text:
Ordinary Council Meeting
MINUTES OF THE Ordinary Council Meeting HELD ON 17 November 2011
The following document is the minutes of the Council meeting held on 17 November 2011. These minutes are subject to confirmation as to their accuracy at the next meeting to be held on 1 December 2011 and therefore subject to change. Please refer to the minutes of 1 December 2011 for confirmation.
Cr Rhonda Hoban (Mayor)
Cr John Ainsworth
Cr Martin Ballangarry OAM (arrived 5.50 pm)
Cr Janet Court
Cr Brian Finlayson (arrived 6.04 pm)
Cr Paula Flack
Cr Michael Moran OAM
Cr Elaine South
Cr Anne Smyth
Michael Coulter (General Manager)
Greg Meyers (Director Environment & Planning)
Paul Gallagher (Director Engineering Services)
Monika Schuhmacher (Minute Secretary)
Presentation to Council of the Acquisitive Art Prize from the Nambucca Valley Bi-Annual Art Exhibition, by:
Jenny Urqhart, President Nambucca Valley Arts Council
Lyn Greentree, Coordinator Art Exhibition
Artist, Phyllis Moss (not in attendance)
Capt Nicole Viles from the Salvation Army offered a prayer on behalf of the Nambucca Minister's Association.
Councillor Smyth declared a non-pecuniary less significant conflict of interest in Item 11.2 (GPC) Nambucca Heads Flood Investigation - Phase 2 Seaview Street Catchment under the Local Government Act as Cr Smyth has a family member who owns property in the catchment area.
CONFIRMATION OF MINUTES - Ordinary Council Meeting
3 November 2011
That the minutes of the Ordinary Council Meeting of 3 November 2011 be confirmed subject to the following:
Cr South – Page 3—Item 8.5 (Macksville Saleyards) Recommendation 3 should read:
That Councillors be advised of the date and time of an audit by both Departments so that they may be available to hear first hand of any problems and that a representative from GJ Kennedy be invited to attend.
NOTICE OF MOTION - CR Hoban
ITEM 5.1 SF1595 171111 Notice Of Motion - South Arm And Mchughs Creek Road - Concerns Raised Over Work Practices & Grading Of Unsealed Roads (Trim SF403)
3218/11 RESOLVED: (Hoban/Moran)
1 That Council note the petition distributed previously under cover of a separate memo.
2 That Council staff be asked to consider the comments and suggestions of residents when formulating future asset management plans and work schedules for the unsealed sections of South Arm Road and McHughs Creek Road.
3 That the Director of Engineering Services be asked to consider the unsealed section of South Arm Road or McHughs Creek Road as one of the sites for expenditure of the $25,000 allocated in the budget for unsealed pavement management testing.
Notice of Rescission
ITEM 6.1 SF1541 171111 Rescission Motion - Planning Proposal Lots 11 & 12 DP 1017408 and Lot 2 DP 514920 Pacific Highway Lower Nambucca
3219/11 Resolved: (Moran/South)
That Council rescind the resolution:
That no further action be taken with this with respect to this planning proposal at this time and that further investigation may be undertaken when staff are able to prepare a Local Growth Management Strategy – Residential Lands.
3220/11 Resolved: (Ainsworth/Moran)
That the following delegations be heard:
(i) Mr Geoff Smyth (Geoff Smyth Consulting)
i) 6.1.... Rescission Motion - Planning Proposal Lots 11 & 12 DP 1017408 and Lot 2 DP 514920
Pacific Highway Lower Nambucca – Supporting Recommendations of Report
ii) Ms Louise Robinson (Nambucca Heads Local Aboriginal Lands Council) Objecting to the waiver
(iii) Ms Leanne Boatfield representing local residents – Objecting to the waiver
(iv) Mr Geoff Smyth (Geoff Smyth Consulting)
11.5 Request for Comment for SEPP 71 Waiver of Masterplan Lot 2 DP 1119830,
Alexandra Drive, Bellwood – Supporting Waiver on behalf of applicants......................... 47
Mr Geoff Smyth addressed Council making the following points in regard to the Notice of Rescission:
· Will clarify some of the background information
· Department of Planning's letter – not opposed, but rather fits in and is part of Council's strategy
· That area has to be sewered particularly for oyster leases
· Development will contribute to cost of sewer
· Council has supported all the way up to now
Notice of Rescission
1 That Council resolve to prepare a Local Growth Management Strategy - Residential Lands and seek advice from the Director General of the Department of Planning and Infrastructure as to what specific matters it requires to be addressed.
2 That following receipt of the Director General's requirements, a brief be prepared and estimates be sought from external consultants so that appropriate funds may be allocated for consideration in the 2012/2013 budget.
3 That the applicant be advised that Council supports the proposal and that it proposes to prepare a Local Growth Management Strategy - Residential Lands during the 2012/2013 budget year which will include the consideration of Lots 11 and 12 DP 1017408 Lot 2 DP 514920 Pacific Highway Lower Nambucca as an urban release area.
Amendment: (Ainsworth/Moran)
1 That Council resolve to prepare a Shirewide Growth Management Strategy - Residential Lands and seek advice from the Director General of the Department of Planning and Infrastructure as to what specific matters it requires to be addressed in the Shire wide strategy.
2 That following receipt of the Director General's requirements, Council consider either preparing the strategy internally or through a consultant following the organisational review.
For the motion: Councillors Court, Ballangarry, Flack, Smyth, Ainsworth, Moran, South
and Hoban (Total: 8)
Against the Motion Nil
The amendment was carried and it became the motion and it was:
3222/11 Resolved: (Ainsworth/Moran)
1 That Council resolve to prepare a Shirewide Growth Management Strategy - Residential Lands and seek advice from the Director General of the Department of Planning and Infrastructure as to what specific matters it requires to be addressed in the Shire wide strategy.
2 That following receipt of the Director General's requirements, Council consider either preparing the strategy internally or through a consultant following the organisational review.
DELEGATIONS continued
Ms Louise Robinson addressed Council in regard to Item 11.5, making the following points:
· CEO for Nambucca Aboriginal Land Council
· Acknowledge Elders past and present
· NHLALC – on behalf of the Aboriginal men – believe they have not had enough consultation because plans do not reflect cultural needs of the area
· The consultation is extremely important to the Aboriginal men in order for the process to go smoothly.
Ms Leanne Boatfield addressed Council in regard to Item 11.5, to a prepared statement which is on File LF4148.
3223/11 Resolved: (Moran/Flack)
That Ms Boatfield be granted an additional five minutes in which to address Council
Mr Geoff Smyth addressed Council making the following points:
· The extension of Marshall Way
· The master plan is about more generic issues of area, not the details that go with the area
· Reports have been between the Department of Planning and Department of Works – certainly be shown to the Aboriginal Land Council
· Bellwood was part of a development control plan
· And included the protection of Aboriginal sites
· The majority of those principles have been achieved hence a more generic plan
· This is not a new release are
· Marshall Way is the last of the areas
· Need to achieve consultation – will not be achieved affectively through the master plan
· Happy for report to go on public exhibition and some reports may need updating as part of the development application
ITEM 11.5 LF4148 171111 Request for Comment for SEPP 71 Waiver of Masterplan Lot 2 DP 1119830, Alexandra Drive, Bellwood
That Council advise the Department of Planning and Infrastructure that it does not support the request for a Waiver of the requirement to prepare a Master Plan due to the lack of a site specific Development Control Plan that has been through the rigours of community input and consultation.
For the Motion: Councillors Ballangarry, Flack and Smyth (Total 3)
Against the Motion: Councillors Court, Finlayson, Ainsworth Moran, South and Hoban (Total: 6)
The motion was LOST.
3224/11 Resolved: (Ainsworth/Finlayson)
1 That Council advise the Department of Planning and Infrastructure that Council would be relying on the generic provisions of the Nambucca Development Control Plan 2010 and any studies and reports prepared by and for the proponents for the assessment of any Development Application should one be lodged.
2 That Council advise the Department of Planning and Infrastructure that should it issue a waiver, the applicant be advised that any application lodged with Council should be accompanied by reviewed and revised reports prepared for the former Part 3A Major Project Application. In particular attention to the recently adopted Lower Nambucca River Flood Study Review 2011, the recently endorsed Bushfire Hazard Maps and Australian Standard 3959 (2009) - Construction of Buildings in Bushfire Prone Land.
3 That Council forward to the Department of Planning the two papers presented at Council's meeting by the Nambucca Heads Local Aboriginal Land Council and Ms Leanne Boatfield.
For the Motion: Councillors Hoban, South, Moran, Ainsworth, Finlayson and Court (Total 6)
Against the Motion: Councillors Ballangarry, Flack and Smyth (Total 3)
3225/11 Resolved: (Flack/Smyth)
LF967, LF2460 and sf15 171111 FILLING ACTIVITIES ADJACENT TO NAMBUCCA RIVER AT LOWER NAMBUCCA, PACIFIC HIGHWAY
That Council receive a report on the outcome of the investigation of filling activities adjacent to Nambucca River at Lower Nambucca, Pacific Highway, as per Cr Flack's Questions with Notice and the Director Environmental Services' responses below:
1 Is the land filling activity occurring on the edge of the Nambucca River at Lower Nambucca occurring on the same land ie either Lots 11 &12 DP 1017408 or Lot 2 DP 514920 Pacific Highway, which are currently the subject of a planning proposal to rezone rural land to residential land and which are the subject of a rescission motion at the next Ordinary Meeting of Council on 17 November 2010?
The Land the subject to the placement of fill material is the same land that the Planning Proposal rescission motion relates.
2 If so, does this activity relate in any way to the planning proposal to rezone rural land to residential land on these same Lots?
Advice from the Landowner will be sought today (17/11/11) as to the purpose of the filling.
3 Does the dumping of soil at this location require consent from Council or any other authority?
A number of standards exist under SEPP (Exempt and Complying Development Codes) regarding earthworks and retaining walls. As such the application of the standards suggests that the filling may not be exempt.
As the site is within the Oyster Growing area it may be a controlled activity under SEPP 62 (Sustainable Aquaculture).
Due to its proximity to the river and oyster leases, any DA would need to be referred to both the Office of Water and DPI (fisheries).
4 What action has Council taken so far to investigate the dumping of soil at this location?
There has been no formal complaint lodged through Council's complaint system. The matter was identified by a member of staff who noticed several piles of material dumped along the ridge of the property which generated the initial investigations.
5 Has Council undertaken an on-site inspection?
Due to the landowners being abroad for several months, Council officers could not gain permission to enter the property. Photos were taken from neighbouring properties and due to the lie of the land Councils officer could not see the area in question.
6 Where is the soil being deposited? Is it on private land, Crown land or both?
Based on aerial photos it is considered that the placement of the soil is within the bounds of Lot 11. An onsite inspection is being carried out today by Council Officers to confirm this.
7 If the activity does require consent from Council and has been undertaken illegally, what action has been taken so far to address the matter?
It may be that the proximity of the fill to the river (within 50mtrs) would trigger the requirement for a Development Application. Pending the outcome of the onsite inspection today a decision will be made as to what action is required.
Should an application have been required the landowner will be given the opportunity to show cause why an infringement notice should not be issued and Issue a stop work notice so no further fill is imported to the site until the matter is fully investigated.
8 For what purpose is the soil being deposited?
As there has been no formal investigation or interview with the owners of the land at this stage, (noting an onsite meeting today) Council does not know the purpose of the fill being deposited.
From air photos from 2000 to 2010 it does not appear that any land has been reclaimed, this will be confirmed at today's onsite inspection.
10 Will the dumping of soil at this location affect flooding patterns in the area?
The subject area may be out of the 1:100 year ARI affected land however this will need to be confirmed.
11 Has the dumped material been screened for contaminants?
Council is unaware but will be confirmed through today's onsite inspection.
There were no Questions for Closed Meeting where due Notice has been received.
General Manager Report
3226/11 RESOLVED: (Ainsworth/Moran)
That the list of outstanding actions and reports be noted and received for information by Council.
ITEM 10.2 SF319 171111 Macksville Showground Committee of Management - Minutes of Annual General Meeting - 26 September 2011
3227/11 RESOLVED: (Moran/Ainsworth)
That Council endorse the Minutes of the Committee of Management for the Macksville Showground's Annual General Meeting held on 26 September 2011 and thank the Committee for their continuing good work over the past twelve months.
3228/11 RESOLVED: (Moran/South)
That the Accountants' Report on Investments placed to 31 October 2011 be noted.
General Manager Report – LATE
ITEM 10.4 SF1609 171111 Grants - Regional Development Australia Fund Round 2 - Inability to Apply
3229/11 RESOLVED: (Hoban/Court)
That Council note the report and write to Minister for Regional Australia, Regional Development and Local Government, the CEO of RDO Mid North Coast as well as the Local Government and Shires Association expressing Council's utter disappointment that the new guidelines for grants for Regional Development Australia Fund Round 2 exclude smaller rural and regional councils accessing the substantial funds made available by the Federal Government and request that the guidelines be reviewed.
Director Environment and Planning Report
ITEM 11.1 SF1589 171111 DEP Applications and Statistical Reports July 2011-June 2012, 2007-2011 and Certificates Received 2007-2011
3230/11 RESOLVED: (Moran/South)
1 That Council note development application statistics and processing times for July 2011-June 2012 compared with July 2010-June 2011.
2 That Council note Occupational Certificate released for the years 2007 to 2011.
3 That Council note the statistical information for Applications and Certificates received by Council for 2007-2011.
ITEM 11.2 SF1148 171111 Contract Regulatory Officer's Report October 2011
3231/11 RESOLVED: (Flack/Ballangarry)
That the report from the Contract Regulatory Officer for October 2011 be received and noted by Council.
ITEM 11.3 SF1589 171111 Outstanding DA's greater than 12 months, applications where submissions received not determined to 4 November 2011
3232/11 RESOLVED: (Ainsworth/Moran)
1 That the list of outstanding development applications (in excess of 12 months old) be noted and received for information by Council
2 That the applications where submissions have been received be noted and received for information by Council.
ITEM 11.4 SF1589 171111 DA's and CDC's Received and Determined under Delegated Authority to 4 November 2011
3233/11 RESOLVED: (Moran/Smyth)
That Council note the Development Applications/Complying Development Applications received and determined under delegated authority.
Item 11.5 was dealt with under Delegations.
Director Engineering Services Report
ITEM 12.1 SF203 171111 2009/10 NSW Water Supply and Sewerage Performance Monitoring Report
3234/11 RESOLVED: (Ainsworth/Ballangarry)
That Council note the release of the 2009/10 Water Supply and Sewerage Performance Monitoring Report by the Minister for Primary Industries and receive and note the summary to address Triple Bottom Line Performance Report, Commentary and Action Plans to address areas of under performance.
Director Engineering Services Report – LATE
ITEM 12.2 SF565 171111 Request for extended duration of temporary road closure of Ridge Street Nambucca Heads between Bowra Street and Estuary Lane
3235/11 RESOLVED: (Moran/Flack)
1 That Council approve the extended duration of the temporary road closure of Ridge Street, Nambucca Heads between Bowra Street and Estuary Lane between 3.30 pm and 9.30 pm on Thursday 1 December 2011 for the Christmas Street Event hosted by the Nambucca Heads Chamber of Commerce.
2 That Council readvertise the Christmas Street Event with the amended hours 7 days prior to the event.
Director Engineering Services Report – LATE
ITEM 12.3 SF593 171111 Nambucca Flood Investigation – Phase 2 Seaview Street, Nambucca Heads
3236/11 RESOLVED: (Flack/South)
That this matter be deferred.
General Purpose Committee REPORT
ITEM 9.1 SF820 171111 Quarterly Performance review - 30 September 2011 (Item 9.1 in the General Purpose Committee)
3237/11 RESOLVED: (Court/Flack)
That Council note the extent to which the Performance Targets have been achieved for the quarter ending 30 September 2011.
ITEM 9.2 LF3123 171111 Nambucca Valley Youth Services Centre (Item 9.2 in the General Purpose Committee)
3238/11 RESOLVED: (Flack/South)
1 That Council's Property Officer consider the proposed licence conditions which have been put to the Nambucca Valley Youth Services Centre (NVYSC) by the Land and Property Management Authority and provide advice to the NVYSC in relation to the proposed conditions.
2 That if possible, Council's Property Officer be available to attend a meeting with the NVYSC and the Land and Property Management Authority to discuss the terms of the proposed licence agreement.
3 That Council act in an advocacy role to support the NVYSC's continued occupation of the building noting the importance of the services they provide and their need for security of tenure to continue to provide those services.
ITEM 9.3 SF1460 171111 Review of Rating Structure (Item 9.3 in the General Purpose Committee)
3239/11 RESOLVED: (Hoban/Ainsworth)
1 The structure of Farmland Category be changed to incorporate the statutory minimum with ad valorem maintaining current yield. Minimum amount to be increased by approved rate peg annually.
2 Council undertake a review of the current farmland criteria to better reflect higher intensity farming pursuits. Future farmland inspections to be assessed on the revised criteria.
ITEM 10.1 SF135 171111 Draft Supplementary State of the Environment Report 2010-2011 (Item 10.1 in the General Purpose Committee)
3240/11 RESOLVED: (Ainsworth/Court)
That the Draft 2010/2011 Supplementary State of the Environment Report be endorsed for submission to the Department of Local Government.
ITEM 10.2 SF1541 171111 Planning Proposal - Lot 1 DP80572 Egan Street Macksville (Item 10.2 in the General Purpose Committee)
3241/11 RESOLVED: (Hoban/Ainsworth)
That given flood modelling results for Macksville are imminent, that Council defer a decision on this proposal until their completion and the applicant be advised in the meantime to consider addressing the outstanding issues raised in the report to the General Purpose Committee.
For the Recommendation Councillors Hoban, Ainsworth, South, Finlayson, Court (Total – 5)
Against the Recommendation Councillors Moran, Ballangarry, Flack and Smyth (Total – 3)
Note: Cr Moran requested that his vote be recorded as against this resolution because he believes we are bringing issues into things when we haven't got them in place.
ITEM 11.1 SF826 171111 Working Trial of Asphalt Zipper (Item 11.1 in the General Purpose Committee)
3242/11 RESOLVED: (Flack/Smyth)
That Council note the trial of the asphalt zipper but await a report on the trial including advice as to all of the costs involved with using the machine.
ITEM 11.2 SF593 171111 Nambucca Heads Flood Investigation - Phase 2 Seaview Street Catchment (Item 11.2 in the General Purpose Committee)
3243/11 RESOLVED: (Flack/Court)
1 That Council adopt the option to retain the existing and approved drainage system within the Seaview Street Stormwater drainage catchment which was constructed to the standards and knowledge of the day.
2 That Council adopt the option to retain the existing and approved drainage system within all other established stormwater drainage catchment areas within the Nambucca Shire Council LGA which was constructed to the standards and knowledge of the day.
3 That in accordance with the Australian Rainfall and Runoff (AR&R) and AUS-SPEC#1 Design Specifications September 2002 future developments shall define a surcharge path for systems even where a 100 year Annual Recurrence Interval flow can be maintained within the system.
4 That in accordance with the Australian Rainfall and Runoff (AR&R) and AUS-SPEC#1 Design Specifications September 2002 future developments are to provide easements within private property over pipe systems and surcharge paths.
5 That the following recommendation be deferred to Council's meeting on 17 November 2011 to allow for definitions to be provided for the terms and categories used:
That Council adopt a drainage design standard in accordance with the Australian Rainfall and Runoff (AR&R) and AUS-SPEC#1 Design Specifications September 2002 for future urban developments as follows:
100 years for the 'major' system in all developments
20 or 50 years for intensely developed business, commercial and
industrial areas
10 years for other businesses, commercial and industrial areas and
intensely developed residential areas
5 years for other residential areas and open spaces
In addition, where a development is designed in such a way that the
major system flows involve surcharge across private property, then
the underground system (both pipes and inlets) shall be designed to
permit flows into and contain flows having an ARI of 100 years from
the upstream catchment which would otherwise flow across the
6 That Council instigate a detailed stormwater management plan for the Valla Beach, Nambucca Heads, Macksville, Bowraville and Scotts Head as funding and resources become available which explore options and opportunities for funding towards flood conveyance improvements.
ITEM 11.3 SF788 171111 Waste and Sustainability Improvement Payment Program (Item 11.3 in the General Purpose Committee)
3244/11 RESOLVED: (Court/Ainsworth)
That Council endorse the proposed projects as options for the allocation of future Waste and Sustainability Improvement Payments with the exception that the Sustainability Officer not be considered at this time.
ITEM 11.4 SF1092 171111 Draft Plan of Management for Part Lot 40 DP 711098 Old Coast Road, Kingsworth Estate (Item 11.4 in the General Purpose Committee)
3245/11 RESOLVED: (Court/Ainsworth)
1 That Council endorse the draft Plan of Management for Part Lot 40 DP 711098 Old Coast Road, Kingsworth Estate and place the document on public exhibition for a period of 28 days seeking public comment.
2 That Council receive a further report on the draft Plan of Management for Part Lot 40 DP 711098 Old Coast Road, Kingsworth Estate at the completion of the public exhibition period.
There being no further business the Mayor then closed the meeting the time being 7.10 pm.
Confirmed and signed by the Mayor on 1 December 2011.


text:
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- Price$484,000
- Beds4
- Baths2
- SQ. Feet1,880
- Acres5.00
- Built1936
879 Oliverea Road, Big Indian
Traditional Swiss Chalet in Big Indian Enter another world of elegant and relaxed European Charm. Approx. 1900 SF 4 BR, 2 BA with Mt. views, just 15 min. from Phoenicia and 5 min. to Belleayre Ski Ctr. This meticulously maintained complex features the Main Lodge, a 2 car garage, a Writer's Studio, a Tool Shed and a Handyman Shed all nestled in nooks and crannies within the slope side portion of the 5 acre wooded property bounded by NYS State Lands. Lodge features custom crafted fieldstone fireplace, a tavern room with full bar and special cook's hooded fireplace in the brand new gourmet kitchen with polished stone counters and S.S. appliances. Unique hand crafted doors moldings and trim throughout the interior and exterior. New roofs on lodge and garage, fully insulated, stained glass windows, brand new 4 BR septic systems, and waterfall feature in campfire area, workshop basement and open sun filled TREK decks and custom stone patio, THIS IS TRULY A ONE OF A KIND DWELLING ready to go with SUPERB attention to detail.
Essential Information
- Price$484,000
- Bedrooms4
- Bathrooms2.00
- Full Baths2
- Square Footage1,880
- Acres5.00
- Year Built1936
- TypeResidential
- Sub-TypeSingle Family
- StyleCHALET
- StatusACTIVE
- FeaturesBAR-DRY, CEILING-BEAMED, CEILING-CATHEDRAL
- ParkingDRIVEWAY, OFF STREET, 2-Car
- # of Garages2
- GaragesGarage Detached
- ViewMOUNTAIN
- Lot DescriptionADJACENT TO STATE LAND, LANDSCAPED, LEVEL, MEADOW, PRIVATE, SLOPE-GENTLY, SLOPE-MODERATE, SLOPE-STEEPLY, WOODED
- RoofASPH SHINGLE
- ConstructionFRAME/STICK, LOG
Additional Information
- ZoningR 1
Community Information
- Address879 Oliverea Road
- AreaSHANDAKEN
- CityBig Indian
- CountyUlster County
- StateNY
- Zip Code12410
- AppliancesDISHWASHER, DRYER, EXHAUST FAN, RANGE, REFRIGERATOR, WASHER
- HeatingFORCED AIR, ABV GROUND FUEL TANK
- CoolingCEILING FAN, CENTRAL
- FireplaceYes
- FireplacesFREE STANDING, STONE
School Information
- DistrictONTEORA CENTRAL
- ElementaryPHOENICIA
Listing Details
- Listing AgentFaye Storms
- Listing OfficeRUTH GALE REALTY
Price Change History for 879 Oliverea Road, Big Indian, NY (MLS® #20192998)
|Price Reduced (from $499,000)
|$15,000 (3.01%)
The data relating to real estate on this website comes in part from the IDX of the Multiple Listing Service of Ulster County, Inc. Real estate listings held by brokerage firms other than Coldwell Banker Village Green Realty are marked with IDX logo and the detailed information about them includes the name of the listing broker.
Copyright © 2019 by the Multiple Listing Service of Ulster County, Inc. All Rights Reserved.
All information provided is deemed reliable but is not guaranteed and should be independently verified.
IDX information is provided exclusively for consumers' personal, non-commercial use and may not be used for any purpose other than to identify prospective properties consumers may be interested in purchasing.

subset: commoncrawl


text:
Heading for the light
Today I ran the New Balance Waterfront 15km in Ajax and, I have to say, I felt great. I ran it as a workout, 12 x 1km hard with 250m "float" where I averaged 2:56 for the 1km repeats and around 3:30/km for the 250m float. That netted me 46:15 for 15km. Cool weather today was a relief after having a couple of my big efforts in September done in hot and humid conditions. Not Doha hot, not even close, but still hot enough to affect pace.
Rock 'n Roll Philly Half on September 15th was a good effort and I have to be pleased with running 65:47 in hot conditions. The course is fast and I can see how one would run fast there under the right conditions. I felt really good that morning and found a nice pack early on and just focused on running with the pack and not looking at my watch.
As we approached 10km my pack had recently split apart and I saw my 10km split on the clock. I was expecting to see 31:00 so when I saw 30:32 I got excited and thought today was going to be alright in the heat. After that I was paying attention to my splits (to stay honest whilst running alone) and after 12km I kept getting slower and slower. I was catching up to a few athletes so it felt as though I was still moving ok but my pace kept suffering.
Seeing 65 on the clock was disappointing but once I saw the full results and talked to other athletes who thought they were going to run much faster it put things in perspective.
On Wednesday I got a knock on the door at 6:30am after having just gone to the bathroom. That meant I had to drink a couple bottles of water and a couple cups of tea to produce a urine sample for the drug testers. After blood and urine samples completed I drove to Guelph for a my longest session of the build, (5km warm up, 26.2km marathon effort, 4km cool down).
During the 26km marathon effort my stomach didn't feel great and I blamed it on the extra fluid I drank that morning. However when I got done my cool down I found out my son had puked up his breakfast that morning. Perhaps I had a little bug too? But I really didn't feel that bad and still averaged 3:11/km for the 26km. I was hoping to average 3:09/km for that session so I really wasn't that far off and it was a little warm.
That's why it was sweet to run 46:15 for 15km today when I had expected to run about 47:00.
STWM should be an interesting race for the spectators. There are a lot more quality Canadian men than usual because of the trials nature of the race. I could see a big pack going out faster than 2:14 pace. Whereas usually there are only 1-4 athletes going out at that pace or faster.
Cam Levins, Tristan Woodfine, Trevor Hofbauer, Thomas Toth, Aaron Cooper, Rory Linkletter, Evan Esselink, Kevin Coffey, Sami Jibril, Chris Balestrini and Dylan Wykes. I might be forgetting someone right now but that is a lot of guys who have potential to run a fast marathon.
STWM OCT 20, three weeks to go!
Coming around
On August 3rd I ran the Beach to Beacon 10km in Cape Elizabeth, Maine. The race is fantastic and I highly recommend it, I had a great time. In fact, I was enjoying it so much I thought I'd take more of it in and stay out on the course a little longer (that's a joke). I ran 30:45 for 10km and truthfully that was about 45 seconds slower than I thought I was going to run. I was rattled by my performance because I just couldn't get it going that day. In the big picture I wasn't too worried because I really felt I was just flat from upping my volume plus, I had another race in 3 weeks to turn the tables.
I started training for the Scotiabank Toronto Waterfront Marathon towards the end of July. Right before I started the build I took a few days off and went fishing with friends in Northern Ontario. As usual I crave some downtime, eating whatever and drinking beer. And then, as usual, I'm craving the hard training even more.
My second race in August was the Crim 10 Miler in Flint, Michigan. Also a very fun race, great atmosphere although not as scenic as Maine. My goal at Crim was to run a faster pace than Beach to Beacon. Luckily I found a good pack to run with so I never looked at my watch. Well, not until after 9 miles. I rolled with this chase pack, faded off from them up the hills around mile 5 and then caught back up with them. And then faded again around 8 miles to finish 14th.
I averaged 3:03/km (faster than Beach to Beacon) and ran my last two km at 3:01/km. Needless to say I was pretty happy with the way Crim went and was pleased to get the Canadian Masters record 49:13 (as long as I get the paperwork done). The nice weather at Crim certainly helped with the fast pace but more than that I was able to push into a gear that wasn't available at Beach to Beacon. Simply getting used to 180km weeks made the biggest difference.
Next up is the Rock & Roll Philly Half on Sept 15th.
On the line Paul boasted that he once led the Crim.
He didn't lead this year but he did get out hard and got in front of me. I think I passed him 100m into the race.
Worst blog ever
Whoa, July just zipped by! It was only after 8pm on July 31st that I realized I hadn't wrote a blog this month. I thought about writing one earlier this month after I raced Edmonton but decided to put it off in hopes I'd have more to share by now… Should have written a blog a couple weeks ago.
Canada Running Series once again put on a stellar event at the LuluLemon 10km in Edmonton (July 7). My expectations going into the race weren't high as it was only 6 weeks after the Ottawa marathon. I took a full week off running and then a very easy week after the marathon. I won the race in Edmonton the year before but I had a lot more time after my Spring marathon.
Early on during the 10km I felt as though I was holding onto the chase pack (Levins, Watson, Ballestrini, Coffey, Rawlings) and wasn't even concerned about Essalink and Chesoo a few seconds up the road. Sometime before the halfway mark I put myself into the mix and started to feel better. My previous 3 races I had talked myself out of it for one reason or another and this time I started to tell myself "it's just an early season race." But I pushed those thoughts out of my head and found the fire to compete.
At 9km there was still Levins, Balestrini, Coffey and myself shooting for the last place on the podium. I started to push the pace hoping to drop anyone, or everyone. As we rounded the corner with less than 50m to go Levins went by me pretty quickly and I looked back, saw Balestrini and knew I had to keep on the gas or get passed. I held onto 4th place and was very pleased with the result. It had been quite a while since I surprised (a good surprise) myself in a race.
Since then my training has progressed well and I'm starting to raise my volume ahead of a fall marathon. In the meantime I have a couple races that I'm really looking forward to:
August 3rd – TD Beach to Beacon 10k
August 24th – Crim 10 Miler
Beach to Beacon will be my first race as a master. Although I like the idea of competing for an added prize money pool my ambitions to do well in the overall race are still my goal.
Ok, time to pack for my flight to Portland, ME tomorrow.
Like some cheese-eating high school boy
I re-watched the Ottawa marathon broadcast. I can't believe how annoying it is to see myself always flicking sweat off my face.
On a hot run earlier this month I started to sweat and, consequently, flick sweat off my face. Then I tried to not touch my face. When a big drop of sweat is sitting on my face it's like an itch and I automatically swipe it. It took all my mental fortitude to not touch my face and the instant something distracted me, (such as looking for traffic) I would wipe the sweat away. I think when I'm focused during a race that if I tried not to touch my face I'd lose focus on trying to go fast.
Anyways, recovery from Ottawa went well and I resumed running after a full week off. My first week was easy and then I started to ease into workouts. I still feel sluggish 5 weeks after the marathon (one week out from Edmonton Lululemon 10km) but each week I see improvement.
Last year I won Edmonton 10km but it was 12 weeks after Boston. 12 weeks and 6 weeks is a massive difference. Thankfully my training has gone smoothly and I hope to have a decent race and a kickstart to the rest of the summer season.
On a run last week I passed a group of high-school boys walking on the rail trail. About 100m later one of them was running beside me. He was wearing his school uniform and did not look like a runner.
Me: "Only 26km left."
High-school kid: "You got something on your hat." And flicked the top of my hat.
Me: "Showing off for your buddies, eh?"
HSK: "Actually, I get five bucks if I run beside you until the top of the hill."
Me: "Hmmm, I'll have to decide if you get to keep the five dollars or not."
And then I picked up the pace for a few strides. He cockily smiled as he kept up. Five seconds later he realized he was in big trouble and the smirk quickly disappeared. I slowed back down and let him regain his composure. At that point we started to go up the hill and I had to slow down.
Me: "Maybe I'll let you get the five bucks."
HSK: Out-of-breath.
Me: "We're crawling now and this is getting boring, I'm going to put you out of your misery" I then took 3 fast steps and realized he had nothing left. I felt for him so slowed way down for the final 20m of the hill.
Me: "Let me know when you've earned the five bucks."
We crested the hill side-by-side.
HSK: "That's…good….thanks"
Ottawa 2019
Last minute blog to maintain posting once a month. Here we go.
Running 2:17:37 at the Ottawa marathon was about four minutes slower than I originally wanted but it wasn't all that bad. Here are the pros and cons.
Training went really well. It was my best build-up since 2016.
I felt comfortable running ~3:10/km early in the race. In the past I've been able to feel a pace and figure out if that pace was sustainable or not.
I got all my bottles and consumed more carbs than planned. I've never done that before.
Ottawa was a really fun race and the crowd support was amazing.
First Canadian came with a good payday. 8th overall paid as well and added some bonus points towards world ranking.
Recovery went well and I felt like going for a run on Wednesday (I didn't, I'll wait a few more days).
I didn't change my planned pace even though it was 17C at the start and got hotter when the sun came out. I was too stubborn.
The humidity made it a tough day and I struggled to push really hard in the final 7km.
It was a little lonely out there, I only passed 2 runners throughout the race and once I caught them I didn't see another runner as they were too far up the road.
Even with the bonus points for finishing 8th my time was too slow to really help with Olympic qualification.
The morning before and after the race the temps were cool and it was calm. I know that doesn't make a difference for the actual race, but it somehow makes it more frustrating to think of what could have been.
On the topic of Olympic qualification. (Edited June 1st after I crunched numbers on IAAF.org again) I feel that it will take about 1120 points to be ranked high enough to make the Olympics. [I still have to calculate how many runners not ranked in the top 80 (3 per country) have achieved an auto standard of top 10 at WWM, top 5 at GL…]. That's just a guess and to be safe I'm aiming for 1125 points. A 2:12:05 gets you 1125 points. If I had run 2:13:48 (1095 points) and finished 8th at Ottawa, the extra 30 placing points would have given me one performance at 1125 points.
Or, just get the auto 2:11:30. Right! haha
Running 2:11:30 doesn't seem likely for me (unless training really improves in the next few months). However, I do feel that sub 2:13 is within my capabilities and that might be good enough if I can get two in that range with some placing points.
My last three races have been my slowest marathons of my life (Boston 2018, STWM 2018, Ottawa 2019) which is starting to screw with my head more than it should. I know why each race was slower than it could have been but I'm still guessing as to where my top-end lies.
I'll have to wait about 5 months to have another crack at the marathon. In the meantime I'll get after some shorter races and chase the Speed River guys to find some turnover.
Weather in Ottawa on Sunday was not ideal.
A few people have already guessed this announcement: I'm planning on running Ottawa Marathon on May 26th!
In my last blog I wrote how I was pulling out of the Hamburg marathon because I was behind in my training. It was an easy decision for me because I knew that if I could get quickly back into training then I'd have plenty of time to prepare for Ottawa. When I was scheduling a Spring marathon my five possibilities were Hamburg, London, Prague, Ottawa and Boston. It made sense to race at the end of April because we planned a Feb/Mar/Apr training camp/vacation and Hamburg had a good pace option for me. But when that date was out the window it was a no-brainer to pick Ottawa.
After a couple of relatively low-key workouts in early April to get back into the groove things started to click again. I had a solid 60′ marathon-effort on April 19th (with coach DST following in a car with bottles) and then a few days ago I ran 4 x 5km and was pumped to have averaged 15:52. I've been running my fartlek sessions and long-runs with the Boulder Harriers and those have gone well, having people to push me has been very valuable. I've averaged about 180km/week the past four weeks and that seems to be my sweet spot these days. With one month until Ottawa I'm happy where my fitness is.
10 years ago I ran my debut marathon in Ottawa (2:17:09). I've always wanted to head back but I've actually only run four Spring marathons since then (2013 Rotterdam DNF, 2014 London 2:13, 2015 Rotterdam 2:11, 2018 Boston 2:25 9th place). It's funny to think how green I was to the marathon in 2009. The thing I remember most is being very curious as to what the final 12 kilometres were going to feel like, (spoiler: it hurt!!). I'd say my race in Ottawa was a success as I hit my goal (qualified for 2009 World Champs) and wanted to race another marathon. I'm heading back to Ottawa with a good feeling about the race.
Some funny things I wrote down 10 years ago in my post-Ottawa blog: The most I ran in a single week leading up to Ottawa was 145km. I only had two days where I ran twice in the same day, (but many days had a second x-trainign session). I only planned racing the marathon 6.5 weeks before race day. Due to breaking my foot the previous November, I only ran 96 of the 168 days leading up to the race and my first running workout was March 29th.
I finally made it to the infamous Magnolia Road last weekend for a long-run. It reminds me a lot of the hilly dirt roads around Iten, Kenya. Even though they are dirt roads the undulation beats your legs up plenty. Great marathon preparation and great views.
Where do we go now?
March flew by and I need to keep my 2019 resolution to blog at least 1 x month. But this isn't a blog to simply check a box, lots to report here.
This month started off really well. On March 1st I did a session that involved 2km hard followed by 1km "float." I did this for 20km straight and averaged 3:19/km for the whole thing (2km @ 3:08/km and 1km @ 3:42/km). This was over 8 weeks before Hamburg and I felt that I was in a great position to have a good marathon on April 28th.
Two weeks later I caught a flu (vaccine didn't protect me from this strain) and was in bed for two days. I missed a big workout (6 weeks out) but only missing one workout isn't a big deal. My first workout after being sick was compromised but I was improving and thought I might as well still race Papa John's 10 Miler in Louisville, KY that weekend (March 23rd).
I flew to Louisville on Friday night and met my parents for dinner. Raced the next morning at 8am and was back on a plane at 3pm. I was only in Louisville for 20 hours.
The race itself wasn't great. I could tell 1km in that I wasn't running with full power and knew I should change my expectations. Those expectations kept getting slower throughout the race and I ended up averaging about 3:10/km for 50:42 (5th place).
Although I know I should have been faster than 3:05/km if I was feeling good it was still a solid effort and a good session to prep for Hamburg. My legs didn't feel sore from the race but I think the downhills started to throw something off in my back.
I decided to get back on schedule right away and do my long-run Sunday. The run itself went well but that afternoon my back was stiff and sore. I didn't think it was anything to worry about and was telling Marie that if everything fell into place I was confident I could have a good race in Hamburg. The next day things fell out of place.
The next morning (March 25th) I ran easy for 15km but I felt off right from the beginning. My back was sore and I felt I was protecting it each stride. After the run it started to tighten up really fast. Luckily I had a massage already lined up.
The massage loosened everything up and I left there feeling pretty good. But 10 minutes later I was in pain again. My friend told me about a good chiro I could get in to see. He concluded that my rib head was out of place and that my vertebrae was also mis-aligned. I had immediate relief but I could tell this wasn't over.
The next afternoon I had brutal back spasms where my muscles around my rib head would tighten up and not let go. I've never felt much relief from ibuprofen in the past but it's been a saviour for me lately. Without it I was pretty much immobile on Tuesday.
Wednesday I could tell that I was actually recovering. No more spasms and the ache was duller. I've run short and easy the past two days but it still hurts to breathe deeply. I've had my rib pop out before. About 13 years ago it happened and I recovered from it pretty quickly once I saw a chiro and had it adjusted. I'm hoping that this can be similar and by early next week be back into normal training.
However, I think that is just a little too late to have a legitimate shot at a good race in Hamburg. Missing a workout 6 weeks out, then having some compromised training and now missing all quality this week and likely just easing back into workouts next week is just too much. I won't be racing Hamburg marathon. A bit of a bummer, but there will be other races.
In hindsight the long-run the day after my race may have been the culprit for putting my back out. Perhaps I had run easy and got proper recovery it wouldn't have gotten so messed. Who knows?
At this point I'm training to qualify for the 2020 Olympics so I only want to race unless I have a good shot and making headway towards qualification. Qualifying for the Olympics is whole other topic. I wasn't going to write about it because I know it will take a long time. But I'll try and break it down without writing a novel.
The IAAF announced the Olympic standards for Tokyo 2020 this month. There is a new way to qualify that is different than past Olympics. This time there is a set of standards that are supposed to capture 50% of the field (2:11:30 for men's marathon). The next 50% will be filled from the world rankings.
With the IAAF aiming to have 80 marathoners in each the men's and women's marathon (there were 155 and 156 in Rio for men and women) the standards will be quite harder for 2020. But not necessarily for Canadians. In 2016 the IAAF standard for men was 2:19:00 and the Athletics Canada standard was 2:12:50. You can most-likely run slower than 2:12:50 and qualify for Tokyo via the IAAF qualifying process.
I calculated that in order to be ranked in the top 80 (3 per country) you'll need to run about 2:14:00 twice, or 2:14:00 and 1:02:29 for the half (you need two results in the past 18 months to have a world ranking). Basically, your average time will need to be about 2:14:00. So a 2:12:00 and a 2:16:00 might be good enough. There are bonus points you can get from placing well at certain races (World Marathon majors and Gold Label races have good points for top 10).
Fot instance, if you place 8th at a Gold Label marathon (STWM and Ottawa are gold label btw) you get an extra 30 points. If you run 2:15:15 and get 30 points that is equivalent to running 2:13:31. So yeah, points can make a BIG difference when trying to get a good world ranking.
Also, if you finish top 5 at a Gold Label marathon or top 10 at a World Marathon Major you get an auto qualifier. If there are many athletes hitting the 2:11:30 standard and placing well at majors/gold label who aren't ranked then you'll need to be up higher than top 80 (3 per country) in the world rankings.
But the big thing for Canadians will be what Athletics Canada decides to do. If they go with the IAAF qualifying route then they might have to compare and choose between athletes who have qualified under different routes. Take this for example:
Athlete A: 2:09:30 (auto standard, only result = no world ranking)
Athlete B: 2:12:45 and 2:13:15 (ranked 65th in world rankings)
Athlete C: 2:14:30 and 1:01:45 (ranked 60th in world rankings)
Athlete D: 2:18:00 (8th at Boston) and 1:03:00 (ranked 75th in world)
Athlete E: 2:14:15 (10th at New York City, only result= no world ranking)
Athlete F: 2:14:00 and 2:15:00 (1st Canadian at STWM and ranked 78th in world rankings)
Here are 6 athletes who have met IAAF qualification for 2020 Olympics. Who do you pick? (Each country can only pick 3 who have met IAAF qualification).
I say you go off of fastest marathon time for those who have qualified, after you add the first Canadian at STWM (AC already announced that that athlete would get auto-selected given they meet minimum IAAF standard). It's too hard to say what is better, a 10th at Boston or 8th at NYC.
And it might not be great to go off of world rankings because there might be a half marathon specialist who isn't great at the marathon. Or, you might get someone who is 2 minutes slower but got an extra 35 points for a certain placing and therefor is ranked higher. Or, an athlete who isn't even ranked because they only have one result but that result is a great marathon time.
Here are the three athletes I would pick:
Athlete F (due to first Canadian at STWM rule already in place)
Athlete A 2:09:30 is the fastest marathon
Athlete B 2:12:45 is next fastest time
I could go on and on about the new world ranking system. But I'll say this: I don't like it. It's too confusing to the average fan and the way placing points are distributed isn't going to be fair.
Boulder has been fantastic. There are so many great places to run here with lots of variety. I often end up driving 10-20 minutes to get to trails but it's worth it. I still have yet to run Mags, but once my back is better it's on the list.
We've been on some pretty cool hikes too. Our favourite was Rocky Mountain National Park. We weren't expecting so much snow because we didn't realize how high we were going. At almost 3000m (10,000 feet) of elevation you're going to get a lot more snow! (As a note: I haven't loaded Louis on my back, lifted weights or run many hills in the week leading up to my back injury because I was prepping for the 10 mile race, and sick the week before that).


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Volunteers are requested to provide a sausage sizzle for the BBQ&A being organised by the Autism Hub. It is on the 30th July '18 with sizzle preparations starting at 5:30 PM and finishing around 7:30 PM.
There is a button in the events area for the evening of 30th July where you can let us know that you can come.
We will be using the barbeque behind the back sliding door and with some luck will have lighting out there. Bring camping lights to be on the safe side!
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Types of flag poles; especially for desk flags, the pole types are as follows.
Desk flags; there are six types namely: Single pole, Double pole, Triple pole, Quartet pole, Type L pole and Type T pole.
Office flags; there are two options: Brass and Chromium flag poles.
Desk Flag Poles Types
Telescopic Flag Poles Types
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Hundreds of aspirant teachers appear in test
Jacobabad, Sindh: Hundreds of aspirant teachers appeared in the test for the recruitment of primary schoolteacher here Sunday. The National Testing Services (NTS) conducted the test in Government KG School and Government Girls High School. There were 648 female candidates registered for the PST test and out of them 516 appeared in the exam, while out of 984 registered male candidates 857 appeared in the test. The result of the test would be announced in a week.


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Rob & Nikita Proposal & Engagement
Despite having lived in NYC for the past five years, I've never been to the New York Botanical Garden until Rob reached out to me to ask about proposing to his then-girlfriend, Nikita. He told me they were both nature people, so the NYBG would be the perfect spot for the proposal. Because they're both from out of town, I visited the Azalea Garden, where he planned to propose, the week before to check out the location and figure out the best spot to pop the question.
Despite some miscommunication that happened with the garden regarding the logistics of it all, the proposal went well and she said yes to a future with Rob and also to a stunning ring! Check out the photos below, and hear the story in Nikita's words on The Knot's How He Asked! (The link will open in a separate window)


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Purpose: Wilms' tumors (WTs) exhibit more than one pattern of differentiation, each of which is associated with distinctive clinical features and treatment responses. Mutations in the WT1 gene are found predominantly in WTs with stromal histology. To better understand the biological and clinical features in different WTs, we have analyzed WTs with and without WT1 mutations for a set of parameters.
Experimental Design: Twenty-two new WTs were analyzed for WT1 mutations by PCR single-strand conformational polymorphism. Five tumors with WT1 mutations and six tumors without WT1 mutations were studied for the presence of WT1 transcripts and protein as well as for the expression of differentiation markers.
Results: Two new WT1 mutations were identified in stromal-predominant tumors, and none were identified in the other histological subtypes. Tumors with WT1 mutations expressed mutant messages, cytoplasmic truncated WT1 proteins, and muscle markers. In contrast, blastemal-predominant tumors without mutations showed nuclear WT1 protein staining. Both tumor types were positive for markers of early-induced mesenchyme and one marker of uninduced mesenchyme, but blastemal-predominant tumors also expressed cytokeratin, suggesting that these are further along the epithelial differentiation pathway.
Conclusions: Our data show that the two-hit inactivation of WT1 is operative in stromal-predominant WTs. Cells without functional nuclear WT1 protein start a faulty differentiation program. In contrast, blastemal-predominant tumors express wild-type WT1 and show early signs of epithelialization. The extensive rhabdomyomatous differentiation and the presence of WT1 mutations may be used as a diagnostic tool to identify a tumor subtype that seems to respond poorly to chemotherapy. These studies provide a foundation for improvement in tumor classification and ultimately for the development of more individualized tumor treatments.
WT3 or nephroblastoma is the most frequent renal tumor of childhood, affecting 1 in 10,000 children, usually before the age of 5 years (1) . Histologically, it mimics various stages of nephrogenesis, indicating aberrant differentiation of a multipotential mesenchymal renal stem cell (2) . Most tumors contain a mixture of undifferentiated blastemal cells, as well as differentiated epithelial and stromal elements. However, heterologous components such as striated and smooth muscle, cartilage, bone, or adipose tissues are observed in approximately 10% of WTs. Tumors with extensive rhabdomyogenesis have been called "fetal rhabdomyomatous type" and occur in younger children and are frequently bilateral (3 , 4) .
Nephrogenic rests, considered to be precursor lesions of WTs, are found in 30–44% of tumors (2) . ILNRs, located deep within the parenchyma and indicating an earlier developmental disturbance, are associated with intralobar WTs containing heterologous elements. ILNR-like WTs are observed in patients with sporadic WT often associated with GUs, anirdia, or Denys-Drash syndrome. PLNRs occur at the periphery of the renal lobe later in renal developmental and are associated with perilobar WTs lacking heterologous elements. PLNR-like WTs are often found in patients with hemihypertrophy and Beckwith-Wiedemann syndrome. Recently, Ravenel et al. (5) identified LOI of IGF2 as a valuable molecular tool for the classification of these two pathological subtypes. LOI and altered expression of IGF2 were found mainly in PLNR-like tumors.
The WT1 gene, a ZF transcription factor, was found to be mutated in 10–15% of sporadic WTs (for a review, see Ref. 6 ). Recently, we identified a high percentage of WT1 germline mutations in stromal-predominant/ILNR-like tumors (7) . Loss of the wild-type allele (LOH) was found in 7 of 13 analyzed cases, confirming Knudson's two-hit hypothesis for this subclass of WTs. All but one were nonsense mutations predicted to result in protein truncation. Only one missense mutation was found in a blastemal-predominant tumor, suggesting that mutations in WT1 may play a minor role in the development of this subtype (7) .
Alternative splicing of the WT1 gene affecting exons 5 and 9 results in the synthesis of four major isoforms of the protein with different nucleic acid binding and transcriptional properties (8, 9, 10) . The WT1 protein has many postulated functions both in regulating transcription and in posttranscriptional RNA processing (reviewed in Ref. 11 ). It is targeted to the nucleus by the presence of at least two nuclear localization signals, one lying within ZF I and one in ZF II and/or III (12) . Reporter construct experiments have identified many putative in vivo target genes for both transcriptional activation and repression by WT1 (13 , 14) . However, significant variation has been found in these experiments, depending on the cellular context and the promoter used to express WT1 (15) . At present, little is known about the in vivo situation. By analyzing WT1 missense mutants, English and Licht (16) have found that activation and not repression by WT1 is critical for growth control. WT1 is predominantly expressed in the developing kidney, where it may play a role in mesenchymal to epithelial transition (17 , 18) . This hypothesis is supported by the findings that in WTs missing functional WT1, the mesenchymal cells can no longer differentiate into epithelia but can take on different fates, either stromal or myogenic (7 , 19) . During early nephrogenesis, a low level of WT1 is first detected in the uninduced mesenchyme and then increases dramatically during induction (20) . Expression then becomes restricted to the posterior part of newly formed epithelium and is limited to the podocytes of the adult kidney. In tumors, WT1 is seen in the malignant counterparts of those elements that express the protein during normal development (that is, blastema and glomeruloid structures) but is absent from stromal cells (21) .
To confirm and extend our previous observations that most stromal-predominant/ILNR-like tumors are caused by WT1 mutations, we have studied more patients for mutations and examined these for the expression of a mutant WT1 transcript and protein. Furthermore, we have studied the expression of a set of specific kidney development/differentiation markers as a first approach to identify a set of useful classification markers for WT.
Patients and Methods
Patients, Tissue, DNA, RNA, and Protein Isolation.
Snap-frozen material from a total of 86 WTs was analyzed for WT1 mutations [64 from a previously published study (7) and 22 reported here]. The parents of the patients were informed about the study and gave written consent. All patients were enrolled in the International Society of Pediatric Oncology SIOP9/GPOH study and received preoperative chemotherapy according to the SIOP9/GPOH protocol (22) . Histological typing of the tumor specimens was determined after chemotherapy by two of us (I. L. and D. H). DNA, RNA, and protein were isolated simultaneously as described previously (23) . Briefly, frozen tissue was finely ground with a mortar and pestol in liquid nitrogen and homogenized in 4 m guanidinium thiocyanate containing 0.5% sodium N-laurylsarcosine, 0.7% β-mercaptoethanol, and 25 mm EDTA (pH 7.0). The homogenate was loaded onto a cushion of 5.7 m CsCl and 0.1 m EDTA (pH 7.0; treated with diethyl pyrocarbonate), and RNA was pelleted overnight at 20,000 rpm. Proteins were recovered from the top part of the tube and dialyzed against 50% glycerol, 50 mm NaCl, 10 mm HEPES (pH 7.9), 0.5 mm phenylmethylsulfonyl fluoride, and 0.5 mm DTT. DNA was isolated from the layer above the CsCl cushion, phenol-CHCl3 extracted, and precipitated. The CsCl was removed from the centrifuge tube, and the RNA pellet at the bottom of the tube was rinsed with 70% ethanol and resuspended in H2O.
Mutation Analysis of the WT1 Gene.
Of the 86 collected tumor specimens, 64 were previously analyzed for WT1 mutations by PCR single-strand conformational polymorphism and direct sequencing (7) . Here we have analyzed 22 additional patients with the same method.
Northern Blot.
Total RNA (10 μg) was loaded on a 1% agarose gel in 4-morpholinepropanesulfonic acid buffer containing ethidium bromide and 0.22 m formaldehyde. Separation was at 30 V overnight. After photography, the gel was rinsed in H2O for 5 min and transferred overnight in 10× SSC to gene screen membranes (New England Nuclear, Dupont). After transfer, the RNA was UV-cross-linked, and the filter was baked for 2 h at 80°C. Preparation of RNA, hybridization probes, and conditions were as described previously (24) .
For the identification of mutant WT1 transcripts, different oligonucleotide primers were used: GW21 (5′-CCGCCTCACTCCTTCATC-3′) and GW25 (5′-ACCGAGTACTGCTGCTCAC-3′); and Sus39 (5′-TCGCAATCAGGGTTACAG-3′) and Sus40 (5′-GTGGGTCTTCAGGTGGTC-3′); P1 and P4 were as described previously (25) . Five μl of cDNA were amplified in a 50-μl PCR reaction containing 25 pmol of each primer, 200 μm each deoxynucleotide triphosphate, 1× Taq DNA polymerase buffer, and 1 unit of recombinant Taq polymerase (Life Technologies, Inc.). PCR cycle conditions were as follows: (a) for P1/P4, 3 min × 94°C; 30 cycles of 94°C × 1 min, 60°C × 2 min, and 72°C × 2 min; followed by a final elongation step at 72°C for 10 min; and (b) for GW21/25 and Sus 39/40, 3 min × 94°C; 30 cycles of 94°C × 1 min, 53°C × 1 min, and 72°C × 1 min; followed by a final elongation step at 72°C for 10 min.
Restriction Enzyme Digestion.
Wild-type or mutant WT1 alleles and transcripts were identified by digesting 15 μl of PCR products with enzymes specific for the mutant site. For digestion with BsiEI (New England Biolabs), MgCl2 was adjusted to 10 mm final concentration. Products were visualized on a 3% NuSieve-agarose gel stained with ethidium bromide.
For immunohistochemistry, the following antibodies were used in this study: anti-collagen I (COL-1; 1:5000; Sigma); anti-collagen IV (CIV 22; 1:50; DAKO); anti-cytokeratin (KL1; 1:10; Immunotech); anti-desmin (D33; 1:250; DAKO); anti-fibronectin (1:2000; DAKO); anti-LamA (4C7; 1:300; DAKO); anti-MyoD1 (5.8A; 1:10; DAKO); anti-myogenin (F5D; 1:30; DAKO); anti-SA (Sr-1; 1:10; DAKO); anti-vimentin (Vim3B4; 1:10; Roche Diagnostics); and anti-WT1 [C19 (1:500); WT-180 (1:100); Santa Cruz Biotechnology).
Immunoperoxidase staining was performed using the Labeled StreptAvidin Biotin kit from DAKO according to the manufacturer's recommendations. After fixation in acetone or a 1:1 mixture of acetone and methanol for 10 min at −20°C and blocking of unspecific sites, sections were stained with the first antibody for 30 min followed by incubation with the biotinylated secondary antibody and the streptavidin-peroxidase reagent. Negative controls consisted of consecutive tissue sections of each case in which the primary antibody was omitted.
Neutralization of WT1 Antibody.
To demonstrate the specificity of the WT180 antibody in immunostaining, experiments were repeated after preincubation of the antibody with an excess of a purified WT1 protein for 1 h at room temperature. A fusion plasmid of GST and WT1 (26) was used for bacterial expression, and purification of the GST-tagged recombinant WT1 protein was performed using GSTrap columns from Amersham Pharmacia according to the manufacturer's recommendations.
Western Blot.
Total protein extracts were separated on 10% SDS-PAGE and transferred to a nitrocellulose membrane (Schleicher & Schuell). The proteins were visualized using enhanced chemiluminescence (Amersham Pharmacia Biotech) as described by the manufacturer's recommendations. The membrane was sequentially probed with three primary antibodies: (a) WT180 (1:500; Santa Cruz Biotechnology); (b) WTC19 (1:1000; Santa Cruz Biotechnology); and (c) α-tubulin (1:500; Sigma).
Cell Culture and Transient Transfection.
NIH3T3 and Cos-7 cells were cultured in DMEM (Biochrom) containing 10% FCS (Biochrom). For transfection, the cells were seeded at a density of 1–3 × 105 cells/coverslip and transfected according to the manufacturer's (Roche Applied Science) recommendations by using 3 μl of FuGENE 6 and 1 μg of plasmid DNA. The culture medium was changed after 16 h, and the cells were maintained for another 48 h to allow optimal expression of the fusion protein. The transfected cells were examined under a Zeiss Axiophot fluorescence microscope.
Plasmid Construction.
By using different pairs of primers, three fragments were PCR amplified from a WT1 cDNA clone named CMV-WIT2F (kindly provided by Dr. Paul Baird, Australia) and cloned into pCRII from the TA cloning kit (Invitrogen). The primers used to generate fragments were as follows: (a) for the WT450 fragment, VS1a (5′-CGAATTCTATGGGCTCCGACGTGCGGGA-3′) and VS5a (5′-CAGTCGACTCAAAGCGCCAGCTGGAGTTT-3′); (b) for the WT380 fragment, VS1a and VS4a (5′-CAGTCGACTCAACCTGTATGTCTCCTTTG-3′); and (c) for WT220, VS1a and VS3a (5′-CAGTCGACTCACCTCAGCAGCAAAGCCTG-3′). The amplicons were sequenced with IRD800-labeled M13 primers on a LICOR automatic sequencer to verify that no mutations had been introduced. All primers contained a start or a stop codon as well as EcoRI or SalI restriction site that permitted subsequent directional in-frame cloning into pEGFP-C1 (Clontech), generating the following plasmids: (a) pEGFP-WT450, representing the wild-type WT1 isoform containing exon 5 and KTS; (b) pEGFP-WT380, representing a truncated WT1 form with a stop codon in exon 9; and (c) pEGFP-WT220, representing a truncated WT1 form with a stop codon in exon 3.
Mutations in the WT1 Gene.
Here we report the analysis of 22 WTs (3 stromal-predominant tumors, 8 blastemal-predominant tumors, 5 tumors with triphasic histology, and 6 regressive tumors). Two new mutations were found in bilateral stromal-predominant tumors. One male patient (patient 9179) with genital abnormalities had a constitutional nonsense mutation in exon 8 (R362X) and one female (MD37) had a nonsense mutation in exon 6 (R295X) in the tumor. No blood DNA was available to test for a constitutional mutation.
Taken together with our previous study (7) , we have now analyzed a total of 86 WT patients and found WT1 mutations/deletions in 21 cases (24%). The mutation frequency was significantly different among the various histological subtypes: mutations were seen in 63% of tumors of stromal-predominant histology; 14% of tumors of triphasic histology; and 6% of tumors of blastemal-predominant histology. Of the 21 mutations, 17 were present in the germline, and 3 of these were deletions in patients with Wilms' tumor, aniridia, genital malformation, mental retardation syndrome. Six of 10 stromal tumors with WT1 mutations showed LOH of 11p13 markers, and 1 tumor had a second somatic mutation.
These results confirm that the presence of WT1 mutations correlates with the specific histological subtype of stromal-predominant or ILNR-like WTs.
Expression and Subcellular Localization of Mutant WT1 Transcripts and Proteins in WTs.
Using our previously described protocol (23) , DNA, RNA, and protein were extracted simultaneously, allowing us to analyze the same material for WT1 mutations as well as for RNA and protein expression. To examine whether tumors with WT1 mutations express WT1, we first performed Northern blot analyses on WTs of different histology. As has been observed previously (21) , tumors with blastemal-predominant histology expressed high levels of WT1, whereas stromal-predominant tumors expressed only low or undetectable levels of WT1. A Northern blot of different WTs probed for WT1 and actin is shown in Fig. 1A⇓ . In the three stromal-predominant WTs 9274, 9177, and 9184, a WT1 transcript with the highest level can be seen in 9274, where a smaller amount of RNA was loaded when compared with the actin signal. It was estimated that a mRNA can be detected by Northern blots if it comprises 0.001% of the total mRNA, corresponding to 50 copies/cell (27) . Therefore, these stromal-predominant tumors contain an amount of RNA within the normal range of a low abundant message. From the right tumor of patient 9200, which shows no signal on the Northern blot (Fig. 1A)⇓ we have performed real-time RT-PCR to quantify WT1 expression level. The expression was 178-fold lower than that in blastemal-predominant tumor 9168 (data not shown). Because tumor 9200 contains a mixture of cells, some of which most likely do not express WT1, we can deduce that this tumor contains about 5 copies or less of WT1 RNA, which explains why, in this case, no transcript can be seen in Northern blots (Fig. 1A)⇓ .
To study whether the detected WT1 RNA corresponds to the mutant transcript, we performed RT-PCR and restriction enzyme digests. Four tumors were analyzed, and in all a mutant WT1 transcript was detected (for tumor 9274, see Ref. 28 ). Fig. 1B⇓ shows an example for patient 9318, where a constitutional mutation (C>A in exon 7) causes the loss of a BsiEI restriction site. The analysis was performed on tumor and blood DNA (DNA-PCR) and tumor RNA (RT-PCR) using two different pairs of primers. As seen in Fig. 1B⇓ , right, the tumor DNA only shows the mutant allele resulting in a 235-bp undigested fragment of exon 7. Amplification of the cDNA and digestion with BsiEI leads to an undigested 771- and 720-bp fragment, containing or lacking exon 5, respectively (Fig. 1B⇓ , left). This fragment represents the mutant transcript. A schematic drawing of the fragment sizes after digestion of the DNA-PCR and RT-PCR product is given below the figure.
Next we analyzed whether a truncated protein is synthesized from these mutant messages. By immunohistochemistry, we studied frozen sections for the presence of a wild-type protein using the WTC19 antibody directed against the COOH terminus and the presence of a truncated as well as a wild-type protein using the WT180 antibody against the NH2 terminus. As described previously (17 , 20) WT1 staining was restricted to the nucleus of podocytes and their precursors in normal embryonic kidney (Fig. 1C⇓ , NEK), demonstrating that both antibodies were specific. Although all six blastemal-predominant tumors showed nuclear staining with both antibodies (tumor 9345 in Fig. 1C⇓ and Table 1⇓ ), no immunoreactivity was seen with the COOH-terminal antibody in stromal-predominant WTs (tumor 9200 in Fig. 1C⇓ and Table 1⇓ ), demonstrating that no wild-type protein was synthesized. Using the NH2-terminal antibody, all five cases with nonsense mutations had cytoplasmic staining in cells with a skeletal muscle morphology. In Fig. 1C⇓ , tumor 9200 shows a positive reaction with the WT180 antibody, and it can be seen that the cells, which were cut longitudinally, show the elongated morphology of muscle cells with small nuclei and staining of the cytoplasm. No reactivity was seen in the surrounding stroma. Most of these tumors had lost the wild-type allele, confirming that only a truncated mutant WT1 protein is synthesized. The specificity of the NH2-terminal WT180 antibody was examined by the use of three different batches and by preabsorption with a purified GST-WT1 fusion protein containing aa 1–181 before incubation on tissue sections. This lead to a greatly reduced staining (data not shown).
Western blot analysis on total tumor protein extracts from two blastemal-predominant tumors (NL and 9168) and three stromal-predominant tumors (9385, 9274, and 9318) showed a full-length protein of Mr 52,000, with the WTC19 and WT180 antibodies only in the blastemal-predominant tumors (Fig. 1D, b)⇓ . No wild-type or truncated WT1 protein could be detected with either antibody in the stromal-predominant tumors (Fig. 1D, s)⇓ . This indicates that the amount of mutant WT1 protein is below the detection limit of the Western blot method, which is not suitable to detect a protein of low abundance (29) , specifically if the cells used for protein extraction represent a mixture of WT1-expressing and non-WT1-expressing cells. In Fig. 1C⇓ , it can be seen that only a few cells in the stromal-predominant tumors are positive for WT1, further explaining why the truncated protein may not be visible in Western blot analysis.
Analysis of Subcellular Localization of Truncated WT1 Proteins in Cell Culture.
To support our findings of a cytoplasmic truncated WT1 protein in tumors, we studied the subcellular localization of EGFP-WT1 fusion proteins of different length after transient transfection in NIH3T3 and Cos-7 cells. The full-length construct contained a cDNA encoding all 450 residues of the WT1 protein including exon 5 and KTS and an NH2-terminal fusion of EGFP (EGFP-WT450). Two WT1 proteins truncated at aa residues 220 and 380 were constructed and designated EGFP-WT220 and EGFP-WT380, respectively (Fig. 2A)⇓ . These corresponded to the naturally occurring mutations in patients 9184 and 9394. EGFP-WT220 lacked the entire ZF domain, whereas EGFP-WT380 lacked parts of ZF III and the entire ZF IV. The nuclear localization signal in ZF I was still present here. The constructs were verified by transient transfection in NIH3T3 cells and subsequent Western blot analysis (data not shown). In contrast to previous studies on subcellular localization of WT1 proteins (12 , 30) , we have directly visualized the EGFP-WT1 fusion proteins in native cells under a fluorescence microscope without fixation. Whereas the wild-type EGFP-WT450 fusion protein was restricted to the nucleus in both cell types, the deletion mutant EGFP-WT380 showed nuclear as well as cytoplasmic distribution (Fig. 2B)⇓ . In contrast, the EGFP-WT220 fusion protein was mostly excluded from the nucleus in NIH3T3 cells and was present in the nucleus and the cytoplasm in Cos-7 cells. Fig. 2C⇓ summarizes the results.
Expression of Muscle Markers in Stromal-predominant WTs with WT1 Mutations.
Previously, four tumors with WT1 mutations were studied for the expression of the myogenesis genes MyoD1, myosin heavy polypeptide (Myh3), and myogenin (MyoG) and found to be positive (19) . To extend these observations, we have studied the expression of additional muscle differentiation markers in our five stromal-predominant tumors with WT1 mutations. The muscle markers desmin, sarcomeric actin, MyoD1, and myogenin were present in the same cells that expressed the mutant WT1 protein but absent in the surrounding stroma. None of these markers were present in the blastemal clusters seen in the blastemal-predominant tumors (data not shown and summarized in Table 1⇓ ).
Analysis of Kidney Differentiation Markers.
To determine whether ILNR-like, stromal-predominant WTs with WT1 mutations can be distinguished from PLNR-like, blastemal-predominant WTs by their differentiation stage, we have studied a set of markers. Markers for uninduced and early-induced mesenchyme were vimentin, collagen I, and fibronectin, and markers for condensed mesenchyme were cytokeratin, laminin A, and collagen IV (31) . Using this set, we could show that cells in both tumor subtypes correspond to early-induced mesenchyme but express one marker of uninduced mesenchyme. The organoid structure of cell clusters in blastemal tumors and the beginning of cytokeratin expression demonstrates that this subclass is already one step further toward epithelial differentiation. Fig. 3⇓ shows a representative example of one stromal-predominant tumor (tumor 9200) and one blastemal-predominant tumor (tumor 9345). Cytokeratin staining is negative in tumor 9200 and is seen only within blastemal clusters of tumor 9354. Vimentin staining is seen in stromal cells of both tumors and in blastemal clusters of tumor 9345, whereas muscle cells in tumor 9200 are negative (cluster of blue-stained cells in the middle of the picture). Staining for the extracellular matrix proteins collagen IV and fibronectin, as well as the membrane protein laminin A, surrounds the entire blastemal clusters in blastemal-predominant tumors, except in one case, where the individual tumor cells were positive (data not shown). In contrast, in stromal-predominant tumor 9200, each individual cell was surrounded by positive staining. Collagen I staining is negative in both tumors.
Table 1⇓ summarizes the immunohistochemical results of five stromal-predominant tumors with WT1 mutations and six blastemal-predominant WTs without WT1 mutations as well as normal kidney specimens. Expression of markers was evaluated only in those cells that were positive for WT1 in both subclasses and compared with the expression of these markers as described in the kidney development database.4
A major focus of this work was to establish diagnostic markers for the classification of clinically distinct subsets of WTs.
Our previous work and data presented in this report demonstrate that WT1 mutations occurred in a high precentage (63%) in one subset of WTs, the stromal-predominant or ILNR-like tumors. Importantly, 11 were present in the germline, and all but 1 were mutations resulting in protein truncation or loss of the entire gene (Ref. 7 ; data reported here). The germline mutations were found in three normal females, one normal male, and seven males with GUs of varying degrees. Interestingly, two males without GU had a tumor-specific mutation. Unilateral tumors developed in eight patients with germline mutations. An important clinical information from these data is that patients with germline mutations can present with unilateral tumors without associated abnormalities, and it should be kept in mind that these patients have a high risk for a contralateral tumor. Six of 10 (60%) stromal-predominant tumors had lost the wild-type DNA allele, and 1 tumor had a second somatic mutation, confirming Knudson's two-hit inactivation model for WT1 in the development of this tumor subclass. In contrast, only one missense mutation in exon 2 was found in 16 blastemal-predominant WTs, suggesting that WT1 mutations play a minor role in the development of this tumor subtype.
Mutant WT1 mRNA was found in stromal-predominant WTs, and in cases where LOH occurred, only mutant transcripts were expressed. To our knowledge, only one other group found a mutant transcript in a tumor with a WT1 alteration (32) . In quantitative RT-PCR and Northern blot experiments, we have determined that the amount of the mutant transcript in different stromal-predominant tumors is between 5 and 50 copies/cell, which is in the normal range of low abundant messages (27) . This could correspond to the amount found in early-induced cells, at a time when WT1 expression starts. Blastemal-predominant WTs contain about 180-fold more WT1 mRNA, which may correspond to the normal range in condensed, epithelialized cells.
In stromal-predominant WTs with a mutation, the WT1 protein could be detected by immunohistochemistry in individual cells only with an antibody that recognizes the NH2 terminus, but not the COOH terminus, demonstrating that no intact wild-type protein was present. The mutant form was aberrantly localized in the cytoplasm, resulting in the absence of a functional WT1 protein in the nucleus. This was confirmed by analyzing the subcellular localization of different EGFP-WT1 mutants in unfixed, native cells. It is interesting that Englert et al. (30) found a WT1 protein truncated at aa 326 localized exclusively in the nucleus of Saos-2 and U2OS cells, two embryonal tumors not derived from kidney. Bruening et al. (12) , who used the same cells as we did (NIH3T3 and Cos-7), also noted a cytoplasmic localization with some constructs. They have deduced that a second nuclear localization signal is present between aa 291 and 350, which is absent in our almost exclusively cytoplasmic construct WT220. Because all truncated proteins still contain the dimerization motif, it is possible that at a time where LOH has not yet occurred, the wild-type protein might be excluded from the nucleus by dimerization with the cytoplasmic truncated protein. This could be the first step in an abnormal differentation of early-induced cells. In contrast, blastemal clusters in blastemal-predominant tumors expressed wild-type WT1 protein in the nucleus. Therefore, one molecular distinction between these two subtypes of WTs is the presence of a nuclear wild-type WT1 protein in blastemal-predominant tumors and its absence in stromal-predominant tumors.
Interestingly, in stromal tumors, the truncated protein was restricted to cells with a rhabdomyomatous phenotype expressing desmin, MyoD, myogenin, and sarcomeric actin. It was described previously that tumors with WT1 mutations may aberrantly differentiate into skeletal muscle (19) . This suggests that the lack of nuclear WT1 protein in multipotential mesenchymal cells results in the failure of a proper epithelial differentiation and alternative differentiation programs; predominantly skeletal muscle as well as fat, cartilage, and bone are turned on. All these structures are not found during normal kidney differentiation and in blastemal-predominant tumors, suggesting that alternative programs are normally repressed. Whether WT1 is this repressor is not known. It should be noted, however, that all stromal-predominant tumors that we have studied were obtained after chemotherapy, and therefore rhabdomyomatous differentiation in response to chemotherapy cannot be excluded as has been proposed by Brisigotti et al. (33) . However, we have studied several untreated stromal-predominant WTs for the presence of muscle differentiation by the expression of desmin and found them positive, suggesting that at least some muscle differentiation is present before chemotherapy.5
During early induction of kidney differentiation, vimentin, collagen I, and fibronectin are expressed, and at the stage of condensation, cytokeratin, laminin A, and collagen IV are expressed (31) .4 A switch from fibronectin to cytokeratin occurs at the stage of mesenchymal epithelial conversion. In stromal-predominant tumors, the cells expressing mutant WT1 and showing a rhabdomyomatous phenotype are at the differentiation stage of condensed cells except for the expression of fibronectin, which is normally turned off at that stage. Cells expressing wild-type WT1 in blastemal-predominant tumors are positive for markers of early-induced and condensed mesenchyme and may be one step further in epithelial differentiation because they start to express cytokeratin. Laminin A and collagen IV staining was not seen around each individual cell as in the case of stromal-predominant tumors but was seen surrounding a cluster of blastemal cells, suggesting that these may be further along the development to a more organoid structure. The expression of laminin A increases when epithelial cell polarization begins (34) . It was shown that laminin A, collagen IV, and heparan sulfate proteoglycan first appear as spots on aggregating cells and then form a semicontinuous sheet on the surface of comma- and S-shaped bodies, suggesting a role in cell aggregation and later in epithelial differentiation (35) . Forced expression of WT1 in mesenchymal NIH3T3 cells leads to up-regulation of E-cadherin, cytokeratin, heparan sulfate proteoglycan, and collagen IV, suggesting that WT1 is a positive regulator of epithelial differentiation (36) . We have shown previously (37) that blastemal-predominant WTs expressing wild-type WT1 are positive for PAX-8, which is turned on when epithelial differentiation begins, further supporting the notion that tumor cells expressing wild-type WT1 are at an early stage of epithelialization.
Taken together, these data show that two distinct subclasses of WTs can be distinguished on the basis of the presence or absence of WT1 mutations and by their associated expression of specific differentiation markers. A similar subclassification was made on the basis of the presence or absence of LOI in PLNR-associated/blastemal-predominant WTs and ILNR-associated/stromal-predominant WTs by Ravenel et al. (5) . This will be the basis to distinguish these two subtypes in future studies, rather than simply placing all WTs into one category. By doing so, it is expected that many of the previously unresolved inconsistencies might be resolved.
Based on the present data, we propose a model for the development of these two distinct subclasses of WTs (Fig. 4)⇓ . Tumor cells in blastemal-predominant/PLNR-like WTs correspond to an early stage of induction/conversion to an epithelial phenotype. Cells arrested at the proliferative stage of early-induced mesenchyme are blocked from further differentiation, possibly by continuous expression of WT1 and/or LOI of IGF2. In contrast, the first event leading to stromal-predominant/ILNR-like WTs is either a germline or early somatic truncation mutation in a multipotential kidney stem cell (Fig. 4⇓ , nucleus marked in red). This is followed by a second hit in a cell, most likely after induction by the ureteric bud, resulting in complete absence of a functional nuclear WT1 protein. Mesenchymal stem cells lacking WT1 fail to epithelialize and stochastically differentiate into other mesenchymal lineages. This ultimately leads to a heterogeneous tumor containing multiple cell types at the time of diagnosis. Microdissection of different cell types in two stromal-predominant tumors revealed LOH in all, indicating that this genetic event occurred in a multipotential stem cell early in tumor development (data not shown). Therefore, cells with a normal stromal appearance are also tumor cells, as has been described previously (38) ; however, they are negative for the WT1 protein. In summary, the two-hit inactivation of WT1 is essential in stromal-predominant/ILNR-like WTs.
Of clinical importance is that stromal-predominant and fetal rhabdomyomatous tumors (which may be the same) respond poorly to chemotherapy, in contrast to most blastemal-predominant tumors (4 , 22 , 39 , 40) . Although patients with stromal-predominant tumors generally do not have a poor prognosis, these tumors tend to be very large at the time of diagnosis. Beckwith et al. (41) described the importance of differentiating between an aggressive tumor and a responsive tumor. Generally, tumors with a predominantly epithelial differentiation have a low degree of aggressiveness, but tumors presenting at an advanced stage may be resistant to therapy, and death rates may be higher than those for more aggressive tumors. None of the patients with a stromal-predominant tumor in the SIOP9/GPOH developed any relapse. Therefore, Weirich et al. (22) proposed to test a reduced or milder postoperative treatment in cases where tumors are completely resected, to avoid the unnecessary side effects of chemotherapy. Data from our work suggest that the presence of WT1 mutations and extensive rhabdomyomatous histology could be used as diagnostic markers for nonresponding tumors. This will allow identification of patients who will benefit from changes in treatment protocols and possibly from the development of novel therapeutic approaches.
We thank all SIOP9/GPOH-participating clinicians for sending us tumor material. We thank Dr. Hans-Dieter Royer for critical reading of the manuscript.
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵1 Supported by the Elterninitiative "Kinderkrebsklinik e.V.," Düsseldorf.
↵2 To whom requests for reprints should be addressed, at Institute of Human Genetics, University of Düsseldorf, Postfach 101007, D40001 Düsseldorf, Germany. E-mail: royer{at}uni-duesseldorf.de
↵3 The abbreviations used are: WT, Wilms' tumor; ILNR, intralobar rest; GU, genitourinary malformation; PLNR, perilobar rest; LOI, loss of imprinting; LOH, loss of heterozygosity; GST, glutathione S-transferase; RT-PCR, reverse transcription-PCR; EGFP, enhanced green fluorescent protein; ZF, zinc finger; aa, amino acid(s).
↵4 http://www.ana.ed.ac.uk/anatomy/database.
↵5 V. Schumacher and B. Royer-Pokora, unpublished observations.
- Received November 4, 2002.
- Accepted February 10, 2003.


text:
Lynyrd Skynyrd "Gimme Three Steps"
Could be the last tour for Skynyrd! Its cool to go back to the beginning for a version of "Gimme Three Steps" and so we go back to March of 1976 at The Winterland Theater for this one. I would guess with a lot of confidence that "Gimme Three Steps" will be on the set list for both gigs later on this month, June 22nd at PNC Bank Arts Center and the very next night June 23rd at Northwell Health At Jones Beach Theater. As it is on many weekends, your participation in The Q Workforce will get you in the running for the big upgrade to the front row! Good luck winning YOUR tickets all weekend long from New York's Concert Station, Q1043!


text:
An ad popped up on craigslist in Vancouver, where it can be as frustrating as in New York City to find a good place. It was a very ordinary request, really: an offer to be a pet dinosaur in exchange for a house.
Here's the ad:
This offer is not for everyone. Those of you who have saved every penny for most of your life to afford a down payment and currently work around the clock to make mortgage payments, I commend you on your efforts, but this post is not for you.
Do you own more than one property? Do you have so many rental homes with no mortgage payments, yet you still feel unfulfilled? Tired of your illegal tenants whining that there are rats in the walls? Have you always wanted your own dinosaur? Now is your chance my friend.
In exchange for one of your properties, I will be your personal dinosaur for one year.
I will be at your beck and call, 24 hours a day, wearing a dinosaur costume. The type of dinosaur is negotiable. I can babysit your children (references upon request), scare the mailman, wash dishes, entertain and impress your guests, and much more. (No sex stuff though, sorry.) I will make realistic dinosaur sounds, eat what the particular dinosaur eats and maybe even sit on a fake dinosaur egg, if you are so inclined. I am well educated, fluent in English and French (as well as dinosaur), can play several musical instruments and have no criminal record or outstanding warrants.
All this and more. This is the only way you will ever have your pet dinosaur, and the only way I will ever be able to acquire a house in Vancouver.
Serious offers only please.
Thank you.
RELATED: Left behind in the Rapture? Brooklyn will suffice
The Village Voice reached out and learned that the ad was actually posted by a couple – Justin, 31, a musician, and Rheanna, 27, a graphic designer – who thought they could better fill the on-call needs of a pet dinosaur owner together. And though they haven't gotten any viable offers, they've started compiling their responses to the ad on a website, including an offer to let them sleep in a Honda Element SUV.
And for the record – Rheanna would prefer to be a Triceratops while Justin says he's more of a T-rex type.