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Strange Bedfellows -- Politics News
News about Seattle and King County government as well as national politics.
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Judge blocks tunnel initiative from fall ballot
A King County Superior Court judge on Monday ruled that an initiative that sought to prohibit city streets from being used to help build the tunnel replacement for the Alaskan Way Viaduct can't go before voters this fall. Judge Joan E. DuBuque said Initiative 101 went beyond the scope of initiative powers because it concerned an […] [Read More]
Classen picks up endorsements from pols
Maurice Classen, the King County deputy prosecutor challenging City Councilwoman Jean Godden, has been endorsed by four local politicians, his campaign said Monday. Those blessing Classen over Godden were King County Councilman Bob Ferguson, state Sen. Margarita Prentice, D-Renton, state Rep. Hans Dunshee, D-Snohomish and state Rep. Roger Goodman, D-Kirkland. In the clubby world of local […] [Read More]
24-hour bar service in Seattle? State not so sure
The state's Liquor Control Board is taking a wait-and-see approach to a request from Seattle to extend bar hours – including the possibility of 24-hour service. On Thursday Mayor Mike McGinn and City Attorney Pete Holmes will say they're beginning efforts to get the city permission to have certain establishments sell beer and liquor after […] [Read More]
Ashton Kutcher Tweets crash City of Seattle blogs
Dude, where's my blog? The City of Seattle's blog network was severely impacted – and in some cases knocked down – because Ashton Kutcher name dropped the Jet City and Mayor Mike McGinn in several Tweets on Friday. aplusk ashton kutcher Seattle responds to Human Trafficking #dna http://post.ly/2JelJ aplusk ashton kutcher I think every mayor should […] [Read More]
Group tries again to shield names in gay rights fight
A conservative group trying to shield the names of more than 100,000 people who signed petitions trying to block a new law expanding gay rights in Washington state is again trying to keep the information from becoming public. Protect Marriage Washington, which lost year lost its argument on broad First Amendment claims before the United […] [Read More]
Study: Seattle among top 'green' cities in North America
Seattle is the fourth "greenest" city in the United States and Canada, according to a study published Thursday by the Economist Intelligence Unit. The examination, commissioned by Siemens, ranked 27 cities. So which places were ahead of the Emerald City? San Francisco, Vancouver, British Columbia, and New York. The study looked at nine areas, including environmental governance, […] [Read More]
Study: Want jobs? Build bike lanes
Building bike lanes is the best way for governments to create jobs through infrastructure spending, according to a University of Massachusetts, Amherst study that included Seattle. Researcher Heidi Garret-Peltier looked at 11 municipalities and concluded: Bicycling infrastructure creates the most jobs for a given level of spending: For each $1 million, the cycling projects in this study create […] [Read More]
Chamber of Commerce: Thumbs down on sick leave plan
The Greater Seattle Chamber of Commerce said Wednesday it does not support a compromise, paid sick leave proposal unveiled by advocates and some business interests at City Hall this week. "Representing nearly 1,700 small businesses in the metropolitan Seattle area, the Chamber is committed to the wellness of employees as well as the vitality of our […] [Read More]
New sick leave idea for Seattle workers
Health advocates and some business interests on Tuesday put forward a new proposal for paid sick leave for people who work in Seattle – one that varies by a company's size and gives employers flexibility. Businesses with up to 49 workers would have to offer up to five days, which would be accrued by hours worked. The […] [Read More]
McGinn transpo chief signs viaduct tunnel EIS
The head of Seattle's transportation department said Monday he signed the final Environmental Impact Statement concerning the tunnel replacement for the Alaskan Way Viaduct, removing one potential obstacle to completion of the controversial project. Peter Hahn, transportation department director, told seattlepi.com he agreed to the document after the state incorporated parts of a city […] [Read More]
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text: Incredible Pencil Animation [VIDEO] By Kat Mykals October 24, 2011 10:43 AM Share on Facebook Share on Twitter I don't know that I have the words for just how freaking awesome this video is! I do know that I lack the patience to have drawn out all of these images. Take a look and be prepared to be impressed! Filed Under: Pencil Art, Vimeo, viral video Category: Humor, Technology, Videos Share on Facebook Share on Twitter More Leave a comment Send to a friend! Print this page Pin it! Reddit This! Share on Tumblr! Best of 106.1 KISS FM Recommended For You Around the Web

subset: commoncrawl


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Powdery mildew cures??
Is there REALLY an effective organic fungicide to fight powdery mildew??
I've tried neem, sulfur dust/spray, and baking soda with absolutely no success.
Don't tell me -- Milk! Why would I think that milk would work?? The same places I've read the above remedies worked are the same places that mention milk!
Sorry for sounding pessimistic. I really do prefer to do things organically, but sometimes it just seems that there aren't any good organic remedies for certain diseases and pests.
I'm not going to pay hundreds of dollars for beneficial insects integrated into my backyard garden every year just to have them go next door.
Anyhow... anybody have any tricks for powdery mildew?
Anything that WORKS would be appreciated.
Kevin, I'm afraid my experiences with powdery mildew don't offer much as far as getting rid of it.
I think a lot depends on having plenty of organic matter in the soil; for plant nutrition of course, but even more so to maintain moisture.
I find powdery mildew takes over when plants are water stressed. Like most people's summers, later it gets, the drier things are.
But by the end of the season at my place, when it really starts to slow down the zucchini, I'm rather relieved!
Understanding why Powdery Mildew appears often helps with control. Sulfur is effective if applied before signs of PM appear while other controls can be effective when properly applied.
Baking Soda (Sodium Bicarbonate) mized one teaspoon per quart of water, Fat Free Milk and water mixed 50/50, Potassium Bicarbonate, and Neem oil have all been effective at controlling PM, but not at curing the problem. Since the spores that cause PM are blown in on the wind there probably is nothing that will cure this problem.
Here is a link that might be useful: About Powdery Mildew
First, you need to understand that you can't cure powdery mildew - once it is present (and you see the symptoms), it's present and the best you can expect is to control the spread. Period.
Fungicides are prophylactic, not curative. They can sometimes prevent the disease, will often limit the spread or contain the disease but they will not eliminate or "fix" existing diseased tissue.
Powdery mildew is most often a cultural ailment. It arises typically in late season when temperatures are warm, soils are dry and humidity typically high. Lack of good air circulation can exacerbate the condition. Not a lot one can do about temps or humidity but making sure the soil is adequately moist and improving circulation can go a long way to slowing the disease. Knowing what plants are most susceptible and treating before you see any signs can help. FWIW, with most woody plants, PM is more of a cosmetic issue than a serious threat to plant health. Also, PM spores cannot live in water - hosing down the plants early in the day (so there is adequate time for the foliage to dry properly to deter other fungal issues) can help as well - you can actually wash the spores off the leaves.
The reason milk is often recommended - and with good reason - is that it is a natural anti-fungal/anti-bacterial agent. It also contains lactic acid, which alters the pH of the leaf surface and makes it inhospitable to the fungal pathogen. This is the same principle behind the use of baking soda as well. btw, it doesn't make any difference what formulation of milk you use - powdered, fat free, high octane - it all works the same.
Yes, any fungicide regsistered for PM, including those home remedies like the milk, baking soda, etc., will help to prevent the spread of fungal spores to unaffected foliage and if applied prior to infection (usually before the symptoms are evident), should prevent the problem altogther.
But do follow good garden sanitation and proper cultural practices as well - all the spraying in the world won't help significantly if you are just welcoming the disease by offering poor growing conditions.


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On the 2014 Fortune 500 list of the largest U.S. corporations, released last week, CVS Caremark Corp. inched upward to No. 12 from No. 13 on the 2013 list. Also making the Fortune 500 again were Textron Inc. (No. 228) and United Natural Foods Inc. (No. 427). more
By PBN Staff
Rear Adm. Michael E. Jabaley will become commander of the Naval Undersea Warfare Center on Friday, relieving Rear Adm. David M. Duryea, during a change of command ceremony. more
By PBN Staff
The Naval Undersea Warfare Center dedicated a new $24.9 million payload research facility on Wednesday. more
By PBN Staff
Textron Inc. – parent company of Bell Helicopter, Cessna Aircraft Co. and Textron Systems – reported that its net income dropped 28.6 percent in the first quarter to $85 million, or 30 cents per diluted share, due in part to restructuring costs incurred by the manufacturer's acquisition in March of Beech Holdings LLC. more
By PBN Staff
The U.S. Navy has awarded a $17.6 billion contract to General Dynamics Corp. and Newport News Shipbuilding for the next block of 10 Virginia-class submarines to be built beginning in fiscal year 2014 and continuing on to fiscal 2018. more
By PBN Staff
The U.S. Small Business Administration announced a partnership with the R.I. Commerce Corporation and its Procurement Techincial Assistance Center to offer Rhode Island small businesses free counseling and training programs on procurement procedures at the local, state and federal level. more
By Eli Sherman
Bell Helicopter, a subsidiary of Providence-based manufacturer Textron Inc., will lay off 325 workers due to the continued decline in defense contracts, the Fort Worth Star-Telegram reported Monday. more
By PBN Staff
South Korea's military is buying one of the top anti-armor weapons developed by the Pentagon, a Textron Inc. system designed to destroy tanks and vehicles. more
By Tony Capaccio
KVH Industries Inc., a provider of guidance and Internet systems for the maritime and defense industries, reported second-quarter net income of $55,000, or zero cents per share, an improvement over the first-quarter net loss of $1.1 million but a 96.5 percent decline compared with net income of $1.6 million, or 10 cents per share, for the second quarter of 2013. more
By PBN Staff
The Governor's Workforce Board has awarded a total of $1.1 million in Industry Partnership grants to eight groups with proposals aimed at addressing the skills gap in Rhode Island. more
By Patricia Daddona


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Cam Wallace - HurryUpandWait
Artist: Cameron Wallace
Twitter: Cameron Wallace on Twitter
Avg Rating: 4.3 ( 4 total votes )
Multi-platinum producer and buzzmaking emcee Cam Wallace has come together with The DJBooth to bring fans his latest street release, HurryUpandWait.
The project features 17 original records (plus one bonus track) from the Houston native, including reader-approved leaks "Saturday" and "Pyramids."
Joining Wallace on the guest tip throughout HurryUpandWait are Le$, Greg Lawrence and Blush. All production comes courtesy of Cameron Wallace himself.
Stream HurryUpandWait
Submit your Rating
First DJ Booth Appearance:
"Saturday" (2011)
Total DJ Booth Features:
5 Songs
Commentsblog comments powered by Disqus
DJBooth TV
Discover the best new songs, videos, and albums added to the Booth.


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About: Ryan Bertroche, born in 1991, in Iowa, USA, is an American model. Bertroche grew up in a farming community in Iowa, his hobbies and interests include riding horses and helping other farmers in his home town. Bertroche attended PV High School in Bettendorf and started modeling shortly after graduation. Bertroche's modeling career has already taken off, posing for photographers such as Bruce Weber, Mariano Vivanco, Alasdir Mclellan, and Carlotta Manaigo ... (more)
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4th Avenue
Boerum Hill, New York, 11217
Commute to Downtown New York
Daily errands do not require a car.
World-class public transportation.
Flat as a pancake, excellent bike lanes.
Score Details
4th Avenue has a Walk Score of 99 out of 100. This location is a Walker's Paradise so daily errands do not require a car.
4th Avenue is a three minute walk from the D 6 Avenue Express, the N Broadway Express and the R Broadway Local at the Atlantic Av - Barclays Ctr stop.
This location is in the Boerum Hill neighborhood in New York. Nearby parks include Thomas Greene Playground, Public School 38 and North Pacific Playground.
Nearby schools include PS 133 William A Butler School, Park Slope Christian Academy and Khalil Gibran International Academy.
The closest grocery stores are Slope Natural Plus, eCosway USA Inc. and Nine One Two Corporation.
Nearby coffee shops include Canteen Delicatessen, The Chocolate Room and Gorilla Coffee. Nearby restaurants include Die Koelner Bierhalle, The KBH (Die Koelner Bierhalle) and Papa John's Pizza.
4th Avenue is near Thomas Greene Playground, Dean Playground and Sixteen Sycamores Park.
There are excellent bike lanes and the terrain is flat as a pancake. 4th Avenue is very bikeable, biking is convenient for most trips. Bike sharing is available from Citi Bike.
Explore how far you can travel by car, bus, bike and foot from 4th Avenue.
4th Avenue is a Rider's Paradise which means world-class public transportation. Car sharing is available from RelayRides, Zipcar and Hertz On Demand.
Rail lines:
D 6 Avenue Express
0.2 mi
N Broadway Express
0.2 mi
R Broadway Local
0.2 mi
4 Lexington Avenue Express
0.3 mi
2 7 Avenue Express
0.3 mi
3 7 Avenue Express
0.3 mi
Q Broadway Express
0.3 mi
5 Lexington Avenue Express
0.3 mi
B 6 Avenue Express
0.3 mi
Port Jefferson
0.3 mi
Bus lines:
B103 Canarsie - Downtown Brooklyn Ltd
0.0 mi
B65 Downtown Brooklyn - Crown Heights
0.1 mi
B63 Bay Ridge - Cobble Hill
0.1 mi
B67 Brooklyn Navy Yard - Kensington
0.2 mi
B41 Kings Plaza - Downtown Brooklyn
0.2 mi
Car shares:
Bike shares:
View all Boerum Hill apartments on a map.
Popular apartment searches include fireplace and furnished. View all New York apartments and rentals.
4th Avenue is in the Boerum Hill neighborhood. Boerum Hill is the 18th most walkable neighborhood in New York with a neighborhood Walk Score of 98.
Moving to New York from another city? View our moving to New York guide.


text:
MLB Baseball - Preview
|Cincinnati Reds vs. Cleveland Indians
The Cincinnati Reds and the Cleveland Indians will both be trying to pick up a win on Thursday when they battle at Progressive Field.
Righthander Homer Bailey will take the mound for the Reds to start this game. Bailey is 3-3 this season with a 3.08 ERA.
The Indians will counter Bailey with Scott Kazmir. Lefthander Kazmir has a 5.94 ERA to go along with a 2-2 record this season.Cincinnati lost its last outing, a 5-2 result against the Indians on May 29. Bettors who backed the Indians at -135 on the moneyline won on the day, and the total score (7) sent UNDER bettors to the payout window. Team records:
Cincinnati: 33-20 SU
Cleveland: 28-24 SU
Cincinnati most recently:
When playing on Thursday are 6-4
Before playing Pittsburgh are 5-5
After playing Cleveland are 5-5
After a loss are 7-3
Cleveland most recently:
When playing on Thursday are 6-4
Before playing Tampa Bay are 4-6
After playing Cincinnati are 6-4
After a win are 6-4
A few trends to consider:
Cincinnati at Pittsburgh, Friday, May 31
Cleveland home to Tampa Bay, Friday, May 31


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My Final State of the Race Report: November 5, 2012
The basic problem we have with calling this election is a dearth of reliable polling data. Gallup and Rasmussen seem to be predicting that the composition of the electorate will be closer to the one we saw in 2010, while almost all of the other pollsters seem to be anticipating a Democrat advantage similar to the one we saw in 2008 — if not even better.
On its face, this idea seems ludicrous. Obama's crowds are much smaller, the enthusiasm levels are almost McCain-like, the early voting numbers have dropped off significantly — but more importantly, you have to look at the Republican side. The biggest reason Democrats had such a huge advantage in 2008 was because Republicans didn't turn out. This time around, the crowds are huge, the energy level is electric and the base is rabid with dislike for Obama. Could Democrats have an advantage over Republicans in turnout? Conceivably, but if so, it should be quite small.
If you want some evidence of this, look at the fact that millions are being spent in states like Wisconsin, Minnesota, Pennsylvania, and Michigan. None of those states were even remotely in play in 2008 and even Bush lost all 4 states both times although the margins were RELATIVELY small. Although it's not the most likely outcome, it seems conceivable that Romney could win all four states. That's no small matter because as a practical measure, Obama probably can't afford to lose any of those states and win.
Additionally, keep in mind that Romney seems to be winning independent voters in most polls, even the ones that are heavily weighted towards the Democrats, by 7-20 points. Simply put, it's hard to imagine Romney losing the election if he wins independents by that much. Obama won independents by 8 and waltzed to victory. George H.W. Bush won independents by 10 when he crushed Dukakis. If Romney wins independents by 7 points, which seems like a safe bet going by most of the polling data out there, hes going to win the election.
Long story short: The most likely outcome seems to be that Romney will win by a large enough margin that Ohio won't matter. The second most likely outcome is that Romney wins and then the third most likely possibility seems to be Obama in a tight race. That being said, it's difficult to say what will happen with a large amount of certainty because unlike 2008 or 2010, we just don't have enough quality polling data to get a reliable picture of the race.
PS: If it does turn out that the polls are off as badly as many pundits seem to think, my best explanation for it would be a polling bubble.
Also see...
Making a man believe that his child doesn't exist for decades is an unusually cruel thing to do. But for Tony Trapani, a secret letter he found after his wife...Read More
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New York Times Slams Romney on Foreign Finances, Ignores Democrats'
The New York Times is aghast that Mitt Romney won't succumb to demands from Democrats and the Old Media that


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Page 1
Separation of splenic red and white pulp occurs before birth
in a LT??-independent manner
Mark F. R. Vondenhoff,*,1Guillaume E. Desanti,†,1Tom Cupedo,‡Julien Y. Bertrand,§
Ana Cumano,†Georg Kraal,* Reina E. Mebius,* and Rachel Golub†,2
†Unite du De ´veloppement des Lymphocytes, INSERM U668, Institut Pasteur, Paris, France; *Department of
Molecular Cell Biology and Immunology, VU University Medical Center, Amsterdam, The Netherlands;‡Erasmus
Medical Center, Department of Hematology, Rotterdam, The Netherlands; and§University of California, San Diego,
La Jolla, California, USA
Peyer's patches, lymphoid tissue inducer (LTi)
cells are crucial in triggering stromal cells to re-
cruit and retain hematopoietic cells. Although LTi
cells have been observed in fetal spleen, not much
is known about fetal spleen development and the
role of LTi cells in this process. Here, we show that
LTi cells collect in a periarteriolar manner in fetal
spleen at the periphery of the white pulp anlagen.
Expression of the homeostatic chemokines can be
detected in stromal and endothelial cells, suggest-
ing that LTi cells are attracted by these chemo-
kines. As lymphotoxin (LT)?1?2can be detected
on B cells but not LTi cells in neonatal spleen,
starting at 4 days after birth, the earliest formation
of the white pulp in fetal spleen occurs in a
LT?1?2-independent manner. The postnatal devel-
opment of the splenic white pulp, involving the
influx of T cells, depends on LT?1?2expressed by
B cells. J. Leukoc. Biol. 84: 152–161; 2008.
For the formation of lymph nodes and
Key Words: lymphoid tissue inducer cells ? organogenesis ? embryo
? chemokines ? stroma ? mouse
The spleen is composed of a branching splenic artery that
eventually ends in venous sinuses. The arterial branches,
central arterioles, are surrounded by a layer of lymphoid tissue,
the white pulp. It consists of T cell areas and B cell follicles,
more or less resembling the organization found in lymph nodes
(LN). The arterial blood ends in a sinusoid system in the area
surrounding the T and B cell zones, thereby forming an ana-
tomical border between the white and red pulp, the marginal
zone. Here, sinusoid spaces formed by lining cells continuous
with the endothelium of the arterioles and reticular fibroblasts
make up a network through which the blood freely percolates
on its way to the red pulp and can be scanned for pathogens
and debris by macrophages and dendritic cells (DC). The blood
runs from the marginal zone through the red pulp cords into the
venous sinuses, enabling the spleen to exert its function as a
filter of the blood by removal of effete RBCs [1–5]. For devel-
opment and organization of lymphoid organs, the members of
the TNF superfamily are crucial. The organogenesis of Peyer's
patches (PP) and LN is dependent on the expression of lym-
photoxin (LT)?1?2and other TNF family members [6, 7].
During fetal and neonatal life, LT?1?2is expressed by
CD45?CD4?IL-7R??CD3–lymphoid tissue inducer (LTi)
cells in PP and LN anlagen [8, 9]. LTi cells are attracted by
nonhematopoietic stromal cells that express the homeostatic
chemokines CCL19, CCL21, and CXCL13, as well as the
adhesion molecules VCAM-1, ICAM-1, and mucosal addressin
cell adhesion molecule-1 (MAdCAM-1) [10–12]. These mole-
cules are thought to be induced by the interaction of LTi and
stromal cells, which leads to LT? receptor (LT?R) and TNFR-I
triggering [13]. Expression of these molecules favors the sub-
sequent recruitment and retention of more LTi cells as well as
other hematopoietic cells [10].
For the proper development of the murine neonatal lymphoid
part of the spleen, indications that LT?1?2is involved so far
only stem from data obtained after birth, which show that
LT?1?2-expressing B cells are required for the induction of
sufficient CCL21, produced by stromal cells in the T cell zone
areas [14]. Considering the important role of LTi cells in the
organogenesis of other lymphoid organs, relatively little is
known about their role in spleen development, although their
presence has been demonstrated in the fetal spleen as early as
Embryonic Day 13.5 (E13.5) [8, 15]. Transfer of in vitro,
IL-7-activated, splenic LTi cells was shown to restore the B
and T lymphocyte (B/T) segregation in spleens of LT??/?mice
[16], suggesting a role for LTi cells in white pulp development.
As these experiments may not reflect the actual interactions
and cellular requirements during fetal and neonatal spleen
development, we studied the role of LTi cells in the developing
splenic white pulp in more detail. Our results show that neo-
natal, splenic LTi cells lack LT?1?2expression and that
LT?1?2, required for correct formation of the splenic white
pulp, is expressed by B cells starting approximately 4 days
after birth. Analysis of the earliest events during spleen devel-
opment demonstrates that compartmentalization of red and
1These authors contributed equally to this work.
2Correspondence: Unite du De ´veloppement des Lymphocytes, INSERM
U668, Institut Pasteur, 25, Rue du Dr Roux, 75724 Paris cedex 15, France.
E-mail: firstname.lastname@example.com
Received September 28, 2007; revised March 13, 2008; accepted March 13,
doi: 10.1189/jlb.0907659
152Journal of Leukocyte Biology
Volume 84, July 2008
0741-5400/08/0084-152 © Society for Leukocyte Biology
Page 2
white pulp areas starts to be regulated during embryogenesis.
By grafting fetal spleens, distinct white pulp areas containing
donor-derived LTi and B cells and red pulp areas harboring
erythrocytes can be observed. Therefore, the white pulp stroma
are already primed before E15.5 to segregate away from the red
pulp areas. Moreover, we suggest that this early phase of
stroma instruction is independent of the LT?R pathway during
murine spleen development.
C57BL/6 mice were purchased from Harlan (Horst, The Netherlands) and from
Charles River (Saint Germain sur l'Arbresie, France), and LT??/?mice were
purchased from Charles River (Maastricht, The Netherlands). All mouse
strains were bred in the animal facilities of the VU University Medical Center
(VUMC; The Netherlands) or the Institut Pasteur (France) and kept under
routine laboratory conditions. The ethical committees of the VUMC and
Institut Pasteur approved all animal procedures.
Timed pregnancies
Mice were mated overnight, and the day of vaginal-plug detection was marked
as E0.5. Pregnant females were killed at different time-points, and embryos
were harvested and frozen in OCT-embedding medium (Sakura Finetek Europe
BV, The Netherlands) or used for dissection.
Cryosections (7 ?m) were fixed in dehydrated acetone for 2 min and air-dried
for an additional 15 min. Endogenous avidin was blocked with an avidin-biotin
block (Vector Laboratories, Burlingame, CA, USA). Sections were then prein-
cubated in PBS supplemented with 5% (v/v) mouse serum for 10 min. Incu-
bation with primary antibody for 45 min was followed by a 30-min incubation
with Fluor-Alexa-labeled conjugate (Invitrogen Life Technologies, Breda, The
Netherlands) when needed. All incubations were carried out at room temper-
ature. Before embedding in polyvinyl alcohol, sections were counterstained
with Hoechst 33342 (Invitrogen Life Technologies) for 10 min. Stainings were
analyzed on a Leica TCS-SP2-AOBS confocal laser-scanning microscope
(Leica Microsystems Nederland BV, The Netherlands), and images were ob-
tained with Leica confocal software. Image processing involved contrast en-
hancement and region of interest selection, which was carried out with Jasc
Paintshop Pro 7.0. Lenses used were dry lenses: 20? (HC PL APO CS 0.7);
Fetal spleens grafted under the kidney capsule were dissected and incu-
bated overnight in 15% saccharose PBS at 4°C prior to cryosectioning.
Sections were analyzed on a Zeiss Axioplan 2 imaging upright microscope with
a Zeiss Axiocam Hrc camera and Zeiss Axiovision 4.2 software.
Whole-mount fetal spleen immunostaining
E15.5 fetal spleens were incubated overnight in 4% paraformaldehyde in PBS
while agitating at 4°C. Spleens were washed twice in PBS 1?, 10% FCS, 0.1%
Triton X-100 solution while agitating during 90 min and incubated overnight
with RMA4-5 (anti-CD4-FITC) antibody at 4°C under agitation. After washes,
spleens were stained with Hoechst 33342 for 90 min, washed, and then
embedded in Vectashield (Vector Laboratories) and analyzed on a Zeiss
Axioplan 2 imaging upright microscope with a Zeiss Axiocam Hrc camera and
Zeiss Axiovision 4.2 software.
Fetal spleen grafts
The fetal spleen harvested from E14.5 to E16.5 C57BL/6 (CD45.2) embryos
was placed under the kidney capsule of Rag2/?c?/?(CD45.1) mice by surgery.
The Rag2/?c?/?(CD45.1) recipient mice [17] were anesthetized by peritoneal
injection of 1.4 mg/g ketamine (Merial, France) and 7 ?g/g xylazine (Sigma-
Aldrich Steinheim, Germany) diluted in PBS. Two hours before the anesthesia,
some Rag2/?c?/?(CD45.1) recipient mice were injected with a total C57BL/6
(CD45.2) fetal liver cell suspension. For negative control, sham-operated
recipients were anesthetized, and their kidney capsule was opened without
grafting a fetal spleen.
For flow cytometry and immunofluorescence, the following antibodies were
used: GK1.5 (anti-CD4 [18, 19]), MECA-367 (anti-MAdCAM-1 [20]), MP33
(anti-CD45, BD Biosciences, Belgium), 6B2 (anti-B220 [21, 22]), 3E2 (anti-
ICAM-1 [23, 24]), and MOMA-2 (a pan macrophage marker) [25]. All of the
antibodies were affinity-purified from hybridoma cell culture supernatants with
protein G-Sepharose (Pharmacia, Uppsala, Sweden) and biotinylated or labeled
with Alexa-Fluor 488, Alexa-Fluor 546, or Alexa-Fluor 633 (Invitrogen Life
Technologies). 429 (anti-VCAM-1, BD Biosciences), A7R34 (anti-IL-7R?,
eBioscience, San Diego, CA, USA), 11D4.1 [anti-vascular endothelial (VE)-
cadherin, BD Biosciences], RMA4-5 (anti-CD4, BD Biosciences), 30-F11
(anti-CD45, BD Biosciences), 104 (anti-CD45.2, BD Biosciences), RA3-6B2
(anti-B220, BD Biosciences), 1D3 (anti-CD19, BD Biosciences), Ter-119 (BD
Biosciences), PK136 (anti-NK1.1, BD Biosciences), RAM34 (anti-CD34, BD
Biosciences), Avas 12?1 [anti-vascular endothelial growth factor receptor
(VEGFR)-2, BD Biosciences], HL3 (anti-CD11c, BD Biosciences), 145-2C11
(anti-CD3ε, BD Biosciences), 7G6 (anti-CD21, BD Biosciences), and A20
(anti-CD45.1, BD Biosciences) anti-VE-cadherin (Alexis Corp., Switzerland) were
used as biotinylated, fluorescently labeled or unconjugated, primary antibodies.
The antibody anti-rat-IgG-tetramethylrhodamine-isothiocyanate (Chemicon Inter-
national, El Segundo, CA, USA) was used as a secondary antibody.
Flow cytometry
LN and spleens were dissected using a stereomicroscope, and single cell
suspensions were made by digestion with 0.5 mg/ml collagenase type IV
(Sigma-Aldrich) in PBS, 2% FBS, for 30 min at 37°C while constantly stirring.
For surface LT?1?2detection, cells were pretreated with 2.4G2 (anti-CD16/
32), supplemented with 5% normal mouse serum for 30 min, and subsequently
incubated with a LT?R-human IgG fusion protein [26] for 60 min. Anti-
human-IgG-PE (Jackson ImmunoResearch Laboratories, West Grove, PA,
USA) was used as a second-step conjugate. Splenocytes from adult LT??/?
mice were used as negative controls. Flow cytometric analysis was performed
on a FACSCalibur using CellQuest software (BD Biosciences). Cells were
negatively gated for TCR-?? to exclude all T cells. 7-Aminoactinomycin D
(Molecular Probes, Eugene, OR, USA) was used to exclude dead cells. For
each staining, negative control stainings were carried out, in which LT?R-
human IgG fusion protein was incubated together with anti-LT? mAb (BBF6),
which prevents the binding of LT?R-human IgG (Alexis Benelux, The Neth-
erlands) to cell surface LT?1?2[8].
For analysis of fetal spleen and fetal spleen grafts, the organs were dissected
using a stereomicroscope, and single cell suspensions were made by dissoci-
ation with a 26-gauge, 3/8-inch needle. Propidium iodide (Sigma-Aldrich) was
used to exclude dead cells. Flow cytometric analyses were performed in an
upgraded LSR (Becton Dickinson, San Jose, CA, USA) using FlowJo software
(Tree Star, Ashland, OR, USA).
RT-PCR, semi-quantitative PCR, and quantitative
real-time PCR
Stromal cells from E15.5 fetal spleen were sorted by flow cytometry on a
MOFLO (Dako, Fort Collins, CO, USA), lysed in RLT buffer (Qiagen, Ger-
many), and frozen on dry ice. Total RNA was extracted with an RNeasy micro
kit (Qiagen), according to the manufacturer's protocol. The cDNA was prepared
from 3 ? 103to 3 ? 106cells using random primers, SuperScript II RT, RNase
OUT in a reaction volume of 24 ?l. PCR reactions (Invitogen Life Technolo-
gies) were performed with Amplitaq Gold (Applied Biosystems, Bridgewater,
NJ, USA) in a reaction volume of 25 ?l.
The primers used were ?-actin, forward: CCGCGAGCACAGCTTCTTT,
?-actin, reverse: CTTTGCACATGCCGGAGC; Vcam-1, forward: GCTCTGG-
GAAGCTGGAACGA, Vcam-1, reverse: TTCATGAGCTGGTCACCCTTGA.
cDNA was diluted at 1/10 and 1/100, and results from the 1/100 dilution were
used to measure intensity of the band using the ImageJ 1.38 application.
Intensity was normalized using the ?-actin transcript levels (100%).
The cDNA samples used for quantitative real-time PCR were obtained by
using RT2PCR Array First Strand kit (SuperArray Bioscience Corp., Freder-
ick, MD, USA) for the RT step. The RT2Profiler PCR Array "Mouse Chemo-
Vondenhoff et al.
Spleen development153
Page 3
kines and Receptors" (SuperArray Bioscience Corp.) and the RT2Real-Time
SYBR Green/6-carboxy-X-rhodamine PCR Master Mix (SuperArray Bioscience
Corp.) were used for real-time PCR quantification. The real-time PCR was per-
formed on a 7300 Real-Time PCR system (Applied Biosystems). The products of
each reaction were checked by 2% agarose gel electrophoresis migration.
CD4?IL-7R??CD3ε–cells from 3-day-old mice intestine previously dis-
sociated by dispase (Gibco-BRL, Grand Island, NY, USA) treatment and
CD4hiLin–(Lin: Gr-1, B220, Ter119, CD11c, CD19, NK1.1, CD3ε) cells from
E15.5 fetal spleen were sorted by flow cytometry on a MOFLO (Dako), lysed in
TRIzol (Gibco-BRL), and frozen at –20°C. Total RNA was extracted according
to the TRIzol (Gibco-BRL) manufacturer's protocol. Oligo (dT)-primed cDNA
was prepared using SuperScript II RT (Invitogen Life Technologies) in a
reaction volume of 20 ?l. PCR reactions were performed with Amplitaq Gold
(Applied Biosystems) in a reaction volume of 25 ?l. The primers used were
retinoid-related orphan receptor (ROR)?/?t, forward: GCCTCCTGAGAGCCT-
CAGG, ROR?/?t, reverse: CACCTCCTCCCGTGAAAG.
Characterization of the developing spleen
Human fetal spleen development was recently characterized,
and already at the 2nd stage of development when the first
lymphocytes start to colonize the organ, distinct areas could be
observed where the homeostatic chemokine CXCL13 was ex-
pressed. The presence of CXCL13 in arterial smooth muscle
cells as well as cells around these arterioles [27] suggested the
interaction of these stromal cells with LTi cells in analogy with
the developing LN and a role in the further development of the
spleen. To further define the earliest events in spleen devel-
opment, we addressed whether these distinct areas could also
be found in murine spleen. Here, fetal spleens from E18.5 were
stained for endothelial, hematopoietic, as well as stromal mark-
ers. Combined staining of tyrosine kinase with Ig and epider-
mal growth factor homology domain 2 (Tie-2/Tek), a marker for
endothelial cells [28], and CD45 revealed the presence of an
artery with a contracted lumen and a vein with an open lumen
in between hematopoietic cells at this time-point in develop-
ment (Fig. 1, A and D). CD45?hematopoietic cells were
remotely located in a ring-like pattern around the artery (Fig.
1B), and the venous blood vessel was immediately surrounded
by hematopoietic cells (Fig. 1C). To define the expression of
MAdCAM-1, a marker for LN stromal organizer cells [29] as
well as high endothelial venules [30], fetal spleens were
stained for MAdCAM-1 in combination with the endothelial
marker VE-cadherin [31]. The results showed that most VE-
cadherin? endothelial cells expressed MAdCAM-1 at high
levels (Fig. 1E and Supplemental Fig. 1, A–C). Stainings of
subsequent sections showed that endothelial cells of large
vessels also expressed ICAM-1, and VCAM-1 was absent or
expressed at low levels (Fig. 1F). In addition, around the artery,
VCAM-1?cells were found. As LTi cells were reported by us
to be present at early stages in developing spleen [8], we
performed additional stainings for IL-7R? and CD4 in subse-
quent sections to detect these cells. Interestingly, CD4?IL-
Fig. 1. Murine fetal spleens at developmental stage E18.5. Immunofluorescence stainings (A–C and E–G) and schematic drawings (D and H). (E–G) Serial
sections. (A) Murine fetal spleen consisting of Tie-2?endothelial cells and CD45?hematopoietic cells (Tie-2 in green; CD45 in red). The outline of the fetal spleen
is indicated in white. A major vein (A and C, arrowheads) and artery (A and B, double arrowheads) can be distinguished at this time-point in development. (D)
Schematic drawing of fetal spleen showing outline and position of vein and artery. (E) High magnification image of blood vessel endothelial cells expressing
MAdCAM-1 (in green) and VE-cadherin (in red). VE-cadherin?blood vessels with the highest MAdCAM-1 expression (yellow) are marked by arrows. Vein
(arrowhead) and artery (double arrowhead) are MAdCAM-1?VE-cadherin?(orange). (F) Expression of ICAM-1 (in red) and VCAM-1 (in green) by endothelial cells
and splenic stromal cells. ICAM-1?VCAM-1?arterial endothelial cells (orange) are immediately surrounded by VCAM-1?stromal cells (double arrowhead, green).
The outline of these cells is indicated in white. Arrowhead indicates a vein. (G) LTi cells expressing IL-7R? (in green) and CD4 (in red) positioned distal to the
artery and VCAM-1?stromal cells, forming a ring-like structure (double arrowhead) that is not found around the vein (arrowhead). (H) Schematic drawing of the
outlines of arterial endothelial cells (i), VCAM-1 ? ICAM-1- stromal cells (ii), and LTi cells forming a ring-like structure (iii). Outward from this structure, blood
vessels with the highest MAdCAM-1 expression are found. Overview image of fetal spleens is shown at 20? original magnification in A. Detailed images of fetal
spleens are shown at 40? original magnification, Zoom 2, in B, C, and E–G. Data are representative of four individual mice.
154 Journal of Leukocyte Biology
Volume 84, July 2008
Page 4
7R??LTi cells were located in a ring-like pattern at the
periphery of the VCAM-1?stromal cells, adjacent to MAd-
CAM-1hiVE-cadherin?endothelial cells (Fig. 1, E–G). The
distinct locations of these cell types are depicted in a sche-
matic drawing (Fig. 1H). To confirm that CD4?IL-7R??cells
were indeed LTi cells, we performed triple stainings for ROR?,
which is indispensable for LTi differentiation [32], IL-7R?,
and CD4. These stainings showed that all CD4?IL-7R??
contained ROR?, indicating that these cells were indeed LTi
cells (Supplemental Fig. 1, D–H and Supplemental Fig. 2).
Few ROR??IL-7R??CD4?cells were found at developmental
stage E18.5 (data not shown).
To see whether a distinct pattern of LTi cells in the prox-
imity of vessels could also be observed at earlier stages of
spleen development, serial sections of E14.5 and E16.5
spleens were analyzed. Therefore, we investigated the vascu-
lature of the fetal spleen in more depth by staining sections of
E16.5 and E14.5 fetal spleens for VE-cadherin combined with
4?,6-diamidino-2-phenylindole (DAPI) and MECA-32 staining.
The latter antibody has been described as an endothelial
marker [33]. The combination of both endothelial markers
allowed us to focus on the artery, which can be distinguished
by expression of VE-cadherin and lack of MECA32 (VE-
cadherin?MECA-32–). At E16.5 in development, we could
identify LTi cells around this VE-cadherin?MECA-32–artery,
again organized in a ring-like pattern. In addition, we found
single LTi cells and small, poorly organized clusters in other
areas of the fetal spleen (Fig. 2, A–D). At E16.5, most of the
vessels were MAdCAM-1?(Supplementary Fig. 1, I–K), sim-
ilar to E18.5 splenic vessels. In addition, VCAM-1?stromal
cells could be distinguished that were organized around the
large VE-cadherin?MECA-32–artery (Fig. 2, B and C). Stro-
mal cells and LTi cells appeared to be intermingled, instead of
strictly separated, as seen in E18.5 fetal spleens. Although at
E14.5, LTi cells could be detected in the fetal spleen in the
vicinity of blood vessels, they were not yet organized in a
ring-like structure, as observed at E16.5 and E18.5 (Fig. 2,
E–H). Strikingly, we could only identify one major blood vessel
in the center of the spleen at E14.5. The lumen of this vessel
was large, and apart from the expression of VE-cadherin, it was
weakly stained for MECA-32 (Fig. 2F). In addition, at this
time-point in development, numerous smaller vessels ex-
pressed MAdCAM-1 and ICAM-1, but VCAM-1 expression in
the fetal spleen was limited at this developmental time-point
(Fig. 2G and Supplementary Fig. 1, L–O). VCAM-1 expression
could be observed in deeper layers of the stomach wall, indi-
Fig. 2. Murine fetal spleens at stages E14.5, E15.5, and E16.5 of development.
Immunofluorescence staining of serial sections from E16.5 (A–D) and E14.5
(E–H) spleens. (A) Double staining for MECA-32 (in green) and VE-cadherin (in
red) counterstained with DAPI (in blue) to visualize nuclei. Most of the smaller
vascular blood vessels appeared to express MECA-32 and VE-cadherin. (B)
Higher magnification image showing a MECA-32?vein that is VE-cadherinlo
(arrowhead) and a VE-cadherin?artery that lacks expression of MECA-32 (dou-
ble arrowhead). (C) ICAM-1?VCAM-1?artery (double arrowhead) surrounded by
VCAM-1?stromal cells (ICAM-1 in red; VCAM-1 in green). (D) IL-
7R??CD4?CD45loLTi cells are organized around arterial endothelial cells
(double arrowhead) (CD45 in green; IL-7R? in red; CD4 in blue). (E) E14.5
spleen (arrow) and stomach (arrowhead) containing MECA-32?VE-cadherin?
blood vessels (MECA-32 in green; VE-cadherin in red) counterstained with DAPI
(in blue). (F) At E14.5, all blood vessels (orange) are double-positive for
MECA-32 and VE-cadherin including a larger blood vessel. The outline of the
larger vessel is indicated in white. (G) At this time-point, most endothelial cells
express MAdCAM-1 (in blue) and ICAM-1 (in red), and VCAM-1 expression is virtually absent in the fetal spleen (in green). (H) In the near vicinity of the larger
vessel, CD45?hematopoietic cells are localized, including IL-7R??CD4?CD45loLTi cells (CD45 in green; IL-7R? in red; CD4 in blue). Images showing
sagital-sectioned fetal spleen (E16.5) or transverse-sectioned fetal spleen and stomach (E14.5) at 20? original magnification. Higher magnification images of fetal
spleens are shown at 40? original magnificaiton and Zoom 2 in B–D and F–H. The outline of major blood vessels is marked in white. (I) On E15.5 fetal spleen,
whole-mount immunofluorescent staining was performed to locate CD4?cells. Data are representative of four (E14.5) or five (E15.5, E16.5) individual mice.
Vondenhoff et al.
Spleen development155
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cating that it was not a result of a detection failure (Supple-
mentary Fig. 1, L and M). To see whether LTi cells indeed
preferentially distribute to certain splenic areas, whole-mount
E15.5 spleens were stained for CD4. These stainings showed
that CD4 cells were clustered and that these clusters were
distributed linearly along the length of the entire spleen (Fig.
2I), indicating that the early compartmentalization of the fetal
spleen may attract LTi cells to periarteriolar domains. In
addition, a limited number of single CD4?cells were found in
the spleen at other areas.
Distinct stromal subsets are present in the
fetal spleen
As LTi cells are localized within special areas of the spleen, we
assumed that these cells are colocalizing with a specific stro-
mal cell subset and that various stromal cell populations could
be expected to exist in fetal spleens. We therefore analyzed
stromal cells present in E15.5 fetal spleen by FACS (Fig. 3A).
After elimination of leukocytes (CD45?) and erythrocytes
(Ter119?), the E15.5 fetal spleen stroma could be separated
into two populations, based on the expression of VE-cadherin.
As can be inferred from immunofluorescent stainings (Figs. 1
and 2), splenic stromal cells that surround arterial vessels lack
endothelial markers such as VE-cadherin. In addition, endo-
thelial cells express VE-cadherin and are CD45–, determined
by FACS analysis (Fig. 3, A and B). To confirm that VE-
cadherin?cells indeed represent endothelial cells, expression
of other endothelial markers was analyzed. Although 17% of
the CD45–Ter119–cells could be distinguished as a population
of CD34?ICAM1?VEGFR2?VE-cadherin?Tie-2?endothe-
lial cells, the remaining 82% of fetal spleen CD45–Ter119–
cells was characterized as CD34–ICAM1–/loVEGFR2–VE-cad-
herin–Tie-2–stromal cells (Fig. 3, A and B). By semiquantita-
tive RT-PCR, we found VCAM-1 transcripts in both subsets
with a probable lowest quantity in the CD34–stromal subset
(Fig. 3C). As LTi cells were found to be associated with
endothelial cells in the spleen, we addressed whether splenic
CD45–cells could produce the chemokines that are known to
attract LTi cells. Quantities of CXCL12, CXCL13, CCL19, and
CCL21b transcripts were determined in CD34–stromal and
CD34?endothelial subsets of CD45–Ter119–cells (Fig. 3D).
The expression levels of CXCL12 and CXCL13 were relatively
similar in both CD45–Ter119–subsets. In contrast, levels of
CCL19 and CCL21b transcript expression were 100- and
1000-fold higher in the CD34?endothelial fraction than in the
CD34–stromal fraction, respectively.
White pulp anlagen is already primed
in fetal spleen
The expression of homeostatic chemokines by CD45–Ter119–
cells and the preferential localization of LTi cells to VE-
cadherin?endothelial cells at the periphery of the MAd-
CAM-1?stromal areas raised the question of whether these
periarteriolar areas constituted the anlagen of the white pulp.
To address whether these designated areas could mature into
splenic white pulp areas, spleens from E15.5 C57BL6 embryos
were grafted under the kidney capsule of adult Ly5.1 Rag2/
?c?/?immunodeficient mice (Fig. 4A). The use of Rag2/
?c?/?alymphoid mice was essential to avoid substantial con-
tamination by host-derived lymphocytes. B cell precursors are
present in fetal spleen at the time of transplantation, and they
are able to differentiate into mature CD19?B220?B cells in
situ. When the grafts were analyzed at 3 weeks after transplan-
tation, they showed good vascularization and had increased in
size, showing that the hematopoietic progenitors present in
E15.5 fetal spleens were able to proliferate and differentiate in
situ. Grafts performed with E14.5 spleen also showed prolif-
eration and differentiation of CD45?progenitors within the
grafted spleens (data not shown).
Analysis by immunofluorescence showed that B cells and
LTi cells colocalized in an area that did not contain Ter119?
erythrocytes, indicative of a B/LTi-containing white pulp area
and a Ter-119-containing red pulp area (Fig. 4A). In Figure
4A, only LTi cells are stained to clearly observe the structure
of the spleen (LTi clusters and vein). Development of these
areas within the grafted spleens indicated that the stroma had
been instructed to retain LTi cells to restricted areas that are
common to B cells. It appeared that the designation of the white
pulp anlagen, containing LTi cells, had already taken place at
E15.5, independently of mature, circulating lymphocytes.
In parallel, splenic grafts were analyzed by FACS to assess
the presence of donor-derived LTi, B, NK, and T cells (Fig.
4B). Depending on the graft, the proportion of hematopoietic
donor cells (CD45.2?) varied, whereas the percent of host
(CD45.1?) hematopoietic cells, containing myeloid and
CD11c?DC, was constant (approximately 10%; Fig. 4B).
Within the pool of donor-derived hematopoietic cells, clear
populations of LTi (CD4hiCD3–IL7R??) and B (CD19?B220?)
cells were seen, whereas some of these B cells expressed high
levels of CD21, potentially representing marginal Zone B cells
(Fig. 4B). Few graft-derived NK cells could be observed (Fig. 4B).
Injection of E15.5 total fetal liver cells from C57BL/6 em-
bryos during splenic transplant permits full reconstitution of
the hematopoietic compartment of the alymphoid recipients. In
these cases, the B and T cells and erythrocytes were clearly
distributed into organized white and red pulp areas. The white
pulp areas showed normal B/T segregation, indicating that T
and B cells could normally enter the white pulp areas (Fig. 4A).
LT?1?2expression during spleen development
LTi cells in developing LN are instrumental for the induction
of homeostatic chemokines in developing LN through their
expression of LT?1?2[34]. According to the literature, the devel-
opment of the spleen requires LT?1?2-expressing cells postna-
tally [14, 35, 36], whereas we showed here that homeostatic
chemokines could already be observed before birth. Therefore, we
addressed at what time-point the earliest LT?1?2-expressing cells
could be observed using FACS analysis of postnatal spleens.
Mesenteric LN (MLN) from the same animals were included as
positive controls. Analysis of Days 0, 2, and 4 and adult wild-type
spleens revealed a gradual increase of LT?1?2on splenic B cells
(Fig. 5A). At Days 0 and 2, only 1% of all B cells expressed
LT?1?2, and this increased to 2.7% at Day 4. In adult spleens,
12.5% of total B cells expressed LT?1?2(Fig. 5B).
In contrast, few LT?1?2-expressing LTi cells could be
detected in neonatal spleens, and LTi cells from MLN
expressed high levels of LT?1?2, in accordance with our
156Journal of Leukocyte Biology
Volume 84, July 2008
Page 6
earlier report (Fig. 5C) [26]. When compensated for a spe-
cific binding (Supplementary Fig. 3), the percentage of
2.2% of all LTi cells, implicating that LT?1?2
are rare (Fig. 5D).
?LTi cells in the spleen varies between 1.5% and
?LTi cells
LT?1?2-dependent white pulp development
As we observed few LT?1?2-expressing cells at Day 2 after
birth, we reasoned that before Day 2, splenic development,
including the formation of the white pulp anlagen, occurred
independently of LT?1?2-mediated LT?R triggering. To con-
firm this, spleens from LT??/?mice taken at Days 2, 4, and 6
after birth were analyzed. In wild-type and LT??/?mice, small
accumulations of B220?B cells could be found at 2 days after
birth around arterioles in the spleen, indicative of undisturbed
formation of white pulp areas. In addition, few CD3?T cells
were seen at this time-point in or near B cell areas in wild-type
and LT??/?animals (Fig. 6A). The effect of the absence of
Fig. 3. The stromal compartment of E15.5 fetal
spleen. (A) At E15.5, the fetal spleen (FS) stroma
were isolated as CD45–propidium iodide (PI)–
Ter119–cells, and two subsets were distinguished
based on CD34 and ICAM-1 expression. Most cells
were CD34–ICAM1–/lo, and a small subset was
formed by CD34?ICAM1?cells. FSC, Forward-
scatter; SSC, side-scatter. (B) These subsets of
stromal cells were analyzed for the surface expres-
sion of VEGFR2, VE-cadherin, and Tie-2. (C)
Semiquantitative RT-PCR analysis of the Vcam1
gene in the E15.5 fetal spleen populations (dilution
with a factor 10). Intensity was quantified to mea-
sure the difference of Vcam1 expression normal-
ized by ?-actin levels. (D) Quantitative real-time
PCR was performed to study the relative expression
of Cxcl12, Cxcl13, Ccl19, and Ccl21b chemokines
from the CD34–ICAM1–(blue histograms) and CD34?ICAM1?(orange histograms) stromal populations. The average of Gadph and hypoxanthine guanine
phosphoribosyl transferase housekeeping gene levels was used for the normalization. Results are obtained from two independent cell sorting, and each sorted
population is represented by a specific histogram.
Vondenhoff et al.
Spleen development157
Page 7
LT?1?2-expressing cells could be observed around Day 4,
when B cell areas were still small in LT??/?, and they had
clearly increased in size in wild-type mice from Days 2 to 4.
In addition, T cell areas were completely absent in LT??/?
spleens at all stages analyzed, and these were clearly
present in wild-type spleen at Day 4 and had further en-
larged at Day 6 (Fig. 6A). Despite the dramatic differences for
the B and T cell populations, we could not find a clear difference
with respect to MOMA-2?macrophages. In fact, the amount of
MOMA-2?cells that were found in close proximity to central
arterioles appeared even higher in LT??/?spleens when com-
pared with wild-type spleens at Day 4 (Fig. 6A, and data not
An additional difference was observed when splenic white
pulp stromal cell populations of wild-type mice were compared
with LT??/?mice. When staining for VE-cadherin, VCAM-1,
and ICAM-1, VE-cadherin-negative stromal cell populations
could be analyzed. VCAM-1?ICAM-1locells were located
directly around the central arteriole. At the periphery of this
white pulp area, VCAM-1?ICAM-1?cells could be seen (Sup-
plemental Fig. 4). When wild-type and LT??/?spleens were
compared, these cellular subsets could be distinguished at Day
2, but the increase in VCAM-1?ICAM-1?at the periphery of
the white pulp area seen at Days 4 and 6 in wild-type was not
observed in LT??/?mice (Fig. 6B). These results indicate a
cation where the marginal zone will develop and is in agree-
ment with earlier observations that LT?1?2/LT?R interactions
are mandatory for homeostasis and proper functioning of mar-
ginal zones in adult spleens [16, 36–39].
?-mediated increase of VCAM-1?ICAM-1?at the lo-
Fig. 4. Early priming of the fetal spleen stroma. LTi cells maintain in fetal spleen graft and colocalize with
B cells. (A) Three weeks after engraftment of CD45.2?fetal spleens under the kidney capsule of CD45.1?
Rag2/?c?/?mice, grafts were analyzed by immunohistology for LTi cells (CD4 in green), B cells (B220 in
white), and erythrocytes (Ter119 in red; FS Graft). The first quadrant represents LTi cells forming ring-like
structures (white ovals) with a vein (arrowhead) devoid of LTi cells. The second quadrant displays LTi cells
and B cells in close contact to each other. The same experiment, supplemented with injection of CD45.2?
fetal liver cells (FL), was used as a positive control for splenic architecture, and most CD4?cells (green) are
T cells (FS Graft?FL Injection). Original blue bar, 200 ?m. (B) Grafts were analyzed for their composition.
Cells were separated between graft-derived (CD45.2?) and host-derived (CD45.1?) and analyzed by flow
cytometry for the presence of LTi (CD4?CD3ε–IL7R??), B (CD19?B220?and B220?CD21?), NK
(NK1.1?CD3ε–), and T (CD3ε?) cells and DC (CD11c?). Data are representative of analysis of three
individual grafted mice.
158 Journal of Leukocyte Biology
Volume 84, July 2008
Page 8
The potential role of LTi cells in white pulp ontogeny has been
a matter of debate, as these cells express the ligand for the
LT?R in developing LN, and signaling through this receptor is
indispensable for normal white pulp development [37, 40].
Although it has been shown that triggering of the LT?R by
LT?1?2-expressing B cells is crucial for white pulp develop-
ment, the possibility remained that triggering of the LT?R by
other cells, such as activated T cells or LTi cells, further
facilitated this process. Here, we show for the first time that the
majority of splenic LTi cells lacks cell surface expression of
the LT?R ligand. It has been shown that fetal and neonatal
splenic LTi cells express the LT? and LT? transcripts [9, 41].
The adult spleen environment, but not the fetal, has also been
proposed to regulate the LT? transcript expression in LTi cells
by TL1A [41]. Our results suggest that the neonatal, splenic
environment does not lead to cell surface expression of LT?R
ligands on LTi cells. An in vitro, dose-dependent IL-7 stimu-
lation is required for an effective LT?R ligand expression by
intestinal LTi cells [10]. We previously detected Il-7 transcript
expression by fetal spleen nonhematopoietic cells [42]. At Day
4 after birth, the time-point that T cell areas start to emerge, a
ligand for the LT?R, most likely LT?1?2, is only expressed by
B cells. Thus, for white pulp development, B cells will give the
LT?1?2-dependent, inductive signal, similar as LTi cells do
during LN and PP formation, and this is in accordance with
earlier observations [14]. This signal is required to attract more
B and T cells to the developing white pulp. In contrast to the
white pulp development into functional T and B cell areas, the
separation of white and red pulp areas occurs before birth and
is independent of LT?-expressing cells
In E15.5 fetal spleen, patches of LTi cells are found
throughout the spleen with a distribution reminiscent of white
pulp areas of adult spleen. Although our results indicate that
LTi cells do not directly contribute to white pulp development,
the localization of these cells, their close association with blood
vessels, and their probable interaction with VCAM-1?ICAM-
1–/lostromal cells at E18.5 are intriguing but might just be a
reflection of the chemokines and adhesion molecules that are
locally expressed.
The Cxcl12 and Cxcl13 transcripts were similarly expressed
within the "stromal" and "endothelial" stromal cell subset. On
the contrary, the CD34?ICAM?endothelial stromal cell subset
expressed higher levels of Ccl19 and Ccl21b transcripts than
the stromal subset. Assuming that these relative transcript
expressions are revealing the protein expression levels, we
propose that the perivascular distribution of LTi cells could be
mediated by the high expression of CCL19 and CCL21 che-
mokines from the endothelial cells. Indeed, the lower expres-
sion of CCL19 and CCL21 by the stromal cells would establish
CCL19 and CCL21 concentration gradients, decreasing from
the vascular endothelial cells to the inner part of the fetal
spleen. The CXCL12 and CXCL13 chemokine expressions by
the splenic stroma probably also participate in the attraction
and retention of LTi cells into the fetal spleen. Early expres-
sion of CXCL13, before the entry of B cells, has been observed
around arterioles in developing human spleen [27]. Thus, expres-
Fig. 5. LT?R-binding cells in neonatal spleen and MLN. LT?R-human IgG
(huIgG) binding to B cells (A and B) or LTi cells (C and D) was analyzed by
flow cytometry at indicated days after birth, and four mice per group were
Vondenhoff et al.
Spleen development 159
Page 9
sion of these homeostatic chemokines most likely contributes to
the periarteriolar localization of LTi cells in the fetal spleen.
Our graft experiments show LTi and B cells colocalizing into
well-defined areas within fetal spleen, illustrating the early spe-
cific capacities of the spleen stromal cells. In the grafts, all B and
LTi cells are donor-derived and have differentiated in situ. Both
spleen stromal populations expressed the transcripts for CXCL13
and CXCL12, important for LTi cell interaction and B cell attrac-
tion and differentiation [11, 43, 44]. Moreover, we observed a
small host-derived CD11c?DC population in these grafts. It was
shown that CD11c?cells can be found in a neonatal spleen, only
a few days after birth [45, 46]. Hence, DC have migrated into the
grafted spleen in response to the chemokines produced.
In our grafts, a typical adult spleen organization was ob-
served when total fetal liver cells were injected previously. The
B/T lymphocytes that differentiated from these fetal liver cells
were attracted by the fetal spleen graft into their specific
domains, reinforcing the capacity of the white pulp anlagen to
express chemokines essential for the development of its archi-
tecture. It was shown that CCL21 could be expressed by two
different genes after an activation of a LT?1?2-dependent or
-independent pathway, whereas the two chemokines conserve
the same chemotactic activity [47]. Most of CCL21 expression
depends on LT?1?2in the adult spleen but not in nonlymphoid
tissue [47]. It is possible that the first chemokine expression
found in the fetal spleen at E15.5 is triggered in a similar
LT?R-independent manner.
Finally, our data show that the fetal spleen stroma are
instructed at an early time-point to segregate lymphoid and
nonlymphoid zones, which correlate with red and white pulp areas
of the adult organ. The graft experiments and the low numbers of
B or LTi cells in the fetal spleen that express the LT?R suggest
that these first driving events are LT?R-independent.
This work was supported by fellowships from Ministe `re Fran-
c ¸ais de la Recherche et de l'Enseignement Supe ´rieur, from
Association pour la recherche sur le cancer (ARC), from ANR,
and from the European Stem Cell Research Program to
G. E. D., a Vici grant from the Netherlands Organization for
Scientific Research (918.56.612) to R. E. M., and a genomics
grant from the Netherlands Organization for Scientific Research
(050-10-120) to M. F. R. V. The authors declare that there is no
conflict of financial interest. We acknowledge A. Louise for cell
sorting and the Flow Cytometry Core Facility (Institut Pasteur,
France) as well as E. Perret for her advice and the Dynamic
Imagery Core Facility (Institut Pasteur).
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individual mice per group.
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Page 10
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Spleen development161


text:
District Eight Design 27.75" Metal Bowl Ceiling Fan Fitter Shade
District Eight Design 27.75" Metal Bowl Ceiling Fan Fitter Shade
Weights & Dimensions
|Product Type: Ceiling fan bowl shade
|Attachment Type: Screw on
|Style: Traditional
|Shape: Bowl
|Color: Black
|Lifestage: Adult
|Theme: Solid
|Primary Material: Metal
|Country of Manufacture: Imported
Still have questions about this product? We're here to help
District Eight Design 27.75" Metal Bowl Ceiling Fan Fitter Shade
About the ManufacturerDistrict Eight Design is an office and workshop located in a refurbished colonial townhouse facing Saigon River. Founded in April 2010, the firm is one of the first in Vietnam to merge design and build. By combining the office and workshop, District Eight Design has the ability to rapidly prototype furniture and lighting while easily producing individual, custom pieces for each design project. Although District Eight Design was originally created to preserve and develop historically significant buildings, warehouses, and objects in Ho Chi Minh City, the firm has rapidly expanded to include a production line of modern industrial furnishings that respond to these environments, both residential and commercial, both here and abroad.
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OUR SKU # NFV101327.75" Metal Bowl Ceiling Fan Fitter Shade is also sometimes listed under the following manufacturer numbers: HGDA153


text:
Map to Park
Ice Park is right in the middle of Fairbanks. An oasis of Ice Art and Kids Park!
Single Block Winners
Single Block World Championships winners Ivan Zuev and Eduard Ponomarenko of Russia.
Open Through Mar 28
Park closes TODAY - on March 28!
2015 Multi-Block Winners
FIGHTER; 1st Place Realistic, BY: Junichi Nakamura Shinichi Sawamura Chan Kitburi Dean Murray; JAPAN, USA
News and Updates
Ice Park Closes Early
ICE PARK CLOSES EARLYGreetings to All! Fairbanks is experiencing a spectacular spring season, this being the third day in a row that the temperature has breached the +50F in the Park, it's just too much for the ice. So we are announcing that today is the last day to play in the Park. 5:00 p.m. - NEWS UPDATE - PINS JUST ARRIVED!Good afternoon! I just got the call from Mr. Brickley. The Pins just arrived this afternoon. The office is working out the logistics of trying to get everyone their souvenir...
Rhonda's Blog
A Day at the Park
The heat of competition is over and like many of the artists and volunteers, I have gone back to my regular day job. I stopped by the Ice Park today to catch up on the support needs from my department, in Web and Media. For me, the big show is the building of the Park, and the competitions in Single Block and Multi Block exhibits. But, for a lot of folks, the Park is the big attraction, with it's train ride tour, and especially for the kids, even the adult kids, the abundance of slides. For ...


text: You are viewing Favorites - 1 of 2 Videos Agent Officers Episode 2: Sunny D (Part 1) By ROCKETCOOLCOMEDY Leaving off from the first episode, we see our two heroes being held captive from the... more »Leaving off from the first episode, we see our two heroes being held captive from the mysterious Optimus Crime. Not only do they have to figure out a way to get out of this sticky situation, but they still have to find out what was in that black indestructible briefcase! Word on the street is that there's a new drug in town thats gotten all the addicts crazed for more. Some call it the new cocaine. Others call it Sunny D. « less Published March 22, 2009 270 views More Info » Full Credits ROCKETCOOLCOMEDY Uploader Stats & Data 22 Funny Votes 1 Die Votes 272 Views Published March 22, 2009 Categories: Web Series Clean Comedy Parody Keywords: Sunny D We See Agent Officers 2 Optimus Crime Up Next: Agent Officers Episode 2: Sunny D (Part 2) AUTOPLAY Up Next In: 10 Mini Golf FAIL The Best of Conan Bowie Sings CAINE on POP!Now Walk Hard - Reefer Follow Facebook Twitter Tumblr Instagram Browse Related Comments Favorites 10+ more View All ▶ Agent Officers 2 2+ more View All ▶ 10+ more View All ▶ ROCKETCOOLCOMEDY Vaguely Related Videos Vaguely RelatedPics & Words Advertisement Advertisement From Around the Web ZergNet


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Possible to Paint Fireplace Trim Gold to Black?
Hi! Sorry if this has been asked already, but I couldn't find it on a Search...
The doors and some of the surrounding grill of our zero-clearance fireplace doors (converted from wood to gas) has GOLD metal trim (80's style!) I was wondering if anyone knows if it's possible to paint that same trim BLACK? If so, what kind of paint would you use and what sheen? I'd probably have to do it in place versus removing all the parts (or at least it'd be easier). I am hoping to update the look without having to replace the whole front panel. Thanks a million for your help!
We did. We used a special high heat fireplace paint. Looks brand new now, no 80's
THANK YOU for the reply!
Did you sand the gold areas first? (If so, just to rough it up OR to actually sand off as much gold as possible?)
Do you remember what brand paint and/or where I should ask? (Home Depot? Sherwin Williams type specialty store? Fireplace store?)
(I just hate to start something and be left a big mess on my hands!!)
Thank you so much again!
I think it would be a good idea to sand the surface first, and even then, there is a risk that some of the new paint will not adhere properly and may need to be redone. The reason I say that is because we had a black wrought iron table pedestal repainted professionally. We were told that they "abraded" the surface first by sanding it down before applying the new paint, which did not involve a change in color.
I would therefore try to make the surface as rough as possible before putting paint on it.


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Air Dubai
These songs sound like they've been percolating in the imaginations of Julian Thomas and Jon Shockness for years in terms of what would make for killer hip-hop songs. "Nevermind the Bollocks..." clearly references the Sex Pistols, but the music is reminiscent of the type of jazz-lounge appropriation employed by Protection-era Massive Attack, while "Warm Day in L.A." conjures a montage of sunset daydreams, and "Suicide Bomber" confronts American fears of the Muslim world by musically embodying those fears. The tracks here flow not just with deft lyrics, but also with intelligent observations and a rich frisson of artistic traditions. Fans of PM Dawn's particular fusion of R&B, jazz and hip-hop will appreciate how well Air Dubai integrates all of its sounds with perfect harmony.
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The Road Ahead With An Innovative Product (re)Search Engine
PriceBaba is a unique product search engine that has a bold ambition to bridge the gap between the online and offline world. PriceBaba began with a belief that people research on the internet, but more often than not, end up making their purchases in the real world.
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We are a Mumbai based Startup building a unique product research engine for the Indian market. It is an enormous task to deliver the right value to our customers and users, enough to keep our engineers on their toes to find innovative solutions. We realize that there is plenty we can do with the reach that we have already built and are looking to work with engineers who have experience in building internet products.Requirements:
We are looking for experienced front-end/full-stack engineers to complement our current team. You would take ownership of all or part of any of the applications built @PriceBaba. This would include, design and architecture of software features, team to understand and influence the roadmap, and implement UI and RESTful resources for new features.
We value autonomy and self-reliance, and above all the ability to ship.
We are looking for a DevOps Engineer responsible for the core operational services that enable engineering to operate at maximum speed. In addition to provisioning, maintaining, and curating the many cloud-hosted servers upon which all the engineering and production services reside, the Engineer will collaborate on the actual engineering and production services including monitoring, tuning, upgrading, and advising where appropriate.
We value autonomy and self-reliance, and above all the ability to ship.
PriceBaba serves over 4 million users a month on its local commerce platform. In a mobile first Indian economy.
PriceBaba is building up its mobile development team to offer a great app experience to its users.
PriceBaba is service over 12 million page views to over 4 million users every month. There is plenty of Ad space to be sold. We are looking at an executive with existing experience in selling ad space in the consumer electronics / mobile phones / telecom / technology sector.Requirements
We are looking for a highly motivated professional to assist in our back office operations. The candidate should be able to work well in a team environment and be self motivated. He/She should be very well organized and have close attention to detail. Importantly he/she must be comfortable accepting responsibility for their tasks.Who Are You?


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Moving On
By Larry McMurtry
(Simon & Schuster, Paperback, 9780684853888, 800pp.)
Publication Date: June 4, 1999
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Moving On is a big, powerful novel about men and women in the American West. Set in the 1960s against the backdrop of the honky-tonk glamour of the rodeo and the desperation of suburban Houston, it is the story of the restless and lovable Patsy Carpenter, one of Larry McMurtry's most unforgettable characters.
Patsy -- young, beautiful, with a sharp tongue and an irresistible charm -- and her shiftless husband, Jim, are adrift in the West. Patsy moves through affairs of the heart like small towns -- there's Pete, the rodeo clown, and Hank, the graduate student, and others -- always in search of the life that seems ever receding around the next bend.
Peopled with a riotously colorful cast of highbrows, cowpokes, and rodeo queens, in its wry humor, tenderness, and epic panorama, Moving On is a celebration of our land by one of America's best-loved authors. Moving On is vintage McMurtry.
Larry McMurtry is the author of twenty-nine novels, including the Pulitzer Prize-winning Lonesome Dove, three memoirs, two collections of essays, and more than thirty screenplays. He lives in Archer City, Texas.
The New York Times
A novel of monumental honesty....Attention must be paid.
Los Angeles Times McMurtry can transform ordinary words into highly lyrical, poetic passages...He presents human drama with a sympathy and compassion that make us care about his characters in a way that most novelists can't.
The Boston Globe There aren't many writers around who are as much fun to read as McMurtry. He is precise and lyrical, ironic and sad.
Saturday Review A Texas-sized book...Mr. McMurtry is blessed with an absolutely solid sense of place. His backgrounds and scenic descriptions are inherent parts of his story, contributing as much to the novel as does the completely natural dialogue.


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Category Archives: Mount Vernon
"Route 66″ gets its kicks in Baltimore, Maryland
And Buz Murdock does geneology research at Pratt Library! "The Mud Nest" Route 66: Season 2, Episode 7 November 10, 1961 An encounter with a rural Maryland family bearing a striking resemblance to him leads Buz (George Maharis) to Baltimore … Continue reading
Stop Lies, Defamation, Ignorance, and Racism
Read and Learn! by Tom Warner (Baltimore Or Less) It started with anonymous notes posted throughout the Enoch Pratt Free Library's Central Library on 400 Cathedral Street. The notes – spotted on bookshelves and, in at least one case, left … Continue reading
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Styrofoam Sermon on Mount Vernon
And seeing the multitudes, He went up on a mountain, and when He was seated His disciples came to Him.Then He opened His mouth and taught them… (The Gospel According to Matthew, 5-7) All Christians know about the Sermon on … Continue reading
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5 votes
Video : Charles Bukowski on the Majority Rule, Democracy and Evil MenSubmitted by DistalDub on Sat, 08/03/2013 - 12:41
Trending on the Web
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"Wisdom is doing everthing the crowd does not do"
I was a big fan of Bukowski a few years ago. He spoke a lot of truth. Thanks for posting.
"It's not pessimistic, brother, because this is the blues. We are blues people. The blues aren't pessimistic. We're prisoners of hope but we tell the truth and the truth is dark. That's different." ~CW
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Deliverance is a free single-player expansion pack for Sierra's Half-Life. In Deliverance you reprise your role as Gordon Freeman in a story set after the end of Half-Life. Voted #8 in the Top Ten Half-Life Add-ons by PC Gamer magazine in May 2000.
Feb 2nd, 2013
No review provided
Nov 29th, 2011
29 votes submitted.
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Ratings closed.


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Lightening Hot Male
Summer_NHMay 28, 2005
I see I have some messages to get caught up on. I have been busy redoing some rooms in my house. I had to do something with this hottie tube, so here's a treat for you gals eyes, LOL! I made a set of tags also.
Below are the links for the tags in 4 zip files. My sig tag is preview. There are 23 tags. I made as many tags as I could with it, LOL!


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Mz_Major: Thursday, June 28, 2007 - 13:24
I wanna be in the video too
Jus saw the Wall to Wall video a few days ago... i was like dang... what is really goin on. How come nobody hears about auditions. I dont have time to get a agent n do this n that. I jus wanna audition. I dont even have to get the lead role i jus wanna dance. He needs to come back out to VA and shoot a video.


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Fart Extinguisher - Fun, Novelty Air Freshener
The novelty fire extinguisher shaped air freshener bottle.
Product Code
Price in GBP
More than 10 in stock.
See all ourNaughty Presents
Why not include these in your basket as well?
- Description
- Delivery
- Reviews
Fart Extinguisher
Fart Extinguisher (ex·tin·guish·er)
A portable device that discharges a fresh, clean fragrance extinguishing the foul smell of a Fart.
Every home should have at least one but for complete safety, keep one in every room.
The novelty Fart Extinguisher is a fire extinguisher shaped can of air freshener, which sprays a fresh garden fragrance perfect for clearing the air of nasty smells.
Directions for use:
- Twist collar to unlock.
- Hold upright.
- Stand at safe distance.
- Aim at the centre of the fart.
- Squeeze lever, sweeping side to side until fart is under control.
Product Details:
- Bottle Size & Volume:
H15 cms (H6 inches). 100ml (3.5floz)
- Extra Information:
Realistic looking mini extinguisher bottle.
The perfect novelty present or stocking filler
Keep yourself safe, never have to suffer a fart again!.
Service rating : Great service. efficient quick delivery, safely packaged. No problems.
Product : A funny gift. Created a good laugh.
Quick and easy transaction, quick delivery
Service rating : It was delivered promptly and I was really pleased I found the site because they had so original gift ideas.
Product : It was the funniest thing I had seen in a long time and was an apt present for my brother and his long suffering wife.
Service rating : Very useful service
Product : Good joke present for grandson
Great delivery time!! Everything there! Good website
The service was good and effective and on time.
Service rating : ...................
Product : .............
Excellent service through a period of none delivery of stock - managing expectations well and delivering pre Christmas in the end
would recommend on that basis - and have
Service rating : Quick and efficient
Product : Funny
Service rating : Fab
Product : Fab


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Terri Bender
No. 6 UIC at No. 3 Cleveland State Wednesday for HL Quarterfinals
Flames defeated the Vikings in the 2014 Quarterfinals
Campbell and Bender Earn All-League Honors
Ruvanna Campbell won her second straight Defensive Player of the Year award
Flames End Regular Season at Cleveland State
Tip-off with the Vikings is set for 1 p.m. CT inside the Wolstein Center
Women's Basketball Begins Ohio Trip in Dayton
UIC (16-11, 6-8 HL) seeks its first three-game win streak since Nov. 30 - Dec. 6
Truiett to Be Honored as UIC Hosts Oakland Saturday
Senior Gisselle Truiett's career will be celebrated prior to the contest
2014-15 Game-By-Game
2013-14 Game-by-Game
2012-13 Game-by-Game
PREP AND PERSONAL: Four-year letterwinner at Pius XI high school in basketball ... helped the Lady Popes win a state championship in 2010, a state appearance in 2011 and three consecutive conference titles (2010, 2011, 2012) ... selected to the All-State team her sophomore through senior seasons and scored more than 1,000 points in her prep career ... averaged 14 points, five rebounds, three steals and three assists per game in 2012 ... competed for the Wisconsin Playground Elite AAU team ... daughter of Terry and Yolanda ... siblings named Lorell, Tamara, Tierra and Taryn ... brother Lorell played basketball at Tuskeegee University and sister Tierra currently plays basketball at Concordia-St. Paul ... enrolled as a Psychology major.


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AboutA web log for all fans of Walt Disney, The Disney Company, and its many products. Using the theories behind Walt Disney's family oriented entertainment and education, this blog is intended to be a safe read for the whole family. Proud to be a part of Quality Will Out Omnimedia. The Disney Blog is not affiliated with The Walt Disney Company in any way. All Disney parks, attractions, lands, shows, event names, etc. are registered trademarks of The Walt Disney Company. The Disney Blog is affiliated with LaughingPlace.com, the number one Disney Fan site on the internet. Please see creative commons license at the bottom of this page for copyright information. All content on this website is my own and does not necessarily represent the views of my employer or anyone else on the planet. The Disney Blog is supported by advertising and some of the links on this site and in product reviews are revenue share affiliate ads. I also do accept product for review and to give away as prizes. However, I compose all my editorial separate from any decision regarding advertising or affiliate links. YMMV.
Tag Archives: synergy
Marvel and ESPN Team Up
Yes, you read that correctly. Disney is doing what it does best – synergy. Marvel and ESPN have come together to create a documentary and a series of video shorts. Both projects combine the style of ESPN's films with Marvel's … Continue reading
Posted in ESPN, marvel Tagged disney, ESPN, marvel, sportcenter, super heroes, synergy Comments Off
How the Walt Disney Company Solved The "Boy Problem"
The most crucial difference between Michael Eisner and his successor at Disney, Bob Iger turns out to be a belief in building around franchises. The buzzword from Eisner was always synergy, code word for cooperation between divisions. Iger came from … Continue reading
Posted in Corporate Tagged acquisition, Bob Iger, business, business strategy, CEO, Corporate, dis, disney, franchise, lucasfilm, marvel, michael eiser, pixar, synergy, the muppets, twdc 1 Comment
ESPN Muppets Crossover in Latest Ad
There's something to be said for synergy. If you add up it's own library, Pixar, Marvel, Lucasfilm and other purchases Disney now owns the rights to well over 25,000 characters. Disney acquired The Muppets early in this string of acquisitions. … Continue reading
Posted in ESPN, Muppets Tagged ad, commercial, ESPN, jim henson, synergy, the muppets, tv 3 Comments
Targeting Young Boys with Phineas & Ferb
It's no surprise that Disney has a hit with the animated TV show Phineas and Ferb. It took the company a while to wake up to the revenue potential for the show, but now that they think they've found another … Continue reading
Posted in Television Animation Tagged Animation, disney, franchise, perry the platypus, phineas and ferb, synergy, tv, tv animation 17 Comments


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Stingers notebook: Stingers add 4 for '14
WILLMAR — Two weeks from the season opener, the Willmar Stingers pushed their roster to 28 with four new signings, including one of the best base stealers in the country.
Illinois State outfielder Sean Beesley is tied for 10th in Division I with 27 steals on the season to go with a .270 batting average and .375 on-base percentage out of the leadoff spot.
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When it comes to children's decor, we always try to combine elegance with a hint of humor. A lot of childhood is about playing and having fun, so we think it's important to try to add a touch of whimsy to their rooms. Here are some picks that are playful and chic at the same time. […]
Design Reveal: Little Man in a Big World
Following the reveal of the baseball room two weeks ago, we now reveal his older brother's geography and travel themed room. Our six-year-old client loves geography, so our first quest was to find a way to incorporate a world map into the room. We decided to make the map the focal point and selected a world map mural with a
It's a Small World After All
More and more we have parents asking us to incorporate world maps and globes into their children's rooms. Whether it's to introduce our world to their little ones or to incorporate a sense of wanderlust, we love finding innovative ways to fulfill this request no matter what the style of the room. This warm-toned Small World Ottoman, from Restoration Hardware,
Design Reveal: Modern Gray and Orange Nursery
When we started working on this nursery, there were three elements that we needed to incorporate—the iconic Hicks Hexagon wallpaper that was already installed in the room, a vintage Dr. Seuss print that the parents-to-be owned, and they wanted to feature a family tree somewhere in the space. Tying together these three disparate elements with the parent's desire for a


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Great Lakes nearing 80 percent ice cover … and rising
Mike Simonson, Wisconsin Public Radio
The National Oceanic and Atmospheric Administration says Lake Erie, Lake Huron and Lake Superior are now almost totally covered with ice.
Monday marked the 22nd straight day of subzero temperatures recorded by the Duluth National Weather Service (NWS), tying a 50-year-old record. NWS representative Steve Gohde expected Tuesday morning would break that record.
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