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Is It Smart For Us To Go Upstream?

I've suggested

that instead of constantly cutting costs, we should consider how to

more money.

However, there's a right way to do that and a wrong way.

The wrong way, IMHO, is proposed in an

this week by Kofi Amoo-Gottfried, chief strategy officer at

FCB Garfinkel New York.

Kofi starts by summing up what he sees as the problem, by using a quote from a Diageo marketer that has gained quite a bit of attention recently: "Agencies unable to prove they are driving value for clients risk becoming little more than dust."

It's an attention-grabbing quote, but even a quick analysis shows it to be somewhat meaningless. Surely

business, in any field, anywhere in the world, faces oblivion if it is unable to prove it creates value?

But I guess it's his solution that I really disagree with. Kofi writes: "The client-agency relationship needs to start way upstream of the communications brief. Clients need to invite agencies into the depths of their business, to share all of their data...

we need to become true general contractors."

Presumably, becoming 'general contractors... upstream' means going into areas beyond marketing.

Sounds exciting. But here's my question. What are we actually going to do, when we start getting involved with areas beyond marketing? Are we really going to get involved with finance? HR? Distribution? Manufacturing?

We just don't have the skills.

Are we really proposing to send a Comms Planner to a finance meeting, to sit alongside the Client's Finance Director, and a couple of guys from Goldman Sachs?

Are we really proposing to send a Copywriter to a meeting about building a new factory, alongside the Client's Head of Manufacturing, and a couple of guys from Balfour Beatty?

It's a joke.

And worse than that, it depreciates what we actually

In an age of commoditisation, marketing (and hence marketing communications) are more important than ever.

Land Rover was once a unique product. Now everyone makes an SUV. Gordon's once had a near-monopoly on gin. Now there are 50 gins.

In fact I'd turn the Diageo marketer's question back onto the client companies themselves: how is the average maker of a vodka, beer, training shoe, mid-size sedan, vitamin, juice, or coffee... or provider of insurance, mortgages, or personal loans... doing anything to drive value for their corporations?

Their products are almost completely undifferentiated. The corporate structures (of large corporations) are almost all identical. Their financing and management techniques do not significantly differ.

It's primarily

that can make the difference.

And yet

80% of CEOs do not trust their marketers, and 70% of CEOs believe marketers are disconnected from business results. (

The truth is that it's not we who need to go upstream, it's our Clients.

The Marketer is able to create far more value for the corporation than the Manufacturing Guy (since most companies are making me-too products), or the HR person, the legal counsel, etc.

Given the importance of marketing, every Marketing Director should sit on their company's board. Hell, every CMO should be sitting right next to the CEO.

And we should make it our mission to help them get there.

Because if they rise - which they deserve to - we rise.

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The ECD Who Doesn't Sign Off Any Work

Very interesting interview this month with Nils Leonard, Chairman and CCO of Grey London.

The bit that really jumps out is that he claims he doesn't sign off any of the work.

Let's rewind. When Leonard first took over at Grey, a few people carped along the lines of 'how can he be an ECD when he's never done any great work as a creative', which is a rather foolish argument, akin to questioning Arsene Wenger's managerial ability on the grounds that he was a mediocre left-back.

Leonard's success is undeniable. Grey London has been utterly transformed under his watch. The agency where people "went to die", and whose creative floor was once known as "Jurassic Park", is now arguably one of the most dynamic in the world. In the last five years, the place has won a shitload of awards, and more than doubled in size.

So what did he do that was so different?

I'm pretty sure I know the answer, but he himself prefers not to tell us. Because it certainly can't be any of the three points he makes in his interview.

The first of these was his decision to go open-plan. As regular readers will know, I'm not a fan. But perhaps Leonard has a new take on it? "We tore down the offices", he says, "and for a reason: it literally is a physical barrier between an idea happening or not if you have to stop outside a door and knock to go and talk to somebody."

Sounds hip, yeah. But if you actually examine it, I reckon this argument is super-weak. I mean... is that really such a huge barrier - a fucking door? Last time I checked, doors do open. And fairly easily, too. I don't recall them being much of a barrier when we had them at DDB London. They certainly never kept any suits out who wanted to come in. Or indeed anyone. They simply knocked, and entered! And once inside, you could actually have a proper chat... which in an open plan office, you can't.

But anyway, whatever the merits of open-plan, this move cannot be the cause of Grey's recent successes (21 pitches won out of 24), since every other agency in London has gone open-plan too. Hence, no competitive advantage there.

His second point is around looking for what he calls 'long ideas' rather than 'big ideas'. This means ideas that people want to spend time with, rather than simply ideas which can support multiple executions. And he's walked the walk here, for example producing a stage show 'The Angina Monologues' for the British Heart Foundation that was also broadcast on TV.

He's phrased it beautifully - "long ideas" - but a commitment to producing longer-form content cannot be the source of Grey's competitive advantage either, since every other agency in town is doing the same.

His last point is around "no sign-off". Leonard explains that a team consisting of a creative, a planner, and "I guess, a suit, or a producer" (he means a suit, but doesn't want to sound old-school) takes ultimate responsibility for the work - not him.

There are arguments both ways here. Yes, it's true that if people know the buck stops with them, they feel a greater sense of ownership, and may create better work. But on the other hand, you could argue it's a mistake to remove the CCO from the process - does it really make sense for the agency's best creative not to be involved in the work?

He's certainly being a little disingenuous by reducing the CCO's role to a mere 'sign-off'. The good ECD's or CCO's or whatever the top person is called in an agency are doing a hell of a lot more than just signing off the work. They're adding to it, improving, finessing... sometimes transforming it.

In any case, once again this can't be the secret of Grey's recent out-performance, since many other agencies in London operate exactly the same system - including the last two where I worked, DDB and BBH - as do many other agencies around the world.

And it's certainly not true that this system is, as Nils Leonard claims, significantly faster. "If you trust people," he writes, "you don’t put barriers in the way and you speed up the process... you’ll be twice as fast as most agencies."

Really? Twice as fast? The ECD gets a day or two max to have their input - sometimes an hour. That's not 50% of the entire strategy/ideation/creative direction/presentation process. It's way, way less.

So what is the real reason for Grey's success, and why does Nils Leonard not tell us, instead making claims for the success of his agency which sound modern and groovy, but which aren't actually any different to what every other agency is doing?

In my view, the major change that has made the difference at Grey since the arrival of Nils Leonard... is the arrival of one Nils Leonard.

Obviously he doesn't say that in the interview, since it would sound horribly immodest (not to mention old-fashioned) to claim that one great creative leader can make the difference. But we all know that they can.

I have no interest in crawling up the bloke's arse, since I'm 10,000 miles away and not planning to go back. But by all accounts he's just very, very good at his job. Highly charming, highly creative, great with clients, great with ideas, great at hiring... and of course, great at PR.

And surely it's this latter quality that explains why in his interview he weaves a compelling story - a parable of modernity and inclusivity - rather than revealing the rather boring and old-fashioned truth.

The Problem With Lying

I'm attending some important research groups on Monday night... and feeling a little worried about them.

Why? Because people often lie.

In the polls, about 33% of people said they would vote Conservative. In last week's UK general election, about 37% did so. In other words, 4% of people simply lied.

Of course, some people think we lie.

The latest annual study conducted by research firm Roy Morgan into the perceived honesty of different professions has placed advertising in 29th place out of 30. Only car salesmen ranked lower.

I've written before about the irony that advertisers are perceived as dishonest, when the truth is that we don't lie to people, it's the people who lie to us.

The great strategist Russell Davies, on leaving advertising, famously described what he wouldn't miss: "Endless focus groups with company car drivers - constantly lying about why they drove the car they did."

He's so right. No one is going to admit in a research group that they drive a certain car because they want people to think they are rich, or successful, or sexy. But surely those motivations are in there.

And I'll never forget an interview that artists Jake and Dinos Chapman gave to GQ magazine. When shown examples of the brothers' work (similar to the picture above right) a selection of GQ readers unanimously claimed that they didn't like their art, because it was "ugly."

The Chapman brothers' response? "They're lying." The brothers reckoned that the real reason the GQ readers didn't like the art was because "they were turned on", and knew it would be socially unacceptable to say so.

Until neuromarketing research works properly, and we can actually see inside people's brains and discover what they are really thinking, rather than what they say they are thinking, we should continue to be suspicious of what people say. Very suspicious.

I Think We Should Abolish The Word 'Creativity'

Wendy Clark, a senior Coca-Cola marketer in the US, is well-known in Agency circles as a force for good - a Client who has supported great work, time and time again.

And I love that - as chair of the Effectiveness jury at Cannes this year - she writes an article that instead of arguing for the primacy of effectiveness, makes a plea for the importance of creativity.

"If you leave creativity behind, you are leaving some measure of effectiveness behind too," she writes.

Very cool.

However, I do have one quibble with her argument.

She develops her theme by making a big play around the word "and", arguing for work that is both creative "and" effective.

And I guess I feel that 'creativity and effectiveness' are not similar concepts that can be linked together with an 'and', like 'fish and chips'.

Because they don't exist on the same plane really, do they? Surely effectiveness is an outcome, and creativity is a means of achieving it?

"Effectiveness is our goal, creativity is our tool." That's how Nigel Bogle always used to phrase it.

In other words, effectiveness is a hole in the wall, and creativity is a sledgehammer.

Let's face it, you could still have a successful advertising campaign by filling media space with a completely literal and uncreative message.

Here are two executions in which the communication is identical.

First, expressed without creativity:

Now, with creativity:

The first execution could still be effective. A lot of people like Wayne Rooney, and a timely and well-bought media placement that reinforces the association between Nike, Rooney and England could help drive affinity for the brand.

But the second execution will be more effective (because more impactful, more memorable, and more cool. Yes, in sportswear, cool matters).

The fact is, we Agency people are not using creativity because it's more fun for us. (Although it is). We are using creativity because it increases the effectiveness of advertising. Creativity is an amplifier, that's all.

The problem we have is that too many Clients think we like creativity for its own sake, and hence they lack trust in our recommendations.

So what if we stopped using the word 'creativity' completely?

The Creative Department would henceforth be known as the Effectiveness Amplification Department, and the Creative Director as the Effectiveness Amplification Director. Creative awards would be called Effectiveness Amplification Awards.

What do you think?

Our Debate About 'Risky' Versus 'Safe' Advertising Is Embarrassingly Amateur

The attitude that we have towards risk in our industry is embarrassingly amateur.

And I'm pointing the finger at both Agencies and Clients here, who tend to fall into opposite but equally naive traps.

The Agency view, most commonly (but not exclusively) heard from Creatives, is that "safe advertising is actually more risky than risky advertising." The theory here is that if advertising is 'vanilla' it "won't cut through" and is therefore likely to be useless. They rail against Clients' "conservatism", and wish their Clients would have 'more vision' or, moving further down the body, 'more balls.'

Many Clients, on the other hand, make huge efforts to minimise risk. The principal method is via research. If a campaign has 'passed' research, they feel it has a higher chance of being successful, i.e. should give them a higher than average return on investment. If a campaign is similar to things that have worked before, they again feel it has a higher chance of working. If an agency has done similar work before, a photographer or director has done similar work before, they will be more comfortable, because they will be 'reducing the risk' and therefore maximising their chance of a high return.

Both attitudes are so, so wrong.

For a more informed view, we need to learn something from the risk professionals - financial investors.

First of all, they are smart enough to realise that every investment carries risk. Indeed, they classify it. Low-risk investments include short-dated US Treasury Bonds - the chance of the United States going bankrupt within the next 30 or 60 days is tiny. A medium risk investment might be shares in a company like General Electric or Boeing. A high-risk investment could be loans to countries considered at risk of default, such as Greece and Argentina.

The difference with advertising is that financial investors understand that risk and reward are highly correlated. The low-risk 30-day US Treasury Bond delivers only a small return - just 0.03% annually, as of last Friday. Boeing's dividend yield is currently 2.45%, GE's is 3.7%. Whereas a higher-risk Greek government 10-year bond is currently yielding 12.7%, and Argentina 7.2%.

The more risk you are willing to take, the higher your potential reward. It's that simple.

And it's the same for every area of life. Asking a hottie out? High risk, high reward. Working for the civil service? Low risk, low reward.

If only we in advertising could understand this simple correlation. The Creatives arguing for high-risk as a 'form of safety' are idiots. High-risk work is high-risk. It's also potentially higher reward, of course. And the Clients demanding that their work be de-risked should not expect that it will also generate high returns. It won't.

Cadbury's drumming gorilla was high-risk (because very weird and unusual - it could have bombed) but when it worked, the rewards were high. A beer ad that just shows hops being farmed and droplets of water glistening on the bottle is low risk (it's not going to offend anybody) but will be low reward.

TLDR: an ad campaign that has a chance of earning a high return will also involve high risk. A low risk campaign can never earn a high return.

Let's Get Classical

Having the right music makes a huge difference to the success of a TV ad. Trouble is, it can come at a huge cost.

I reckon we're ignoring an infinite supply of amazing yet affordable music - classical.

Hit songs of the last 50 years certainly bring a lot to the party. First off, they're often great pieces of music - that's why they became hits.

And secondly, because people know what a well-known song is 'about', it can amplify the meaning of an ad. Examples: John Lewis 'Always A Woman', Chrysler 'Made In Detroit' (feat. Eminem).

Then there's the sheer fame factor too - recognition and memorability are important (and heavily tracked) aspects of an ad's success.

But these pluses come at a cost. Have you noticed how the price of concert tickets has shot through the roof? That's because artists aren't making what they used to from record sales. And I reckon the cost of music for ads has been another casualty.

A big track by a big artist can cost anywhere from $200,000 to $500,000. It's not uncommon to be quoted six figures for some obscure 60's soul track nowadays.

But there is an alternative.

I've just made a TV ad using classical music as the soundtrack, and it's made me realise what a relatively untapped resource we have here.

Classical music is a hell of a lot cheaper - it's out of copyright, so there's no cost for the publishing rights, also the recordings themselves are extremely cheap to acquire.

And because you have hundreds of years' worth of music to draw from, there will always be a piece - probably a famous piece - that will reflect the mood you want for your ad.

But won't it make my ad seem old-fashioned? I hear you ask. Well, that depends. Some classical music does seem very twee to our ears now. But some sounds more modern than most of today's pop music.

If somehow those arguments have failed to convince you of the merits of classical music, here's the clincher: Jonathan Glazer loves it.

How Long Does It Really Take To Crack A Brief?

We need to talk about time.

Self-evidently, we are being given less and less time to crack briefs nowadays.

And I doubt that's going to change.

So we're going to need to work quicker, smarter... all of that.

But also, I think we need to do a better job of explaining to everyone else the role that time plays in the creative process.

Because so far, we haven't explained it very well at all.

Most Planners, Suits and Clients still think that engaging a Creative is much like engaging a builder. They describe the job, and then ask how long we think it will take.

For the builder, that's easy. If he can place 1000 bricks a day, then he knows that a 5000-brick wall will take 5 days.

But the creative process doesn't work that way.

A creative team might have the job licked in a day. (Something the builder could never achieve).

On the other hand, they might work on it for a week, and not crack it. (Let's define 'crack it' as 'come up with a solution that the CD approves'). In other words they might do a whole week and get all their work rejected, ending up with absolutely nothing. Which would be the equivalent of the builder working for a week and failing to have a single brick in place at the end of it.

Surely this illustrates that to even ask us the question 'how long do you think will this take' is to misunderstand the nature of what we do.

But they're not going to stop asking. So a better answer to give them might be something that's phrased more in terms of 'confidence interval'.

I estimate that a typical team, given a typical brief, has a 20% chance of cracking it in a day. (Obviously a more senior or better-than-average team would have a higher chance, and a more difficult brief means a lower chance).

After three days, I reckon the chance of cracking the brief goes up to about 50%, and after five days (i.e. one week), I'd say the team has a two-in-three chance of cracking it.

I estimate that when given two weeks on a brief, the typical team has a 90% chance of cracking it. But from then on, the crack rate rises very slowly. If they haven't cracked it after two weeks, they probably never will.

So what do you think? Does this tally with your experience?Am I being too generous? Too stingy? Oh, and do you have any tips for working quicker and/or smarter...

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Beyond Open Plan - American Agency Introduces 'Open Play'

US advertising agency Flair Loop, based in Madison, Wisconsin, has announced a radical re-vamp of how its creatives work together… and sit together.

As of April 1st, the shop has removed all desks and chairs from the creative department, and replaced them with thousands of brightly-coloured plastic balls to create a seating plan it calls ‘open play’.

“My kids love playing in those ball pits,” commented CEO Terry Friendly, explaining the rationale for the move. “So I’m sure our Creatives will too.”

Friendly denied that the purpose of the change is to cut costs. “Just as with the switch to open plan fifteen years ago, we’ll actually have the same number of Creatives in the same space, so quite evidently we’re not going ‘open play’ to save money."

"You have to bear in mind that although our product is creativity, for reasons that are too complicated to go into right now, only 20% of our staff are Creatives. This move shows that I'm willing to do whatever it takes to make us a more creative company, short of actually hiring more Creatives."

“I feel it will be a big improvement,” said Account Manager Sally Dazzle. “At the moment, when I go talk to a Creative, I’m having a conversation with someone who knows how to create advertising whereas I don’t, so it’s not a level playing field. With the Creatives floundering in a sea of plastic balls, I will feel more at ease when I’m talking to them.”

However, Planning Director Steven Glasses, while applauding the innovation, cautioned that it should not be seen as a revolution in how the agency functions. “Let’s not forget that the average brief has to spend six weeks in Planning, to ensure we have incorporated every possible angle into the single-minded proposition, and every single suggestion from all the various clients involved. Only then does the brief spend a couple of days in the creative department for them to crack an idea. So the fact that the Creatives are now working in a giant ball pit is not going to make a huge difference to the quality of work that emerges.”

Art Director Matt Hair agreed that ‘open play’ would have little effect on his working habits. “When I’m in the office, I’m just looking at Facebook or watching YouTube anyway,” said Hair. “Everyone knows that creative thinking requires peace and quiet, so when we get a brief, my partner and I already have to go outside to a cafe, or find a park bench we can work on. I guess we’ll just carry on doing that.”

Link to full story here.

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Can You Solve Any Problem By Asking One Simple Question?

The early lumber industry in Europe and North America would float logs along rivers to transport them to the sawmill. Masses of individual logs were driven downriver like huge herds of cattle.

But sometimes, the timber would stack up and cause a logjam.

Men called 'log drivers' were tasked with unblocking the jam. Interestingly, they soon learned that there was often a 'key log', whose removal would free up the entire logjam. Thus their goal became to locate and remove this key log.

Similarly, progress on an advertising project often grinds to a halt. (There are disagreements, there is lack of clarity, there is confusion about goals, or methods, or strategy. Whatever the reason, the project or meeting has stalled).

I often find myself wondering what the key log in an advertising logjam is, and how it can be located.

If a log driver can clear a logjam by locating a single log, and prodding it with a peavey hook, the advertising practitioner - I believe - ought to be able to identify the source of a an advertising logjam by asking a single question.

But what is that question?

One of the best is a question I picked up from a smart Client of mine, who one day when a meeting had run aground in a morass of confusion, simply asked: "What problem are we trying to solve here?"

Another good one, best used when everyone disagrees on what to do, is to ask: "What is the most interesting aspect of this brief?"

If you have a sharp question that you've found helps resolve situations, please share it in the comments below...

A Few Of Jeremy Craigen's Greatest Unawarded Achievements

I've been wanting to write something about my former boss, DDB legend Jeremy Craigen (left).

But when top London ad director Ed Morris (right) penned this brilliant and heartfelt piece about Jez on Facebook, I asked him if he'd let me reproduce it.

Ed writes:

It's Jeremy Craigen's leaving drinks next week. He deserves a drink alright and here's why:

1. He lived through globalisation. Agencies got bought by global holding companies and we all started having to write ads with no words in, or lip-sync 20 different versions for "all markets". If you weren't happy to be global you got kicked out quick. All accounts were centralised and run by extremely political "account barons." D&AD faded and Cannes rose. The Gunn report had the final word. It was a tough time for writers; art directors fared well. Everyone wanted big pictures that traversed all culture and language barriers. English creativity lost all of its nuance and tone. Press advertising died. It was a tough time for Creatives, with nothing but change, disruption and suddenly having to present your work in Spanish.

2. As Creative Director he survived the Antipodean invasion. Most forget this moment, it lasted a couple of years or so. It was when it became instantly fashionable for a management team to kick out the UK Creative Director in favour of someone no one had ever heard of from somewhere no one had ever heard of. It was a tough time to survive. Most of these new Creative Directors were shit to be honest, and ruined creative output and culture within agencies all over London. If you were a Creative Director through that time you had to be fucking good to survive it. Management teams everywhere were making big stupid hires. Many great UK Creative Directors were kicked into oblivion throughout this time.

3. He lived through and survived merger mania. This came throughout the late 90's and early 00's with the slow post 80's downfall in revenue. If your agency wasn't making enough money you merged with one that was. Or you did it anyway just to get bigger. Initially successful on paper, these rash moves ruined agency cultures and ripped creative departments apart. Agencies (like the one I was at - Lowe) eventually suffered badly for it. Jeremy survived merger mania. A tough time. I remember being at BST and it merging with GGT which then merged with TBWA all in the space of 5 months. It was a ridiculously unsettling and insecure time. Can you imagine keeping the work good through that shit?

4. He lived through and survived The Digital Revolution. It was a revolution for the world, but a living nightmare for anyone that got labeled as a "Traditional CD", or just anyone with a very good TV reel. This was very tough. There wasn't a day for anyone in high creative office when you weren't under threat of losing your job to a supposedly "Digitally Savvy Creative Director" most of whom (back then) just turned out to be shit really. Again though, massively stressful and turbulent time. I remember having to DEFEND myself to management for having "the best reel in London" at this time. Ridiculous, and the industry was critical and quick to blame and accuse.

Overall and throughout, Jeremy Craigen managed to bring consistency to his output and his agency against the odds, and through probably the most inconsistent time in the business that has ever been.

People forget about it, kids say it was easier back then, they're talking shit. It was tough, it took a creative genius just to hold on to your job, let alone make the work great. I rode those tough moments, not as well as Jeremy, but enough to know how hard it was and appreciate what he did.

Well said Ed.

Make Videos Or Die

With SXSW here again, it's an appropriate time for us to take a moment and ponder what the future of our business might be.

Okay, stop. The answer is so obvious I'm surprised it even took you that long.

The future of our business is video.

When people use the word 'Content', what do they mean? Videos.

What are consumers doing on Facebook nowadays? Watching videos (hey, it helps that they just play without me even having to click on them!)

What's the world's Number 3 website, after Google and Facebook? YouTube. Where people go to watch videos.

And guess what? People are still watching a hell of a lot of TV, aka moving pictures, aka videos.

Please note, I'm not decrying the innovative/tech stuff that comes out of SXSW. Like this Tinder idea where a woman who may or may not be a robot starts talking to you, and it's actually a promotion for a new sci-fi movie. I'm just placing my own bet on which tech trend is going to have the most profound effect on our industry. And I'm betting screen-agnostic video.

So far so good - agencies have a long history of producing moving pictures. But the problem is, our expertise is in making expensive videos, infrequently, whereas the big demand from brands today is to make inexpensive videos, more frequently.

We need to grow this expertise and we need to grow it quickly.

BBH is often ahead of the game - they've just set up Black Sheep Studios to do this.

I see more and more agencies doing something similar. Good. I do believe, as I wrote in the title of this piece, that we need to start making videos or die. Well, 'die' is a strong word. But we'll definitely wither, if we lose this huge slice of work to other providers.

The effect on us Creatives could be massively positive - it's an opportunity to start doing a new job, that will genuinely be super-fun.

Here's what I mean - these videos need to be cheaper, so we need to mash some of the existing roles together. The two basic skill-sets are 'being creative' and 'getting shit done'. So I see one role that is a combined Creative/Director/Editor and another role that is a combined Producer/Account Handler. Two roles.

So your new work day could involve thinking up ideas, shooting them yourself, and then editing them.

If that became my job, I think I'd quite like it.

Would you?

Living The Dream Is Actually Shit

There's a perception that advertising Creatives would really rather be doing something else.

That we are people who failed at writing novels or screenplays, or didn't have the stomach to pursue the uncertainties of life as an artist.

That a career in advertising is a creative compromise.

It's certainly true that, back in the day, every Creative was writing a novel. Then it was screenplays. Now I hear of quite a few Creatives developing apps or games.

But I get the sense it's happening less.

Even though advertising's glamour rating has been on the slide since about 1987, I get the impression we're all more dedicated to it.

Am I right?

Or are you still hankering for something else? If so, what? Let us know, in the poll below.

What Is Your Dream Job?

Ad Creative
None of the abvoe
Poll Maker
P.S. I have what I think is good news. My own alternative occupation was always 'Writer'. But when I did actually have six months off and wrote a book, I didn't really enjoy the experience.

I was sitting at home (well, mostly lying in bed) writing all day. And it was just a bit, well... lonely. Sure, you can go out to a cafe, but you're still not interacting with people.

If you're someone that loves the stimulation of agency life, as I do, you may find that 'living the dream' is actually shit, as I did.

The fact is, many creative professions are solitary. Artists, writers, composers, actors (when not working), directors (when not working), and musicians (when not working) are all just sitting at home on their own.

If you're the sort of person that craves solitude, then it's fine. But most people who work in ad agencies are not that sort of person.

So would you really rather be doing your dream job, or are you happy doing what you're doing?

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How Big Should The Packshot Be?

Disputes about the size of a logo are the most tedious conversations you can possibly have. Personally I'm glad when a brand has a fixed rule about logo size; it eliminates the debate.

But no one seems to have a fixed rule about packshots.

The general principle at many companies seems to be 'just make it as big as possible'.

In the above ad, you can almost hear the Client cursing the limitations of the media space, for unfairly constraining the size at which his product can be displayed.

Of course, some find a clever way around that.

Well, at least it wasn't horizontal.

Actually there's a surprising degree of variation in pack sizes. Apple packshots, for example, vary from subtle sizes like this:

 To whoppers like this:

This is the smallest packshot I've ever seen. Lovely ad, for Peugeot 106. Apologies for poor scan quality.

This one I reckon is the biggest. It's an Armani ad from 1983. Shame about those little bits of black space around the shoulders of the bottle. Otherwise it would have been perfect!

In terms of what the ideal packshot size is, I honestly don't think it's that hard to work out. The principles are the same as with any art direction (*disclaimer: I'm not an art director).

You simply need to have a clear vision of the order in which you want a consumer to view the different elements of your ad.

In the example below, it was obviously felt that consumers should see LeBron James first, since he's been made the biggest element. (Correct decision, I'd say. It's LeBron that is going to hook people in).

Next, the art director wanted people to read the headline. So that's the second biggest element. Then the shoes (third biggest) and finally the tagline.

Art directors: is this what you actually do, or am I just making it up?

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Forget 'Is It An Art Or A Science' - Is Advertising Actually More Like A Religion?

The quickest way to ruin a meeting, it's been said, is to ask everyone around the table how they think advertising works.

Because no two individuals would agree.

And frankly, this is an embarrassment.

Can you imagine plumbers sitting around, arguing about 'how plumbing works'? Lawyers may disagree on the facts of a case, but they all have a very clear understanding of what the law is. Knife-makers agree how knives work. And bakers all know how bread is made.

Almost unbelievably, some of our theories directly contradict each other. For example, some argue it is essential that advertising has impact - if you don't 'cut through the clutter', your message won't be heard. But others argue that the brain works mostly by Low Involvement Processing - we process marketing messages at least as much when we are paying little attention to them as we do when we consciously take them on board, so cut-through is irrelevant.

I was intrigued, therefore, as to whether 'the answer' would appear in the book that uber-strategist Paul Feldwick has just published called 'The Anatomy of Humbug'.

It's billed as "a book that isn't about how advertising works, but about how people think advertising works."

My copy hasn't arrived yet, but I have to say that the interviews Mr Feldwick has done to promote the book - while fascinating - have left me seriously depressed.

I suppose I've long cleaved to the hope that although we may disagree right now about how advertising works, there will be an answer 'one day'.

Mr Feldwick seems to imply not.

While according to one reviewer "he sidles very close to answering the fundamental question" he eventually concludes "there isn't an answer" and admits that "the book supports multiple points of view."

Most worryingly of all, he talks about the need to 'respect other people's beliefs'.

And that, my friends, is not the language of science, or of art... but of religion.

Was Oreo's Super Bowl Tweet A Waste Of Time?

If you follow the social media scepticism of Bob Hoffman over at The Ad Contrarian, you might want to watch the above video.

But if you haven't got time to watch it, here are the highlights:

Mark Ritson, an English guy who is professor of marketing at Melbourne Business School, calculates that the famous Oreo Super Bowl tweet ("You can still dunk in the dark") - which according to one online publication "won the Super Bowl" - was in fact seen by only 64,300 people. Compared with the over 100 million who saw the TV ads.

Next point. For most big brands, only 2-3% of their customers have liked them on Facebook. "If social media is supposed to be a conversation, it's a bloody quiet conversation," says Ritson. "97.5% of their customers aren't listening."

He cites the case of Australian bank NAB, who have six people in their social media team, but in the previous week, only 276 people had engaged with the brand via Facebook. Out of a total customer base of 12 million. When properly rounded, that's an engagement rate of 0%.

Those are the facts. So what do we do about them?

This is where I part company with Ritson.

He reckons brands "aren't welcome" on social media, and that for most marketers, "social media is mostly a waste of time." He suggests the NAB social media team switch to other duties.

I don't see it the same way.

What if the early television advertisers had been told that people wanted to watch the programmes, not ads, and that they should give up?

Let's say that a brand's current Facebook strategy is not working. The solution is not to quit. The correct response is to find a way to make it work.

Globally, Facebook has 1.35 billion monthly active users. And Twitter has 284 million.

Get on it, people.

Do Ideas Have Different 'Shapes'?

This is one of my favourite ads of all time. It was created by Ben Nott and Adam Hunt at Saatchi & Saatchi London.

(Incidentally, Adam now runs an Asian tapas restaurant in Bondi called Mamasan; it's great and you should totally go there if you're in the neighbourhood. But I digress).

One of the interesting features of the ad is that it has a very distinct 'shape'.

The first three-quarters of the message, while somewhat visually engaging, and somewhat semantically intriguing, are basically pretty straight. The payoff - and what a payoff it is - comes at the end.

So you might say that the 'shape' of the ad is something like this:

(I haven't marked them, but I'm hoping it's clear that the x axis represents Time, and the y axis Reward for the Viewer).

It's very hard to imagine this idea being structured any differently. The first three-quarters has to be quite dry and scientific, and the payoff (as with many jokes) has to come at the end.

So does that mean the idea is intrinsically that shape?

Let's look at another one. The famous Sony 'Balls' ad is kinda entertaining all the way through. There's not really a boring intro part, or a boring 'product bit' at the end, and there's no particular climax at any point either. Something like this?

Then there's a type of ad I like to call the 'Stepper'. This year's Snickers Super Bowl ad works like that. There's entertainment from the beginning, and it just keeps ramping up, with each gag topping the last.

Sometimes there is information that you just have to get across. The 'Climate Name Change' social idea by agency Barton F. Graf 9000 starts out with a lot of serious factology, before unleashing the humour of its central conceit. And once again, I'd argue that this is the only way this idea can work. Giving a shape something like this:

Finally, there's a type of idea that works almost the opposite to that. Entertainment first, and then product message. Example: Little Caesars 'Arm Cast'.

Personally, I reckon that no one shape of idea is actually better than any other. Success comes from recognising what shape your idea is, and executing it in that style.

But what do you think? Any validity to my shape theory? And do you have a favourite shape, or is there one that you hate?

Death, Destruction, and Negativity: Why This Year's Super Bowl Ads Are So Good

It seems like what was once a daring media strategy - to release your Super Bowl ad before the Super Bowl actually airs - has become the new normal.

So I've seen most of this year's ads already.

It's a good crop - big spectacle, big emotion.

And I don't think it's a coincidence that many of the best ones show death, destruction and negativity.

For example, in this great ad - I won't reveal the brand name, because spoilers - catastrophe strikes the entire planet.

This year's Budweiser 'dog and horse' epic - albeit not as good as last year's - contains a scene in which the pup is menaced by a drooling wolf.

In the GrubHub ad, a flying burrito smacks people in the face. Fairly comical, but also undeniably violent.

Even in the Bud Light ad - which basically consists of a guy whooping and hollering like he's just won a car in a game show - there is a moment where he (metaphorically) dies.

I've written before about the power of negative thinking, so I don't want to repeat myself.

Suffice to say that judicious use of negativity helps give a brand depth and relatability, as well as providing a great springboard for engaging creative.

In other words, it's the difference between this...

...and this.

Dear Introverts: You're Shit-Out-Of-Luck, Aren't You?

I love that quote; it's flattering to us ad types, isn't it?

But recently I've been wondering whether it has an unwanted flip-side.

I know a ton of advertising creatives, and I agree with Banksy that the vast majority of us are bright, creative and ambitious. And that's good.

But are we too much of a type? Are we failing to attract the "slow and self-obsessed" who could make a valuable contribution, but are put off by... something?

There's quite a lot going on in Banksy's quote, but I guess part of it is about people who think quick and shallow versus people who think deep and slow.

We mostly get the former. An advertising agency is a tyranny of quick-wittedness. Nearly all the people at the top are the quick-witted type. But do they sometimes get there at the expense of others who might be better than them, but just have a different personality?

I suspect Banksy is also making an observation about a personality difference that is horribly over-simplified, but which at least has the advantage of being well-known and easily understandable: introverts and extroverts.

You'd have to agree that the vast majority of people in advertising are extroverts.

Our industry prizes those who are energetic, articulate, confident, and sociable. And I'm not just talking about Account Handlers, but Creatives too. Especially if you're to reach CD level, you nowadays simply have to be energetic, articulate, confident etc.

I worry this is a problem. We might be discarding, or at least failing to properly promote, some fantastically talented Creatives just because they don't slap people on the back, or crack jokes.

There's a best-selling book by Susan Cain called Quiet: The Power of Introverts in a World That Can't Stop Talking, which makes the point that most modern institutions are geared toward extroversion, which means introverts are underrated, under-used and even demonised.

And nowhere is the issue more acute - I'd guess - than in ad agencies.

So come on, introverts, let's hear from you. Anonymously, obviously. Do you find it difficult, being an introvert in advertising? Do you think extroverts have an unfair advantage? And do you ever take your headphones off?

A Book About Poison Gas

I have a book out and it would be remiss of me - as an ad guy - not to plug my own product.

So I will.

100 Ideas That Changed Advertising is really a history book, charting the development of advertising from the earliest posters (perfectly preserved advertising messages have been found on the walls of Pompeii) to today's online media behemoths like Facebook and YouTube.

I took history A-Level at school, I enjoy history, and I believe we can learn a lot from it.

So what can we learn from the history of advertising?

The main thing I learned, in writing it, is that the history of advertising is a history of innovation - not only that, but of remarkably rapid innovation.

For example, the first cinema in the United States - Vitascope Hall, in New Orleans - opened in 1896. Filmed ads were being produced as early as 1897.

The first commercially licensed radio station in America went on air in 1920. In 1922, the first radio advertising was broadcast.

Twitter launched in 2006, and by 2010 was running advertising.

There are a couple of useful take-aways, I'd suggest, from this trend.

First, don't be afraid to innovate.

In 1921, a group of investors declined to put money into radio, notoriously predicting that “The wireless music box has no imaginable commercial value. Who would pay for a message sent to no one in particular?” Right.

When the first TV ad aired, on July 1st 1941, there were only 7,500 TV sets in New York City. The ad was for Bulova watches and showed a watch-face superimposed over a map of the U.S., while a voiceover claimed that “America runs on Bulova time.” The whole medium must have seemed incredibly shonky, compared to the sophistication of print advertising at that time. People must have wondered if it would ever take off. Well, it did.
Similarly, people questioned whether YouTube would ever make money from advertising, since few believed there would ever be a wide audience for videos of other people's cats. I know.

So what's today's frontier? Mobile? Probably. Don't be afraid to go there.

As I worked through the chronology of our industry to write the book, I started to feel that advertising is like a gas; it seeps in everywhere. Whatever new medium is invented that captures a modicum of human attention, someone will find a way to put an ad there.

And that led to my second take-away. In contrast to the ever-evolving media and technology landscape, there is one thing that hasn't changed, and will never change - the nature of our responsibilities, as advocates for the brands who buy this space.

Another gas metaphor captures it with pungent brilliance. It was American ad-man George Lois who said of successful advertising: "I think advertising should be like poison gas. It should grip you by the throat, it should bowl you over, it should knock you on your ass.”

Is Advertising Really A Business, Or Is It Actually Just A Giant Game?

This is the time of year when the predictions for the future of advertising are published.

Words like integration, big data, and real-time marketing are bandied about, and it all sounds horribly serious.

Yes, advertising is a business - a serious and important one, with high stakes.

But to do it well, we may actually be better off treating it as a game.

As evidence, I'm citing a sci-fi novel called Time Out Of Joint by Philip K. Dick.

The protagonist of this tale is highly skilled at an extremely serious activity - predicting where Lunar rebels will target missile attacks.

But the authorities use drugs and a stage-set to convince him that he is living in suburban America in 1959, a cosy existence where his only job is to enter a local newspaper competition called "Where Will The Little Green Man Be Next?"... which in reality, is predicting the missile strikes.

The key point for our purposes is that the protagonist is more effective at a serious job when he treats it as a game.

I firmly believe it's the same for ad Creatives.

My heart always sinks when the suits come in and explain that this is a really important project, and we mustn't screw it up. Or if the Client gives a speech about "how much is riding on this."

That kind of talk stifles creative people.

Because ironically, serious business success (in a creative business) is best achieved by treating it as a game.

"The creation of something new is not accomplished by the intellect but by the play instinct," said Carl Jung.

I love that.
Oh, by the way, I have a book out.

It's called 100 Ideas That Changed Advertising. Available in Australia, the UK, USA, and no doubt other countries that have internet or bookshops. Call to action: buy it now!


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Ernest B. Ashba

Ernest Bruce 'Ernie' Ashba Sr., 72, of Wabash, IN, and having family in Huntington, IN, died Sunday, Jan. 20, 2019, at 5:55 p.m., at his daughter's home in Fort Wayne.

Mr. Ashba retired from the Indiana Department of Transportation in 1995 after more than 20 years of employment. He was also a truck driver and drove a bus for the Dallas Winchester Senior Center in Wabash.

He was born March 10, 1946, in Anderson to Andrew Jackson and Inez Lucille Redding Ashba. He married Mary Bryant in Wabash on Sept. 14, 1979, and she survives.

Additional survivors include a daughter, Lora (Grant) Acker, of Fort Wayne; two sons, Bruce Ashba and Eric (Amy) Ashba, both of Fort Wayne; eight grandchildren, Derek (Amanda) Hinrichsen, of Columbia, MO; Brittany (Mike) McNally and Kyleigh (Justin) Chambers, both of LaFontaine; Justin Acker and Mackenzie (Dereck) Warner, both of Fort Wayne; Damien (Jocelyn) Ashba, of Huntington; Matthew (Alexa) Owen, of Amboy; and Nathan Owen, of Bunker Hill; and 15 great-grandchildren.

He was preceded in death by a daughter, Rebecca Ashba.

Calling is Wedneday, Jan. 23, 2019, from 5 p.m. to 8 p.m. at Grandstaff-Hentgen Funeral Service, 1241 Manchester Ave., Wabash.  A funeral service will be Thursday, Jan. 24, 2019, at 2 p.m. at the funeral home, with Dr. Kent Young officiating.

Memorials are to the 700 Club or the Animal Shelter of Wabash County.

The memorial guest book may be signed at

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September 14, 2012

Feature Friday - DIY Rock Candy

Ever since I saw THIS POST at Mom's Crafty Space I have been dying to try my hand at making rock candy with the boys.

Last week we gave it a shot.

It takes about a week for it to turn into the rock candy.  We followed Heidi's instructions.

I also did a little more research since I saw that some people had some trouble with it working.  One post recommended putting the sticks an inch above the bottom of the glass to avoid having the glass itself get crystalized.  It did anyway, but it's easy to wash afterwards because the hot water dissolves the sugar.

One post I found recommended covering the jars lightly with saran wrap.  I was worried that because the top of liquid got hard that it wasn't working, but it actually was.  You could see the sticks accumulate more crystals throughout the week.  You can't really see that from this picture, though.

After a week we broke through the top layer and pulled out the rock candy.

The color fades a lot from the original color.

The boys were super excited to pull out the candy after watching it all week.  It was an exercise in patience for them.  (Just ignore that lunch hadn't been wiped off Baden's face yet in this picture :)

And my little ham Rex was amazed as well.

If you want to try your own, you can find Heidi's tutorial HERE.

Thanks for stopping by.

- Can't find the original source of an image?  Susan just showed me how, directing me to THIS POST.  Awesome.

- I don't have a serger, but if you do and you want some tips, Palak just did an awesome series HERE on using a serger.

  1. Just to let you know...still loving the next button. There is also a subscribe button. My blog reading life has just gotten easier and you will get credit for each post.

    1. same here- its been one day of blog reading with the next button, and i love it. and so glad it benefits you (and all the other bloggers) too!

    2. That subscribe button is priceless :)

  2. I remember doing this as a kid with my mom! Something cool you can do along the same lines for winter is make snowflake crystals. You use Borax and pipe cleaners in the shape of a snowflake and it grows crystals--they turn out really pretty!

    Another fun activity I remember doing with crystals is growing them on charcoal briquets using bluing.

  3. Thanks so much for the shout out Cheri!

  4. I wanted to second what gccmom said. Thank you for sharing the next button feature. My blog reading has never been easier. :-)

  5. We did this for science camp (just brought them after I had made them the week before) and some of ours worked awesome and some not. Glad to see a better tutorial for when I try again if I do!

  6. Yay, I'm so glad that it worked out for you guys! Rex's sweet little face is priceless lol! My boys were actually just talking about doing this again too. Thanks for sharing! :)

  7. Does Rex' shirt say "Dude, I'm right here!" Hahahaha! Thanks for sharing the survey results. I'm excited for what you have ahead.

  8. Awesome! I'm going to try this Monday with my kids. They are out of school and we need a project! Thanks for sharing!

  9. Thank you for sharing this and information about google reader...I am a very new baby blogger, and I had no idea. I use google reader all.the.time. Glad you shared. I always love ur ideas. (:

  10. That's cool that it worked!! We tried it this summer, and it didn't work out. I didn't cover it with the Saran wrap, but it got kinda gross and weird-stinky..

    Maybe a good fall break craft..

  11. Thank you Thank you for the tip about the serger!! I just got mine a week ago and I'm still trying to figure things out. This just answered SO many questions I had!!

  12. Don't try this if you have an ant problem...don't ask me how I know ;)

Thank you so much for leaving a comment! If you are asking a question and you do NOT have your profile linked to your email address, I will respond in the comment section, so just come back to check it later. Thanks!

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Home Sweet Anywhere Blog Tour

We are happy to host a stop on the blog tour for Home Sweet Anywhere by Lynne Martin.  Keep reading for a summary of Home Sweet Anywhere, an exclusive excerpt, and the book trailer.  Home Sweet Anywhere launches on April 15, 2014.

An enchanting account of how one couple sold everything to fulfill a dream of living abroad—one country at a time
Reunited in love after 35 years and suffering from a serious case of preretirement wanderlust, Lynne and Tim Martin made a life-altering decision: they sold their house and possessions and hit the road to live internationally full-time. Now tethered to nothing but their suitcases, each other, and their next exotic location, they’ve never looked back. From sky-high pyramids in Mexico to monkeys in Marrakech, this delightful, inspiring memoir is a romantic tale of derring-do for grown-ups and a road map for anyone who dreams of turning the idea of life abroad into a reality.
Twitter: @Lynne1Martin
The cold winds and icy rain of the Cliffs of Mohar did not help a nasty little cold that had started to overtake me when we arrived in Ireland. It increased in intensity as we worked our way down the Irish coast. Tim booked a room at The Lodge, a charming B & B near the center of Kenmare, in County Kerry. The owner, Rosemary Quinn, an attractive young woman whose family lived in a large wing of the lovingly maintained old building, greeted us and showed us to our comfortable room. I snuffled and coughed, feeling miserable. “You’ll be needing a little help with that cold. Have a seat and I’ll be back in a minute with just the thing,” she said.
I felt so rotten that I obeyed and watched while Tim unpacked our essentials. Soon, Rosemary returned with a small silver tray. I knew immediately what she had in mind: a pot of hot water, a plate with a slice of clove-studded lemon, and a flowered china bowl holding dainty silver tongs and sugar cubes sat on the tray. The business end of this graceful presentation contained a man-size tumbler with a big slug of Jameson’s Irish Whiskey! She quickly combined two sugar cubes, the lemon slice with cloves, and the whiskey, and poured in hot water to the brim. “This will fix you right up,” she said. “You’ve lived in Ireland, so surely you’re familiar with good old fashioned hot whiskey! Slainté.” She handed me the potion.
I certainly remembered that Ireland’s wild, wet weather made frequent stops for a hot whiskey a necessary part of any walking expedition. A pub was never hard to find. I’m not so sure about the concoction’s actual curative powers, but once the Jameson’s hit its target, I didn’t really care that I was sick. The Irish cure for cold weather and sniffles is much more fun than Nyquil.

Word verification stinks--- but spammers are worse. Thank you for your patience!


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Saturday's with Stef: A first date......

12:27 AM

Dating. Such a foreign word and concept to me.
In fact, I have never been asked out FORMALLY...Up until this week.

Thursday night I went on my FIRST official date. I am not going into details, for I don't kiss and tell. But I will tell you, I came home smiling.

I think about how far I have come. I remember last year, I was crying and crying over the ex. Today, I am free from the chains of that relationship and his addiction. POINT IS: I AM FREE.

I actually was VERY nervous. So many questions went through my head.

"What if he doesn't like me?"

"What if conversations runs out?"

Lots of "What if's....."' in my head.

I don't know if there will be a second date. However in my personal opinion, not to toot my own horn, but he'd be one lucky guy if you ask me!

Okay I tooted it...TOOT! TOOT!

It felt so good to laugh, smile and have a conversation.

It felt so good to finally say "YES" to a guy who a is talented, smart, and not to mention CUTE.

Anyways, I came home and thought of what life will be like a year from now.

It made me excited.

I know I so badly want a family, but honestly, I look at girls who pop out kids left and right and have babies with men they even haven't dated a year, and I am thankful I am waiting until I find that one guy who I am definitely sure that I am sure about. Does that make sense? I get it, men can change...but I like to think God has someone REALLY special for me. Someone who will want me until death-do-us part. The one man who makes all the other men look like scum that gross at the bottom of the sea.

Do I make sense at all?

Anyways...Point is. I went on a date. I put myself out there. I opened up. I took a risk....

I am glad I did.

It made me feel fearless.

It made me thankful that all those times I have given up on myself, I didn't.

I think I may say "Yes" more....

*Crosses fingers for a second date*

Until then...

I vow to live my life with open arms, an open mind, an open heat, open options, and an open soul...

Marta said...

Well, you went out on a date and that's a start! He'll probably is thinking of taking you on a second date, why not?

Desiree @ Mano y Metal said...

whoohoo! Glad you are venturing out there again!
Cheers to your this date and many more!

Happy Saturday!

Nikol said...

It's a great feeling when you finally meet that someone. I can not imagine my life without my husband. Take risks, love, feel, jump in and breathe!! You looked beautiful I am sure and just remember... we kiss alot of frogs before one turns into a prince.

SoCal Sweetheart said...

Both OF YOU ARE WAY CUTE! and Im glad I got to be there on the side!!!! looking in from a distance... I am truly happy for you <3 and I KNOOOOWWWWWWWW there is a second date happening!! or did it! hehehehehe ;) Love you so much

I wanna be fierce said...

Well all right now missy. Cheers to your date night. Haven't been on one in a while myself. Heres to a second date. Keep us posted.

So happy for you Stef! You go girl! I know it is scary but you have to tear down those walls and let someone in or else you will never be happy! Kiah

Ashley said...

Yay! Glad you went home smiling!

Beauty on a College budget said...

Love that first date feeling! Good luck with this new man! I'm keeping my fingers cross for you.


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International Institute of Communications

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Jean-Jacques Sahel

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Jean-Jacques Sahel

Jean-Jacques Sahel has been involved in international government and regulatory affairs for over 15 years in both the private and government sectors. He was appointed Managing Director of ICANN’s Brussels office in July 2017, to lead the organisation’s corporate strategy and coordinate its operations across the European region. Since 2014 he has led ICANN’s strategic plan for outreach, support and engagement with governments, private sector, and user groups throughout Europe, and worldwide for civil society.

Before joining ICANN, Mr Sahel headed government and regulatory affairs for Skype, then digital policy at Microsoft for Europe, Middle-East & Africa regions. He had started his career in the City of London, before spending several years in the UK Government, leading in particular its international telecommunications policy.

Ex officio, Mr Sahel has chaired the UK Chapter of the International Institute of Communications (IIC) since 2009 and was a member of OSAB, the Spectrum Advisory Board of UK communications regulator Ofcom for 2 terms until 2016. He has authored articles and research in both mainstream media and academic publications particularly on Internet policy and governance.

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Int J Biol Sci 2007; 3(4):198-204. doi:10.7150/ijbs.3.198

Research Paper

Overexpression of Selenoprotein H Reduces Ht22 Neuronal Cell Death after UVB Irradiation by Preventing Superoxide Formation

Kamel E. Ben Jilani, Jun Panee, Qingping He, Marla J. Berry, Ping-An Li Corresponding address

Department of Cell Molecular Biology, John A. Burns School of Medicine, University of Hawaii, Honolulu, HI 96813, USA

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Ben Jilani KE, Panee J, He Q, Berry MJ, Li PA. Overexpression of Selenoprotein H Reduces Ht22 Neuronal Cell Death after UVB Irradiation by Preventing Superoxide Formation. Int J Biol Sci 2007; 3(4):198-204. doi:10.7150/ijbs.3.198. Available from http://www.ijbs.com/v03p0198.htm

Selenoproteins have been shown to exhibit a variety of biological functions, including antioxidant functions, maintaining cellular redox balance, and heavy metal detoxification. UV irradiation-induced damage is partially mediated by increased oxygen radical production. The present study is designed to examine the antioxidative effects of human selenoprotein H (hSelH) after brief period of UVB irradiation on the murine hippocampal neuronal cell line Ht22. Ht22 cells were stably transfected with the hSelH gene or with MSCV empty vector and exposed to UVB irradiation with or without the presence of serum. The results showed that cell viability was significantly higher in hSelH-transfected cells compared to the MSCV vector-transfected cells after 24 h of recovery with or without the presence of serum in the media. Further studies revealed that while the number of superoxide anion (O2˙-) positive cells was increased following a 7 mJ/cm2 of UVB irradiation and 5 h of recovery, overexpression of hSelH significantly reduced superoxide production. These results suggest that hSelH overexpression protects cells from UVB irradiation-induced cell death by reducing the O2˙- formation.

Keywords: selenoprotein H, ultraviolet B irradiation, reactive oxygen species, superoxide anion, cell viability.

1. Introduction

Ultraviolet (UV) irradiation, especially UVB irradiation (290-320 nm), leads to premature skin aging, suppression of immunity, and skin cancer [1]. In neuronal cells, UV irradiation induces apoptosis; therefore, many research groups have used this model as a tool to study the intracellular mechanisms of apoptosis [2-7]. UV irradiation induces cell death via a variety of cellular mechanisms. These include induction of nuclear DNA damage, activation of cell surface death receptors such as the tumor necrosis factor (TNF) superfamily, and formation of reactive oxygen species (ROS) accompanied by mitochondrial dysfunction and release of pro-apoptotic factors [8, 9]. UV irradiation has been demonstrated to produce reactive oxygen species (ROS) such as superoxide anion (•O2) radical, hydroxyl radical, hydrogen peroxide and the highly reactive and destructive oxidant, peroxynitrite (ONOO-) [8-10].

To date, 25 selenoprotein genes have been identified in human. Known biological roles of selenoproteins include antioxidant enzymes, enzymes involved in thyroid hormone metabolism, and proteins functioning in selenium transport and heavy metal detoxification [11-16]. These include several well-known peroxidases and reductases such as glutathione peroxidases (GPxs), phospholipid hydroperoxide glutathione peroxidases (PHGPXs) and thioredoxin reductases (TRs), as well as other lesser characterized selenoproteins such as selenoprotein H (SelH) [17, 18]. Furthermore, selenoprotein P (SelP) may also function as a neural survival factor [19, 20]. Emerging evidence suggests that SelP may also possess antioxidant activity [21, 22]. For example, SelP has been shown to function as a protectant against diquat-induced liver necrosis and lipid peroxidation in the rat [23, 24]. SelP is also protective against peroxynitrite-mediated oxidation and nitration [25]. The antioxidative effects of several other selenoproteins such as Gpx, PHGPXs and TRs have been well characterized [26, 27].

The function of SelH is largely unknown. Recently, it was demonstrated that overexpression of Drosophila melanogaster SelH (dSelH) significantly increased viability in embryos by decreasing lipid peroxidation [28]. The objectives of this study were to investigate whether human Selenoprotein H (hSelH) protects neuronal cells against UVB irradiation-induced cell death and if so, whether the protective effect may associate with ROS production prior to observable cell death. We first determined cell viability following a 7 mJ/cm2 of UVB irradiation injury and 24 h of recovery in the murine hippoccampal neuronal cell line Ht22, transfected with the hSelH gene in Murine Stem Cell Virus (MSCV) or the empty retroviral vector. We then measured O2˙-production after 5 h of recovery in hSelH or empty vector transfected cells using the oxidized dihydroethidine detection method [29, 30]. Our results are consistent with the concept that the neuroprotective effect of SelH is associated with its ability to reduce ROS production.

2. Materials and Methods

hSelH transfection procedures

The plasmid containing the gene encoding hSelH was purchased from Invitrogen (Carlsbad, CA, USA). The target gene was subcloned into Murine Stem Cell retroviral vector pMSCVpuro, purchased from BD Biosciences (Palo Alto, CA, USA). The packaging cells RetroPack PT67 (BD Biosciences) were plated in a 60 mm plate at 70% optical confluency 12 h before transfection. Cells were then transfected with 2 μg of hSelH-carrying retroviral vector or empty vector using Fugene method (Roche, Indianapolis, IN, USA). Culture medium was aspirated from cultures after 4 h of transfection. PT 67 cells were washed twice with PBS, and grown for 24 h in 3 ml complete medium. The cells were then selected by puromycin. The concentration of puromycin was optimized to kill the untransfected control PT67 cells in 7-10 d. The transfected PT67 cells that survived the puromycin selection stably produced virus. For viral infection of the target cells, Ht22 cells were plated 12 h before infection at ~70% optical confluency. The medium from transfected PT67 cells containing virus were collected, filtered through a 0.45 µm filter, and added to the Ht22 cells in the presence of 4 μg/ml Polybrene. The virus-containing medium was replaced after 24 h of incubation. After infection for 48 h, the target cells were subjected to puromycin selection. The selection lasted for ~10 d until the uninfected control cells were killed. The cells that survived the selection made up the hSelH-transfected Ht22 cell line (hSelH-Ht22) and MSCV-transfectant (MSCV-Ht22) cell lines.

Detection of mRNA overexpression on hSelH by Real time PCR

Total RNA was extracted from hSelH-Ht22 and MSCV-Ht22 cells using RNeasy Midi Kit (Qiagen, Valencia, CA, USA). cDNAs were synthesized from total RNA using the SuperScript III First-Strand Synthesis System (Invitrogen), and PCR amplification of target genes were carried out using the Platinum SYBR Green qPCR SuperMix-UDG kit (Invitrogen). The following specific primers were used to amplify hSelH, mSelH, and mGAPDH (house keeping gene) cDNAs:

All PCR reactions were performed in duplicate 20 µl reaction volumes and detection was carried out with a LightCycler 2 real-time PCR machine (Biorad, Hercules, CA, USA).

Cell maintenance and treatment

Murine hippocampal Ht22 neuronal cells were infected either with the MSCV empty vector or with hSelH in MSCV expression vector. For propagation, cells were fed with Dulbecco's Modified Eagle Medium (DMEM) with 10% fetal bovine serum (FBS), 2 mM glutamine, and 200 mM streptomycin/penicillin (Invitrogen) and then cultured at 70%-75% relative humidity in 5% CO2 at 37˚C. Media was renewed every 3 d. For cell viability assays, cells were seeded in 48-well cell culture plates (Corning, Aton, MA, USA) 48 h before treatment. The cells were allowed to reach 80 % optical confluency before UVB treatment.

UVB irradiation

Cells were seeded in 96 or 24 wells plate and propagated to reach 80% cell confluency. Prior to UVB irradiation, cells were washed twice with cold PBS to remove residual serum and dead cells. Cells were placed in serum-containing media (10%) media or without serum for 1 h of incubation in normoxic incubator at 37˚C. Cells were then, submitted to different doses of UVB irradiation from a 20 watt MEABS-25 bulb (Ultra-Lum, Inc., Claremont, CA, USA), which emits most of its energy (60 %) with the UVB range (290-320 nm) with an emission peak at 310 nm band. The irradiation dosages given were 7 mJ/cm2, 11.5 mJ/cm2, and 23 mJ/cm2. After UVB irradiation, cells were returned to the incubator for 5 h or 24 h of recovery at 37˚C.

Measurement of the cytotoxicity

Cell viability was assessed using the Trypan blue exclusion assay as described previously [25]. Briefly, viable and dead cells were discriminated and counted using 0.4% Trypan blue staining with a hemacytometer (Sigma, Saint-Louis, MO, USA). Dead cells were counted as those stained blue by the Trypan blue stain, whereas live and apoptotic cells were those that had excluded the stain.

Detection of superoxide production

MSCV-Ht22 and hSelH-Ht22 cells were exposed to a 7 mJ/cm2 of UVB irradiation in the absence of serum. Production of superoxide anion was assessed using dihydroethidine (HEt) method [31-33] after 5 h of recovery following 7 mJ/cm2 of UVB irradiation, because no frank cell death is seen at this time. HEt was purchased from Molecular Probes (Eugene, OR, USA). Stock solution of HEt (1 mg/ml) was prepared by dissolving in DMSO and stored, protected from light at -20˚C. One µl (1 μg) of HEt was added to 500 μl of cultures 15 min before UVB irradiation. HEt is a cell-permeable fluorescent probe widely used to detect intracellular superoxide anion. It is proposed that the reaction between superoxide and HEt results in the formation of a two-electron oxidized product, which binds to DNA and leads to the enhancement of red fluorescence after being oxidized by superoxide [31, 33, 34]. Twelve microscopic images at the magnification of 20X were captured and the number of oxidized HEt cells, which represent superoxide production, were counted and presented as percentage of total counted cells. Because HEt gives a light background staining, only cells stained with strong bright red color were counted as oxidized HEt-positive cells.

Statistical analysis

All experiments were performed at least in triplicate, repeated two to three times and presented as mean ± SD. ANOVA followed by Scheffe's test was employed for statistical analysis (GraphPad Prism software, SAS, San Diego, CA, USA). A p-value of < 0.05 was considered significant.

3. Results

hSelH infection

hSelH was stably infected into Ht22 cells using MSCV vector. The success of this infection was determined by detection of hSelH mRNA levels using quantitative real-time PCR. The mSelH was a low copy number gene in Ht22 cells. The hSelH mRNA level was increased by about 30 folds compared to endogenous mSelH mRNA level in hSelH-Ht22 cells (Table 1). In control MSCV-Ht22 cells, the murine SelH (mSelH) mRNA level, is comparable to that in SelH-Ht22 but the hSelH mRNA is undetectable.

 Table 1

Detection of mRNA overexpression on hSelH by Real time PCR. Both human SelH (hselh) and mouse SelH (mselh) genes were measured in hSelH-Ht22 and MSCV-Ht22 cells. Each target gene was amplified in duplicates. The table shows a significant drastic increased level of hSelH in Ht22-hSelH cells compared to controls (means ± SD, P < 0.0001).

Target genesMSCV-Ht22SelH-Ht22
hSelHNon-Detectable0.557 ± 0.017*
mSelH0.0149 ± 0.00190.0163 ± 0.0005

Effects of variable UVB irradiation duration on cell viability

In order to identify an optimal time of UVB exposure, we exposed MSCV-Ht22-transfected and hSelH-Ht22-transfected cells to 0 mJ/cm2, 7 mJ/cm2, 11.5 mJ/cm2, and 23 mJ/cm2 of UVB irradiation. Cell viabilities were measured after 24 and 48 h of recovery. Cell viability after 24 h of recovery, was reduced from 100% in non-radiated control to an average of 50%, 40% and 6% in 7 mJ/cm2, 11.5 mJ/cm2, and 23 mJ/cm2 of UVB irradiation, respectively (Figure 1). Cell viabilities were further decreased after 48 h of recovery. Thus, the average survival rate dropped to 19%, 4% and 0% following 7 mJ/cm2, 11.5 mJ/cm2, and 23 mJ/cm2 of UVB irradiation, respectively (Figure 1). This experiment showed that 7 mJ/cm2 of UVB irradiation resulted in approximately 50% cell death at 24 h of recovery, which was determined to be the optimal length of injury. Therefore, a 7 mJ/cm2 of UVB irradiation was used for the remaining studies.

 Fig 1

Variable UVB irradiation exposure parameters on cell viability in MSCV (empty vector) transfected Ht22 cells. The irradiation doses given to the cultures were 0 mJ/cm2, 7 mJ/cm2, 11.5 mJ/cm2, and 23 mJ/cm2, in DMEM medium with serum, respectively. The percentage of cell viability was determined after 24 and 48 h of recovery using the Trypan blue exclusion assay. About 100%, 50%, 40% and 6% of cell viability were observed at 24 h of recovery 0 mJ/cm2, 7 mJ/cm2, 11.5 mJ/cm2, and 23 mJ/cm2 of UVB irradiation, respectively. Cell viabilities were further significantly decreased to 21 %, 4% and 0% at 48 h of recovery at the indicated UVB doses in the Figure. Experiments were performed in triplicate and presented as means ± SD. *Significantly different from non-radiated control at 24 h (P < 0.05) at the indicated UVB dose. #Significantly different from 24 h of recovery at 48 h of recovery (P < 0.05).

Int J Biol Sci Image (Click on the image to enlarge.)

Effects of hSelH overexpression on cell viability after UVB irradiation

Both MSCV-Ht22 and hSelH-Ht22 cells were exposed to a 7 mJ/cm2 of UVB irradiation in the presence or absence of serum. The cell viabilities were examined at 24 h of recovery and the results were compared to MSCV and hSelH transfectants without UVB irradiation. As shown in Figures 2 and 3, MSCV-Ht22 cells exposed to 7 mJ/cm2 of UVB irradiation in the presence of serum exhibited 48% cell viability compared to controls, as expected. However, the cell viability was significantly increased in cells overexpressing hSelH. That is, hSelH overexpression resulted in appreciatively 80% cell survival compared to controls and this was found to be significantly different than that observed with vector control (P<0.0001). In the absence of serum, cell survival rate decreased to about 70% even without UVB irradiation, with no major difference between the MSCV-Ht22 and hSelH-Ht22 cells. Cell survival rate was further decreased to 23% in MSCV-Ht22 cells after a 7 mJ/cm2 of UVB irradiation. However, with the overexpression of hSelH, cell survival only decreased to 52% (P = 0.0004) when cells were exposed to 7 mJ/cm2 of UVB irradiation without supplementation of serum. Figure 2 shows representative photomicrographs taken from MSCV-Ht22 and hSelH-Ht22 cells treated with or without UVB irradiation in the presence or absence of serum. Figure 3 summarizes the cell viability data collected from three independent experiments.

 Fig 2

Photomicrographs showing effects of hSelH overexpression cell viability in Ht22 cells treated with 7 mJ/cm2 of UVB irradiation followed by 24 h of recovery. Increased cell debris was present in UVB radiated MSCV-Ht22 cell compared to non-treatment control cells. More viable cells were observed when hSelH transfected Ht22 cells were exposed to UVB irradiation. Images were taken at 20X magnification with a Zeiss LSM5 laser-scanning confocal microscope. Bar = 10 μm.

Int J Biol Sci Image (Click on the image to enlarge.)
 Fig 3

Effects of hSelH overexpression on Ht22 cells exposed to 7 mJ/cm2 of UVB irradiation with or without serum in media. Cell viability was examined after 24 h of recovery and presented as percentage (mean ± SD). Overexpression of hSelH protected cells from UVB-induced damage. *Significantly different from MSCV control, P < 0.0001 and #Significantly different from MSCV transfectant, P = 0.0004, in the presence or in the absence of serum, respectively.

Int J Biol Sci Image (Click on the image to enlarge.)

Overexpression of hSelH reduces superoxide formation

In order to determine the effects of UVB irradiation and overexpression of hSelH on free radical production, we measured superoxide production using the oxidized HEt method. Since a majority of MSCV-transfected cells die after 24 h of recovery, we selected an early recovery time point (5 h) for detection of superoxide formation. A 7 mJ/cm2 of UVB irradiation resulted in 48% of oxidized HEt positive stained cells in MSC-Ht22 after 5 h of recovery. In contrast, the percentage was decreased to 16% (P = 0.0017) in hSelH-Ht22 cells (Figure 4). A set of fluorescent photomicrographs from MSCV-Ht22 and hSelH-Ht22 cells that were exposed to 7 mJ/cm2 of UVB irradiation is shown in Figure 5.

4. Discussion

Previous published studies on neuronal cells have shown that UV irradiation induced apoptosis is associated with activations of capspase, calpain, JNK, and the mitochondrial apoptosis pathway [4-7]. UV irradiation provokes oxidative stress through the generation of ROS and to induce apoptosis by activation of both receptor-mediated and mitochondrion-initiated cell death pathways [8, 9, 35, 38]. In this study, O2- production increased significantly after 5 h of recovery following a 7 mJ/cm2 of UVB irradiation. This is consistent with the results published by Maglio and colleagues who observed a 5-fold increase in superoxide production in the skin after UV irradiation [39]. Under physiological conditions, superoxide dismutase (SOD) reduces O2˙- to H2O2 that is further reduced to H2O by catalase or glutathione peroxidase. However, excess production of O2˙- leads to formation of the highly toxic hydroxyl radical, •OH, in the presence of metal or peroxynitrite (ONOO-) in the presence of nitric oxide (NO). Hydroxyl radicals and ONOO- are potent oxidants of proteins and DNA, and alter the function of key enzymes through nitration and nitrosylation. Increased ROS in the mitochondrion induces formation of a mitochondrial permeability transition (MPT) pore. The MPT allows release of proapoptotic proteins that subsequently activate a cascade of proteases, ultimately leading to cell death [40, 41].

The selenoprotein family contains 25 members in the human proteome [13]. A majority of published studies have been focused on the expression and functions of GPxs, TRs, iodothyronine deiodinases, and SelP. Studies have shown that these enzymes possess antioxidative effects, in addition to their roles in male fertility, thyroid hormone metabolism, heavy metal detoxification and neurological function and development [20, 26, 42]. For example, glutathione peroxidase acts as peroxynitrite reductase, preventing oxidation and nitration reactions resulting from peroxynitrate [17, 18]. Similarly, SelP protects low-density lipoproteins (LDL) against oxidation (traps ONOO- and protect astrocytes from various injuries including irradiation and tert-butyl hydroperoxide-induced oxidative stress (10, 21, 22, 27]. The functions of most of the recently identified selenoproteins, including SelH, have not been elucidated.

To study whether overexpression of hSelH is capable of protecting cells against UVB-induced oxidative stress, we first transfected the murine hippocampal Ht22 neuron cell line with the hSelH or the empty vector MSCV. As shown in Table 1, the hSelH mRNA level was 30 fold higher than the endogenous mSelH mRNA level in hSelH-Ht22, suggesting a high transcriptional rate of the transfected hSelH gene. We then exposed the cells to a 7 mJ/cm2 of UVB irradiation. Overexpression of hSelH reduced cell death caused by UVB irradiation. This result is consistent with a previous publication showing that overexpression of a selenoprotein homologous to glutathione peroxidase blocks UV-induced cell death [43]. We finally measured O2˙- production in cultured cells at 5 h after a 7 mJ/cm2 of UVB irradiation. The results showed that overexpression of hSelH inhibited O2˙- formation in neurons in the early recovery phase after irradiation. Concurrently, the percent of oxidized HEt-positive cells detected at 5 h of recovery was correlated with the percent of cell death observed at 24 h of recovery, suggesting increased superoxide production might be the underlying cause for the observed cell death. Our findings are in accordance with a previous study reported by Morozova and colleagues [22]. In that study, it was demonstrated that silencing the expression of Drosophila melanogaster SelH (dSelH) gene significantly reduced the total antioxidant capacity and embryo viability. In contrast, overexpression of dSelH aided cell survival by reducing lipid peroxidation in vivo [28].

It is not clear through what mechanism(s) hSelH reduces ROS production. A parallel study carried by Panee and colleagues has shown that overexpression of SelH sustains the total glutathione (GSH) level in Ht22 cells during oxidative stress (Panee et al., manuscript in preparation). GSH plays multiple roles in the protection of cells from ROS, electrophiles and xenobiotics [44, 45]. More interestingly, SelH overexpression upregulated the expression of genes involved in antioxidant defense (Panee et al., manuscript in preparation) and phase II detoxification [44]. Therefore, it is likely that SelH overexpression decreases the ROS level and sustains cell viability in Ht22 cells after UVB irradiation via these pathways.

Taken together, our study shows that overexpression of hSelH protects cells against UVB irradiation-induced cell death by ameliorating ROS formation in the early recovery period. The results are in line with previous publications showing that selenoproteins possess antioxidative capacity.

 Fig 4

Percentage of oxidized HEt positive cells in MSCV-Ht22 and hSelH-Ht22 cells after 5 h of recovery follow a 7 mJ/cm2 of UVB irradiation. hSelH overexpression significantly reduced oxidized HEt positive cells compared with MSCV-Ht22 cells. *Significantly different from MSCV-Ht22, P = 0.0017.

Int J Biol Sci Image (Click on the image to enlarge.)
 Fig 5

Representative photomicrographs showing oxidized HEt positive staining in MSCV-Ht22 (A) and hSelH-Ht22 cells (B) after 5 h of recovery follow 7 mJ/cm2 of UVB irradiation. Arrows denote oxidized HEt cells which indicating the formation of superoxide. Images were captured at 20X magnification with a Zeiss LSM5 laser-scanning confocal microscope. Bar = 10 μm.

Int J Biol Sci Image (Click on the image to enlarge.)

This work was supported in part by NIH R01NS40302, NIH/G12RR03061, NIH/RR16453, and Hawaii Community Foundation. We thank Sarah Timtim and Andrew Sue-Ako for their active participation in this work during their summer research training, Wanyu Liu for technical assistance, and Drs. Rick Bellinger, Peter Hoffman and Steve Seifried for helpful discussions.

Conflict of interests

The authors have declared that no conflict of interest exists.

1. Melnikova VO, Ananthaswamy HN. Cellular and molecular events leading to the development of skin cancer. Mutation Res. 2005;571:91-106

2. Licastro F, Sarafian T, Verity AM, Walford RL. Inhibition of polymerases-alpha and -beta completely blocks DNA repair induced by uv irradiation in cultured mouse neuronal cells. Biochem Biophys Res Commun. 1985;132:929-933

3. Subrahmanyam K, Rao KS. Ultraviolet light-induced unscheduled DNA-synthesis in isolated neurons of rat brain of different ages. Mech Ageing Dev. 1991;57:283-291

4. Klums D, Schwartz T. Molecular mechanisms of UV-induced apoptosis. Photodermatol Photoimmunol Photomed. 2000;16:195-201

5. McCollum AT, Nasr P, Estus S. Calpain activates caspase-3 during uv-induced neuronal death but only calpain is necessary for death. J Neurochem. 2002;82:1208-1220

6. McCollum AT, Estus S. Ngf acts via p75 low-affinity neurotrophin receptor and calpain inhibition to reduce uv neurotoxicity. J Neurosci Res. 2004;77:552-564

7. Berglund CM, Radesater AC, Persson MA, Budd Haeberlein SL. UV-induced apoptosis in sh-sy5y cells: Contribution to apoptosis by jnk signaling and cytochrome c. J Neurosci Res. 2004;78:580-589

8. Kulms D, Schwartz T. Independent contribution of three different pathways to ultraviolet-B-induced apoptosis. Biochemical Pharmacol. 2002;64:837-841

9. Kulms D, Schwartz T. Molecular mechanisms involved in UV-induced apoptotic cell death. Skin Pharmacol Appl Skin Physiol. 2002;15:42-47

10. Roussyn I, Briviba K, Masumoto H, Sies H. Selenium-containing compounds protect DNA from single-strand breaks caused by peroxynitrite. Arch Biochem Biophys. 1996;330:216-218

11. Burk RF, Hill KE. Selenoprotein P: An extracellular protein with unique physical characteristics and a role in selenium homeostasis. Annu Rev Nutr. 2005;25:215-235

12. Kohrle J. Selenium and the control of thyroid hormone metabolism. Thyroid. 2005;15:841-853

13. Kryukov GV, Castellano S, Novoselov SV, Lebanov AV, Zehtab O, Guigo R, Gladyshev VN. Characterization of mammalian Selenoproteomes. Science. 2003;300:1439-1443

14. Rayman MP. The importance of selenium to human health. Lancet. 2000;356:233-241

15. Saito Y, Takahashi K. Characterization of selenoprotein P as a selenium supply protein. Eur J Biochem. 2002;269:5746-5751

16. Saito Y, Sato N, Hirashima M, Takebe G, Nagasawa S, Takahashi K. Domain structure of bi-functional selenoprotein P. Biochem J. 2004;381(Pt 3):841-846

17. Sies H, Sharov VS, Klotz LO, Briviba K. Glutathione peroxidase protects against peroxynitrite-mediated oxidations. A new function for selenoproteins as peroxynitrite reductase. J Biol Chem. 1997;272:27812-27817

18. Sies H, Arteel GE. Interaction of peroxynitrite with selenoproteins and glutathione peroxidase mimics. Free Radic Biol Med. 2000;28:1451-1455

19. Hill KE, Zhou J, McMahan WJ, Motley AK, Atkins JF, Gesteland RF, Burk RF. Deletion of selenoprotein P. alters distribution of selenium in the mouse. J Biol Chem. 2003;278:13640-13646

20. Hirashima M, Naruse T, Maeda H, Nozaki C, Saito Y, Takahashi K. Identification of selenoprotein P fragments as a cell-death inhibitory factor. Biol Pharm Bull. 2003;26:794-798

21. Steinbrenner H, Alili L, Bilgic E, Sies H, Brenneisen P. Involvement of selenoprotein P in protection of human astrocytes from oxidative damage. Free Radic Biol Med. 2006;40:1513-1523

22. Traulsen H, Steinbrenner H, Buchczyk DP, Klotz LO, Sies H. Selenoprotein P protects low-density lipoprotein against oxidation. Free Radic Res. 2004;38:123-128

23. Burk RF, Hill KE, Awad JA, Morrow JD, Kato T. et al. Pathogenesis of diquat-induced liver necrosis in selenium-deficient rats. Assessment of their roles of lipid peroxidation by measurement of F2 isoprostanes. Hepatology. 1995;21:561-569

24. Valentine WM, Hill KE, Austin LM, Valentine HL, Goldowitz D, Burk RF. Brainstem axonal degeneration in mice with deletion of selenoprotein P. Toxicologic Pathology. 2005;33:570-576

25. Arteel GE, Mostert V, Oubrahim H, Briviba K, Abel J, Sies H. Protection by selenoprotein P in human plasma against peroxynitrite-mediated oxidation and nitration. Biol Chem. 1998;379:1201-1205

26. Imai H, Nakagawa Y. Biological significance of phospholipid hydroperoxide glutathioneperoxidase (PHGPx, GPx4) in mammalian cells. Free Radic Biol Med. 2003;34:145-169

27. Yant LJ, Ran Q, Rao L, Van Remmen H, Shibatani T, Belter JG, Motta L, Richardson A, Prolla TA. The selenoprotein GPX4 is essential for mouse development and protects from radiation and oxidative damage insults. Free Radic Biol Med. 2003;34:496-502

28. Morozova N, Forry EP, Shahid E, Zavacki AM, Harney JW, Kraytsberg Y, Berry MJ. Antioxidant function of a novel selenoprotein in Drosophila melanogaster. Genes Cells. 2003;8:963-971

29. Muranyi M, Li PA. Hyperglycemia increases superoxide production in the CA1 pyramidal neurons after global cerebral ischemia. Neurosci Lett. 2006;393:119-121

30. Muranyi M, Ding C, He Q, Lin Y, Li PA. Streptozotocin-induced diabetes causes astrocyte death after ischemia and reperfusion injury. Diabetes. 2006;55:349-355

31. Bindokas VP, Jordan J, Lee CC, Miller R. Superoxide production in Rat hippocampal neurons: selective imaging with hydroethidine. J Neurosci. 1996;16:1324-1336

32. Murakami K, Kondo T, Kawase M, Li Y, Sato S, Chen SF, Chan PH. Mitochondrial susceptibility to oxidative stress exacerbates cerebral infarction that follows permanent focal cerebral ischemia in mutant mice with manganese superoxide dismutase deficiency. J Neurosci. 1998;18:205-213

33. Zhao H, Kalivendi S, Zhang H, Joseph J, Nithipatikom K, Vasquez-Vivar J, Kalyanaraman B. Superoxide reacts with hydroethidine but forms a fluorescent product that is distinctly different from ethidium: potential implications in intracellular fluorescence detection of superoxide. Free Radic Biol Med. 2003;34:1359-1368

34. Fridovich I. Editorial commentary on "Superoxide reacts with hydroethidine but forms a fluorescent product that is distinctly different from ethidium: potential implications in intracellular fluorescence detection of superoxide" by H. Zhao et al. Free Radic Biol Med. 2003;34:1357-1358

35. Ho JN, Lee YH, Park JS, Jun WJ, Kim HK, Hong BS, Shin DH, Cho HY. Protective effects of Aucubin isolated from Eucommia ulmoides against UVB-induced oxidative stress in human skin fibroblasts. Biol Pharm Bull. 2005;128:244-1248

36. O'Donovan P, Perrett CM Zhang X, Montaner B Xu Y-Z, Harwood CA McGregor JM, Walker SL Hanaoka F, Karran P . Azathioprine and UVA light generate mutagenic oxidative DNA damage. Science. 2005;306:1871-1874

37. Shiu CT, Lee TM. Ultraviolet-B-induced oxidative stress and responses of the ascorbate-glutathione cycle in a marine macroalga Ulva fasciata. J Exp Bot. 2005;56:2851-2865

38. Valencia A, Moran J. Reactive oxygen species induce different cell death mechanisms in cultured neurons. Free Radic Biol Med. 2004;36:1112-1215

39. Maglio DHG, Paz ML, Ferrari A, Weill FS, Czerniczyniec A, Leoni J, Bustamante J. Skin damage and mitochondrial dysfunction after acute ultraviolet B irradiation: relationship with nitric oxide production. Photodermatol Photoimmunol Photomed. 2005;21:311-317

40. Green DR, Reed JC. Mitochondria and apoptosis. Science. 1998;281:1309-1312

41. Green DR, Kroemer G. The pathophysiology of mitochondrial cell death. Science. 2004;305:626-629

42. Chen J, Berry MJ. Selenium and selenoproteins in the brain and brain diseases. J Neurochem. 2003;86:1-12

43. Shisler JL, Senkevich TG, Berry MJ, Moss B. Ultraviolet-induced cell death blocked by a selenoprotein from a human dermatotropic poxvirus. Science. 1998;279:102-151

44. Lu SC. Regulation of hepatic glutathione synthesis: current concepts and controversies. FASEB J. 1999;13:1169-1183

45. Maher P. The effects of stress and aging on glutathione metabolism. Aging Res Rev. 2005;4:288-314

Author contact

Corresponding address Correspondence to: Dr. Kamel E. Ben Jilani, Department of Cell Molecular Biology, John A. Burns School of Medicine, 651 Ilalo Street, BSB 217, Honolulu, HI96813, USA; E-mail: kameledu

Received 2006-11-27
Accepted 2007-2-5
Published 2007-2-11