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STARTING Aug. 27th – some fun! Want a challenge? Want to go to our September event, but need an extra ticket? We're having a scavenger hunt and you can win tickets! All you need to do is look on our website and find (at least) three salmon (it's a fish image). We've added the salmon images Sunday, August 27th and if you find three (3) salmon (images) just tell us where you found them on our website by posting on Facebook or Instagram or twitter and you're in the running to win a single ticket! AND if you find ALL eight (8) images you're in the running for 2 free tickets! OK, ready, set get looking & remember, this special challenge for tickets starts August 27, 2017 at noon and ends September 10th, so don't wait too long to enter! What a great gift for a friend to come together to this great Salmon Bake fundraiser, Sept. 23, 2017, and it supports the Washington County Kids. #WashCoKids
Eclipse Reset on what's important
When you experience something like a total eclipse of the sun, something that will not happen (the way it did) for another 100 years, you often have the opportunity for a "reset" or re-grounding on life. The word "totality" itself is something to consider. Are we living in that zone? Are we doing what we want to be doing. Are we doing what's important? As we ruminate what's important steps ahead into the forefront and what's extraneous falls away.
What's important to you? What's important to Washington County Kids (org) is to work as hard as we can to ensure that kids are fed, have the opportunity to learn, play and grow and to succeed. We support summer learning, food and nutrition, early learning, and before and after school programs by supporting and expanding the programs that do the most good. Some programs are doing an awesome job, but they're serving 20 kids when there are 200 waiting.
We hope you will consider attending the Washington County Kids fundraiser special event – a Salmon Bake – on September 23, 2017 at the Helvetia Winery. What a wonderful beautiful rustic setting. But, what's the event for? What does it mean? It's more than just food and fun (although there will be great food, music, and friends). It's about celebrating the success of working together in the community and of making a difference in our own backyard. Many of our partners on the mission to do good will be there. We hope you will be too.
Summer hunger – libraries fill the gap, NYT's article
We thought you might want to see this article on how libraries are filling the gap on hunger by feeding kids who would normally be fed at school. Because it's summer there are no meals. Check it out – worth reading.
Should kids go hungry just because it's summer? Washington County Kids works hard to help make sure that kids are fed all year.
We sent the invites – did you get one?
We sent the invites this morning! Did you get yours? If you didn't receive an invite to the Salmon Bake Fundraiser for Washington County Kids, please let us know! We are hard working volunteers, but maybe we missed you! You can drop into our FB or Twitter page and tell us! Or email email@example.com and let her know. We are SO excited about this event. It is a lovely venue, out in the vineyards and under the great trees. The food will be superb and there's music, a silent auction, award winning wines. Can't wait! Here's a link to tickets. #WashCoKids (that's our hashtag:)
***Oh, and here's a flyer invitation as a PDF (almost forgot!)*** WashingtonCountyKids_SalmonBake_FLIERinvite
WCK Salmon Bake Fundraiser Announced!
Support The Washington County Kids – SEPT. 23, a great afternoon and early evening of FUNdraising: Savor salmon, toast the best of Washington County's success stories (supporting kids & families) at Helvetia Winery and meet the folks who make a difference in Washington County all while supporting WCK. Sample award winning wines and tuck into an extravagant outdoor salmon buffet authentically prepared by a Native American family. There'll be a silent auction and live music to enjoy under giant trees in a beautiful rustic setting.
Tickets are on sale now! Follow us on FB, and Twitter to stay in touch! Looking forward to seeing you there!
***link to downloadable PDF invitation***
July WCK e-newsletter – 07/17
The July e-News is out! Enjoy the Washington County Kids (WCK) e-newsletter online. And be sure to let us know if you'd like to be on the mailing list. Enjoy!
Fall Fundraiser date: Sept 23
Mark your calendars for September 23, 2017 – Washington County Kids announces a special event gathering. We hope you'll join us! Celebrate a year of good works and continue the conversation about how to best support kids and families in Washington County. Join WCK's celebration (details coming soon!) and enjoy good food, good company, a silent auction (with art, wine and more), a delightful evening with sumptuous food, music and much, much more! It's all for the kids.
Yay – the first donation was received today for the silent auction – ooh, a large photograph. Here's a mini version of it.
If you have something to donate, please reach out to Rachel G. Thank you!
Partner Profile: Meet Rachel Parker
Melissa Habit, a recent (wonderful!) WCK volunteer, did a great story based on an interview with the amazing Rachel Parker of the Boys and Girls Club. Have you seen it? It's in the Washington County Kids' eNews this month, but here's a quick link to read it.
UPDATED: 09/28/17 – Congrats and best wishes to Rachel Schultz on her recent marriage and promotion!
New WCK website
If you're on this page you know we have a new Washington County Kids (WCK) website. (Yay!) Launched over the last few days, it's still us, but we're moving into a format that allows us to tell the stories in a better way, with more pictures and details of kids and families that need your help. Tell us how you like it! And today we'll release WCK's e-newsletter with lots of news plus a wonderful "Partner Profile" by our amazing WCK volunteer, Melissa Habit interviewing equally amazing E. D. Rachel Parker . Here's a peek. Watch for it!
Fall fundraiser – Dinner announced by WCK
For those who love the fall and its opportunity to regroup – we have the perfect venue! It's the Helvetia Winery and (rumor has it) a salmon bake fundraiser complete with live music, awesome food, a silent auction and more! Save the date, will you? Oh, Sept. 23rd! We're looking forward to seeing you. More coming!


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The Durango was built by Durango Systems, Inc in San Jose, CA. It came with a 8085 processor running at 5 MHz, 64K memory as standard and could be expanded to 128K in the multiuser version.
The F-85 was marketed as a portable computer with integrated 180 cps dot matrix printer, two floppy disc drives and a 9" monitor. Well, only very strong users could carry it ;-)
The Durango ran a proprietary operating system, DX-85, as well as CPM. DX-85 had multiuser extensions and the business applications were supported by a proprietary ISAM handler. In addition to the system user, 4 users could hook up via video terminals on serial connections. These users could run programs in their allocted memory space on the F-85. Some interprocess communications were available.
More information from Chuck Guzis who was involved in the development of the Durango operating system:
� The basic system came with 64K of parity-checked DRAM; an optional 64K
or 128K of parity-checked DRAM was optional. When the additional memory
was added, a bipolar mapping RAM was also included that divided the 64K
memory space of the 8085 into 64 1K pages. Any page of physical memory
could be mapped to one or more page locations.
� Only an IEC power connector and 9 pin DIN CRT connector and 37-pin DIN
external floppy connector were present on the basic unit. However, options
included an IEEE-488 interface, 4-port serial interface and
async/sync/bisync serial communications, so the number of external
connectors could vary.
� The standard floppy configuration was 2 100 tpi 5.25" group-coded
floppy drives, each storing 980KB on a DSDD floppy. The data encoding was
Durango-proprietary. Early systems interfaced to the then-new Shugart 14
inch Winchester drive via a modified IEEE-488 protocol. Capacity of these
drives varied from 7 to 40Mb. Later systems replaced one of the floppy
drives with a 5.25" hard drive of between 7 and 40MB. An optional 2-drive
floppy external unit was available as an option; the F-85 could support up
to a maximum of 4 floppy drives.
� In addition to the extensions mentioned above, the integrated
dot-matrix printer could be upgraded from a single-pass 2 monospaced font
mode to a multipass high-quality print mode with downloadable font sets.
In this case, the printer ribbon was changed to mylar film instead of linen
and the carriage drive, printhead and electronics were upgraded. The
printer could feed standard perforated forms or be switched to a friction
feed for printing on standard stationery.
� The CRT was a 9 inch unit that was placed on top of the floppy drive part
of the system.
� The standard operating system was called DX-85M, a multi-user multitasking
system, which interfaced to up to 4 additional users by means of terminals
(made by Beehive) connected to the serial multicomm interface. It included
most standard file management capabilites as well as an integrated ISAM
file system. It was not necessary to regenerate the system when the
hardware configuration changed; the necessary extensions were loaded at
boot time.
� There were two busses used, one in the main system card cage, which was
loosely based on the Intel Multibus and one used in the smaller auxiliary
card cage located behind the floppy drives, which was a proprietary 100-pin
� The standard programming language was a compiled version of BASIC called
Star-Basic. Almost all applications and utilities were written in this
language, which compiled source code to byte codes.
� CP/M 2.2 was available as an option. There was a project to include MP/M 2.0, but was discontinued when Durango began work on its Unix-based Poppy
16-bit system.
� A standard set of business applications was available including GL, AP, AR,
Inventory, Payroll, Word Processing and a spreadsheet.
� Durango's philosophy was to provide the customer with a complete system
with all needed applications and customer support included.
� Around 1980, a marketing change was made and the F-85 was re-named the 800; a cost-reduced version with a slower printer (really just a different set of firmware) was called the 700 and the multitasking model with extra
memory and high-resolution printer was called the 900. In fact, any model
could be changed to any other in the field.
� Sometime in 1982, Durango merged (or was purchased by) Molecular Systems,
which subsequently became insolvent and was dissolved in 1984. At that
time, Durango was attempting to market its 16-bit 80186/80286
dual-processor Unix/MS-DOS system, but could not compete against the new
IBM-compatible PCs that had been introduced.
I was involved in the development of the operating system; I wrote the ISAM
file manager and the floppy disk drivers. I was in charge of the Star
Basic language and was responsible for its design and implementation. I
also designed the bank-switching mechanism for the add-on memory. I also
did the CBIOS adaptation for CP/M 2.2.

subset: commoncrawl


text:
Content Count192
Donations195.00 USD
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About wrcbonk
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- Birthday 04/22/1972
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Roseville, CA
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The making of Zilla Z
wrcbonk replied to Arif's topic in Nissan RB ForumBeautiful engine bay, looking forward to updates.
Fuel swirl pot
wrcbonk replied to Fairlineguy's topic in Nissan RB ForumAs Neverdone and Seattlejester indicated there are pros and cons to each setup. I went with an external system. It has been working without issue for 4+ years in my RB25DET/S30. There is enough room back there to mount everything and I used the existing factory wiring (hot Green wire and Black chassis ground). Both the low pressure supply clicker style pump and the Walbro EFI pump work great and don't get hot. I have never had the fuel cut in a corner (street use only). The Walbro fuel pump whines under normal operating conditions. My understanding is that the Bosch is more expensive and quieter, spend the money. I am using the small BC Broncos swirl pot which doubles as a filter. Internal pumps are industry standard because the fuel cools the pump, but you must modify your tank. No problem if you know how to modify a thin steel tank, go for it (and many people on this site have those skills), but imagining you will find someone or a shop to modify your fuel tank maybe wishful thinking. Regardless you must have a properly sized return line to keep everything in balance. I replaced the 1/4-inch line with a 5/16ths and used the upper 3/8ths vent port on the tank to return the fuel. TonyD points out you can use the fuel vent line (5/16ths or 3/8ths) as your return that runs from the from the back near the tank to the engine bay, as long as you are willing to reconfigure your fuel vapor system after that.
Enjukuracing oil pan
wrcbonk replied to JamesRB25280zx's topic in Nissan RB ForumOk, I will fall for this, how about pm Pat1...
inner tie rod question
wrcbonk replied to zombie-nsk's topic in Brakes, Wheels, Suspension and ChassisRead the Amazon reviews for more information on how to install the Rare Parts inner tie rods. https://www.amazon.com/Rare-Parts-RP26575-Tie-Rod/dp/B000CK8XTS
Fuel Tank & Evap Design
wrcbonk replied to NylonAdmiral's topic in S30 Series - 240z, 260z, 280zI had the same concerns. I traded a few emails with Tony D and came up with the following. I kept 13 plumbed from the tank to the two way valve in the engine bay. The blow part of the valve (the end where excess fuel vapors are expelled from the tank) is plumbed to my intake. The Suck end of the valve is plumbed to a small air filter (a low pressure fuel filter I had laying around, whatever it does the job) ensuring no particulates get into my fuel system. In the past I had issues with fuel tank vapor pressure forcing fuel out my filler causing havoc. I have had no issues with the present plumbing and I live in HOT Sacramento. Bonk 2 cents
Car stalls under Hard Acceleration / Boost
wrcbonk replied to Klmykvl's topic in Nissan RB ForumNot sure what kinds of boost level you are running with the stock ECU. I have hit Fuel Cut several times running the stock ECU with too much boost pressure ~13 lbs or more). It's Nissan's way of saving the engine. I can't afford the engine management computer I want at the moment so I run modest boost pressure, ~ 9lbs, and use Royal Purple Octane Booster if I want to rev it out. The stock ECU timing table is set for 97 octane, it pulls timing and dumps fuel when the knocks start coming from fuel starvation or inadequate octane. my 2 cents
~73 Turbo L28 to N/A RB25 240z Project~
wrcbonk replied to ~KnuckleDuster~'s topic in S30 Series - 240z, 260z, 280zKnuckleDuster, You find the best stuff, that angled shifter would save me a lot of headache, modify my stock unit. Didn't see it in the Nismo shop online. Do you have a part number? Thanks Bonk
Help with rb20det swap into 76 280z
wrcbonk replied to claymann's topic in Nissan RB ForumIf your engine comes with a harness in good shape making the connections is straight forward. It will save you a big chunk of change. Remember you are only talking about the engine harness, not the whole car. You are mainly just giving the harness power and ground for ignition and fuel. The key is to go one circuit at a time and use quality electrical supplies (solder, heat shrink, good wire, good relays (Bosch)). The only reason I bought a harness from WS was because the harness that came with the engine was trashed and I made it worse being a dumbas@. 2 cents
Swami's Orange 260z
wrcbonk replied to Swami's topic in S30 Series - 240z, 260z, 280zCame back around to this thread after losing touch. The sterileness of your engine bay is amazing, especially the electrical. Look forward to more updates. Bill
~73 Turbo L28 to N/A RB25 240z Project~
wrcbonk replied to ~KnuckleDuster~'s topic in S30 Series - 240z, 260z, 280zI was admiring your glovebox gauge holder. Do you remember where you got it? Thanks
My custom fuel setup
wrcbonk replied to 240zdan's topic in Fuel DeliveryThat SS strap is very nice, cool fuel delivery setup.
RB CAS issues with Haltech (how to bench test?)
wrcbonk replied to grenadesandham's topic in Nissan RB ForumThis is of interest to me because I was planning to buy the Haltech Platinum Pro ECU 55001 for my Series 1 R33. Is the 550001 the model you have? Do you have the stock ecu? You could try to use the stock computer to get the car running. I was able to start my engine first crank using the stock ECU (that is after we fixed all the fuel leaks that started as soon as I charged the pump). Have you emailed Haltech yet? I emailed them in the past and It took a few days, but they got back to me with good information. Will be watching your progress. Bill
EMS or Coils?
wrcbonk replied to wrcbonk's topic in Nissan RB ForumThanks guys, I will look into testing my coolant temp sensor. Doing some other research I found that another benefit of EMS is adjustment of spark dwell, which can be optimized for the LS coils.
EMS or Coils?
wrcbonk posted a topic in Nissan RB ForumAfter reading the thread Coil Suggestions I have some follow up questions for the RB group, but didn't want to hijack Blob1620's thread. I have a very stock Series 1 rb25det swap in my s30: stock ecu, STOCK COILS, throttle body, no downpipe, 2.5-inch exhaust, freddy plenum, the intercooler is probably 50% bigger than stock, stock fuel rail, injectors, and FPR. I am currently running the boost at the stock max of 7-8 psi through a manual controller and I have a wideband. Naturally, my engine stubbles from @4500 to 6000 where AFRs plummet to 10 and the engine runs roughly. Common wisdom (Hybridz and SAU) states the ecu is retarding timing and adding fuel to preserve the engine from knock as US fuel is lower in octane. This is not a new story to the group. There are numerous changes that can be made to add power and I intend to start making those changes, but my first priority is to smooth out my rev band. I am more interested in smooth running than power at the moment. My question is this. Should my first step be to obtain a programmable ems allowing me to tune for US fuels or is replacing the coils more important? Put another way, is the rough running more a function of poor ignition performance or the fact that the fuel and timing maps are locked and meant for higher octane fuel? Thanks Bill
Golden RB 240z
wrcbonk replied to rk2334's topic in S30 Series - 240z, 260z, 280zCool project, a true hybrid, can't wait for more updates and eventual tuning.


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Busy Girl Bakes: Banana Bread
Banana bread is such an easy way to satisfy a cake craving – plus it contains fruit, so is clearly one of our five-a-day, right?! What we love about this cake is that it's pretty much throwing everything together, mixing it up and then baking to perfection. This Mary Berry recipe has a final finishing flourish of a drizzled honey and sugar topping – delicious.
Perfect for using up those bananas at the bottom of the fruit bowl, banana bread is an ideal weekend snack or energy boosting work treat to beat the 4pm slump. Give it a go!
This banana bread – technically called Banana and Honey Teabread – is from Mary Berry's Baking Bible, our current go-to cookbook for delicious teatime treats from the queen of cosy baking.
Easy Banana Bread Recipe
- 225g self-raising flour
- ¼ level teaspoon freshly grated nutmeg
- 100g butter
- 225g bananas (approx. 2 medium bananas)
- 100g caster sugar
- Grated rind of 1 lemon
- 2 large eggs
- 6 tablespoons thick pale honey
For the topping
- 2 tablespoons honey
- Crumbly sugar for sprinkling (we used demerara)
- Pre-heat your oven to Gas 3/160C (fan 140C). Grease a 2lb loaf tin and then line the base with baking parchment.
- Measure the flour and nutmeg into a large mixing bowl. Rub in the butter using your fingertips until it looks like fine breadcrumbs.
- Peel and mash the bananas. Stir them into the flour mixture, along with the sugar, lemon rind, eggs and honey.
- Beat the mixture well then pour into your prepared tin and make sure the surface is level.
- Bake for around 1hr 15mins – until a skewer into the centre of the cake comes out clean. (If it's browning during the end of cooking, cover loosely with tin foil.)
- Leave to cool in the tin for about 5 mins then turn out and peel the parchment off the base. Leave to cool on a wire rack.
- Make the topping: warm the honey in a small pan or microwave and brush over the top of the cooled banana bread. Sprinkle with the sugar.
- Eat and enjoy!
That's it – all that's left to do is tuck in!
Get baking
Trying out this banana bread recipe? Share a pic of your finished cake with us – @tweetjanes.


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Home / Punch Out
Mike Tyson Tries to Beat Himself in Punch-Out!!
Mike Tyson playing Mike Tyson's Punch out is a surreal moment. Sure, Jimmy Fallon is far from original (this bit… Read More »
People just don't be knowin
They don't! I've been saying Iron Mike Tyson is the greatest HW Boxer of my generation. Is he as prolific as Ali?… Read More »


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Please login with Facebook to see your result
This quiz is using FaceApp's advanced technology - find it in the app store!
What creature do you become when you're hangry? Find out today!
Show more
No one likes being hungry, that's just a fact, but some of us don't deal with it very well at all. We become our worst possible selves! If you can relate, continue now!


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'Buffet Rule' fosters shared sacrifice
Despite being labeled as a hypocrite by some a few weeks back because of Berkshire Hathaway's failure to pay its taxes, Warren Buffet's reputation as a millionaire who loves paying taxes lives on strong in President Barack Obama's newly proposed "Buffet Rule." According to talking points obtained by the Huffington Post, the "Buffet Rule" is a simple principle which states, "No household making more than $1 million annually should pay a smaller share of its income in taxes than middle-class families pay." For the fair-minded, it's hard to argue against the "Buffet Rule." It is the most practical manifestation to date of the "shared sacrifice" narrative Democrats and Republicans alike have been espousing since the recession of 2008.
For too long now, the burden of our country's debt fell on the shoulders of the middle class and the poor, who have seen higher taxes and deflating public services. For a long time, it looked like this was going to continue being the case, despite the loud protestations of people like Buffet and Netflix CEO Reed Hastings, two millionaires who practically publicly begged Obama to raise their taxes in order to help with America's growing debt. People like Buffet and Hastings were some of the only public figures actually trying to make shared sacrifice a reality, even though it seemed like their efforts were falling on deaf ears.
Which is why it is so great to see that Obama actually took note of such agitation. Under the "Buffet Rule," millionaires will no longer be taxed at absurdly minuscule rates. Instead, they will finally start helping the middle class and poor shoulder the burden they have been sagging under for so long. The United States is the country that gave these millionaires and billionaires the opportunities to succeed and become rich. It is only fair they start giving back.
We are sure cries of socialism will resound from Obama's detractors once he brings the "Buffet Rule" to them. However, this is not socialism — it is a request for the rich to do what is right for the country that made them. Too many people are suffering because of American debt. It's time that suffering end. The rich should be excited to help right our capsizing economic boat. The United States is a democracy, which means that we, the people, are all in this together. A high economic class should not be reason to shirk your responsibility to this country.
Please note All comments are eligible for publication in The Daily Targum.


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by S.W. Basics
(Product code: PC0172)
Beautiful results for...
This S.W. Basics Exfoliant is pure kitchen-cabinet awesomeness. They clever peeps in Brooklyn take organic oats and almonds, grind them until they're not too rough but not too soft, add mineral-rich sea salt harvested from coasts nearby, and voila: a simple, nourishing, super-effective exfoliant that leaves your skin soft and glowing
More info
How it works
Made with just three ingredients is gentle, yet oh so effective. The organic oat flour, organic almond flour and sea salt work together to gently work away any excess on the skin, getting right down into pores without causing irritation, leaving you with lovely glowed skin.How to use it
How to use it
Blend a small amount with water or pour into wet hands. Gently scrub onto face, using small circular motions. Leave on for 1-5 minutes, rinse with warm water.
What's inside
- Maine sea salt
- Organic oats
- Organic almonds
Beauty duty
S.W. Basics support small, family farms in a bid to create the most effective products in the most responsible way - we salute!
- Maine sea salt
- Organic oats
- Organic almonds


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Hi, it looks like you're new. Welcome to Voat!
Voat is a censorship-free community platform where content is submitted, organized, moderated and voted on (ranked) by the users.
Users can earn a percentage of our ad-revenue share for the content they submit.Take a tour
Archived Deep State is in a Panic Fearing Putin Shared with President Trump Devastating Information Not Yet Reported (thegatewaypundit.com)
submitted ago by Scrooblemeyer
[–] 0fsgivin 3 points 2 points 5 points (+5|-3) ago (edited ago)
That's the thing though...
There is already plenty to put the Clintons in prison. Putin isn't going to offer anything new or insightful.
In fact if he does...it will be less credible than what we already know simply because of the source.
No Clinton is going to jail any time soon... This shit is just LARP bullshit.
A dupont raped his own daughter...and got house arrest. Clinton ain't going to prison.


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International shipping Secure payment
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One of the most popular tapes on the market and designed especially for lace based system. Although the tape has a blue colour to it, when applied it goes on clear and has a nice matt finish.
Pro Flex II tape has been on the market for yesrs, proving a trusty alternative to other tapes on the market at the moment. A double sided tape, that is clear when peeled off, when used correctly, this tape can give you a hold time of around a week.
Reported to be the strongest tape on the market for Hair Replacment Systems.
Reported to be the strongest Tape on the market for Hair Replacment Systems. The ultrahold tape gives you all the benefit of ultrahold glue, but now in a new tape roll.
Block title
Quick hints on how to get the parting on your wig laid nice and flat


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Students in grades K–12 are invited to create their own Google Doodles inspired by the theme "If I could travel in time, I'd visit…" With materials and Museum educators on hand to help out, the Philadelphia Museum of Art is the perfect place to doodle in the final week of the competition!
The Philadelphia Museum of Art is delighted to partner in this year's Doodle 4 Google competition. This summer, 10 Pennsylvania State Finalists in the Doodle 4 Google contest will have their doodles exhibited in the Museum, in the Education Corridor. In addition to seeing his or her artwork shown at the Museum as well as on Google's homepage, the winning doodler will take home a $30,000 college scholarship and a $50,000 technology grant for their school.
This event is free (does not include Museum Admission). Pick up a Doodle 4 Google pass at any Visitor Services Desk on the day of the program.
After all doodles have been submitted to Google by March 23, a team of Googlers and then guest judges—such as Katy Perry, Phineas and Ferb creator and executive producer Jeff "Swampy" Marsh, and recording artist Jordin Sparks—will help choose the top doodles. Then on May 2, one winner from each grade will be chosen by a public vote. First prize will be awarded on May 17 and the winning doodle will be showcased on google.com on May 18.
Sunday, March 18
Noon–3:00 p.m.
Philadelphia Museum of Art,
Great Stair Hall Balcony
26th Street and Benjamin Franklin Parkway
Philadelphia, PA
Philadelphia-area children and families
Philadelphia Museum of Art Educators
For more information, visit the Event Facebook page:


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Plantation shutters in Portsmouth, supplied and fitted by trained and professional shutter fitters. Call us for a free consultation today.
Plantation shutters offer high levels of privacy and light control in any sized room as well as an additional layter of insulation which is ideal for maintaining the temperature of the room.
- Choice of 64mm, 89mm and 114mm slat
- Full height, tier-on-tier, café style or on track system
- Hand crafted wood and Sante Fe Nevada PVC
At JPB Blinds we guarantee to offer a highly competitive price, first-class fitting and a wide range of options to make your plantation shutters perfect.


text:
New Research In
Physical Sciences
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Featured Portals
Articles by Topic
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- Agricultural Sciences
- Anthropology
- Applied Biological Sciences
- Biochemistry
- Biophysics and Computational Biology
- Cell Biology
- Developmental Biology
- Ecology
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- Genetics
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- Population Biology
- Psychological and Cognitive Sciences
- Sustainability Science
- Systems Biology
Breast cancer classification and prognosis based on gene expression profiles from a population-based study
Communicated by Patrick O. Brown, Stanford University School of Medicine, Stanford, CA, May 14, 2003 (received for review December 13, 2002)
Comprehensive gene expression patterns generated from cDNA microarrays were correlated with detailed clinico-pathological characteristics and clinical outcome in an unselected group of 99 node-negative and node-positive breast cancer patients. Gene expression patterns were found to be strongly associated with estrogen receptor (ER) status and moderately associated with grade, but not associated with menopausal status, nodal status, or tumor size. Hierarchical cluster analysis segregated the tumors into two main groups based on their ER status, which correlated well with basal and luminal characteristics. Cox proportional hazards regression analysis identified 16 genes that were significantly associated with relapse-free survival at a stringent significance level of 0.001 to account for multiple comparisons. Of 231 genes previously reported by others [van't Veer, L. J., et al. (2002) Nature 415, 530-536] as being associated with survival, 93 probe elements overlapped with the set of 7,650 probe elements represented on the arrays used in this study. Hierarchical cluster analysis based on the set of 93 probe elements segregated our population into two distinct subgroups with different relapse-free survival (P < 0.03). The number of these 93 probe elements showing significant univariate association with relapse-free survival (P < 0.05) in the present study was 14, representing 11 unique genes. Genes involved in cell cycle, DNA replication, and chromosomal stability were consistently elevated in the various poor prognostic groups. In addition, glutathione S-transferase M3 emerged as an important survival marker in both studies. When taken together with other array studies, our results highlight the consistent biological and clinical associations with gene expression profiles.
Breast cancer patients with the same diagnostic and clinical prognostic profile can have markedly different clinical outcomes. This difference is possibly caused by the limitation of our current taxonomy of breast cancers, which groups molecularly distinct diseases into clinical classes based mainly on morphology. Microarray technology with its ability to simultaneously interrogate 10,000-40,000 genes has changed our thinking of molecular classification of human cancers (1). Two major reports have described the use of microarrays to assess the molecular classification of human breast cancer and have defined new subgroups based on expression that are relevant to patient management (2, 3). Sorlie et al. (2) investigated 51 carcinomas from a single patient cohort with locally advanced T3/T4 breast cancer with node involvement treated with primary chemotherapy. van't Veer et al. (3) studied 78 cases of patients with sporadic cancer all under the age of 55 with no lymph node involvement and not treated with adjuvant chemotherapy.
The tumors in both studies could be partitioned into two majors subgroups based on their estrogen receptor (ER) status as suggested by others (4, 5). Additionally, these expression cassettes could provide a refined estimate of prognosis, perhaps beyond those clinical indicators currently available to us. Sorlie et al. (3) identified a luminal subgroup (subgroup A) of ER-positive tumors associated with the best outcome. van't Veer et al. (3) addressed this problem, by investigating a narrow subset of node-negative breast cancer patients. They found 231 genes significantly associated with disease outcome as defined by the presence of distant metastasis at the 5-year mark. Van de Vijver et al. (6) provided a validation of the van't Veer predictor applied to 234 new patients from the same institution and using the same array platform.
In the present work, we have undertaken a population-based study from a regional cancer center where there are 350 new patients a year referred in from a population of 1.5 million. Over a 2-year period, 700 new cancer cases were seen, and of these 700 cases we analyzed 99 cases representative of the population. The overall survival of this group of 99 cases, adjusted for standard prognostic factors of tumor size and nodal status, is comparable to that of the 700 patients selected from the cohort seen in the years 1993-1995. Because patients were treated within existing standards of practice in those years, there was variation in patient management, reflecting the heterogeneity in clinical presentation of breast cancer.
Materials and Methods
Clinicopathological Characteristics of Breast Cancers. Tumor samples from 103 patients with primary local breast carcinoma were accessed from the John Radcliffe Hospital from January 1993 to December 1994. Four of the 103 samples were excluded from the analyses because of technical difficulties, leaving a total of 99 breast tumors. A detailed list of all samples and clinical and histopathological data for the patients is in Table 2, which is published as supporting information on the PNAS web site, www.pnas.org). All of the tumor samples were invasive ductal carcinomas; 46 individuals were node negative and 53 were node positive. Almost all of the patients received adjuvant treatment after surgery, consisting of radiotherapy (80 patients), chemotherapy (32 patients), and endocrine therapy (78 patients) according to accepted practice guidelines at that time. The chemotherapy regimen for the majority of the patients consisted of six cycles of cyclophosphamide, methotrexate, and 5-fluorouracil, and the endocrine therapy consisted of tamoxifen for at least 5 years after surgery. All tumor samples had been flash-frozen and stored at -80°C. All of the tumor samples contained >50% of tumor cells based on frozen sections adjacent to the selected samples.
Relapse-free survival (RFS) was defined as the interval elapsed between the date of breast surgery and the date of diagnosed further episode of breast cancer, whether the breast cancer was classified as a recurrence or second primary, and whatever the histology. Breast cancer survival (BCS) was defined as the interval elapsed between the date of breast surgery and the date of breast cancer-related death (documented from hospital records). ER status was determined by using ligand-binding assays and immunohistochemistry. Grade was determined by using the Elston-Ellis grade system (7).
RNA Extraction and Probe Preparation. Isolation of RNA was performed by using the TRIzol method (Invitrogen) according to the manufacturer's instructions. RNA quality from each tumor biopsy was assessed by visualization of the 28S/18S ribosomal RNA ratio on 1% agarose gel. Total RNA was linearly amplified by using a modification of the Eberwine method (8, 9). Total RNA from the Universal Human Reference (Stratagene) was amplified and used as reference for cDNA microarray analysis. The cDNA microarray chips consisted of 7,650 total features and were manufactured at the National Cancer Institute microarray facility.
A detailed protocol for RNA amplification and cDNA probe labeling and hybridization is available at http://nciarray.nci.nih.gov/reference/index.shtml. genepix software (Axon Instruments, Union City, CA) was used to analyze the raw data, which were then uploaded to a relational database maintained by the Center for Information Technology at the National Institutes of Health.
Data Analysis. Images of all of the scanned slides were meticulously inspected for artifacts, and aberrant spots and slide regions were flagged for exclusion from analyses. Log (base 2) ratios for each spot were calculated as follows. In each channel, signal was calculated as foreground median minus background median. If the signal was <100 in any single channel, the signal value in that channel was set to 100. If the signal was <100 in both channels, the spot was flagged as unreliable and not used in any further analyses. Also, if >50% of the pixels in the foreground in either channel reached the saturation threshold, the spot was flagged and not used in analyses. For all remaining (nonflagged) spots, a log ratio was calculated as log2[(red signal)/(green signal)]. The log ratios were then normalized within each array by subtracting from each the median log ratio value across the spots on the array. The channel-specific intensity data and normalized log ratios of all 99 experiments are available in Tables 3-5, which are published as supporting information on the PNAS web site.
The first phase of the analysis was to compare expression profiles between specimens segregated according to values of standard prognostic variables. In particular, we considered the following comparisons: tumor grade 1 or 2 vs. 3; tumor size ≤2 cm vs. >2 cm; age <50 years vs. age ≥50 years (menopausal status); node negative vs. node positive; and ER-vs. ER+. These comparisons were made by parametric t tests using the statistical software splus (SPLUS 6.0 Professional, Insightful, Seattle). To control for multiple comparisons, we reported as significant genes only those that reached significance at level P = 0.001. Testing 7,650 probes at this significance level, we expect that the average number of spuriously significant (false positive) results will be eight or less.
Cluster analyses were conducted to search for natural groupings in the profiles. Before clustering, a screening procedure was applied to eliminate genes showing minimal variation across the set of 99 specimens. Specifically, for each gene, the 5th and 95th percentiles of the ratios were calculated. If the ratio of the 95th to 5th percentile was <3 that gene was not included in the cluster analysis. This process left 706 probe elements for the cluster analyses. Hierarchical agglomerative clustering using the statistical package brb-arraytools software (available at http://linus.nci.nih.gov/BRB-ArrayTools.html) was applied to these normalized log ratios by using both compact linkage and average linkage and both Euclidean and one minus Pearson correlation distance metrics. Normalized log ratios were median-centered within each gene for all of the cluster analyses. The clustering results obtained by using compact linkage with one minus Pearson correlation distance applied to the 706 probe elements appeared by visual inspection to yield the most distinctive clusters (remaining blinded to any clinical or outcome variables), and hence this was the clustering algorithm used for the unsupervised cluster analyses based on these probe elements (Fig. 1). The presence of significant clustering was assessed by applying the global test of clustering proposed by McShane et al. (10). The same techniques were applied for the clustering analyses using gene subsets sets derived from the van't Veer and Sorlie studies (Figs. 3, 4, 5, 6 which are published as supporting information on the PNAS web site).
Survival comparisons among clusters resulting from unsupervised cluster analysis were made by using Kaplan-Meier estimation and Cox proportional hazards regression. To assess univariate associations of individual genes (log ratios) with survival, Cox proportional hazards regression methods using the splus software were used.
Classification of Tumors Samples Based on Clinical/Pathologic Characteristics. Clinical parameters such as ER status, nodal status, tumor size, tumor grade, and menopausal status of the patient affect the behavior of breast cancers. We asked whether these clinical/pathologic characteristics were associated with differential gene expression. Parametric t tests identified 606 probe elements of the 7,650 elements represented in our array that could segregate ER+ and ER-breast tumors (Table 1, P < 0.001). The detailed gene list is in Table 6, which is published as supporting information on the PNAS web site. Within this list were genes previously known to be estrogen responsive or associated with ER status such as LIV-1, TFF3, GATA 3, c-myb, and BTG2 (11-14). A total of 137 probe elements distinguished high-grade and intermediate/low-grade breast tumors (Table 1, P < 0.001), including genes involved in cell cycle progression such as topoisomerase II α, MCM2, BUB1, and proliferating cell nuclear antigen (PCNA) (15-17). The detailed gene list is included in Table 7, which is published as supporting information on the PNAS web site. Few genes, however, discriminated tumor size (3 probes, P < 0.001), nodal (11 probes, P < 0.001), and menopausal status (13 probes, P < 0.001). This finding suggests that ER status has a strong association with gene expression, and tumor grade has a moderate association. However, there is no strong evidence that nodal and menopausal status of the patient or tumor size is associated with the expression profiles of the tumors.
Unsupervised Clustering Identifies Natural Groups Corresponding to Breast Lineage Markers. Using an unsupervised hierarchical clustering approach, we sought to define natural subclasses of breast tumors as determined by gene expression profiles. We performed the unsupervised clustering on the 706 probe elements selected as exhibiting high variability across all tumors (see Materials and Methods). Included in this list are genes corresponding to the standard prognostic parameters, such as ER and HER-2/neu status. Application of the global test of clustering and reproducibility measures of McShane et al. (10) showed borderline statistically significant evidence for clustering (P = 0.057), and the robustness indices suggested (robustness >75%) that the most reproducible clustering structure was evident when the dendrogram was cut at two clusters. Our results show that the tumor samples could be confidently separated into two main groups primarily associated with ER status as determined by the ligand-binding assay and confirmed by immunohistochemistry (Fig. 1). The entire cluster diagram including gene expressions is included in Fig. 7, which is published as supporting information on the PNAS web site. Eighty-two percent of the tumors in the left main branch are ER-, and 88% of the tumors in the right main branch are ER+ by immunohistochemistry. This finding corroborated the earlier analysis (Table 1) that the major factor discriminating the expression phenotype is ER status. As expected, the ER-cluster had a higher percentage of high-grade tumors than the ER+ cluster (Fig. 1).
The dendrogram further branched into smaller subgroups within the ER+ and ER-classes. Although our sample size was not large enough to establish high reproducibility of smaller subgroups, we were able to locate in our dendrogram several subgroupings previously identified in other studies. We describe here what we observed in our data relative to those clusters that have been reported by others. Within the ER-cluster are tumors with "basal"-like expression characteristics as defined by higher gene expression of keratin 5, keratin 6, metallothionein 1X, and fatty acid binding protein 7 as reported (Table 8, which is published as supporting information on the PNAS site) (2, 18). Furthermore, they exhibited higher expression of the secreted frizzled-related protein 1 (SFRP1) and the oncogene c-kit and lower expression of fibronectin 1 and mucin 1. Basal 1 subgroup was differentiated by higher expression of matrix metalloproteinase 7 and cell growth-related genes such as topoisomerase II α, mitotic feedback control protein Madp2 homolog (MAD2L1), cell division control protein 2 homolog (CDC2), and PCNA, suggesting a signature for a high proliferation rate (Table 8). In contrast, the basal 2 subgroup was distinguished by higher expression of many components of the transcriptional factor AP-1 such as c-fos, c-jun, and fos B, as well as by overexpression of activating transcription factor 3, caveolin 1 and 2, hepatocyte growth factor, and transforming growth factor β receptor II. A subgroup distinct from the basal-like groups in the ER-subset is defined by a high rate of HER-2/neu overexpression (HER-2/neu: 5/7 tumors). This HER-2/neu subgroup was further distinguished from the basal-like subgroups by the higher expression of MDR1, S100 calcium-binding protein P, fatty acid synthase, RAL-B, RAB6A, fibronectin 1, and syndecan 1 and lower expression of c-kit and c-myc.
The ER+ subgroup showed differential expression of genes associated with ER activation such as LIV-1, trefoil factor 3, neuropeptide Y receptor Y1, keratin 8, GATA3, and X-box binding protein 1. These are also genes that define the ER+ cluster as having "luminal" characteristics as defined (2, 18). Moreover, the ER+ cluster could be further segregated into three smaller subclasses: luminals 1, 2, and 3 (Fig. 1).
We then asked whether the array-derived tumor groups demonstrated any differences with respect to survival. When the basal/neu (predominantly ER-) and the luminal-like (predominantly ER+) cluster were compared by Kaplan-Meier and Cox regression analysis, the luminal-like subgroup had a significant advantage in both RFS and BCS (Fig. 2 A and B). The three subclasses within the basal/neu (ER-) cluster appeared to have similarly poor survival characteristics (Fig. 2 C and D). By contrast, the three subgroups within the luminal-like (predominantly ER+) cluster showed distinct differences in survival (Fig. 2 E and F). Luminal 1 had the best outcome with an 80% 10-year RFS. This luminal 1 subgroup was also correlated with lower-grade tumors and was further characterized by differential higher expression of c-kit, hepatocyte growth factor, insulin-like growth factor-binding protein-3, ATF-3, and components of the AP-1 transcriptional factor such as c-fos, c-jun, fosB, and jun-D, and as expected by lower expression of cell growth related genes such as topoisomerase II α, mitotic kinesin-like protein-1, PCNA, CDC2, BUB1, and MAD2L1 (Table 8). This was significantly different from the luminal 2 subgroup that had the worst outcome with a 10-year RFS of 40% (P = 0.022 luminal-like 1 vs. luminal-like 2). This subgroup showed higher expression of a protein tyrosine phosphatase type IVA member, tumor necrosis factor receptor-associated factor 3, RAD21, and BRCA1-associated protein 1 (BAP1) and lower expression of FGFR1, CXCR4, ATF-3, and vascular cell adhesion molecule 1. The luminal 3 subgroup had an intermediate survival outcome of 60% at 10 years. Intriguingly, although some of the ER+ tumors were HER-2 overexpressors (6 of 66), they were dispersed among the luminal subgroups.
In the Sorlie et al. study (3), tumor classification was performed based on the 456 cDNA clones (427 unique genes) in their "intrinsic" gene list. To assess how these genes could perform in classifying our tumor set, we sought the overlap between either the intrinsic gene list and our cDNA array or the overlap between the intrinsic gene list and the 706 variably expressed probe elements in this study. We found 332 (285 unique genes) that overlapped with the full set of probe elements on our cDNA array and 105 (96 unique genes) probe elements that overlapped with our 706-gene list. Interestingly, in both situations hierarchical cluster analysis segregated our tumors into two distinct subgroups mainly based on their basal (predominantly ER-) and luminal (predominantly ER+) characteristics (Figs. 5 and 6).
The luminal-like tumors were further segregated into at least two (possibly three) smaller subgroups, which may correspond to luminal A, B, and C subtypes. Additional analysis revealed that 100% of the specimens in the luminal A subtype in the 332-gene cluster and 77% in the 105-gene cluster were assigned as luminal-like 1 tumors in our unsupervised cluster analysis (based on the 706-element cluster, Fig. 1). In contrast, the specimens in the luminal B and C subtypes were assigned either as luminal-like 2 or luminal-like 3 tumors.
Although the luminal A subtype showed a favorable clinical outcome when compared with other luminal subtypes, this difference was not statistically significant.
Identification of Gene Clusters Associated with Survival. Our database includes the survival data of all patients studied with a median follow-up of 6.1 years. To determine the genes associated with improved RFS, we performed Cox proportional hazards regressions on each of the 7,650 probe elements on our array. A group of 485 different probe elements were identified that could separate RFS in the 99 patients with a P value <0.05, but only 16 probe elements were significant at the more stringent P < 0.001 level used to control for multiple comparisons (Table 9, which is published as supporting information on the PNAS web site). We discuss results for the larger list of 485 so that our results may be compared with results of previous studies. Of the genes reported to be associated with prognosis in breast cancer, only PCNA and topoisomerase II α were found to be elevated in the poor prognostic group (HER2-neu/basal-like). Interestingly, this gene list did not include the ER, Her-2/neu, or p53. This is consistent with the findings of van't Veer et al. (3), who found no association between HER2 and ER expression levels and survival in the node-negative cases included in their study.
In the van't Veer et al. data set, 231 genes were noted to be prognostic for RFS in the node-negative breast cancer patients. We asked first whether any of these potentially prognostic genes were included in our 7,650-element array. We observed that 93 probe elements representing 56 unique genes overlapped, and based on their expression levels hierarchical cluster analysis we could separate our patients into two distinct subgroups. When Kaplan-Meier analysis was performed, a statistically significant survival difference was seen between these two groups (P = 0.03, Fig. 3). This finding demonstrated that a subset of the genes identified by van't Veer et al. to be prognostic in untreated node-negative patients could be confirmed to have an association with clinical outcome in an independent cohort of treated individuals with mixed nodal status. To identify a minimal number of the most important prognostic genes, we sought the overlap between our optimal survival list of 485 probe elements and the 231 genes in the van't Veer et al. prognostic gene set. This overlap survival gene list consisted of only 11 unique genes represented by 14 probe elements. As expected (because these 14 elements were among those selected because of their observed significant univariate association with survival), these 14 elements separated our patients into two major groups, showing a significant difference in survival (as visualized in Fig. 4). Intriguingly, 5 of the 11 unique genes, RFC4, MCM6, MAD2L1, BUB1, and CKS2 appear to be involved in DNA replication and chromosomal stability, and all were up-regulated in the poor prognostic group. This finding suggests that differences in replicative potential distinguish the prognostic groups.
Microarray analyses on breast cancers have identified gene expression profiles able to separate tumor classes associated with patient survival (1). Perou et al. (18) and Sorlie et al. (2) showed that the expression profiles primarily distinguished ER+ from ER-tumors and called them luminal and basal subtypes because of their respective luminal and basal characteristics. van't Veer et al. (3) had similar observation but extended this to gene expression (or genetic profile) associations with survival in an untreated, node-negative cohort.
Here, we present an analysis on 99 tumors from node-positive and node-negative patients, the majority receiving adjuvant treatment according to accepted practice guidelines at the time of the diagnosis. Our results were significant in their concordance with those of the earlier studies despite the differences in patient populations, treatments used, and technology platforms used. Thus, our results provide supporting evidence for the prognostic importance of genes identified in previous reports on a completely independent patient cohort with an independent microarray platform. We found that the ER status of the tumor was, indeed, the most important discriminator of expression subtypes and that tumor grade was a distant second. Other clinical features, namely lymph node positivity, menopausal status, and tumor size were not strongly reflected in the expression patterns obtained with the 7,650-feature microarray in this investigation. This finding confirms that ER biology plays a central role in breast carcinogenesis defining the configuration of the final tumor. Furthermore, investigation of gene expression in primary tumors may be unlikely to identify a set of genes whose expression reliably correlates with lymph node metastasis. This finding is consistent with data showing that only a small fraction of cells in a tumor mass have metastatic potential (19, 20). The genetic signature from this metastatic fraction would be "diluted" by the signals from nonmetastasizing cells.
Similar to the findings of Sorlie et al. (2), unsupervised hierarchical clustering analysis segregated the tumors into two main clusters based on their basal (predominantly ER-) and luminal (predominantly ER+) characteristics. Furthermore, within each of these clusters we could identify smaller subgroups that were characterized by distinct gene expression signatures involving potential different oncogene-specific pathways. A HER-2/neu subgroup was characterized by higher expression of the oncogene her-2/neu and higher expression of genes involved in the ras pathway such a Ras-related GTPases, RALB, and RAB6A. Convergence of neu and ras pathways in breast cancer tumorigenesis has already been documented (21). In contrast, basal 1 and 2 subgroups were characterized by higher expression of the oncogenes c-kit, c-myc, and SFRP1. SFRP1 is a modulator of Wnt signaling. Recently, aberrations of Wnt signaling were reported to be involved in the pathology of various human neoplasms (22). Activation of the Wnt signaling pathway appears to lead to the cytosolic stabilization of a transcriptional cofactor, β-catenin, that can regulate the transcription from a number of target genes including the cellular oncogene c-myc. In breast carcinoma, SFRP1 expression has been associated with loss of ER and the presence of lymphoplasmocytic reaction around the tumor associated with a more aggressive disease (23, 24). Furthermore, basal 1 type exhibited higher expression of genes involved in cell cycle and growth such as PCNA, CDC2, and BUB1 whereas the basal 2 type showed higher expression of transcription factors such as c-fos and ATF3, expression signatures that both could be modulated by c-myc (21). These data raise the possibility that Myc, which is amplified in 15% of breast cancers, may have a more important role in determining the expression profile of a breast cancer than previously thought.
Moreover, as noted in earlier studies, our basal and luminal subgroups also showed the expected differences in survival with a better outcome in the luminal group. More interestingly, we found that the 231 genes described by van't Veer et al. (3) as separating survival groups in node-negative untreated patients may have distinct prognostic capabilities in a more heterogeneous population of node-positive/negative patients treated with adjuvant therapy. Using the 93 probe elements, 56 unique genes, from the van't Veer prognostic set represented in our microarrays, we could easily separate our 99 patients into two prognostic subsets. This finding appears to confirm the importance of some subset of these 56 genes as bona fide prognostic markers. In particular, the overlap between the van't Veer 231 genes and our 485 probes associated with survival (at P < 0.05 level) was 14 probe elements representing 11 unique genes. Five of the 11 genes in this set are involved in cell replication and chromosomal stability and were up-regulated in the poor prognostic setting, suggesting a molecular mechanism for this clinical outcome. An intriguing question is what might be a "minimal" set of genes necessary to establish a reproducible prognostic classification. In leukemia, with well-defined genetic changes, gene profiles segregate with particular translocations (25). In our breast cancer series the major groupings follow previously defined signaling pathways such as ER+, ER-, and c-erbB2/ras. Our study design did not permit us to relate BRCA1 and BRCA2 status to these expression patterns.
Finally, gene profiles that relate to prognosis may help define new therapeutic targets. In our study, cell cycle regulation is clearly important and suggests continued use of antiproliferatives is a rational approach. However, the melanoma tumor antigen PRAME was highlighted and should be further investigated in breast cancer as a potential tumor antigenic target. Also the glutathione S-tranferase pathway, well recognized to have a role in drug resistance, was associated with poor outcome and appeared to be strongly correlated with survival in both our study and that of van't Veer et al. (3).
This work was supported in part by Fonds National de la Recherche Scientifique Grant Ext. 260 V6/5/2-ILF 14773 (to C.S.), the National Cancer Institute, and the Genome Institute of Singapore.
↵∥ To whom correspondence should be addressed. E-mail: gisliue{at}nus.edu.sg.
Abbreviations: ER, estrogen receptor; RFS, relapse-free survival; BCS, breast cancer survival; PCNA, proliferating cell nuclear antigen.
- Received December 13, 2002.
- Copyright © 2003, The National Academy of Sciences
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Sorlie, T., Perou, C. M., Tibshirani, R., Aas, T., Geisler, S., Johnsen, H., Hastie, T., Eisen, M. B., van de Rijn, M., Jeffrey, S. S., et al. (2001) Proc. Natl. Acad. Sci. USA 98, 10869-10874.pmid:11553815
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Gruvberger, S., Ringner, M., Chen, Y., Panavally, S., Saal, L. H., Borg, A., Ferno, M., Peterson, C. & Meltzer, P. S. (2001) Cancer Res. 61, 5979-5984.pmid:11507038
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West, M., Blanchette, C., Dressman, H., Huang, E., Ishida, S., Spang, R., Zuzan, H., Olson, J. A. J., Marks, J. R. & Nevins, J. R. (2001) Proc. Natl. Acad. Sci. USA 98, 11462-11467.pmid:11562467
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McShane, L. M., Radmacher, M. D., Freidlin, B., Yu, R., Li, M. C. & Simon, R. (2002) Bioinformatics. 18, 1462-1469.pmid:12424117
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In the past, the Chinese people believed in dragons and they represented the emperors as well as the symbol of power, self-confidence, luck, protection. Chinese dragon is differently seen from other dragons in other parts of the world, or better said, as they are perceived in other cultures. This negative image was possible especially through the films that were made that presented them as very dangerous and bad.
Chinese dragon transformed in an inspiration for Chinese dragon art and the result was a real piece of art as many people are very talented. The dragon is a very important symbol in China and in case you visit it, you will observe it on some buildings. Chinese dragon art has developed covering many things as pictures, tattoos, statuettes, fashion and so on. As you can see, Chinese dragon became a great source of inspiration for many fields and has spread in many parts of the world. More, Chinese dragon symbolizes wisdom, power, it is celebrated by feast and art, so would be very intersting to see such an event.
Chinese dragon is also a great idea of a souvenir in case you visit China or you can decorate your home with a piece of Chinese dragon art. The good thing is that you do not have to travel up to China to buy a dragon, as they have spread in so many parts of the world, so you can buy right from your city. There are many colors of Chinese dragon and also several types. Chinese dragon was the subject of many legends that became famous and very known with the passing time. Chinese dragon is present in so many traditions and it is very important for the Chinese people. Chinese dragon art has advanced a lot and there are so many talented people that will impress you with what they can do. It seems as they would do some magic, so interesting is the result of having an object or a tattoo inspired by Chinese dragon art.
Click here to Buy Chinese Dragon Products!


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What are the main differences between prime and sub-prime applications?
Essentially, there is no difference between them. With both prime and sub-prime applications, the mortgage adviser must look carefully at lenders' product criteria, to see which lender and product matches their client's needs.
The two types of application start to diverge when a mainstream lender has fairly standard credit criteria and makes extensive use of credit scoring (for example, a few products designed for bulk lending) and a sub-prime lender offers many products to cater for specialist needs. Here, the specialist lender is always trying to strike a balance between security and risk. For example, if the LTV is low, specialist lenders may be happy to allow a higher degree of adverse credit, or, if the LTV is relatively high, the lender may want to see full documentary proof of earnings rather than allowing self-certification.
The resulting portfolio of products will mean the adviser dealing with a non-mainstream client must look more carefully at the lender's criteria than when dealing with a mainstream applicant.
How much adverse credit does it take to make an applicant sub prime?
The idea of adverse and sub-prime credit is not a description of a certain kind of applicant per se. It is a by-product of the growth of credit scoring and credit referencing used by mainstream lenders to, in effect, make the lending decision for them. Before the mid-1990s, the sub-prime mortgage market did not exist because anyone with a past history of credit problems could simply not get a mortgage at all. Since then, a number of lenders have been willing to take a higher level of risk than high street lenders, and have produced products for borrowers who have had credit problems in the past.
The classic categories for grouping such borrowers is by asking how many county court judgements (CCJs) and arrears they have, and whether they have been bankrupt or are involved in an individual voluntary arrangement (IVA) with their creditors. However, there are many other factors that will cause borrowers to be declined by credit scoring systems. It is a matter of advisers getting a good feel for what sort of applicant the various specialist lenders can cater for, and then matching them to each client.
Why are interest rates for sub-prime mortgages higher than for mainstream?
This is the simple economics of risk and rewardš that is common to all financial services products. A good analogy is that of motor insurance or private health insurance. Those with a history of claims, or a lifestyle that is known to be high riskš will have to pay a relatively higher premium for cover. Similarly, those with a history of credit difficulties are classed as high risk, so lenders need to charge a premium to cover that.
What are the most common mistakes made on sub-prime applications?
These will be the same for specialist lending as they are for mainstream. For example, not supplying the specified documentation, missing out questions or even the applicant not signing the form. Also ' and this is not something the mortgage adviser can always guess in advance ' applicants sometimes forget or gloss over their history, without realising that further credit status searches will come up with a different history of addresses, or more overall outstanding adverse credit than the applicant originally stated.
Why do most sub-prime applications go through packagers?
Most specialist lenders use highly experienced underwriters to make the lending decisions. In order for them to do their job effectively, they need a stream of properly completed applications, with documentary back-up where necessary. By using specialist and experienced packagers to filter applications from individual brokers, lenders can be sure they will receive well-packaged applications.
How can advisers ensure a quick decision on sub-prime applications?
We always work on the principle that, in specialist lending, a quick decision is more important to applicants than any other service element. This is just as necessary when declining applications as it is when making an offer. In short, there are a few golden rules to remember:
• Do not ignore questions on the form that look awkward for the applicant to complete in the hope the lender will not notice. It will just cause delays.
• Do not send in extra information on one topic, in the hope this will compensate for a lack of information in another area. Again, all this will do is cause unnecessary delays.
• If there are mitigating circumstances for the borrower's past problematic credit history include a short written explanation.
• Title insurance can speed up applications considerably, so it is worth getting to know about it, and which lenders specify it.
• If speed is important to your clients, and you need a decision and/or completion quickly, then place the application with a lender who is known to you either by experience or reputation as one that can guarantee a quick turnaround.
Stuart Aitken is director of credit at Southern Pacific Mortgage Limited (SPML)


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Create beautiful wedding memories in Tucson
Radisson Suites Tucson offers beautiful venues for wedding ceremonies and receptions. Walk down the aisle and say your vows surrounded by the scent of orange blossoms in the picturesque Orange Grove. Then, join your guests for a reception in the Catalina Ballroom or Reddington Room, where you'll enjoy fresh, gourmet dishes from your customized catering package. The indoor facilities can accommodate up to 240 guests and host other wedding-related events, such as a cocktail hour or rehearsal dinner. We provide a specially designed event website that offers information about the hotel and allows your guests to book their overnight stay at a discounted price.
Contact for weddings, quinceañeras, social events and corporate parties
Nicole Nevin, Catering Sales Manager
Phone: +1 (520) 239-1135
Nicole Nevin, Catering Sales Manager
Phone: +1 (520) 239-1135
Se Habla Español


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How to Care for NJ Inground Pools
NJ inground pools require a great deal of maintenance to keep them running efficiently. While it might seem overwhelming, once you break it down, you will see it is not as difficult as it seems. There are a few common mistakes new pool owners make that can hurt their structures, but if you know these mistakes ahead of time, you can avoid making them, giving you more time to enjoy your pool.
Watch the Chemistry
The chemistry of the water is the most important part of your maintenance schedule. With improper levels of chemicals, you could not only ruin your structure but also put the health and wellness of you and your family at risk. It is important to check the chemical levels in the water at least twice a week during the months during which it is actively used and once per week during the
months it is not used but still has water in it.
Clean the Walls
The walls of your pool will normally become laden with algae and bacteria. If you let them sit without frequent cleanings, they can become more difficult to clean and cause the water to become toxic. In order to stay ahead of the buildup, you should clean the walls at least once per week. If you begin to notice calcium buildup on the tiles, you should call a professional for help as soon as possible.
Repair Broken Parts
While a broken drain or suction component of your pool might not seem like an emergency, they need to be repaired or replaced right away. It is vital for the proper operation of your pool, as well as its safety. If you have a broken fence or rail around your structure, it is just as important to have it fixed as it is to have a part inside the pool fixed. The security surrounding your structure is important to keep everyone safe when you are not around.
NJ inground pools are a beautiful addition to any home. In order to keep yours as beautiful as it was the day it was installed, you need to practice proper maintenance. It is important to keep a careful watch on the level of chemicals in the water to keep everyone safe. In addition, the cleanliness of the walls and the proper function of all the parts of your pool are vital to its safety and longevity.


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Business in Action, 9th Ed.
Chapter 13: The Art and Science of Marketing
"After starting a business, your next task is finding ways to acquire and retain customers.
"Despite its reputation as the underdog of the social media world, Pinterest outranks Twitter, Snapchat and Linkedin in popularity, according to this 2018 study.
"The average American adult is expected to spend nearly 3.
"WSJ's Joanna Stern "bumps into" Steph Korey [photo, left] in the elevator and asks about the luggage company's push into physical retail, her advice for raising capital and her travel habits.
"As you're brainstorming a new content marketing campaign, the word "humble" may not come to mind.
"Behavior plays a major role in new targeting options on LinkedIn.
"Buy online, pick-up in store is often heralded as the future of retail: Customers shopping on their own terms, as efficiently as possible.
"You have crafted your business.
"[In August], cashierless store Zippin opened its doors to customers in San Francisco for the first time, beating Amazon Go to become the first cashierless store in the city.
"[Recently,] a ribbon was cut in Gluckstadt, Mississippi.
"If good brands don't communicate exactly what the company does up front, they quickly make themselves synonymous with those products or services.
Tamara Kleinberg observes fellow customers complaining to her but not to the customer service agent and asks if your business is experiencing the same phenomenon.
"Do you advertise on Facebook?
"Spying on your ecommerce competition can help with product pricing and marketing strategy.
"With the right mix of speed, timing and guts, smart founders can profit hugely from their much, much, much larger rivals' misfortune.
"Ever wondered why it's called "7UP"?
"Retailers and manufacturers are putting more dollars into front-line efforts in the stores themselves, says Paco Underhill, founder of Envirosell and author of Why We Buy: The Science of Shopping.
"I've been chronicling the slow demise of B&N for years now, watching the company bleed out, drop by drop, until it has become a shell of its former value.
"Video is the marketing juggernaut that continues to grow year over year.
"This [past] fall, Nordstrom opened a new store in West Hollywood, Calif.
"Find out why the smartphone will be crucial for retailers in 2018 and beyond with the first part of a brand new slide deck from BI Intelligence called The Future of Retail 2018.
"This is an interesting tactic from the retailer, and one that makes a lot of sense from a behavioral perspective.
"In August, the Interactive Advertising Bureau (IAB)'s release of a buyer's guide for podcast advertising signaled that advertisers and marketers are beginning to take podcasting seriously as a marketing channel.
"Some smaller retailers will tug at shoppers' heartstrings during the holidays, trying to create an emotional experience or connection that a big national chain might not provide.


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Price: £6.50
Quantity: 1 available
Faber Gallery series. Price clipped. 11 colour plates tipped-in. Colour plate tipped-in on cover. Creasing along edges of wrapper. Sunning to wrapper.
Title: Venetian Paintings.
Categories: Arts: Original Works and Prints,
Edition: First edition
Publisher: London:, Faber & Faber,: 1949.
Binding: Paperback
Book Condition: Very Good in Very Good DJ
Weight: 1.00 Item
Seller ID: 3735
Keywords: Art History Italian Art Orginal Paintings


text: 0 0 Anyone selling a Louis Vuitton brushstroke camouflage wallet, or know where I can buy one??? Men Louis Vuitton Back to BopTalk Share Facebook Twitter Google+ Tumblr LinkedIn Mail ty_babcock posted 3 years ago Replies Your Reply Please login first to submit. Sort by: Newest First Oldest FirstRandomMost VotesRecently Active Post Reply Showing 0 results