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Ellis Veryard Items4 textbooks were found
An Account of Divers Choice Remarks, As Well As Geographical, As Historical, Political, Mathematical, Physical, and Moral Taken in a Journey Through ... Isles of Sicily and Malta ... As Also, a VoyaRentList Price:$34.75
An account of divers choice remarks, ... taken in a journey through the Low-Countries, France, Italy, and part of Spain; with the isles of Sicily and ... Levant: ... Illustrated with divers figures.RentList Price:$33.75
An account of divers choice remarks, taken in a journey through the Low-Countries, France, Italy, and part of Spain: with the isles of Sicily and ... with divers figures By E Veryard, MDRentList Price:$34.75
An Account Of Diverse Choice Remarks As Well Geographical As Historical, Political, Mathematical, Physical And Moral, Taken In A Journey Through The Low Countries (1701)RentList Price:$33.95
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It can hardly be called good news, bad news, because the latest earnings report was almost entirely filled with bad news. The only really positive thing about it was the commentary offered by Greg Garland, the company's CEO.
Unlike last quarter, in which he offered much more detail, Garland pretty much just repeated the same sentiments that Phillips 66 has seemed to have made into a mantra:
"We achieved record safety performance and refining utilization rates, started up the Freeport LPG export facility and returned $2.3 billion to shareholders through dividends and share repurchases. However, fourth-quarter financial results were disappointing, and reflect challenging market conditions.
We expect to generate additional free cash flow and create shareholder value as we complete major growth projects in 2017. Our portfolio of refining, midstream, chemicals and marketing assets positions us to capture opportunities across the value chain. We remain committed to operating excellence and growing our higher-valued businesses, while maintaining a strong balance sheet and disciplined capital allocation."
One thing remains clear. Phillips 66 places a very high value on the satisfaction of the company's shareholders.
Management seems determined to increase shareholder value by raising its dividend and buying back shares. However, in order to do so while also meet the company's capital spending requirements, Phillips 66 will need additional cash on hand.
That goal appears to be cautiously doable.
The company expects to see results by the end of the year from both its CPChem petrochemical plant and its Freeport LPG export terminal. Additionally, some of the capital constraints on the company would be alleviated somewhat by consolidating DCP Midstream into one entity. That would also be good for the company over the long term.
All of these factors combined add up to 2017 looking like a good year. They will also provide a hedge against potential refining margin weakness if the current refining environment doesn't improve during the year.

subset: commoncrawl


text: 95 mins R The Owl and the Pussycat | 1970 Comedy A call girl is attracted to an intellectual clerk. 95 mins R Director: Herbert Ross Writers: Buck Henry, Bill Manhoff Stars: Barbra Streisand, George Segal, Robert Klein | See full cast & crew » Awards: 2 Golden Globe nominations. | full awards » Jump to Related Articles Photos Cast & Crew 2Awards Synopsis Cast Barbra Streisand ... Doris Actor George Segal ... Felix Actor Robert Klein ... Barney Actor Allen Garfield ... Dress Shop Proprietor Actor Roz Kelly ... Eleanor Actor Jacques Sandulescu ... Rapzinsky Actor Jack Manning ... Mr Weyderhaus Actor Grace Carney ... Mrs Weyderhaus Actor Barbara Anson ... Ann Weyderhaus Actor Kim Chan ... Theater Cashier Actor Stan Gottlieb ... Coatcheck Man Actor Joe Madden ... Old Neighbor Actor Full Cast & Crew Director Herbert Ross Director Full Cast & Crew Writers Buck Henry Screenplay Bill Manhoff Play as Source Material Full Cast & Crew Company Rastar Productions Production Company Full Cast & Crew Synopsis A call girl is attracted to an intellectual clerk.


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Recent Posts
John Mahoney shows his 12" tall 3D printed figure, and gives painting tutorial.
Italian ceramic artist Mara Colecchia's latest series of collectable arts combine cute and dark elements and turn baby skull into the Merry Skulls.
John Mahoney showed 25 drawing at Hive Gallery Line Attack 4 - The Art of The Doodle show. 15 drawing sold at the opening night.


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Helping healthcare organizations meet CMS requirements.
Is your Emergency Management team ready to meet the new Centers for Medicaid and Medicare Services (CMS) standards for emergency preparedness? The time to start evaluating your emergency preparedness program is now, before financial penalties affect your healthcare organization.
On Sept. 16, 2016, CMS published a new rule, Emergency Preparedness Requirements for Medicare and Medicaid Participating Providers and Suppliers, in the Federal Register. Rule requirements must be met by Nov. 15, 2017. Battelle can help you cut through the confusion and develop a plan that will ensure the safety of your patients, fully meet the new CMS regulations, and avoid potential financial penalties.
Battelle has extensive experience in helping healthcare facilities assess their capability to respond to emergency situations, especially ones that require coordination with other hospitals, government officials and first responder agencies. We developed a standardized CMS Emergency Preparedness and Response Assessment (snapshot assessment) based on the published CMS regulations and deeming entity emergency preparedness standards, such as The Joint Commission, Det Norske Veritas-Germanischer Lloyd and Healthcare Facilities Accreditation Program.
Our team is also available for a range of follow-up services, including modifying or developing emergency preparedness plans, developing implementation plans, training and planning community-based disaster drills.
Our two-day onsite snapshot assessment uses a coaching/mentoring process in partnership with your Emergency Management team. Here's what you can expect:
The Battelle snapshot assessment leverages our emergency preparedness expertise to provide effective, affordable and easy-to-implement assessment and planning tools for healthcare organizations of all sizes.


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Downloading and installing apps can be a painful process as we all know. However, Google could be looking at a way to change that by allowing you to potentially "stream" some applications on Android, reports The Information. This could be achieved if the user has consistent data connectivity on tap, so it could be slightly limited in its approach, although the idea is pretty novel.
It is being rumored that Google's acquisition of a cloud based service named Agawi last year has been crucial in these new plans. This service specialized in bringing cloud apps to mobile devices and has also been associated with NVIDIA in some capacity. It is assumed that Agawi was involved in the NVIDIA Grid streaming service, which was a breakthrough in the gaming industry.
Agawi's expertise will supposedly be used with Google Search, where users can simply search for a service/app etc and start using it right there. But instead of opening the browser version of the app, it would open in a condensed app like format.
So Google certainly has some grand plans for the Android platform, if this report is anything to go by. It will be interesting to see if third party developers will be allowed to bake in support or if this service will be limited to Google's native services like Sheets, Docs etc.
Source: The Information – Requires subscription
Via: Droid Life


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Is Sugar Any Good For You At All?
If you think this article might absolve you from eating chocolate, think again!
Here's why a study says some sugar is good, and what you can do to adjust your diet.
What exactly is sugar?
Sugar is a carbohydrate present in most foods. Its role is to provide us with energy. Sugar is usually added to foods that are calorific like sweets, chocolate, cakes, pastries, fizzy drinks and shop bought coffees because in general people prefer the sweet taste.
Sugar goes by many names including fructose, honey, molasses, hydrolysed starch, corn syrup, maltose, sucrose and glucose.
How much?
The NHS says we should eat not more that 5% of our calorie intake from sugar each day. That's about 30 grams for the over 11s.
If you're wondering about fruit juice, well, it's a good choice and counts as one of your five-a-day, but because it's in liquid form it can stick to your teeth more easily. Breaking down fruit into a juice releases its sugars, so drink it with food and not after you've brushed your teeth.
Is sugar good at all?
There's some new research from the Science Signalling journal that highlights a good relationship between a certain sugar and non-alcoholic fatty liver disease.
Too much sugar can result in non-alcoholic fatty liver disease (NFLD). It's thought one in three people in the UK are in the early stages of NFLD and there is no drug treatment available for it.
When sugar enters the liver, it's stored as trigcerides fat which leads to the disease. However, a form of sugar called trehalose blocked fructose sugar, one that causes NFLD, from entering the liver of test mice.
After blocking the sugar it also aided in breaking down and flushing out fat inside liver cells by triggering 'autophagy' a process whereby the liver cell thinks it is starving and begins to eat the fat it's carrying.
Trehalose is a natural sugar found in plants (and insects), but before you rush out to buy it, remember this study was conducted on lab mice, not humans.
Why is sugar bad?
Obesity is a growing concern. The World Health Organisation (WHO) labels it a pandemic and says sugary, fatty and salty foods are in part to blame for the rise in obesity.
Eating too much sugar increases your weight and causes obesity. Obesity then causes health issues like heart disease, stroke and diabetes.
Heart disease
Heart disease is the leading cause of death in the UK killing 73,000 people every year.
A high sugar diet causes obesity, high blood pressure and puts a strain on the heart. When arteries narrow and become clogged with cholesterol, it's difficult for blood to pass through. When a piece breaks free it can block arteries and cause a heart attack.
Stroke is the third biggest killer in the UK. A stroke happens when arteries that feed the brain get blocked. Starved of oxygen the brains begin to die. It's a life-threatening condition and causes long-term disability. 110,000 people have a stroke every year in England alone.
Diabetes occurs when blood sugar levels are too high and the body becomes resistant to energy releasing insulin - the body's fuel.
Teeth rot when plaque builds up and creates acid. Sugary foods are quickly turned into energy and release plaque acids at the same time. When plaque builds up the acids can dissolve tooth enamel and create holes in your teeth which are called cavities.
It's thought a diet high in sugar triggers thrush, a vaginal and oral yeast infection. Yeast feeds on sugar so by cutting it out you may suffer less.
How to cut down on sugar
It's not always easy to cut out sugar because there are so many foods available that contain high amounts.
It doesn't help that sugary foods appeal to our sweet tastebuds and with manufacturers adding more in a bid to get our attention, sweet tasting healthy foods like tomatoes and bananas fall by the wayside. Here's how to get back on track:
- Have a clear out of sugary treats. Start by cutting down to one day and then stop buying them.
- Read nutritional labels on processed foods to check the sugar levels. If a form of sugar appears near the beginning of the ingredients list it is high in sugar. Search out the 'Carbohydrates of which sugar' line.
- Stop buying fizzy or sports drinks and fill up with water instead. If you don't like the taste, buy carbonated or add a splash of fruit juice. Fizzy drinks can often contain caffeine too, something to be aware of in case you get withdrawal headaches.
- When you're out shopping don't choose a sugary hot drink; tea or flat white coffee is the healthier option. Some shop bought coffees can have over 300 calories of sugar.
- If you use tinned fruit choose fruit in juice rather syrup - remember syrup is sugar.
- Breakfast cereals can be a great source of iron, fibre and minerals. Make yours count by dropping those Frosties in favour of Bran Flakes with some soft fruit such as strawberries.
- Cut out sugar from your tea and coffee
- 22.5g per 100 grams is high in sugar
- 5g per 100 grams is low in sugar
Make an informed decision by choosing the option with the least amount of sugar. Add flavour with pepper and herbs such as sweet basil or coriander.
Given that we're used to high sugar diets cutting out the white stuff can be a struggle, but every gram you manage to avoid improves your health.
If you stick to preparing fresh foods, or even eating only half of that processed lasagne with a head of fresh broccoli instead, the health benefits are huge.
Sadly not all sugars are the helpful type identified in the study. For now, it's best to cut right back on sugar consumption and save it for a treat.


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You bought your V10 because its reliability and value appealed to your senses; so why would you be okay with second best when it comes to great quality auto parts? Car aficionados appreciate the a brand you can trust built into each Chevrolet, and know that only the highest quality repair V10 parts will suffice when your car or truck needs fixing. Anyone who owns a Chevrolet knows that when it comes to maintaining their vehicle only the most superior replacement parts will do. While it may not be difficult to locate replacement parts for Chevrolet vehicles, these days the necessity for nothing but the highest quality are required to repair your automobile.
Believe it or not, but for a long time there was no such thing as power steering. If you suspect your Chevrolet V10 power steering pressure hose has developed a leak, start the car and turn the steering wheel a couple of times. If you see a puddle of fluid, then you know there's a problem. Without a power steering pressure hose, it becomes very difficult to maneuver your vehicle... especially at low speeds. That's because the power steering pressure line on your Chevrolet V10 takes the hydraulic pressure generated by the power steering pump and sends it to the steering gear box. There, the motions you make when you are steering are assisted by the pressurized fluid... all thanks to the power steering pressure hose.
Make all your future car parts purchases at carpartsdiscount.com and save time and money on all your upcoming repairs. At Car Parts Discount, we have live customer service members here with enough expertise to help you purchase the appropriate Chevrolet V10 part for your project or repair. Bring your V10 back to life with the best new or rebuilt parts from the experts at Car Parts Discount. Need 1987 Chevrolet V10 Power Steering Line - Pressure parts? We've got them right here.


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Question: The direction of any magnetic field is taken to be
The direction of any magnetic field is taken to be in the direction that a compass points. Explain why this means that magnetic field lines must leave from the north pole of a permanent bar magnet and enter its south pole.
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View Solution:
Answer to relevant QuestionsSuppose a long, straight, current- carrying wire had a current from west to east. If it were immersed in a vertically upward magnetic field that was stronger on its west side than on its east side, what would be the initial ...(a) As you start in the very middle of Fig. 19.3b and move horizontally to the right, what happens to the magnetic field spacing as indicated by the iron filing pat-tern? What does this imply in terms of the magnetic field ...A permanent bar magnet (# 1) is vertically oriented so that its north end is below its south end. The north end of this magnet feels an upward magnetic force of 1.5 mN from an identical vertically oriented magnet (# 2) that ...In a mass spectrometer, a doubly charged ion having a particular velocity is selected by using a magnetic field of 100 mT perpendicular to an electric field of 1.0 kV/m. This same magnetic field is then used to deflect the ...A wire carries a current of 10 A in the +x-direction. (a) Find the force per unit length on the wire if it is in a magnetic field that has components of Bx = 0.020 T, By = 0.040 T, and Bz = 0 T. (b) Find the force per unit ...
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MLD 2012 Montagu Allan QF: Regina Pats vs. Brynäs IF
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Originally Posted by
- Peplinski and Franzen are more comparable as they are the physical beasts of the 4th lines. Believe it or not, their career averages for adjusted ESP/GP are the same, which makes Peplinski a much better producer because 1) he didn't play with Datsyuk and Zetterberg or anyone similar, 2) adjusted points are biased against him and for Franzen, and 3) his record is based on 50% more games. Both are big, strong, hard to move guys, but Peplinski had no quit, and Franzen apparently has had lots of quit lately. Peplinksi can fight, take faceoffs and play other positions if needed. Franzen can't. As much as Peplinski is the more attractive producer, he is, in particular, a much more attractive 4th liner.
Peplinski's estimated linemates (by players with whom he most frequently shared a point on an even strength goal).
1980-81 Willi Plett Eric Vail
1981-82 Jamie Hislop Lanny McDonald
1982-83 Lanny McDonald Kevin LaVallee
1983-84 Kent Nilsson Steve Tambellini
1984-85 Kent Nilsson Steve Tambellini
1985-86 Joel Otto Joe Mullen
1986-87 Joel Otto Joe Mullen
Although his totals were usually relatively low, meaning that these were his most common linemates but not his linemates all or even most of the time, necessarily.
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The star of this show is Cari Lekebusch, the veteran Swedish producer and honcho of the seminal Hybrid Sound Architectures label (later known as H. Productions). He'll be playing his deep and driving beats atop Long Island City's Ravel Hotel, with Basic NYC's Sleepy & Boo lending top support.
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Ask about the over 50 other support services that are available! Are you travelling and need a boardroom? As a Telsec Business Centres client you can take advatage of over eight hours of free boardroom space and facilities per month in any of over 500 Business Centre locations in 40 countries.
Telsec also has many services that may be of interest as well.
Locations Home >> GB >> Nottingham >> Cumberland House
Office Space and Virtual Offices
Located in: Nottingham,
35 Park Row,
Location Details:
This modern building is nestled in the heart of Nottingham and is easily accessible to the M42 and East Midlands Airport. The Cumberland House is in close proximity to Park Row, right off Maid Marion Way, and public transportation. Our centre offers 65 fully furnished and serviced offices, with excellent conference and boardroom facilities.
Office Space Services: Featuring furnished offices, meeting rooms and more with business center services.
Virtual Office Services: Featuring live telephone answering, advanced telecommunications and business address mail receipt.


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The Federation Science Council is a sub-group of the Federation Council which concerns itself with scientific research and policy within the United Federation of Planets.
Circa 2267, the Federation Science Council temporarily reclassified the planet Mestiko from class M to class L. (TOS - Mere Anarchy eBook: The Centre Cannot Hold)
In the 2360s, Hekaran scientists presented their claims that warp drive was damaging subspace to the Federation Science Council. The Council dismissed these claims until the Hekarans proved their theory in 2370 by creating a subspace rupture. This led to the Science Council's recommendation to the full Council that a Warp 5 "speed limit" be established. (TNG episode: "Force of Nature")
Council Members
2270s Edit
- Vulcan- T'Nuri (Chairwoman)
- Betelguse- Chuu'iik Hru'uith
- Ithen- Danga Sitru
- Makus III- Konahr Lutet
- Regulus III- Maria Moi
External linkEdit
Federation Science Council article at Memory Alpha, the wiki for canon Star Trek.


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Choose JOY
JOY is a choice. It doesn't belong to a lucky few whose lives seem to go off without a hitch. No! It's there for the taking for all of us...despite and in spite of the set-backs and pitfalls that are part of the human condition. Choosing JOY becomes our way of exercising our will to get up every time we fall. We are the captains of our ships, the drivers of the bus and we emerge victoriously every time we choose JOY!
Wake-up to JOY and let it spread like wildfire! Every purchase of one of our mugs from our JOYFUL line helps support at-risk kids. Start with JOY and let it ripple outwards!
Each cup is designed to remind you to think positively, to find joy in the smallest of spaces and to remember to pass joy on.
When giving as a gift, this gift gives in two ways:
1. To the recipient of the cup
2. To the children who will benefit in myriad ways.
With every Cup of Joy ordered, you will receive:
1. An instant download print of the saying that matches the mug, as pictured above!
Print on beautiful paper and frame (frame is not included). It's our little way of saying "thank you" for supporting this mission. If you are giving your Cup of Joy as a gift, you can print, frame (with your own frame) and present to the recipient right away! Want to print one for yourself, your mama and your neighbor Jeanine? By all means! We get more JOY when we spread it around!
2. Along with your printable, you will also receive a printable gift card (square) that shares the backstory on the "Cups of Joy" mission, how it will benefit children and how it's truly a gift that gives back! We suggest printing this on durable cardstock and adding to your greeting card. Ordering ahead of time and giving the gift in person? Simply tuck the card inside the mug!
About Our Cups of Joy
100% made in America!
This sturdy white, glossy ceramic mug is an essential to your cupboard. This brawny version of ceramic mugs shows its true colors with quality assurance to withstand heat in the microwave and put it through the dishwasher as many times as you like, the quality will not be altered.
• Ceramic
• Dishwasher safe
• Microwave safe
• White, glossy
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4.92 as a percent
Convert from decimal to percent. Here is the answer to the question: 4.92 as a percent or how to convert 4.92 into a percent equivalent.
Use the decimal to percent calculator below to write any decimal as a percentage.
How to convert from decimal to percent
Let's see this example:
We want to write the decimal number 4.92 in percent form. So, to convert this value to percent, we just multiply it by 100.
➥ There is an ease way to accomplish this:
So, 4.92 is equivalent to 492% in percent form.
See also:
Decimal to Percent Calculator
Sample Decimal to Percent Calculations


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The Top 10 Video Game Athletes of All Time
4. Paste (Bases Loaded)
New Jersey's true hero isn't Newark mayor Cory Booker, but a fictional homerun champ who led the most potent offense in Bases Loaded: Paste. If you toss a pitch that doesn't end up a foot off the plate, there's a good chance that New Jersey's—and Bases Loaded's—number one slugger will deposit it into the nosebleed seats.
Bases Loaded isn't a stat heavy game. In fact, you can't keep track of them at all. But the default numbers that are included for hitters, batting average and homeruns, are all that's needed to elevate Paste into the baseball video game pantheon.
Check the numbers: Paste has a remarkable 60 homeruns… 60! Sure, in the aftermath of Flax Seed Oil and Canseco Milkshakes, 60 dingers reads tame, especially when Barry Bonds and Mark McGwire both managed to hit 70 round-trippers. But back in the late '80s, this number was still holy and, Jaleco showed respect to what many see as the real homerun record by having Paste's tater total not equal or surpass Roger Maris' 61.
These home runs are accompanied by an insane a .467 batting average. His season average is what many major leaguers can't achieve during spring training. Paste cranks out hits—many of which are stinging line drives—at an almost 50 percent clip, which means he's making contact a lot. That's extremely hazardous for pitchers when an opponent is popping the ball out of the park 60 times in a season.—Next: Number 3 >


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I recently started dating a guy and he's pretty cool. However I've picked up on something that he does which annoys me and worries me a little. Sometimes when we are talking about something, he will stop texting mid conversation or wish me a goodnight without acknowledging what I say. Usually the topic won't even be something that should be upsetting so I don't know if he gets busy or what. Sometimes if ) e stops texting during the day, he will come back and start talking about something else. I am already having some difficulty getting used to the way he texts since he texts very grammatically correct with barely any haha or lol so I usually end up feeling like he's mad at me or whatever. If any one can shed some light on why he says goodnight mid conversation etc, I would appreciate it a lot! Thank you :)
Why does he cut conversation short or stop responding all of a sudden?
Have an opinion?
What Guys Said 2
Playing hard to get
Doesn't want to spoil his prestige.
Could be a serious person.
Could be moody person
Maybe he suck in texting and it takes so much effort for him that's why he pay attention to his grammar.
He is busy or not interested.00|00|0
My friends have said that since he's a medical school student, he's going to be a different texter than what I'm used to so I don't know. Maybe you're right with playing hard to get though? After our first date he told me that he's gonna be a dime a dozen and I'm going to lose interest in him really quick
Lose interest in him? hmm that's lame to say from a medical student.
If he was a confident guy he might be using this as an excuse to show you that he is not good enough to be with you which lead me to my last point '' He is not interested ''.
What Girls Said 1
He either gets bored or he is doing something else.00|00|0
Learn more
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Cadillac ATS Diesel Confirmed: 2012 Detroit Auto Show
The Cadillac ATS compact luxury sports sedan may have leaked onto the net earlier today, but the representatives of the GM luxury brand did manage to keep one big secret… a secret.
At the car's official unveiling on the eve of the Detroit Auto Show, GM North America President Mark Reuss confirmed that along with three gasoline engines the ATS will be available with a diesel powerplant. Reuss didn't provide any details on exactly what that diesel engine could be, nor did he say when it would arrive. Instead, he told the assembled journalists that the ATS will get a diesel powerplant "within its lifecycle" suggesting the oil-burner is still a long way off.
GALLERY: 2013 Cadillac ATS
See AutoGuide's complete 2012 Detroit Auto Show Coverage Here


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The web team at Definitive watched the segment on 60 Minutes a few days ago on "regenerative medicine" and it was great!
How exciting to know that the future promises to be so helpful in fighting pre-mature death due to heart disease, kidney disease and other ailments. Regenerative medicine is helping wounded soldiers recover from awful injuries and presents a brighter healthier future for everyone.
Check out this video from the segment on Regenerative Medicine on 60 Minutes by clicking here.


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A long time customer (1 of my first) sent me this email 3 hours after picking up her cake:
"Just wanted to contact you because I think this is the first time that I've been dissatisfied with your cake. The iPad cake was supposed to look similar to the cake that was emailed to you not have a screen shot print. It had the apps 3d on the cake & honestly that's what I was expecting to see. I'm thinking that I could have gotten that done at the local market bakery, I know the contract states that it won't be exact but to be honest I figured that lent more to "this cake will not be a 100% replica of the photo" for liability sake. I just can not believe I just spent $90 and 1hr getting a screenshot cake."
And this was my response:
"Hi. It's unfortunate that you were dissatisfied because I do value you as a customer. I sure wish you had said something before you left. This was clearly a misunderstanding and I should have thought to ask whether you wanted the buttons "3D"? Then I would have quoted you more like$150 (probably more) instead of $90 because of the hours of extra work that would have taken. And yes, while you could have gotten a cake with a screenshot at a local market bakery, I am actually a little offended at the comparison. I make all my cakes and icing from scratch with no added chemicals or unnatural ingredients . I bake each cake to order to ensure optimum freshness. I use quality ingredients. Supermarket cakes are mass produced from mixes with hardly any natural ingredients, shipped and most often decorated frozen, and then sit around in a case for weeks until purchased. I realize in this economy $90 is a lot to spend on anything but let me break down the cost for you. The ingredients alone (cake and decorating materials including packaging) is about $20. The screenshot (including travel time and gas used to purchase) another $20. I spent about 3 hours just 2 bake, make icing, and decorate your cake (how much do you get paid an hour?). I under value my time @ $15 per hour so that's another $45. That's already a total of $85 and I haven't even included the gas and electric I use. So yes it is definitely more expensive to purchase a cake from me than the supermarket. I know you may have paid less for your cakes in the past, but the cost of ingredients have gone up significantly. Also the cost for my time has increased. I do hope this helps you to understand why I charge what I do. Like I said I value your business and definitely appreciate your patronage over the years. I never want my clients to be dissatisfied so tell me what you think is a fair resolution?"
Do you guys think the response was overkill? Should I offer a refund or discount of a future cake?
This scenario is always interesting. You put in the time and effort, they take the cake, they SERVE the cake, then they complain about the cake. You are pretty exact about what went into the making of the cake and personally I think it is okay. You give them an opportunity to help resolve the issue so I would wait for their response. I think what this teaches us as cake people is to be more thorough in eliciting what the client really wants and expects. While this can be upsetting, it can be helpful to improve future cake orders.
I do think your response was a little overkill. I looked at you ipad cake photo and yes it does look like one large edible photo placed on a cake. I know it took you many hours to bake, ice, purchase the photo however if the customer showed you a photo and expected a decorated ipad cake, with cut out images then I think she deserves alittle refund. If I were that customer, I am not sure I would be coming back for another cake, especially after that long email. Just my opinion though. Was the ipad photo she showed you alot different?
I agree, I think you went into to much detail in the email. It sounds kind of whiny and like you are just trying to justify why you charge what you do. The customer doesn't care how much materials are or how much you get paid an hour. I know we talk about wanting to educate the cake muggles a lot, but I don't think a dissatisfied customer is an appropriate time to do this.
Not to be harsh, but I think I would have been disappointed too. If I gave you a picture of a cake with 3D "apps" on the iPad screen, and I got an edible image instead, I would feel cheated, unless you had specifically told me "I cannot or will not be doing these apps in "3D", it will be an edible image, is that ok with you?" You HAVE GOT to be clear with someone on what they want, and get a signed order form and or contract outlining the details of the order.
All that said, I think it is good that you asked her what she thought was a fair resolution.
While I completely understand your frustration with this customer and agree that there is nothing wrong with explaining why the cake cost what it did, I do think that your tone toward her should have been much more polite. Instead of saying that you are offended by her, you could have simply said "You're right that a grocery store cake would have also had an edible image. However, I'm sure you will agree that the quality and taste of my cakes are much higher than a grocery store cake would be." And the remark about "How much do you get paid per hour" was completely inappropriate based on the e-mail that you received. It didn't sound like she was attacking you in her e-mail, and since you are the professional, your tone should have been even more respectful. That doesn't mean you can't still say what you feel, you just need to watch the way you say it. Especially if you truly do value her has a customer and want her to order again. I would have to say the likelihood of her ordering from you again is very low with the reponse you sent to her. You've got some great work posted, so I'm sure you don't get dissatisfied customers often. Sorry you're going through this though, and I hope that things go better for you in the future!
I think it was also a little overkill because from her email I don't think she really has a problem with the cost of your cakes. After all she is a repeat customer. It seems to me that her problem this time is that she thought your concept was wrong and is dissatisfied with the way you executed that concept.
That said I think you were right to tell her how the 3d buttons would affect the price and why and definately it was good to ask her what kind of recompense (? not the right word) she thinks would be fair. I just hope she doesn't think a free cake would be it. Sometimes people are ridiculous.
I think part of the problem, too, is that the image is too small for the cake. The screen on the iPad does not have a white border; the desktop fills the screen. The cake should not have had the white border between the image and the black, so it looks less realistic.
I am sorry you are having this problem and hope you and your customer can come to a mutually agreeable decision.
While I agree with everything you wrote to her and certainly wish I could repeat the same thingto customers at times...I probably wouldn't have gone into that much detail...as someone pointed out..Customers don't care how much each cost breakdown is..Give me a price and I will either pay it or not...
You did the right thing about asking her what she feels is a fair resolution but unfortunetly you may have offended/annoyed her and I doubt she will come back..JMO
I agree with secretagent. the white area on the cake is just weird. I would not have been happy either.
Something I learned in my advertising days is that when writing an email to someone who just annoyed you...don't put their email address in the "To" section.
Feel free then just to write out an email and get everything out of your system...then chill for a bit, walk around...and then write the professional response in another email. This allows you to vent your frustration but maintain your professional composure.
I can understand both sides in this situation...for future reference, it's always best to include as many decorative details as possible in written confirmations (esp. when doing a cake based on a design that's been sent in). That way you're in the clear if they don't like something that they've approved.
Wow...don't think she'll be a repeat customer anymore...but at least she'll be educated in how underpaid cake decorators are! Sorry but you went into WAY too much detail and the 'how much do YOU get paid an hour' was insulting. Plus, I agree with the poster that said the image did not fit the cake. It looks like you hurriedly slapped on an image...no matter how long it took to bake etc. Not seeing the image she sent it's hard to see where the communication lapsed but regardless it wasn't something she expected and your email probably didn't help.
If I were you and I wanted to continue her business, I would write her another letter stating that you are sorry but your evil twin had taken over and written that email and that you indeed are sorry she was unhappy and that next time you will be sure to clarify any instructions from you on cake design...plus you'll be happy to discount her next purchase.
I'm sorry to say that I agree with the others. The first 3 lines and the last 2 lines would have been enough. Good luck on this. I hope this can be resolved with both you and the customer being happy.
Sorry, I'm probably in the minority here, & might get trashed for it, but I think the response you sent your customer was extreme overkill.
Your customers email did not come across as disrespectful, it was one of disapointment. She was expecting the cake to look a certain way, it didn't, and she was expressing her disapointment that it was not what she expected for the money she paid.
This could have been handled in a more professional and tactful way so both sides would be satisfied. Sending a flaming angry reply to your customer, and adding "how much do you make an hour" in such a sarcastic way, was not the best idea.
I honestly think your customer who you say had been a returning customer, will not return again unless some miracle solution is come too.
I also think there is a high chance she'll be offended enough, she'll go out of her way to pass on your angry response to her circle of friends. Word of mouth can be deadly to a business.
I worked in retail 30 plus yrs, and one thing you learn fast when dealing with so many different types of customers, is to treat all customer with respect at all times. They can be the most obnoxious customer in the world (and you might want to bop them a good one) but you have to deal with thier complaints with a level head.
If you fly off the handle with your complaining customer, you lose. You lose the customer, and you lose more customers via the angry customer.
I would be interested in seeing the photo she requested you work from...if the photo was all done in fondant, with fondant cut-out apps, and she asked you to replicate the cake, then you certainly should have charged for, and done, the replica, or given her the money-saving option of an edible image...it would have been her choice. I can see how she would be disappointed if the cake she asked you to make had hand-cut icons done out of fondant.
I would just write her, apologize that you did not realize she wanted the cake to look exactly like the photo (again, it would be helpful to see the photo she sent you) and offer at least 50% refund.... Because I have seen the iPad cakes with the fondant cutout apps and it does look a lot different than the cake with the edible image screen shot.....
Your cake is fine, but if it wasn't what she ordered, then it wasn't fine for HER...you know what I mean? Sorry this happened to you.
Oh yeah, kiss her goodbye. Sorry! That was uncalled for. You come vent here, but you should knock yourself over with kindness to them. That's just a sheetcake with an edible image on it. Nothing more. I see she did mention 3d apps, so she was expecting that.
Thanks for all your responses. Did I mention I wrote that email at 2:00 in the morning after having not slept for 24 hours So yeah, after re-reading it I do agree with you all. I did refund her 50% and can only hope she forgives my lapse in judgment.
Thanks for all your responses. Did I mention I wrote that email at 2:00 in the morning after having not slept for 24 hours So yeah, after re-reading it I do agree with you all. I did refund her 50% and can only hope she forgives my lapse in judgment.
Hopefully, there won't be a next time. If there is, don't respond when you are exhausted and not in a good frame of mind. Since she is a repeat customer that you want to salvage, I might send her a handwritten note, apologizing for your error/miscommunication regarding the design, thanking her for her order, and offering her an incentive to order from you again, like 10% off her next order or a free dozen cupcakes. I would do this in addition to the 50% refund you already issued. You want to keep good repeat customers happy. This might help.
Whenever I need to send an email to address a situation that could potentially turn confrontational, I always have someone else read it over before I send it. It is very easy to misread the intent or tone of email due to the lack of nonverbal cues, so structuring your narrative correctly is extremely important, especially in professional communications.
I agree that a handwritten note would help mend fences, maybe even a free unsolicited 6-pack of cupcakes (the cost to you would be minimal if you just make a little extra batter from another order).
You absolutely said what everyone has wanted to say at one time or another! No filter, I have that problem too :0). When I am upset I write/type a draft of what I'm going to say back, revisit it in 24 hrs & see if I feel the same (sleep or lack there of, kids aggravating each other or me), then maybe depending on particulars run it by someone who I feel would give me their realistic opinion. Then couple days later (if at all) send it out. If you feel you were harsh with her, tell her so - people appreciate honesty and a true apology is often accepted! Of course a true apology with some delicious cupcakes.....now who can refuse that!!! Best of luck to you, I hope it all works out!!!
Yep I agree with most posters. I too would be upset if I got the cake she did. I am going to get slammed for this but..... even for an edible image it wasnt very nice.
Plus, she wasnt unreasonable.. .I think your resopnse was.
While we all understand the no sleep part.... that isnt her problem lol. Next time wait until morning, and see how much your response changes
[quote="motherofgrace"]Yep I agree with most posters. I too would be upset if I got the cake she did. I am going to get slammed for this but..... even for an edible image it wasnt very nice.
I agree - edible images should stay at the grocery store.
You're going to need some thicker skin.
I have seen I phone cakes done with an edible image before, and they were nice. But that was with fondant... And i thin kthat makes a big difference when you are doing a cake like this.
$90 was way too much for that cake - sorry. And your response was argumentative and rude, not professional at all.
cycakes, mother of grace, brevity, & wykdgud, this thread was over and done with 6 days ago. The OP has stated what she has done to recitfy the situation. Lets leave it at that.


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Alexander Bernhard
About me
I am a strategic generalist who is also able to jump in and execute on a good idea. I can see the big picture but at the same time be realistic to meet tactical milestones to launch large project from vision to reality I have worn many hats in my career – FMCG purchaser, project developer, manager, digital specialist and strategist. And as a result I have a unique ability to manage multiple-disciplinary projects and to navigate complex challenges. Overall my biggest strength would be a deep sense of ownership for every project I take on. I love mentoring others and work as a part of an executive team to achieve our common goals. I love making new professional acquaintance, so feel free to reach out if you want to get in touch to talk about technology, business or a cool new project.
Oslo, Norway
Product management
Project management


text:
See what he wrote below:
"AMAZING! So this young man gave me N500 (Five Hundred Naira)! He is a cleaner in a company but he's so hungry for success that he bought my book (both the soft and hard copies!), read it and took the advice I gave in one of the chapters, where I said if you wanna meet an 'important' or successful person (in your opinion), TAKE A GIFT NO MATTER HOW SMALL! All he had was 500 bucks and he gave it! Of course I gave him my personal number and now he can call me anytime and we can build a relationship from there! I am so glad that there are still a lot of young people out there who not only respect knowledge but understand that there is power in its application!#SecretsOfTheStreetsBook#EntertainmentSecrets #kingoftalk.


text:
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The molecular chaperone Hsp90 (heat shock protein 90) is a promising target in cancer therapy. Preclinical and clinical evaluations of a variety of Hsp90 inhibitors have shown antitumor effect as a single agent and in combination with chemotherapy. Current Hsp90 inhibitors are categorized into several classes based on distinct modes of inhibition, including i) blockade of ATP binding, ii) disruption of cochaperone/Hsp90 interactions, iii) antagonism of client/Hsp90 associations and iv) interference with post-translational modifications of Hsp90. The different functions of Hsp90 isoforms and the isoform selectivity of drugs need further investigation. The correlation of cell surface Hsp90 with cancer metastasis and the emerging involvement of Hsp90 inhibition in cancer stem cells have become exciting areas that could be exploited. Therefore, the aim of this review is (1) to summarize the up-to-date knowledge of mechanistic studies and clinical prospect of currently available Hsp90 inhibitors, (2) to enhance our perspectives for designing and discovering novel Hsp90 inhibitors, and (3) to provide an insight into less-understood potential of Hsp90 inhibition in cancer therapy.
The molecular chaperone Hsp90 (heat shock protein 90) was initially identified as one of the highly conserved heat shock proteins involved in the stress response (Ritossa, 1996; Westerheide and Morimoto, 2005). Hsp90 is a highly abundant protein, constituting about 1–2% of total proteins under non-stress conditions in most tissues (Welch, 1991; Welch and Feramisco, 1982). It possesses common molecular chaperone functions, i.e., assisting protein folding and preventing aggregation of non-native proteins (Wiech et al., 1992). Over the past several years, the application of global analysis has extended Hsp90 clientele to more than 200 proteins, covering almost all the cellular processes (Falsone et al., 2005; McClellan et al., 2007; Zhao et al., 2005). For instance, these client proteins include transmembrane tyrosine kinases (Her-2, EGFR), metastable signaling proteins (Akt, Raf-1 and IKK), mutated signaling proteins (p53, v-Src), chimeric signaling proteins (Bcr-Abl), cell cycle regulators (Cdk4, Cdk6), and steroid receptors (androgen, estrogen, and progesterone receptors) (Kamal et al., 2004). Many of these client proteins are mutated and/or overexpressed in cancers (Pearl et al., 2008) (Didelot et al., 2007).
Because early studies have shown that the function of cytoplasmic Hsp90 is essential for normal cell viability and growth (Borkovich et al., 1989), it was difficult, if not impossible, to consider Hsp90 as a potential therapeutic target. However, since geldanamycin (GA) was demonstrated to possess potent anti-cancer effects through inhibiting Hsp90 (Supko et al., 1995; Whitesell et al., 1994), a great deal of efforts have been devoted to this area and a diversity of Hsp90 inhibitors have either been identified or synthesized (Schulte, 1998; Whitesell et al., 1994; Cheung et al., 2005 ).
The feasibility of targeting Hsp90 for cancer therapy is well supported:
Although the mechanisms underlying the tumor selectivity of Hsp90 inhibitors are not fully understood, Hsp90 has become validated as a potential target in cancer therapy. Furthermore, preclinical and clinical evaluation of a plethora of Hsp90 inhibitors have already shown promising results as a single agent and/or in combination with chemotherapy (Solit and Chiosis, 2008).
Based on the crystal structures of yeast Hsp90 (Ali et al., 2006) and Grp94 (Hsp90 isoform in mammalian endoplasmic reticulum) (Dollins et al., 2007), Hsp90 exists as a homodimer, each monomer consisting of three highly conserved domains: an N-terminal ATP-binding domain (25 kDa), a middle domain (35 kDa) and a C-terminal dimerization domain (12 kDa) (Pearl and Prodromou, 2006). In eukaryotes, the N-terminal and middle domains are connected by a charged linker (Terasawa et al., 2005). The N-terminus of Hsp90 contains a specific ATP binding pocket (Dutta and Inouye, 2000). The major role of the middle domain is to discriminate various types of client proteins to adjust the molecular chaperone for proper substrate activation (Hawle et al., 2006). The C-terminal dimerization domain strengthens the weak association between the two N-terminal domains of the Hsp90 dimer (Terasawa et al., 2005). The C-terminal domain of eukaryotic Hsp90 has a conserved pentapeptide (MEEVD) implicated in binding to the tetratricopeptide repeat (TPR) domain of cochaperones, such as Hop (Hsp organizing protein) and Sti1 (stress-inducible protein 1, yeast homologue of Hop) (Pearl et al., 2008; Terasawa et al., 2005).
The "open" state of the Hsp90 dimer, with its two N-termini separated, can capture client proteins (Richter et al., 2008). ATP binding triggers the closure of the ATP pocket "lid" and brings the N-termini close to each other, resulting in the formation of a compacted, ring-shaped Hsp90 dimer (Richter et al., 2008; Wandinger et al., 2008). These conformational alterations lead to a "closed" state to "clamp" client proteins inside (Terasawa et al., 2005). The ATPase activity of Hsp90 itself drives the chaperone cycle (Kamal et al., 2004).
The Hsp90 multi-chaperone system has been extensively studied in the maturation of steroid receptor on yeast Hsp90 (Scheibel and Buchner, 1998). The chaperone cycle begins with a newly synthesized or misfolded steroid receptor binding to Hsp70/Hsp40 complex, associated with the "open" state Hsp90 via the bridging cochaperone Hop that interacts simultaneously with Hsp90 and Hsp70 (Pearl et al., 2008). Hop not only binds to the C-terminal MEEVD motif of Hsp90, but also connect with the N-terminal region of Hsp90, preventing the Hsp90 N-terminal domain association. Hop therefore inhibits the ATPase activity and promotes client transfer from Hsp70 to Hsp90 (Prodromou et al., 1999; Richter et al., 2003; Terasawa et al., 2005). Upon ATP binding to Hsp90, Hop is replaced by p23 and immunophilins, converting the intermediate chaperone complex into the mature complex (Neckers, 2003). Another cochaperone, Aha1 (activatior of Hsp90 ATPase), associates with the middle domain of Hsp90, facilitating conformational adjustments to favor ATP binding (Meyer et al., 2004). Both Aha1 and immunophilins stimulate the ATPase activity of Hsp90 (McLaughlin et al., 2002; Meyer et al., 2004; Panaretou et al., 2002). Upon ATP hydrolysis, the correctly-folded client protein is released from Hsp90 (Terasawa et al., 2005). Recent studies have shown that the mechanistic basis of the Hsp90 chaperone cycle are conserved in yeast and human, although a slower turnover rate was observed with human Hsp90 (Richter et al., 2008).
Protein kinases are the largest class of Hsp90 clients, with a similar chaperone cycle (Caplan et al., 2007; Pearl, 2005; Terasawa et al., 2005). However, minor differences are noticed with the cochaperones involved (Caplan et al., 2007). Cdc37 was originally discovered in yeast as an essential cell cycle protein (Reed, 1980), and later it was proved to be a kinase-specific cochaperone of Hsp90 (Pearl, 2005). The Hsp70/Hsp40 complex first prepares a newly synthesized or misfolded protein kinase for interaction with the N-terminal domain of Cdc37, followed by recruitment of Hsp90 to the complex with the help of Hop (Arlander et al., 2006; Mandal et al., 2007). The C-terminal side chain of Cdc37 associates with the "lid" of Hsp90, which closes the N-terminal ATP binding pocket (Roe et al., 2004). Crystallographic studies revealed that the insertion of Cdc37 C-terminus to the Hsp90 N-terminal ATP pocket inhibits the ATPase activity of Hsp90 and prevents its N-terminal dimerization (Siligardi et al., 2002). This holds Hsp90 in an "open" conformation in the intermediate complex, allowing later client loading (Roe et al., 2004). Although the release of Cdc37 C-terminus from Hsp90 N-terminal clamp is required for the transition of the "open" to the "closed", mature conformation, Cdc37 could stay in the complex by interacting with the client protein (Roe et al., 2004). Other cochaperones, such as p23 and Aha1, may be required as well (Wandinger et al., 2008). More details of kinase maturation in the complex remain to be understood.
Benzoquinone ansamycins, represented by geldanamycin (Figure 1) (Workman et al., 2007), were the first class of natural Hsp90 inhibitors to be discovered and substantially studied. Geldanamycin, a natural occurring antibiotic, was originally isolated from Streptomyces hygroscopicus as early as 1970s (Messaoudi et al., 2008). Structural and biochemical studies demonstrated that GA is a competitive inhibitor of ATP binding to Hsp90 (Roe et al., 1999). Binding of GA in the N-terminal ATP pocket restrains Hsp90 in its ADP-bound conformation and prevents the subsequent "clamping" of Hsp90 around a client protein (Blagg and Kerr, 2006; Neckers, 2006), resulting in ubiquitination and proteasomal degradation of the client (Mimnaugh et al., 1996). This N-terminal ATP pocket has distinctive characteristics in comparison with most other nucleotide-binding proteins (Prodromou et al., 1997), which explains the selectivity of GA.
Although GA exhibited potent anti-cancer activities in preclinical in vivo studies, it was determined to have little clinical potential mostly due to the high hepatotoxicity observed in animal models (Neckers et al., 1999). As a result, this has encouraged the search for GA derivatives that maintain similar anti-cancer activities but with better toxicological properties. 17-AAG (17-allylamino-17-desmethoxygeldanamycin; tanespimycin, KOS-953) (Fig.1) (Solit et al., 2007; Workman et al., 2007), and more recently 17-DMAG (17-dimethylaminoethylamino-17-demethoxygeldanamycin) (Fig.1) (Smith et al., 2005; Workman et al., 2007) and IPI-504 (17-allylamino-17-demethoxygeldanamycin hydroquinone hydrochloride) (Fig.1) (Sydor et al., 2006; Workman et al., 2007) were therefore synthesized for further evaluation (Pacey et al., 2006).
A considerable amount of understanding has been obtained from the clinical experience of 17-AAG (Workman et al., 2007). 17-AAG entered Phase I trials in 1999 (Banerji et al., 2005; Pacey et al., 2006) and several intravenous formulations have completed Phase I testing (Heath et al., 2005; Ronnen et al., 2006). Early signs of therapeutic activity have been seen in melanoma, breast cancer, prostate cancer, and multiple myeloma (da Rocha Dias et al., 2005; Grbovic et al., 2006; Modi et al., 2007; Pacey et al., 2006; Solit and Rosen, 2006). Phase II clinical trials for 17-AAG are currently ongoing (Heath et al., 2005; Ronnen et al., 2006), which mainly focus on tumor types hallmarked by specific Hsp90 chaperoning targets, such as leukemia expressing Bcr-Abl and Her-2 positive breast cancer (Dai and Whitesell, 2005; Pacey et al., 2006). 17-AAG was recently administered in combination with trastuzumab in patients whose disease progressed following trastuzumab treatment; this trial has demonstrated promising anti-tumor activity and acceptable toxicity (Modi et al., 2007). However, several drawbacks of 17-AAG, including low water-solubility, instability in solution, and low oral bioavailability (McCollum et al., 2008; Messaoudi et al., 2008) may become the obstacle to further clinical application.
In addition to 17-AAG, other GA derivatives have been developed for clinical use as well. Currently 17-DMAG, a more water-soluble analogue of 17-AAG, has entered Phase I and Phase II clinical testing, and displayed higher oral bioavailability, lower toxicity, and increased stability compared with 17-AAG (Hollingshead et al., 2005; Ronnen et al., 2006). Another water soluble hydroquinone hydrochloride analogue of 17-AAG is IPI-504 (Didelot et al., 2007). IPI-504 is in Phase I and Phase II clinical trials to evaluate its potential for treating cancer that has become resistant to therapy with tyrosine kinase inhibitors, such as Philadelphia chromosome-positive chronic myelogenous leukemia (CML) (Peng et al., 2007).
Radicicol (Fig. 1) (Workman et al., 2007) is a macrocyclic natural antibiotic initially isolated from the fungus Monocillium nordinii and Monosporium bonorden (Delmotte and Delmotte-Plaque, 1953). Like GA, radicicol was also shown to compete with nucleotide for N-terminal ATP pocket of Hsp90 (Roe et al., 1999). Similarly, this interaction between radicicol and Hsp90 constrains the chaperone in its ADP-bound conformation, leading to destabilization of Hsp90 client proteins (Mimnaugh et al., 1996). Radicicol displayed anti-cancer activity in vitro but not in vivo, which was explained by its chemical and metabolic instability (Pearl et al., 2008; Soga et al., 2003). Hence, synthetic efforts have been directed to generate radicicol derivatives with improved stability and in vivo efficacy (Proisy et al., 2006). Several oxime derivatives and cycloproparadicicol have been developed and shown to possess anti-tumor activity in preclinical animal models, as well as tolerable toxicity (Shiotsu et al., 2000; Soga et al., 2003; Yamamoto et al., 2003).
Novobiocin (Fig. 1) (Workman et al., 2007), a coumarin antibiotic isolated from Streptomyces species, was found to bind to Hsp90 at a newly suggested C-terminal ATP binding site with relatively weak activity (Marcu et al., 2000a). Inhibition of Hsp90 by novobiocin induced similar cellular responses as N-terminal inhibitors, i.e., destabilization of a range of Hsp90 client proteins such as Her-2, Raf-1 and p53 mutant via the ubiquitin-proteasome pathway (Allan et al., 2006; Marcu et al., 2000b; McConkey and Zhu, 2008). An allosteric regulation between the C-terminal and N-terminal domains of Hsp90 has been suggested, such that the interaction of ligands with one site might be affected by occupancy of the other site (Garnier et al., 2002; Marcu et al., 2000a; Marcu et al., 2000b). Two related coumarin antibiotics, chlorobiocin and coumermycin A1, also bind to the C-terminus of Hsp90 and have improved activity compared with novobiocin (Burlison and Blagg, 2006; Marcu et al., 2000b). Although a series of novobiocin analogues have been synthesized and screened for inhibitory activity against cancer cell proliferation (Burlison et al., 2006; Le Bras et al., 2007), currently available crystal structures have not yet confirmed the presence of such a second ATP binding site (Ali et al., 2006; Dollins et al., 2007; Shiau et al., 2006).
New natural product scaffolds are being discovered and tested. A recent example is the isoflavone derrubone (Fig. 1) from the Indian tree Derris robusta (Hadden et al., 2007). Derrubone was demonstrated to disrupt the interaction of Hsp90 and Cdc37 with heme-regulated eIF2a kinase (HRI), a Hsp90 client kinase, and exhibit antiproliferation activity in human breast cancer cell lines (Hadden et al., 2007). A green tea polyphenol catechin, epigallocatechin 3-gallate (EGCG) (Fig.1), was shown to inhibit the transcriptional activity of aryl hydrocarbon receptor (AhR) through a mechanism involving direct binding of EGCG to the C-terminus of Hsp90 (Palermo et al., 2005). It remains unclear whether EGCG could inhibit Hsp90 function through this direct binding. These findings may provide new natural product scaffolds to facilitate the development of novel Hsp90 inhibitors.
Thus, encouraging clinical responses have confirmed the potential of targeting Hsp90. However, binding of these ansamycin drugs not only prevents ATP binding but also induces a stress response through the release, activation, nuclear localization and trimerization of heat shock factor-1 (HSF-1) (Kaur and Ralhan, 2000), a transcription factor that binds heat shock elements (HSE) to increase the mRNA and protein levels of Hsp70 (Whitesell et al., 2003). This stress-responsive up-regulation of Hsp70 is believed to reduce the Hsp90-targeted drug efficacy by inhibiting apoptosis signaling (Kaur and Ralhan, 2000; Schmitt et al., 2006). Furthermore, these ansamycins are P-glycoprotein (Pgp) substrates (Tsuruo et al., 2003). Interestingly, a very recent study suggested that HSF-1-mediated stress induction, such as accumulation of Hsp70, may play a more important role in resistance to 17-AAG than drug efflux by Pgp, since depletion of Hsp70, but not P-gp inhibition increased the sensitivity of 17-AAG resistant cells to 17-AAG (McCollum et al., 2008b). These findings encourage the investigation of combining Hsp90 inhibitors, particularly of the ansamycin class, with abrogation of Hsp70 induction to enhance the clinical efficacy of Hsp90-targeted therapy (McCollum et al., 2008).
The search for natural Hsp90 inhibitors has been accompanied by a search for synthetic small molecule inhibitors that potentially possess more specific targeting and better pharmacological profiles. The availability of crystal structures of the Hsp90 N-domain and the development of structure-based design and high-throughput screening (HTS) assays have prompted successful exploitation of a range of new synthetic scaffolds that have Hsp90 inhibitory capacity (Huth et al., 2007; Messaoudi et al., 2008; Park et al., 2007).
The first group of these was empirically designed based on the purine scaffold (Figure 2) (Chiosis, 2006; Solit and Chiosis, 2008), which mimics the unique shape adopted by natural nucleotide ligand inside of the N-terminal pocket of Hsp90 (Chiosis, 2006; Chiosis et al., 2001). The favorable interaction between PU3 and the ATP pocket of Hsp90 was visualized in crystal structure, and consistently the similar biological effects of PU3 with geldanamycin were observed (Wright et al., 2004). PU3 thus became a starting point for the expansion and the improvement of clinically applicable purine-scaffold Hsp90 inhibitors, where efforts are directed toward either position C8 or C9 of purine (Chiosis, 2006). These synthetic derivatives could be roughly categorized to 8-benzyl (He et al., 2006; Vilenchik et al., 2004), 8-phenylsylfanyl (He et al., 2006; Llauger et al., 2005), 8-(7′-substituted benzothiazolothio) (Zhang et al., 2006), and 9-benzyl purine derivatives (Kasibhatla et al., 2007). Major improvements of purine-scaffold Hsp90 inhibitors include insensitivity to multi-drug resistance (Rodina et al., 2007), favorable water solubility, oral bioavailability, and metabolic stability (Biamonte et al., 2006; He et al., 2006; Kasibhatla et al., 2007; Zhang et al., 2006). For instance, PU-H71 and PU-DZ8 are currently under advanced preclinical investigation (Chiosis and Tao, 2006). CNF-2024, an orally available 9-benzyl purine derivative with low nanomolar potency, has entered Phase I clinical trials, tested in chronic lymphocytic leukemia, advanced solid tumors, lymphomas, and more recently in advanced breast cancer (Chiosis, 2006).
Pyrazole (Fig.2) (McDonald et al., 2006; Solit and Chiosis, 2008) is another important class of synthetic small molecules that has been identified (Cheung et al., 2005; Sharp et al., 2007a). A molecule of 3, 4-diaryl pyrazole (CCT018159) binds deeply into the N-terminal ATP pocket of Hsp90, as shown in crystallographic studies (Cheung et al., 2005). Similar with GA, a decrease in Hsp90 client protein levels (Raf-1 and Cdk4) and an increase in Hsp70 expression were observed in CCT018159-treated cancer cells (Cheung et al., 2005). Structure-based design generated more potent pyrazole amide CCT0129397 and isoxazole CCT0130024 (Sharp et al., 2007b); and an optimized analogue NVP-AUY922 has just entered clinical trials (Brough et al., 2008).
Finally, a novel family of short peptide derivatives composed of the three core amino acids, Phe-D-Trp-Leu, at the center of the molecule was recently reported to inhibit ATP binding to Hsp90 (Orosz et al., 2006). They killed various cancer cell lines, but not normal cells in vitro and did not show toxicity in vivo (Orosz et al., 2006). A few of these peptide derivatives were reported to cause a marked inhibition of multidrug resistance in human MDR1 gene-transfected mouse lymphoma cells (Molnar et al., 2007).
Hsp90 requires a series of co-chaperones to assemble a super-chaperone complex for its function. These co-chaperones bind and leave the complex at various stages to regulate the chaperoning process (Neckers, 2003). Arresting the chaperone cycle at these stages by targeting different cochaperone/Hsp90 interactions is likely to achieve similar consequences with the direct inhibition of Hsp90 (Gray et al., 2008; Zhang et al., 2008). Blockade of ATP binding seems to be the most direct and the simplest way to manipulate Hsp90, but its intrinsic non-selectivity for the Hsp90 clientele may limit its further application. Therefore, a potentially more specific approach is to develop drugs that block the interaction between Hsp90 and cochaperones (Pearl et al., 2008). Availability of crystal structures of co-chaperone/Hsp90 interactions plays a critical role in this process.
Cdc37 has a specialized and indispensable role in the maturation of a kinase sub-population of clients, including receptor tyrosine kinases such as epidermal growth factor receptor (EGFR), non-receptor tyrosine kinases Src and lymphocyte-specific protein tyrosine kinase (Lck), and intracellular serine/threonine kinases such as Raf-1and Cdk4 (Gray et al., 2008). Cdc37 acts as an adaptor, loading these kinases onto the Hsp90 complex, thereby facilitating their maturation (Pearl, 2005; Silverstein et al., 1998; Smith et al., 2008; Vaughan et al., 2006). Depletion of Cdc37 using RNAi in human colon cancer cells diminished the association of kinase clients with Hsp90 and decreased the levels of these clients including Her-2, Raf-1, Cdk4, and Akt, leading to reduced cell proliferation (Smith et al., 2008). Another noteworthy observation that silencing of Cdc37 did not induce Hsp70 up-regulation suggests that targeting Cdc37 might be advantageous compared with some Hsp90 inhibitors that induce Hsp70 accumulation (Smith et al., 2008). In addition, a recent study demonstrated that more extensive and sustained depletion of kinase clients and potentiated cell death could be achieved through combining Cdc37 silencing with the Hsp90 inhibitor 17-AAG (Smith et al., 2008).
A potential alternative strategy to direct Hsp90 inhibition was therefore brought up. The inhibition of Cdc37/Hsp90 interaction would probably represent an approach that may offer greater specificity and an improved side-effect profile (Gray et al., 2008; Gray et al., 2007). Indeed, our group has recently reported that celastrol (Figure 3A), a quinone methide triterpene from Tripterygium wilfordii Hook F, exhibits anti-pancreatic cancer activity both in vitro and in vivo through the disruption of Cdc37/Hsp90 association and the subsequent degradation of Hsp90 client proteins (Zhang et al., 2008). Celastrol was identified to block Cdc37 interaction with Hsp90 by molecular modeling (Zhang et al., 2008), taking advantage of the available crystal structure in which Cdc37 associates with the N-terminal domain of Hsp90 by inserting its C-terminal side chain into the mouth of the ATP binding pocket (Figure 3B) (Prodromou et al., 2000; Roe et al., 2004).
Assembly of Hsp70/Hsp90 complex is achieved by associations of their C-terminal tails with two independent TPR domains of Hop, TPR1 and TPR2A, respectively (Yi and Regan, 2008). These interactions, especially the one between TPR2A of Hop and the C-terminal MEEVD motif of Hsp90, are essential for function of this multi-molecular complex (Chen and Smith, 1998; Scheufler et al., 2000). Recently, a designed TPR module, CTPR390+, which binds to the Hsp90 C-terminus with higher affinity and more specificity than TPR2A, was shown to be capable of competing with endogenous TPR2A for Hsp90 binding and thereby to prevent the formation of Hsp70/Hsp90 complex, triggering Her-2 degradation with consequent inhibition of breast cancer cell proliferation (Cortajarena et al., 2008). Furthermore, a high-throughput AlphaScreen assay was developed to identify novel small molecules that hinder the Hsp90/TPR2A interaction and the identified compounds were shown to inhibit the growth of breast cancer cells, which was correlated with a reduced level of Her-2 (Yi and Regan, 2008). Remarkably, inhibition of Hsp90 through this way did not induce Hsp70 expression (Cortajarena et al., 2008; Yi and Regan, 2008). However, it is evident that these novel compounds will have modest selectivity for tumor versus non-tumor cells.
In addition, recent work revealed that whilst the primary binding site of Hop is the C-terminal MEEVD peptide of Hsp90, binding also occurs at additional sites in the C-terminal and middle domains of Hsp90 (Onuoha et al., 2008). The feasibility of targeting Hop/Hsp90 interaction still needs to be addressed.
According to crystallogical studies, the N-terminal domain of Aha1 interacts with the middle segment of Hsp90 and triggers the ATPase activity of Hsp90 (Meyer et al., 2004). Similar to Cdc37, Aha1 knockdown was recently reported to decrease the activation status of Hsp90 clients and a synergistic interaction was observed between Aha1 depletion and 17-AAG in inhibition of cancer cell growth, which indicates that modulation of Aha1 might be another potential therapeutic strategy (Holmes et al., 2008).
Inhibition of client/Hsp90 interactions offers the ultimate selectivity, but little is known about the molecular basis for these interactions (Pearl et al., 2008). The key to targeting the client/Hsp90 interaction is the ability to study the structure and biochemistry of the molecular complexes. An Hsp90/Cdc37/Cdk4 complex has been purified and its three dimensional structure has been determined by electron microscopy (Vaughan et al., 2006), providing the first structural view of the interaction between a client protein and Hsp90 (Ali et al., 2006; Pearl et al., 2008; Vaughan et al., 2006). However, most of the details of the client/chaperone interactions are still unclear, and therefore, the strategy of targeting these associations is more challenging.
Shepherdin, a peptidomimetic, was designed to specifically block the interaction between Hsp90 and the anti-apoptotic client survivin (Plescia et al., 2005). However, shepherdin can interact with the ATP pocket of Hsp90 as well, and affects a range of Hsp90 clients in addition to survivin (Pearl et al., 2008). Although shepherdin did exhibit anti-leukemia activity in animal models (Gyurkocza et al., 2006), its apparent interaction with the ATP pocket of Hsp90 and the effect on a range of Hsp90 clients suggests that it may have a different mode of action.
Post-translational modifications, such as hyperphorphorylation, S-nitrosylation and reversible hyperacetylation, have been thought to be involved in regulating chaperone function of Hsp90 through affecting co-chaperone association and/or ATP binding (Neckers, 2007). The post-translational modifications of Hsp90 may open up a wide range of opportunities to indirectly interfere with its activity (Wandinger et al., 2006).
Several studies have correlated histone deacetylases (HDACs) with Hsp90 chaperone function. HDACs, in concert with acetyltransferases (HATs), control reversible acetylation of lysine residues on Hsp90 (Minucci and Pelicci, 2006), which regulates Hsp90 activity. Lysine K294 in the middle domain of Hsp90 has been recently identified to be an important acetylation site (Kovacs et al., 2005; Scroggins et al., 2007). Identification of other acetylation sites and understanding of their functional significance will require more effort (Scroggins et al., 2007). A very recent work identified p300 as one of the HATs involved in acetylating Hsp90 (Yang et al., 2008). Hyperacetylation of Hsp90 has been reported after treatment with a variety of pan-HDAC inhibitors, such as LAQ824 and LBH589 (Bali et al., 2005; Kovacs et al., 2005). HDAC6, among the 18 HDAC family members identified so far, is unique in that it deacetylates non-histone proteins, such as Hsp90 and α-tubulin (Bali et al., 2005). Specific knockdown of HDAC6 using RNAi enhanced the degree of Hsp90 acetylation (Rao et al., 2008) and hyperacetylation of Hsp90 by either pan-HDAC inhibitors or HDAC6 knockdown was associated with a reduced binding of ATP and/or co-chaperones to Hsp90, thus promoting the degradation of Hsp90 client proteins (Rao et al., 2008). Also, HDAC1 may contribute to deacetylation of Hsp90 (Nishioka et al., 2008). MS-275, a novel synthetic selective HDAC1 (and not HDAC6) inhibitor (Minucci and Pelicci, 2006) induces apoptosis of acute myeloid leukemia (AML) cells expressing mutant fms-like tyrosine kinase 3 (FLT3) (Nishioka et al., 2008). MS-275, by inducing hyperacetylation of Hsp90 in these leukemia cells, inhibits the direct interaction between Hsp90 and FLT3, followed by its proteasome-dependent degradation (Nishioka et al., 2008). Overall, the hyperacetylation of Hsp90 was shown to inhibit binding of ATP, co-chaperone p23 and client proteins to Hsp90, directing the latter to polyubiquitylation and proteasomal degradation (Bali et al., 2005). Therefore, inhibition of either one of the two HDAC family members would probably be associated with hyperacetylation of Hsp90 and consequent impairment of Hsp90 chaperoning function. In addition, siRNA-mediated depletion of HDAC6 improved the affinity of 17-AAG for Hsp90 through enhancing the degree of Hsp90 acetylation (Rao et al., 2008). Moreover, co-treatment with Hsp90 inhibitors and HDAC inhibitors may produce synergistic effect, as suggested by the work on LBH589 and 17-AAG in CML and AML models (George et al., 2005).
A new type of inhibitor, tubocapsenolide A (TA) (Chen et al., 2008) was recently isolated from Tubocapsicum anomalum and shown to have potent anti-cancer activity in various human cancer cell lines (Hsieh et al., 2007). It was suggested that TA rapidly and selectively induced thiol oxidation of Hsp90 and Hsp70 respectively, thereby inhibiting the chaperone activity of the Hsp90/Hsp70 complex; indeed the TA-induced effects could be prevented by N-acetylcysteine, a thiol anti-oxidant. The TA activity was at least in part attributed to proteasomal degradation of Hsp90 clients such as Cdk4, cyclin D1, Raf-1, Akt and mutant p53 (Chen et al., 2008). This finding may open up another window for the targeting of Hsp90.
It is known that phosphorylation negatively regulates Hsp90 chaperoning function (Yang et al., 2008) and disruption of a phosphatase responsible for Hsp90 phosphorylation resulted in inhibition of Hsp90 function (Wandinger et al., 2006). However, the role of site-specific phosphorylation in modulating Hsp90 function has not yet been clarified (Yang et al., 2008).
The two major isoforms of Hsp90 in humans, Hsp90α and Hsp90β, encoded by two distinct genes, share approximately 81% sequence homology (Eustace et al., 2004; Passarino et al., 2003). And both contain the same three highly conserved domains (Pearl and Prodromou, 2006; Prodromou and Pearl, 2003; Soti et al., 2002). Still, it has become increasingly clear that Hsp90α and Hsp90β play different roles (Pearl et al., 2008). While Hsp90β is constitutively expressed at high abundance in most tissues, Hsp90α is typically inducible in response to various cellular stress conditions (Sreedhar et al., 2004).
Although the unique functions and underlying molecular basis of the isoforms remain to be elucidated, the distinct ability to activate some client proteins and the in vivo isoform selectivity of different inhibitors have been described (Chan et al., 2008; Millson et al., 2007). Activation of certain Hsp90 clients, such as v-Src, was more efficient with Hsp90α, rather than Hsp90β (Millson et al., 2007). Geldanamycin and radicicol were reported to interact with Hsp90α and Hsp90β with similar efficiency, whereas some of the purine-scaffold drugs, such as PU-H71, were more effective against Hsp90α (Chan et al., 2008). It is still unclear whether the isoform specificity of any Hsp90 inhibitor would be more beneficial in a particular cancer type (Chiosis, 2006).
Another cytoplasmic variant in mammals, Hsp90N, completely lacks the N-terminal ATPase domain and appears to be membrane associated (Grammatikakis et al., 2002). Higher eukaryotes possess a distinct endoplasmic-reticulum isoform, Grp94 (Ni and Lee, 2007) and a mitochondrial isoform, Trap1 (Felts et al., 2000). Neither clientele nor the biological role of these isoforms is currently known.
Over the past years, most attention has been given to the study of the intracellular Hsp90. However, a pool of Hsp90α has been described to be loosely attached to the cell membrane and facing the extracellular space (Thomaidou and Patsavoudi, 1993), which was associated with tumor cell invasion (Eustace et al., 2004; Sidera et al., 2008; Tsutsumi et al., 2008). A range of conditions, such as serum starvation, hypoxia, high concentration of glucose, as well as oxidative stress have been shown to trigger the extracellular localization of Hsp90α (Eustace and Jay, 2004; Li et al., 2007; Liao et al., 2000). It remains unknown how Hsp90α reaches the cell membrane and/or the extracellular environment (Sidera and Patsavoudi, 2008). One possible explanation is that Hsp90α might be secreted via an exosome pathway, as it was reported to be present in exosomes (Clayton et al., 2005). Although Hsp70, p23, Cdc37 and Hip are present in the extracellular space of tumors as well, their function there has not been explicated (Eustace and Jay, 2004; Shin et al., 2003). In addition, whether ATP binding and hydrolysis are required for an extracellular chaperone function of Hsp90α remains to be determined.
Cell surface Hsp90α was described to serve as a molecular chaperone of matrix metalloprotease 2 (MMP-2), an extracellular enzyme essential for cell invasion, to assist the maturation of this enzyme (Eustace et al., 2004; Passarino et al., 2003). Very recently, cell surface Hsp90α was also shown to specifically interact with the extracellular domain of Her-2 in vitro and in vivo (Citri et al., 2004; Sidera et al., 2008; Xu et al., 2001). This surface interaction is necessary for Her-2 activation and heregulin-induced heterodimerization with ErbB-3, which in turn mediates signal transduction pathways via MAPK and PI3K-Akt, leading to cytoskeletal actin re-arrangement essential for cell motility (Sidera et al., 2008; Sidera and Patsavoudi, 2008).
Inhibition of cell surface Hsp90α with antibodies or cell-impermeable inhibitors could block cell motility and invasion in vitro and metastasis in vivo (Tsutsumi and Neckers, 2007). A recent report identified a potent cell-impermeable Hsp90 inhibitor, DMAG-N-oxide, which is a polar derivative of 17-DMAG and currently in clinical trials (Tsutsumi et al., 2008). Although DMAG-N-oxide lacks the growth inhibitory ability of cell-permeable Hsp90 inhibitors, it displays significant anti-invasion activity in vitro and anti-metastasis activity in vivo (Tsutsumi et al., 2008). Moreover, it was described that the selective blockade of cell surface Hsp90α with a monoclonal antibody, mAb 4C5, could lead to specific disruption of the extracellular Hsp90α/Her-2 interaction, in turn preventing Her-2/ErbB-3 heterodimerization, actin reorganization, and consequent cell invasion (Sidera et al., 2008). Interestingly, disruption of the extracellular Hsp90α/Her-2 interaction did not lead to a reduced level of Her-2 protein on the cellular membrane; instead, it significantly reduced the phosphorylation of Her-2, suggesting that this extracellular interaction is different from the previously characterized intracellular interaction (Sidera and Patsavoudi, 2008). These findings suggest that blockade of cell surface Hsp90α may have clinical benefit in limiting cancer cell invasion and metastasis (Li et al., 2007; Sidera et al., 2004; Stellas et al., 2007).
Recently K69 was identified as one of the acetylated lysine residues on cell surface Hsp90α hyperacetylated by pan-HDAC inhibitor LBH589; and hyperacetylation actually stimulates not only the extracellular localization of Hsp90α, but also promotes its association with MMP-2, resulting in an increased tumor cell invasion (Yang et al., 2008). This study further demonstrated that exposure of breast cancer cells to a novel antibody against the acetyl-K69 Hsp90α could markedly inhibit in vitro invasion compared with the inhibitory effect of an anti-Hsp90α antibody that non-specifically recognizes both acetylated and unacetylated Hsp90α (Yang et al., 2008).
Taken together, these intriguing findings raise the possibility that cell surface Hsp90α plays an important role in modulating cancer cell invasion and metastasis that is independent of the intracellular Hsp90 function, providing a novel extracellular drug target for metastatic cancer therapy (Sidera and Patsavoudi, 2008; Tsutsumi et al., 2008).
It has become increasingly evident that cancer probably is initiated from and maintained by a small sub-population of undifferentiated, tumorigenic cells called cancer stem cells (CSCs) (Gonzalez-Sarmiento and Perez-Losada, 2008). Production of the main mass of the tumor may be attributed to this minor population of CSCs through a particular process of continuous self-renewal and differentiation (Gonzalez-Sarmiento and Perez-Losada, 2008). Thus, CSCs have come into sight as a potential target of cancer therapy (Hassane et al., 2008; Rottenberg and Jonkers, 2008).
Based on an in silico screen of public gene expression data, celastrol, a novel Hsp90 inhibitor reported by us (Zhang et al., 2008), was recently discovered to eradicate acute myelogenous leukemia stem cells through simultaneous inhibition of NF-κB-mediated survival signals and induction of oxidative stress (Hassane et al., 2008). A number of studies have documented a correlation between Hsp90 and NF-κB. Cdc37 and Hsp90 together are required for constitutive and inducible activation and nuclear localization of NF-κB (Broemer et al., 2004; Chen et al., 2002) and the expression of the Hsp90α gene was regulated by NF-κB (Ammirante et al., 2008). Therefore, a regulation loop is likely to exist between Hsp90 and NF-κB. Moreover, overexpression of Her-2, an Hsp90 client protein, was shown to increase the breast cancer stem cell population both in vitro and in vivo (Korkaya et al., 2008). The Wnt/β-catenin pathway is one of the pathways with a critical role in CSCs (Malanchi et al., 2008; Reya and Clevers, 2005) and β-Catenin has been suggested to be regulated by IKK that has been shown to be controlled by Hsp90, connecting the Wnt pathway to Hsp90 (Lamberti et al., 2001).
The putative spheroid-forming breast CSCs usually display resistance to chemotherapeutic drugs (Wright et al., 2008). Surprisingly, a synergistic effect was observed when breast CSCs were exposed to the Hsp90 inhibitor 17-DMAG simultaneously or after drugs such as doxorubicin, cisplatin or etoposide (Wright et al., 2008). Also, the Hsp90 inhibitor IPI-504 was shown to induce a dramatic suppression of leukemia stem cells in mice with Bcl-Abl-T315I-induced CML; in a sharp contrast, analysis of bone marrow from wild-type mice treated with IPI-504 showed that this drug did not affect normal hematopoietic stem cells (HSCs) (Peng et al., 2007). Finally, a growth inhibitory effect and degradation of Hsp90 client proteins including EGFR, Akt and MAPK by 17-AAG in a glioma stem cells in vitro was reported as well as reduction of the tumor-initiating capability of these cells in an orthotopic glioma model (Sauvageot et al., 2008). These intriguing findings may implicate an association between Hsp90 and CSCs properties through one or more complicated pathway(s), which merit further investigation.
The most attractive advantage of targeting Hsp90 is the combined impact on many oncogenic pathways involved in multiple steps of carcinogenesis and cancer progression, as Hsp90 inhibition eventually leads to the ubiquitin-proteasome degradation of a large population of oncogenic client proteins (Workman, 2004). This review not only provides an up-to-date overview of mechanistic studies and the clinical prospect of currently available Hsp90 inhibitors, but also wishes to enhance the perspective both for designing and discovering novel inhibitors as well as to provide insight into less-understood and under-represented potential of Hsp90 in cancer therapy (illustrated in Figure 4).
Most studies have focused on the discovery or synthesis of Hsp90 inhibitors specifically acting against the ATP binding site at the N-terminus of Hsp90. Further evaluation of these Hsp90 inhibitors and development of new inhibitors belonging to this class remains of great interest. However, there are considerable possibilities for inhibition of the super-chaperone system in other ways. Targeting co-chaperone/Hsp90 interactions should receive more attention, as this approach is likely to improve the specificity of Hsp90 inhibition compared with direct inhibition of ATP binding. Another appealing advance is derived from the interference with post-translational modifications of Hsp90, where a majority of the efforts have been put into the hyperacetylation of Hsp90 by HDAC inhibitors. Disruption of client/Hsp90 interactions was suggested to provide the highest selectivity (Pearl et al., 2008). While the basis for antagonizing client/Hsp90 associations is the structural and biochemical understanding of the interactions, the reality is that even nowadays little is known about these interactions (Pearl et al., 2008). Besides, the initially-raised benefit of Hsp90 targeting is the simultaneous impact on multiple oncogenic pathways, which could be eliminated by only focusing on a specific client/Hsp90 interaction. Therefore, the strategy of targeting these client/Hsp90 associations is not only highly challenging, but also need to be further validated. A few studies have been performed to compare the properties and functions of distinct Hsp90 isoforms, but most details remain to be understood. The different functions of Hsp90 isoforms and the isoform selectivity of Hsp90 inhibitors will require further investigation. The inhibition of cancer metastasis by cell-impermeable Hsp90 inhibitors is an interesting area for research as well as specific targeting of cancer stem cells. Since the importance of CSCs in resistance to chemotherapy has been recognized, the evaluation the effect of Hsp90 inhibitors on CSCs properties would provide useful insight and clinical perspective for the future.
Since dose-limiting toxicity is likely to be an important issue when Hsp90 inhibitors are used as single agents, an alternative approach for the application of Hsp90 inhibition is to combine Hsp90 inhibitors with other therapeutic agents to enhance the efficacy with lowered dose-limiting toxicity. This perspective is substantiated by a large number of preclinical and clinical evaluations of Hsp90 inhibitors that have shown promising results in combination with chemotherapeutic drugs or other agents (Solit and Chiosis, 2008). 17-AAG and 17-DMAG are being investigated in preclinical, Phase I and II trials for breast cancers in association with trastuzumab, a Her-2 monoclonal antibody (Didelot et al., 2007). For example, a Phase I dose-escalation study has indicated that co-administration of trastuzumab and 17-AAG is well tolerated and exhibit anti-tumor activity in patients with trastuzumab-refractory Her-2 positive breast cancer (Modi et al., 2007). A recent Phase I study with 17-AAG plus paclitaxel was conducted in patients with advanced solid malignancies to determine the recommended Phase II dose of these two drugs (Ramalingam et al., 2008). Another Phase I dose-escalation trial using 17-AAG and irinotecan suggested that 17-AAG potentiated the anti-proliferative activity of irinotecan possibly by depleting checkpoint kinase (Chk1) (Tse et al., 2008). Co-administration of 17-AAG and proteasome inhibitors such as bortezomib has also been reported to produce synergistic anti-cancer effects, which might result from the concomitant increase in protein misfolding and impairment of proteasome-dependent clearance (Whitesell and Lindquist, 2005). IPI-504 was demonstrated to be more effective when combined with imatinib than when used alone in prolonging survival of mice with Bcr-Abl positive leukemia cells (Peng et al., 2007). A recent report indicated cisplatin-mediated abrogation of the heat shock response through inhibition of HSF-1 activity that may, at least in part, contribute to the synergistic effect of GA and cisplatin (McCollum et al., 2008a). An interesting observation by our group is that an Hsp90 inhibitor and glycolysis inhibitor synergistically inhibited tumor growth in a pancreatic tumor model, possibly by preferentially targeting hypoxic tumor cells (Cao et al., 2008).
In summary, the targeting of Hsp90 for anti-cancer therapeutics has a potentially bright future. Further progress in the development of Hsp90 inhibitors and a deeper understanding of the Hsp90 characteristics further strengthen its promise in cancer therapy.
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"Staff should be more inquisitive regarding to the..."
About: Whipps Cross University Hospital Whipps Cross University Hospital E11 1NR
Posted by Anonymous
I want to the A&E for a Chest problem that is affecting me for quite a long time and as a smoker I am very worried about my health, I explained that to the person at the front desk and they said they won't allow me to see a Doctor because it's not Urgent and I have to go and see my GP book an appointment with him, it is in 2 weeks time, I couldn't wait for so long, so I went to France and They did the necessary tests on the same day (blood pressure, blood and urine test). Result chest pain angina.
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Press committee honoring 4 journalists
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I very dissatisfied with the the changes to the mobile app. They are not intuitive, don't allow you to easily go back and forth between screens and don't seem have to addressed the trouble with the vocabulary lists. Still very buggy. :(
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Give Books / Chronicle Giveaway
Last week I got an early Christmas present from my friends at Chronicle Books. They're starting a holiday campaign called #GiveBooks, and asked if I wanted to pick myself any 10 books from their archives! Lucky me right? Well, lucky you too because I also get to send all 10 books to one of you! How does that sound?
Here are my picks:
"David Choe" by David Choe
Tartine: The Boxed Set
"Imprint" by Daniel Eatock
Masters of Fashion Illustration
"Rift" by James Jean
"Map As Art" by Katharine Harmon
Saul Bass – A Life in Film and Design
100 Years of Fashion Illustration
"Seventy-nine Short Essays on Design" by Michael Bierut
So who wants 'em?


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LUXEMBOURG (Reuters) - German Finance Minister Wolfgang Schaeuble said on Thursday he was confident that euro zone ministers would agree on a next disbursement of loans to Greece and was hopeful the International Monetary Fund would be on board.
"For Greece, as before, I am confident that we will reach a deal about the payment of the next tranche today," Schaeuble said on arrival at the meeting.
"We have found a way which hopefully will allow the IMF management to propose a programme to its board," he continued, adding that he did not believe it amounted to a substantial change of the programme.
Germany's parliament has insisted on IMF participation.
"We cautiously believe that the goal of the programme will be reached and that Greece will again have access to the finance markets at the end of the programme," Schaeuble said.
Reporting By Philip Blenkinsop


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