% CO2. When the cell monolayer reached 80% confluency, the medium was replaced with fresh medium with or without TCDD. TCDD was administered to a final concentration of 10 nM by adding 60 μl of a solution of 2 μM of TCDD in DMSO to 12 ml of medium. In the control treatments, cells were exposed to 12 ml of medium to which 60 μl of DMSO had been added. The final concentration of 10 nM TCDD was sub-cytotoxic and has been used in previous omics studies
A separate study - prior to this one - with additional replicates was done to establish the suitability of the normalization approach based on PL signals. This study involved cell counting. In short, the cells were detached from the flasks by trypsinization and counted using a hemocytometer (Neubauer chamber). The results are given in Table S2, Additional File
Extraction of the apolar and polar fractions
The metabolomics study was performed on two different fractions of the cells, one apolar fraction containing the apolar metabolites (membranes and intracellular lipids), and one polar fraction, containing the polar and semi-polar intracellular metabolites. To extract these, the disrupted cells were centrifuged (4°C) and the pellet (apolar fraction) and supernatant (polar fraction) were separated and processed.
Apolar fraction
The pellet was re-suspended in 1 M NH4Ac and freeze-dried. Once dried, the residue was extracted three times with 1 ml of CDCl3, after which the organic solvent was evaporated under N2 flow. The dried extract was dissolved in 1 ml of CDCl3; an aliquot (0.6 ml) was used for NMR analysis and the remaining sample was stored at -20°C for GC-MS analysis.
Polar fraction
The supernatant containing the polar intracellular metabolites was freeze-dried. The pellet was re-suspended in 1 ml of 100% CD3OD, dried with N2 and re-suspended in 50% CD3OD/50% D2O. The solution was centrifuged to remove the proteins (in the pellet) and the supernatant was dried under N2 flow. The dry residue was dissolved in 1 ml of D2O. Some of the sample (350 μl) was stored at -20°C for UPLC-TOF/MS analysis. To the remainder, 1 M phosphate buffer (90:10; v: v; pH = 7) in 99.95% D2O was added and used for NMR analysis.
Analysis of the polar and apolar fractions
NMR analysis
The samples were analyzed by NMR just after extraction. Some of these samples were also used for a follow-up study with the aim of determining the stability of the different metabolites over time. These samples were stored at -20°C in phosphate buffer (pH = 7) for 9 months.
The 1H NMR spectra were recorded at 400.13 MHz at 300.0 (± 0.05) K on a Bruker Avance 400 narrow bore using a 5.0-mm probe. The spectrometer settings were 2s relaxation delay; 1024 scans for the polar fraction and 128 scans for the apolar fraction, with four dummy scans in all cases; a spectral width of 5000 Hz; a 60 degree pulse; acquisition in 16 K data points. Prior to data analysis, the raw NMR data were subjected to a squared sine bell filter (shifted 1/2 pi), zero-filling to 128 K data points, Fourier transformation and phase correction.
NMR raw data and the corresponding metadata are provided as an additional file (see Additional file
Additional file 2
Metadata and NMR raw data: ZIP file containing: -All NMR raw data (in JCAMP-DX format ".dx") -The metadata on NMR (polars & apolars), LC-MS and GC-MS data sets (as Tab delimited files ".txt").
Click here for file
GC-MS analysis
A miniature scale BF3-mediated methanolysis method for derivatizing fatty acids (for GC-MS) was used. In brief: 132 μl of the apolar fraction was dried under nitrogen. To this dry extract, 40 μl of 0.5 N NaOH in MeOH was added. The capped vial was placed in a pre-heated oven at 65°C for 30 minutes after which 50 μl of BF3 (20% in methanol) was added. After closing it again, the vial was put in the oven for three minutes at 65°C; 0.2 ml of iso-octane was added. The vial was again closed and put in the oven for two minutes at 65°C. After cooling, the volume was adjusted to 1 ml with a saturated NaCl solution. This was shaken firmly and the content was left to separate into two phases. The vial was centrifuged at 2800 rpm and the upper phase was carefully collected. For GC-MS analysis, 75 μl of the iso-octane phase was transferred into a new vial and kept at -20°C pending analysis.
A Trace GC 2000 series gas chromatograph interfaced with a Trace MS Plus mass spectrometer (Thermo Finnigan, San Jose, CA) was used for GC/MS analysis. Two μl of sample was injected using a PTV-Splitless injection. For the separations, a RTX-5 column (length 10 m, internal diameter 0.18 mm, stationary phase film 0.20_m; Alltech, Breda, The Netherlands) was used with helium as the carrier at a constant flow of 1.3 ml/min. The GC temperature program was: 2 min at 80°C; increase temperature at 5.5°C min-1 to 185°C followed by 3.5°C min-1 to 290°C. This latter temperature was kept for 12 min and afterwards it decreased by 120°C min-1 to 150°C. The GC-MS interface temperature was 280°C; the source temperature was 200°C. Mass scanning in EI (electron impact) mode was carried out for the range of 50-500 m/z at a scan time of 0.4 seconds. The detector voltage was set to 500 V and the setting of the EI ionization source was 70 eV. All data were collected consecutively in one analysis series to minimize chromatography differences and the injection sequence was randomized according to Vos and co-authors
Metadata for GS-MS files are provided as an additional file (see Additional file
UPLC-TOF/MS analysis
The fraction containing the polar metabolites was stored for a maximum of 4 months at -20°C (pH = 5) before analysis by UPLC-TOF/MS. UPLC-TOF/MS samples were diluted twice with D2O prior to analysis; formic acid was added to a final concentration of 0.1%. The injection sequence was randomized according to Vos and co-authors
The mobile phase consisted of water, acetonitrile and formic acid (A:100/0/0.2 and B: 0/100/0.2). Gradient elution was performed at 0.4 ml min-1. The initial eluent composition, 100% A, was held for one min after which the composition was changed to 85% A and 15% B in 14 min. Afterwards, the composition of B was increased to 30% in 10 min and subsequently increased again to 90% in three min; this composition was maintained for five min. The injection volume was 10 μl. The effluent of the LC system was interfaced directly with the TOF-MS, which was operated in positive mode polarity. A stable spray was obtained with a capillary voltage of 2.5 kV, a source temperature of 120°C and desolvatation temperature of 350°C. The desolvatation and cone gas flows were 600 and 50 L h-1, respectively. The cone voltage was 50 V. Spectra were collected in centroid mode from m/z 80 to 1500, with a scan duration of 0.2 s. Accurate masses were obtained after lock mass correction. The mass spectrometer was operated in W mode with the Dynamic Range Enhancement turned on and the resolution was 8500 (FWHM).
Metadata for LC-MS files are provided as an additional file (see Additional file
LC-nanomate-Orbitrap analysis
Liquid Chromatography-Mass Spectrometry (LC-PDA-MS)
The system consisted of an Accela U HPLC system (pump and auto sampler) equipped with an Accela PDA with a one cm light Pipe flow cell (ThermoFisher Scientific) and coupled to a LTQ/Orbitrap hybrid mass spectrometer (Thermo Fisher Scientific) that was equipped with a chip-based nano-electrospray ionization source (Triversa NanoMate (Advion BioSciences). The sample (10 μl) was separated on a 150 mm × 2.1 mm UPLC BEH-C8 column with 1.7 μm particles (Waters). The flow rate was set at 0.4 ml min-1. Isopropanol (60 μl min-1 of 100%) was added between PDA and NanoMate via a T-junction to the LC flow to ensure a stable nanospray. The sample was loaded with 100% eluent A (H2O/0.1% formic acid). The gradient was the same as that used for the UPLC-TOF/MS analysis. The PDA detector, placed between the analytical column and the NanoMate, was programmed to acquire data every second from 210 to 600 nm with a resolution of 1 nm and 2 Hz sampling rate. The NanoMate source was operated in the positive ionization mode with a HD_A_384 chip with a spray voltage of 1.4 kV and 0.4 psi N2 gas. The NanoMate was used in the LC coupling mode with fraction collection. The total flow (460 μl/min) was split using capillary tubing for MS spray (480 nl/min) and for fractionation (459.5 μl/min). Fractions were collected by the NanoMate from retention time 0.5 to 19.5 min in 3 s fractions (23 μl) in a 384 well plate (twin tec, Eppendorf). The plate temperature was set at 10°C. After collection, extra isopropanol (5 μl) was added. The plate was sealed to prevent evaporation of the fractions. These fractions were used in the MSn experiments. Parallel to the fractionation, the MS spray from the NanoMate was used to record a full FTMS scan (m/z 80-1000) with resolution of 15.000 (at m/z 400). Automatic tuning was carried out on m/z 566.89. The full AGC target was set to 30000. The Orbitrap was externally calibrated in positive mode using sodium formate clusters in the range m/z 158.69-1110.79.
MSn by direct infusion
The system consisted of a LTQ/Orbitrap hybrid mass spectrometer (Thermo Fisher Scientific) equipped with a chip-based nano-electrospray ionization source (Triversa NanoMate, Advion BioSciences). After manually selecting the masses of interest, recorded in the full scan MS, the fractions containing these masses were subjected to MSn fragmentation in positive mode. Delivery of the sample to MS: The NanoMate used a pipette tip to take the sample from a selected well and infused this sample to the MS via the spray nozzle on the chip. NanoMate parameters were: spray voltage 1.7 kV and 0.6 psi N2 gas. The mass spectrometer was tuned on m/z 566.89. The full AGC target was set to 30000. The Orbitrap was externally calibrated in positive mode using sodium formate clusters in the range m/z 158.69-1110.79. For performing the MSn experiments in MS1 a window of 1 m/z was used to isolate the mass of interest. The resolution used in the MS1 scan event was 60.000. For all dependent scan events (MSn) a resolution of 15000 and normalized collision energy 35% was used.
MSn description:
MS 1 Full scan with limited m/z range, m/z of interest ± 0.5 m/z
MS 2 fragmentation of most intense m/z of MS1
MS 3 fragmentation of five most intense fragments in MS2
MS 4 fragmentation of five most intense fragments in MS3
MS 5 fragmentation of three most intense fragments in MS4
Following the sample infusion and MS analysis, the pipette tip was ejected and a new tip and nozzle were used for each sample, thereby preventing any cross-contamination or carry-over.
Data analysis
NMR data analysis
Pre-processing of the data and alignment
A visual inspection of the technical replicates (4-fold) of each sample was carried out to ensure a high degree of reproducibility. The NMR data were pre-processed and aligned using a program developed in-house. This program was described in the Materials and Methods section of the paper by Lommen et al.
The aligned fingerprint data in the form of generated spreadsheets were normalized using factors obtained from the scaling on the phospholipid signals of the 1H NMR spectra of the apolar fraction (see RESULTS section).
Statistical analysis
The normalized spreadsheets of both datasets were subjected to statistical analysis using Genemaths XT
Identification of metabolites
The identification of relevant signals from the NMR data was carried out by using commercially available standards and/or from literature and databases, such as HMDB
MS data analysis
Pre-processing of the data and alignment
A visual inspection of the technical replicates (4-fold) of each sample was carried out to ensure that high degree of reproducibility. The GC-MS and LC-MS data were pre-processed and aligned using MetAlign
LC-MS data
The aligned fingerprint data of the polar fraction dataset, in the form of generated spreadsheets, were normalized using the phospholipid scaling factors obtained from the 1H NMR spectra of the apolar fraction (see RESULTS section).
GC-MS data
The identified peaks of GC-MS chromatograms were manually integrated. This fraction was normalized by scaling to the raw values of the integrals of docosahexaenoic and arachidonic methyl esters (see RESULTS section).
Statistical analysis
The same procedure as described in the NMR section was used.
Identification of metabolites
A method of facilitating further analysis and identification was accomplished using GM2MS, an application of MetAlign that re-creates "new chromatograms" that contain only the peaks exported from the PCA selection
Authors' contributions
AP, JK, J van D and DJ participated in the design of the study and overall discussion; CH helped to set up the methodology for
The Netherlands Toxicogenomics Centre, NTC (
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The transcriptional signature of dioxin in human hepatoma HepG2 cells
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Use of metabolically competent human hepatoma cells for the detection of mutagens and antimutagens
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Chemical fingerprinting for the evaluation of unintended secondary metabolic changes in transgenic food crops
Aryl hydrocarbon receptor-mediated induction of the cytosolic phospholipase A(2)alpha gene by 2,3,7,8-tetrachlorodibenzo-p-dioxin in mouse hepatoma Hepa-1c1c7 cells
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Agmatine, the bacterial amine, is widely distributed in mammalian tissues
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Regulation of polyamine biosynthesis by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)
Polyamine metabolism and function
Comparative Toxicogenomic Analysis of the Hepatotoxic Effects of TCDD in Sprague Dawley Rats and C57BL/6 Mice
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) alters the mRNA expression of critical genes associated with cholesterol metabolism, bile acid biosynthesis, and bile transport in rat liver: A microarray study
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Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin on lipid synthesis and lipogenic enzymes in the rat
Alterations of hepatic acetyl-CoA carboxylase by 2,3,7,8-tetrachlorodibenzo- p -dioxin
Temporal and Dose-Dependent Hepatic Gene Expression Patterns in Mice Provide New Insights into TCDD-Mediated Hepatotoxicity
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Low-dose dioxins alter gene expression related to cholesterol biosynthesis, lipogenesis, and glucose metabolism through the aryl hydrocarbon receptor-mediated pathway in mouse liver
Aryl-hydrocarbon receptor is an inhibitory regulator of lipid synthesis and of commitment to adipogenesis
Interaction of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) with induced adipocyte differentiation in mouse embryonic fibroblasts (MEFs) involves tyrosine kinase c-Src
Loss of stearoyl-CoA desaturase-1 function protects mice against adiposity
Acylcarnitines in intermediary metabolism
Propionyl-L-carnitine: Biochemical significance and possible role in cardiac metabolism
Effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and fasting on body weight and lipid parameters in rats
Characterization of the hyperlipidemia in guinea pigs induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin
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Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin on lipid profiles in tissue of the Fischer rat
Comparative ability of TCDD to induce lipid peroxidation in rats, guinea pigs, and Syrian golden hamsters
Studies on the Role of Lipid Peroxidation in the Acute Toxicity of TCDD in Rats
Oxidative stress in liver and brain of the hatchling chicken (Gallus domesticus) following in ovo injection with TCDD
Regulation of cellular glutathione
Glutathione: Toxicological Implications
Cytotoxicity and mitochondrial dysfunction of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in isolated rat hepatocytes
The effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on oxidative enzymes in adipocytes and liver
Inhibition of TCDD-induced lipid peroxidation, glutathione peroxidase activity and toxicity by BHA and glutathione
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Taurine reduces the secretion of apolipoprotein B100 and lipids in HepG2 cells
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text:
Villas-Boas tipped to again follow in Mourinho's footsteps, United shading City and much more in the world's first football podcast with an Asia-Pacific perspective
Will Andre Villas-Boas make the same mistake and follow Jose Mourinho's footsteps by taking over at Inter Milan after the sacking of Claudio Ranieri? Are Manchester United winning the battle on and off the field versus Manchester City, with the benefit of [arguably] more favourable refereeing decisions? And as the A-League finals begin, what is the verdict on the regular season performance of childhood friends and fellow marquee men - Harry Kewell [Melbourne Victory] and Brett Emerton [Sydney FC]?
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Is AVB destined for the Inter Milan post?
- Man Utd go three points clear after controversial win over Fulham
Neil Humphreys, award-winning football author, columnist and satirist
Mayur Bhanji, ex-BBC & Real Madrid TV Reporter, Football Asia Producer
Richard Lenton, Premier League magazine show host, Mio Stadium
Jason Dasey, Premier League Host & Executive Producer, Astro SuperSport


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William W.wrote a review of Mammut Ophir Rope Bag on November 9, 2016
Simplistic Design
Familiarity: I've put it through the wringer
I am thrilled with this bag, it is simple and rugged. It easily will fit 80m ropes and hasnt shown any significant wear after 2 months of being strapped to the outside of my climbing pack. The only down side I have encountered is the integrated tarp could be a bit larger.


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July 26, 2017
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Was I Dreaming? Publix Icing Recipe
Decorating By pugmama1 Updated 12 Oct 2010 , 6:32pm by CWR41
Over the last weekend I saw (or at least thought so) a thread that talked about the Publix icing. There were a couple of recipes that called for a cup of boiling water. I didn't save it and would love to get the recipes. Anyone out there know what I am talking about?
Quote by @%username% on %date%


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Hello! So this is my first blog... It;s not much, just letting everybody know that I just managed to fix my disk with ome cling film and an exfoliating make-up removing wipe (?). So, if there is an article on the wiki I think isn't here then I'll make it or edit it! <3 Don't you love the miracles of boredom?
~You've Been Silenced Today by User:Sirens of Oakvale :D 01:53, July 3, 2011 (UTC)