I don't want Louisville shaking in its boots. I want them comfortable, expecting to blow us out like we're Arizona (a team that arguably shouldn't even have made the tourney). I want them thinking we're a poor man's UNC, with our poor man's Hansbrough (Suton) and our poor man's Lawson (Lucas).
You can predict anything you want, but the Cardinals aren't going to hang a hundred on us. We're the real thing. They have to go from a 12 to a 2.
I hope they're ready for the culture shock.
Two more things I wanted to say. I wish you guys could have been walking campus (okay, not campus, but the MAC/Grand River bar route) after the game. It was awesome. The cops were out on horseback, and everyone was very happy.
The other thing I wanted to say was, the only free throw we missed ended up being an extra possession for us (and led directly to Morgan's game-tying slam with 1:44 to go). I almost feel like Summers did it on purpose.
Also, I love this school. I'm so proud to be a Spartan, even though I'm graduating. And I'm so proud of this team. They have yet to let me down in the final minutes when the game is seemingly a coin toss. GO GREEN!
I haven't been that tense during a basketball game since '00 during the Iowa State game. I was pacing back and forth… nervous… anxious… tense. It was aweful!
This group is a really clutch group of guys. They don't have the mental concentration to dominate a game the whole way through yet, but they do know how to finish. If these sophomores stay around, I think the sky is the limit for them!
BTW – I think Chris Allen is playing really well in the tourney; not spectacular but solid. He's played good D, hasn't been a TO machine, and his presence is forcing the other team to respect his shooting. That gives me hope for next season, as well as for tomorrow!
Solid recap kj considering it was almost 2 am. I thought you would forget about Summers put back at the end to cut the lead to 3. Good thing Suton trims his finger nails or we could have been in even bigger trouble. Officiating was terrible for both sides. I wanted to jump through my plasma and choke the ref that called the tripping at the end. Also, what was up with Gus Johnson last night? I lost a lot of respect for him last night. To start rumors about Izzo being under appreciated was completely unprofessional. This team is a less talented version of the 2000 team. The past 2 games were very similar to the sweet sixteen, elite eight games in 2000. Both teams have (had) the onions to win close games and refuse to lose. Regardless of what happens on Sunday this season has been a great success.
A few more thoughts.. Izzo called some masterful plays out of time outs. Whatever halftime adjustments he made also worked considering we only had a few turnovers in the 2nd half. Remember when we were talking about Raymar going pro early after his freshman season? That seams like decades ago. He has to have a double double on Sunday for us to have a chance. Everyone is talking about Louisville being a terrible match up because they force turnovers. Thoses are forced using a press which we usually do a good job with, giving us some open looks. Watch for Louisville to tone down their press and maybe use some zone.
Yeah, the crowd was overwhelmingly Cards fans. I was surrounded by red. Many of them made disparaging comments about our anemic offense throughout the game. Needless to say they are not at all worried about Sunday's game. Hopefully some of this overconfidence will carry over to the Louisville team.
I couldn't believe it when Johnson said that. Izzo feels like he isn't appreciated around campus? PSSSSH! Either someone was blowing smoke up Johnson's wazoo or he flat out made that up. That would be so out of character for Izzo. But that's what happens when national media talk about anything. They just appear clueless.
Forgive me for bringing this up, but we're kind of proud of this win.
Your comment "I think I'm correct in saying that this team still has yet to lose a game that's within a single possession with 2-3 minutes to go" made me go check the box scores.
When Penn State visited East Lansing, MSU closed to within three at the 1:52 mark, two at 30 seconds, then just one with 12 seconds left. When Andrew Jones only converted 1 of 2 foul shots, Kalin Lucas had the ball in hand with a chance to tie or win with 11 seconds left, but he missed a jumper.
Sorry about bringing back that memory but good luck tomorrow.
We have been pretty clutch in close games. I think Morgan just is not back in basketball shape yet. He missed a lot of time and lost a lot of weight – undoubtedly some of it muscle mass.
Two keys to winning the turnover battle in the second half – Lucious and Morgan were sitting for pretty much all of it and they were primary culprits on the turnover front. Lucious has a habit of driving and then no-look passing to where he thinks an MSU player should be, only it is straight to an opposing player. He has to keep the dribble alive then he's down around the foul line so he can back it back out himself. When he picks it up there they double team him, he panics, and throws it away.
The officiating was terrible at the end of the game. Two ticky tack fouls on us and I think one before that on them. It was like they thought we were fouling on purpose there at the end. I don't know what they saw on that last foul on Collins where (thankfully) he missed the front end. It worked out for us but the refs should not have that much impact on the end of a game.
So, does anyone know what happened to Delvon Roe? He played barely, if at all, in the second half.
You've got me on a technicality there. Truth be told, your boys were up on us pretty big with several minutes to go. Only a series of missed FTs let us back into it. So I view that game a little differently; it wasn't a back-and-forth single-possession game down the stretch.


text:
Renters beware: 14 things to look out for in that lease
Buy Curious
What to know if you're looking to buy a place in Forest Hills
This four bedroom colonial-style house in Forest Hills Gardens is listed for $1,980,000.
Rutenberg New York
April 26, 2018 - 10:00am
By Leah Hochbaum Rosner
Looking for a neighborhood that's a mix of urban and suburban and still under an hour by subway from Manhattan? Try Forest Hills, Queens. The neighborhood is storied but unpretentious, with a bustling center and leafy side streets, and a dizzying variety of houses and apartments, from humble co-ops to storybook mansions.
In this week's Buy Curious, Howard Glick of Citi Habitats, Edith Nanazia of BOND New York, and Karina de Rojas-Kosinov of Mdrn. Residential tell you everything you need to know about the area.
What can you tell me about Forest Hills? What does it cost to live there? What kind of real estate is available?
"Forest Hills offers a unique juxtaposition: cosmopolitan chic clothed in the bucolic serenity of a suburb," Glick says. "As such, it has always enjoyed steady and enduring popularity, almost flying under the radar as other neighborhoods have taken spikes in popularity with their development and subsequent price inflation."
Forest Hills is bordered by Rego Park, Middle Village, Kew Gardens, and Flushing Meadows Corona Park.
The Long Island Expressway runs along the northern border. Union Turnpike forms the southern one. Grand Central Parkway is the eastern edge, and 67th Avenue, Selfridge Street, and 102nd Street make up the western boundary. Fancy Forest Hills Gardens, just south of the Long Island Rail Road station, is a private development of single-family houses built in the early 20th century. Nearby Van Court and Cord Meyer in northern Forest Hills are also known for their low-density, residential vibe.
Who lives here?
As of the 2010 census, Forest Hill's population was 86,364. It's 58.3 percent white, 24.2 percent Asian, 12.4 percent Hispanic, and 2.5 percent African American.
The neighborhood has many Jewish residents, and Cord Meyer and neighboring Rego Park are home to one of the largest populations of Bukharian Jews in the world outside of Israel.
Professionals priced out of Manhattan and Brooklyn are among those drawn to the neighborhood.
"People are attracted to the neighborhood for its convenient access to Midtown, its sophisticated yet comfortable atmosphere, and the extremely large, well-priced apartments," Glick says.
Many young families have also moved to Forest Hills because of its school district, de Rojas-Kosinov says.
What types of housing are available?
The housing stock in Forest Hills is a mix of houses and co-op buildings. According to Glick, "the neighborhood developed most actively from the 1930s through the 1950s, accounting for the proliferation of large prewar buildings that define the look of the area. The neighborhood skyline is punctuated by some high-rise development that occurred in the late '50s and early '60s." Those latter buildings are a mix of rentals and co-ops, with a few condominiums sprinkled in.
Then, there are the houses. They are mostly Tudors and colonials, including detached, semi-attached, and attached homes. Forest Hills Gardens is characterized by formal mansions—mostly Tudors and colonials. The neighboring Van Court area has similar, but smaller and more modestly priced homes. And the Cord Meyer section is also made up of single-family houses, including Cape Cods and Tudors, plus new, bigger houses, many built to accommodate large Bukharian Jewish families.
What's pricing like?
The average sales price thus far in 2018 is $667,402, according to appraiser Jonathan Miller of Miller Samuel. That's up 21.1 percent from 2017 when the average price was $551,234, and up 58 percent from 2013, when the average price was $422,311.
A search of StreetEasy shows 258 properties currently for sale in Forest Hills, ranging from a $4,599,000 seven-bedroom house at 36 Wendover Rd. to a $145,000 studio co-op, #6D, at 76-26 113th St. Glick says that in his experience prices for co-op studios typically begin at around $175,000; one bedrooms go for around $225,000; and two bedrooms cost $325,000.
As for condos, pricing for studios begins at around $480,000, one bedrooms start at $550,000, and two bedrooms start at $800,000.
"Modern buildings with better amenity packages price appreciably higher, with one bedroom units opening at over $1 million," he says.
Houses in Forest Hills start at around $1 million for attached homes. Detached homes go for as much as $3 to $4 million. Glick says that pricing in the Gardens is "typically higher, with the more stately properties regularly going for four or more million dollars."
What's the transportation situation?
Forest Hills is served by four subway lines (E, F, R, and M), all of which run along Queens Boulevard. You can get to Midtown via the E and F express trains in 25-35 minutes.
The QM4 express bus runs to Midtown in about an hour. And the LIRR can get you to Penn Station in about 15 minutes.
What is there to do?
It's not trendy, but there's plenty going on in the area. The Westside Tennis Club is a private tennis club with 38 courts and a junior Olympic-sized swimming pool. The club is home to Forest Hills Stadium, a 14,000-seat outdoor tennis stadium and concert venue.
There's also the Community House, located in the Forest Hills Gardens section, which has an indoor pool, and offers an array of classes for kids and adults, including martial arts, water aerobics, yoga, and Zumba.
Forest Hills is also flanked on two sides by major city parks. Forest Park is an athlete's delight with softball and baseball fields, tennis, bocce, and handball courts, and a 110-acre golf course, while Flushing Meadows Corona Park, the largest park in Queens, contains the Queens Zoo, the Queens Museum, the New York Hall of Science, and Citi Field.
What's the restaurant/nightlife situation?
"Forest Hills offers a concentrated and eclectic mix of restaurants, mostly centered along Austin Street and Queens Boulevard," Glick says.
He's partial to Cabana, which offers "Nuevo Latino" food; Tuscan Hills, which serves up Northern Italian eats; Forest Hills Station House, a gastropub with lots of craft beers on tap; and La Boulangerie, a French bakery where the pastries "taste as if they were produced closer to the Seine than the East River," according to Glick.
Nanazia is a fan of Jack and Nellie's, a wine bar with a rustic vibe; Nick's Pizza, known for its brick oven pizza; and Buenos Aires Tango Bar & Grill, which has live tango shows on the weekends.
How about grocery stores?
Forest Hills has plenty of large supermarkets to choose from, including a Foodtown, a Key Food, and a C-Town. There are also a number of green grocers in the area, and Austin Street is home to the seafood store Forest Hills Fishery. Forest Hills also has its own farmers market every Sunday. It's held near the post office, and includes free cooking demonstrations and activities for kids, in addition to all the fruits and veggies.
Curiosity piqued? Check out these houses and apartments for sale in Forest Hills:
51 Wendover Rd.
This fully detached four bedroom house in Forest Hills Gardens (also shown at top) is listed for $1,980,000. The first floor of the colonial style brick house has a large living room with a working fireplace, a dining room looking out over the garden, a kitchen, and a half-bath. The second floor has the master bedroom with a walk-in closet, another bedroom with a full en-suite bathroom, and a walk-in closet. The third floor has another two bedrooms and a half bathroom. There's also a renovated basement with a full bathroom, five storage closets, a storage room, and laundry. Taxes are $955 a month.
94 Groton St.
Also in Forest Hills Gardens, this 3,885-square-foot five-bedroom, four-and-a-half bath Tudor is listed for $3,800,000. The first floor features a living room with a gas fireplace and restored arched French doors, a dining room with sliding doors that lead to a patio, an eat-in kitchen with custom cabinetry, and a sunroom. The private quarters are spread out over the two upper levels. There's also a finished basement with a home theater, a children's play area, a gym room, a full bathroom with a steam shower, a storage room with customized shelves, and a laundry room. The house also has central air conditioning and a one-car detached garage. Taxes are $1,193 a month.
105-24 67th Ave. #5C
Listed for $185,000, this studio co-op has stainless-steel kitchen appliances, renovated closets, a windowed bathroom, and wood floors. The building is pet-friendly, with a live-in super and a laundry room. Maintenance is $416 a month.
109-19 72nd Rd. #2G
Priced at $749,000, this 699-square-foot, one-bedroom condo has high ceilings, oversized windows, solid oak flooring, and an in-unit washer and dryer. It's in the Sunrise, a pet-friendly building with a doorman and an elevator.
70-25 Yellowstone Blvd. #5Q
This one-bedroom co-op has an updated kitchen with new cabinets and stainless steel appliances as well as a renovated bathroom. It's listed for $299,000. The apartment is in Gerard Towers, a cat-friendly, 25-story building with a 24-hour doorman, a seasonal heated pool, a fitness center, a children's playroom, bike and storage rooms, and valet parking. Maintenance is $677 a month.
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Brick Underground articles occasionally include the expertise of, or information about, advertising partners when relevant to the story. We will never promote an advertiser's product without making the relationship clear to our readers.
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redpajama_set_name: RedPajamaCommonCrawl


text:
Q: problems when writing document () of own package I am trying to build my own package with Rstudio, and I have problems with the documentation.
             devtools::document()
I got the following error:
         Updating jcext documentation
         Loading jcext
         Error in FUN(X[[i]], ...) : subscript out of bounds

I am a bit lost,and not sure where the error could be (maybe I'm missing something in the NAMESPACE or DESCRIPTION...any idea?
My NAMESPACE was created and looks like:
      # Generated by roxygen2: do not edit by hand

        export(AddDate)
        export(CalculateCWT)
        export(CalculateConstant)

Any suggestion will help,

redpajama_set_name: RedPajamaStackExchange


Tucked in a picturesque valley south of DeSoto, MO, Camp Ne-O-Tez is a non-profit Christian camp serving primarily school-age students.
The Camp is situated on 140-plus acres of woods, with several cabins and supporting facilities that accommodate over 100 people. Ne-O-Tez hosts a number of age-based camp sessions every summer where kids have fun, the Bible is taught, Jesus is lived out in the lives of the staff, and friendships are formed that last a lifetime.
To learn more about Camp Ne-O-Tez, order your copy of the book, History of Camp Ne-O-Tez, which recounts the years 1963 to 2013 of Camp's history. Mail your request with a $10 check made payable to Camp Ne-O-Tez for each copy of the book you would like to 3761 Three Oaks Drive, Bridgeton, MO 63044.
Have a question about Camp Ne-O-Tez? First check the list of frequently asked questions.


Thanks to all of you for the amazing response to Wednesday's "Vacation Gone Awry" post. The best part was all the stories you guys sent me of your own hilarious vacation gone awry moments. They had me laughing out loud a number of times. Keep them coming! Most of the time during my "Vacation Gone Awry", I was in three different kind of outfits. I wore nice-ish outfits when we did something non-beachy, cover-ups for when we did the beach thing, and kid friendly outfits for activities like Disney.
I thought I would start with my nice outfits, because I LOVE these first two dress so much. I feel like they deserve individual posts, but I'm running out of time because one of them is on SUPER DUPER SALE -eek!
Am I the only one who is loving the oversized shirt dress look?!?! They are comfy and cute at the same time…I'm in. But, I originally fell in love with this dress because of all the pretty colors!
What's not to love about this dress? It's pretty, and classic, and comfy. #winwinwin. And guess what?!?! It's on SUPER DUPER SALE!
Ok, it's official, HUE leggings are my new official travel pant! I have never been so comfortable while traveling!
After we arrived, I kept my travel outfit on, it was so comfy. I pitched the poncho and kept the tank on. I brought the chambray shirt in case it got chilly.
I repeated this trick when we decided to go shopping in Laguna Beach and then go to dinner. I'm not a travel blogger, but here are three things we loved in Laguna- Simple was my favorite boutique, Carmelitas had amazing margaritas, and Nick's was the best place we ate during our entire trip.
Stay posted for the rest of the outfits.
I love the Madewell siesta top and am so sad it sold out in my size! It looks fabulous on you.
On a side note, I've never shopped at UO before. What brands do you like/recommend from there? I like stuff that holds up and doesn't pill, and I always avoided UO because I thought it would pill/unravel on me. But I notice you shop there, so I'm curious now!
If you liked the siesta top, I bet you would love the other two tops I linked just as much. I want them both -ha! I really like UO. I think they have a great price point. Plus, the t-shirts I have purchased have not pilled or holed. They are owned by the same company as Anthropologie and Free People. The brands I like are Ecote, Project Social T, Silence + Noise and Sparkle & Fade. I hope this helps.
What size are you wearing in the Madewell dress? I ordered a small and it looks a bit muumuu-ish on me. Have not washed it. Thanks!
I am wearing an xs. I hope this helps.
I don't know the style name. My friend just found the dress at TJ Maxx though!


For any business, driving traffic usually means paid advertising, either through Google Adwords or Facebook Ads. The cost per click of Adwords is generally more expensive as a person's search term gives a stronger purchase signal than clicking on your ads by chance on their news feed.
This paradigm is changing after Facebook's announcement to deprioritize public content and focus on content that's liked by people in your network. As a business, big or small, this change means your team has to rethink how your business uses Facebook for marketing. This forces businesses to engage users with a content-first strategy. The more engaging and relevant the content of your post is, the more likely you will get likes and have a wider outreach.
Naturally this means there will be less impressions especially for small businesses with a smaller fan base. To maintain the top line, businesses must make sure that this traffic converts. We have rounded up some tips on how to do just that. Remember that growing an online business is always a juggling act of attracting traffic and converting them once they come to your site.
The more engaging and relevant the content of your post is, the more likely you will get likes and have a wider outreach.
1. This is stating the obvious but the latest Facebook News Feed change means your team needs a more meaningful and engaging content. This is the time where quality trumps quantity. Reflect on your brand messages. Look at your previous posts and see what has worked well. What kind of content are your customers responsive to?
2. Encourage your fans to prioritize your posts on their news-feed. If your ad content is engaging, relevant and always refreshed with new product and promotions, your customers are more likely to like, share and click on your ad. This makes it more likely that your content will appear on their news feed when they come back to Facebook.
3. Produce more live-videos. Facebook values live video interactions more than regular posts.
Look at your previous posts and see what has worked well. What kind of content are your customers responsive to?
2. In the digital world filled with instant gratifications, nothing kills the desire to buy more than slow page speed. Do a speed audit of your site and make sure that your store is blazing fast.
3. Relevant and meaningful content also extends to your website. Personalization is the closest to making someone feel special when they visit your store. The misconception is it's impossible to personalize for first-time visitors. This is not true. You may not know if they like a black t-shirt or red jeans but they are not totally anonymous. You can create a personalized experience based on what information you have on first-time visitors such as location, season, time of day, day of week or device used. Dig deep into your data and understand what works well with first timers. For return visitors, their browsing history is your goldmine. What they have clicked on and bought tells a lot about their intent and preferences. Next time they come back, you have a golden opportunity to show that you understand them by offering relevant banners, products and promotions. This is a good introduction of web personalization and some ideas how to get started.
Personalization is the closest to making someone feel special when they visit your store.
At DataCue, we offer a simple yet powerful personalization tool for every type of visitor. We automatically work with information about a user, regardless of whether they have visited your site before, to recommend the products they are most likely to convert. Since our product is an API, the flexibility is yours to integrate our recommendations anywhere on the website. If this sounds interesting, drop us a message for a free diagnostic and demo. Or you can see a quick demo of our algorithm here.


The philosophy of the Little Fawn Nursery is to provide children with a safe, secure and stimulating environment. This environment motivates children to learn and develop through discovery.
Since 2010, the Tk'emlúps te Secwépemc Education Department started the Secwépemc Language Immersion Program at Little Fawn Nursery – this means the children and staff spend the morning learning and using the Secwépemc Language. The nursery staff is guided by fluent speakers, language teachers and cultural experts to plan, organize, and teach the Secwepemc language.
The Little Fawn Nursery will be a full day immersion program in the near future. We want our children to have the chance to hear and learn the language – to raise fluent speakers.
Why Does Immersion Work? Language immersion works well for two reasons. First, it creates a meaningful environment where language is used purposefully. It is much more useful to be able to say in the language, "I am thirsty; can I have some water?" rather than, "one, two, three" or "red, blue, green". No English is used at all!


Placenta growth factor (PlGF), a dimeric glycoprotein with 53% homology to VEGF, binds to VEGF receptor-1 (Flt-1), but not to VEGF receptor-2 (Flk-1), and may function by modulating VEGF activity. We previously have showed that PlGF displays prognostic value in colorectal cancer (CRC) but the mechanism remains elucidated.
Overexpression of PlGF increased the invasive/migration ability and decreased apoptosis in CRC cells showing Flt-1 expression. Increased migration was associated with increasing MMP9 via p38 MAPK activation. Tumors grew faster, larger; with higher vascularity from PlGF over-expression cells in xenograft assay. In two independent human CRC tissue cohorts, PlGF, MMP9, and Flt-1 expressions were higher in the advanced than the localized disease group. PlGF expression correlated with MMP9, and Flt-1 expression. CRC patients with high PlGF and high Flt-1 expression in tissue had poor prognosis.
PlGF/Flt-1 signaling plays an important role in CRC progression, blocking PlGF/Flt-1 signaling maybe an alternative therapy for CRC.
Colorectal cancer (CRC) is the second leading cause of death from cancer in Western countries  and the third most common cancer in Taiwan . The fact that tumor growth and metastasis rely on angiogenesis has been widely accepted . Increased angiogenesis in the primary tumor of CRC has been associated with poor prognosis and relapse of disease [4, 5]. Previously, we have demonstrated that PlGF expression was up-regulated in CRC tissue. Immunohistochemical staining analysis showed that PlGF was expressed mainly in tumor cells and Flt-1 was expressed in tumor cells as well as in endothelial cells. The extent of up-regulation correlated with disease progression and patient survival . We further demonstrated that the preoperative serum placental growth factor levels were higher in CRC patients and could be used as a prognostic indicator for recurrence and survival of CRC . However, the underlie mechanism of PlGF and its receptor (Flt-1) regulating the CRC carcinogenesis remains unknown.
Placental growth factor (PlGF), a dimeric glycoprotein with 53% homology to VEGF [7, 8] binds to VEGF receptor-1 (Flt-1), but not to VEGF receptor-2 (Flk-1), and may function by modulating VEGF activity . In addition to the angiogenic effect, PlGF expression has been reported in renal cell carcinomas, thyroid and germ-cell tumors , as well as the meningiomas . It also has been shown that in human gastric cancer, breast, renal, and lung (non-small cell) cancer, PlGF was over-expressed and displayed prognostic value [12–15].
In contrast to the expression of both Flk-1 and Flt-1 in endothelial cells, Flt-1 is widely expressed in many non-endothelial cell types, including hepatocytes, bone marrow progenitor cells, monocytes, macrophages, neural cells, vascular smooth muscle cells, and various tumor cells [16–19]. Therefore, in addition to its role in angiogenesis, Flt-1 might mediate a variety of hitherto unappreciated biological functions, such as liver regeneration, inflammatory process and cancer metastasis [18, 19].
Here we provide evidence that, in addition to the angiogenesis, PlGF/Flt-1 signaling in colorectal cancer cells can promote CRC invasion through a p38-MMP9 pathway and the association with the poor prognosis of CRC patients was validated by two different CRC cohorts.
This study was approved by the Research Ethics Committee of the National Taiwan University Hospital (201107063RC).
Human 293 T cells and colon cancer cell lines SW480, HCT116, HT29, and LoVo were obtained from the American Type Culture Collection (ATCC, Manassas, VA). They were cultured in DMEM with 10% fetal bovine serum and 1% Penicillin/Streptomycin. Cells were grown at 37°C in a 5% CO2 atmosphere within a humidified incubator.
Colorectal cryosections were prepared from colorectal cancer surgical samples which were collected from September 2000 to June 2003 after obtaining the written informed consent, following the guidelines set forth by the Research Ethics Committee of the National Taiwan University Hospital. All tissues were freshly frozen or immersed in optimal cutting temperature compound (OCT) (Ames Company, Elkhart, IN), and kept at −80°C until use. Clinical staging of cancers was determined based on the UICC-TNM classification. Stages I and II were collectively termed the localized disease group (n = 47) and stages III and IV as advanced disease group (n = 33).
The p38 MAPK inhibitor SB203580 and the MMP9 inhibitor were purchased from Merck KGaA (Darmstadt, Germany), and Zeocin from Invitrogen (Carlsbad, CA). Polyclonal rabbit antibodies against phospho-JNK, phospho-ERK 1/2, phospho-p38, and total JNK, ERK1/2, and p38 were purchased from Cell Signaling Technology (Danvers, MA). Other antibodies used were mouse monoclonal antibodies to FLAG M2 (α-FLAG, Sigma Chemical Co., St. Louis, MO), rat polyclonal anti-CD31 (BD Biosciences, San Jose, CA), rabbit polyclonal anti-PlGF (C-20) (Santa Cruz, CA), anti-Factor VIII (Biomeda Corporation, Foster, CA), anti-cleaved Caspase-3 from Cell Signaling Technology (Danvers, MA) and anti-MMP-9 (Abcam, Cambridge, UK). Recombinant PlGF was purchased from R&D Systems, Inc. (Minneapolis, MN).
The human PlGF expression vector was generated by PCR amplification of full length of PlGF cDNA and inserted into the multiple cloning site of pcDNA4/TO-flag-strepII N1 Vector (Flag-StrepII tandem tag at the N-terminus, derived from Invitrogen pcDNATM4/TO vector). HA-tagged human Flt-1 expression vector, pCMV-TAG-Flt-1 (HA-Flt-1), was generated by PCR amplification of Flt-1 cDNA and insertion into the multiple cloning sites of pCMV-TAG vector (Invitrogen, Carlsbad, CA). Negative control siRNA, siFlt-1, sip38 (p38α), and siPlGF were purchased from Ambion Inc. (Austin, TX). LoVo cells were transfected using Lipofectamine 2000 (Invitrogen, Carlsbad, CA). For generating the stable cell lines (LoVo, SW480, HT29 and HCT116), selection with Zeocin (100 μg/ml) began one day after transfection and maintained under same Zeocin condition.
Total RNA from cell lines and tissue was isolated using an RNA extraction kit (Qiagen Inc., Valencia, CA), according to the manufacturer's instructions. For reverse transcription, 1 μg of total RNA was transcribed using the iScript cDNA synthesis kit (Bio-Rad, Hercules, CA). Quantitative PCR was performed in a DNA Engine Opticon 2 (Bio-Rad, Hercules, CA) using iQ SYBR Green supermix (Bio-Rad, Hercules, CA) with human GAPDH as an internal control. Primers sequences used for RT-PCR were as the following, PlGF: Forward-5′-TGCGGCGATGAGAATCTGC-3′, Reverse-5′-AGCGAACGTGCTGAGAGAAC-3′; Flt-1: Forward-5′-AGCAGGTGCTTGAAACCGTAG-3′, Reverse-5′-GTCGCAGGTAACCCATCTTTT-3; MMP9: Forward-5′-GGGACGCAGACATCGTCATC-3′, Reverse-5′-TCGTCATCGTCGAAATGGGC-3′.
Cells were lysed with NP-40 lysis buffer and centrifuged at 15,000 rpm for 20 minutes at 4°C. The supernatant was assayed for protein concentration (Bradford). Equal amounts of protein (150-250 μg/lane) was added to Tris-Glycine SDS sample buffer (Invitrogen, Carlsbad, CA) and separated on 4-12% gradient Tris-Glycine gels (Invitrogen, Carlsbad, CA). Following electrophoresis, proteins were electro-transferred to polyvinylidene difluoride membranes and blocked with 5% bovine serum albumin in TBST (Tris-Buffered Saline and Tween 20). Membranes were then incubated with specific primary antibody overnight, washed, then incubated with appropriate secondary antibody conjugated to horseradish peroxidase, and developed using ECL (PerkinElmer Life Sciences). Membranes were stripped and re-probed with anti-total MAPK (for MAPK antibodies) or anti-actin to confirm equal protein loading.
Frozen sections (8 μm thick) were stained by using the NoVo Link Polymer Detection System (Leica, Biosystems Newcastle Ltd, UK), followed by AEC substrate kit (Vector Laboratories Inc. Burlingame, CA), according to the manufacturer's instruction. Tissues were counterstained with Mayer's hematoxylin. Isotype antibody was used as the staining negative control.
Concentrations of PlGF in cell culture medium were quantified using a Quantikine PlGF immunoassay (R&D Systems, Inc., Minneapolis, MN), as previously described . Concentrations of PlGF were expressed as pg/ml of protein.
Apoptosis was quantified by staining with the Annexin V-FITC kit (Strong Biotech Corporation, Taipei, Taiwan), data collection by flow cytometer (Becton Dickinson) and analyzed by FlowJo 7.2 software. For tissue sections, immunohistochemical staining with cleaved caspase 3 antibody was used for apoptosis analysis.
The invasive activity of the cancer cells was examined using a membrane invasion culture system in which a polycarbonate membrane with 8-μm pores (Millipore., Billerica, MA) coated with Matrigel (R&D Systems, Inc., Minneapolis, MN) at 5 mg/mL was placed between the upper and lower wells of a membrane invasion culture system chamber. 5 × 104 cells were placed into each upper well of the chamber. After incubating for 48 hours at 37°C, cells that had migrated through the coated membrane were removed from the lower chamber with 1 mM EDTA in PBS and dot blotted onto a polycarbonate membrane with 3-μm pores. Blotted cells were stained with Giemsa (Sigma Chemical Co., St. Louis, MO), and the number of cells on each blot was counted under a microscope at a magnification of × 50. Each experiment was performed three times, and each sample was assayed in triplicate. The migration assay was also performed using the same procedure, except without the Matrigel coating.
Cell growth was measured using MTS [3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt] in the form of the CellTiter 96 Aqueous One Solution Cell Proliferation Assay kit (Promega, UK), according to the manufacturer's instructions. Briefly, 1,500 cells/well were plated in 96 well plates and allowed to proliferate for two days. The OD 490 correlated with cell density. All assays were repeated thrice.
LoVo PlGF expression and mock (vector only) stable cells in log phase were trypsinized and washed twice with 137 mM NaCl, 5 mM KCl, 4 mM NaHCO3, 0.5 mM EDTA, 0.1% (w/v) glucose. 106 cells in 100 μl PBS were injected subcutaneously into the backs of 8-week-old SCID mice. The body weight and tumor size were recorded twice per week. After 14 weeks, the mice were sacrificed. Tumor size was measured and further studies were performed on the tissue.
Using light microscopy at 200X magnification, the vascular counts were measured for the tissue section staining with CD31. The three areas with the highest number of discrete microvessels were chosen for analysis and the region with the highest microvessel counts was selected as the final result for that case . Any immunoreactive endothelial cells that were separate from adjacent microvessels were considered to be countable vessels.
The colorectal cancer patient gene expression data was available on Gene Expression Omnibus (GEO) with accession number GSE17536. Expression data were analyzed using GeneSpring GX software (Agilent Technologies). High and low PlGF/Flt-1 expression was defined according to the median expression level in each group.
Statistical differences between groups were analyzed by Student's t test or Mann–Whitney U test. Data was expressed as means ± standard errors (SE). Correlations between PlGF and MMP-9 expression levels were analyzed by Spearman's correlation coefficient. A p value of 0.05 was considered to be statistically significant. The body weight difference along with the time between the LoVo-PlGF and control group, in terms of group effect, time effect, and their interactive effect, were analyzed using the mixed model. Based on fit statistics for Akaike information criterion (AIC) and Bayesian information criterion (BIC) criteria, the repeated measures were modeled using the first-order ante dependence for the covariance structure .
When we arbitrarily used SW480 expression as 1, Flt-1 expression in LoVo cells was 7.4, and 5971 for the Flt-1 overexpression in 293 T cells; Flt-1 was not detectable in the negative control and barely detectable in both HT29 and HCT116 cell lines (Figure 1A). In contrast, PlGF was expressed (within folds of change) in the four CRC cell lines (Figure 1B) by quantitative PCR.
Expression and biological function of PlGF and Flt-1 in CRC cell lines. Quantitative PCR levels of Flt-1 (A) and PlGF (B) in CRC cell lines. The expression of SW480 was arbitrarily defined as the reference for comparison. (C) PlGF recombinant protein (10 ng/ml) increased the invasion ability in LoVo but not HT29 and HCT116 cells; (D) stable overexpression of PlGF increased the migration ability in LoVo and SW480 cells, but not in HT29 and HCT116 cells. PlGF level in the LoVo-PlGF (PlGF overexpressioin) cells was validated by quantitative RT-PCR (E), Western blot (F) (anti-PlGF at 200 ng/ml) and ELISA (G). The migration (H) and invasion (I) abilities increased in LoVo-PlGF cells as compared to LoVo-pcDNA, respectively. (Neg: negative control; Pos; positive control. * indicated as P < 0.05).
Exogenous PlGF significantly increased the invasive ability of LoVo cells, which expressed the highest Flt-1 levels of the cell lines tested by up to four fold. Conversely, HT29 and HCT116, cell lines in which Flt-1 was virtually undetectable, did not respond to exogenously added PlGF (Figure 1C). To further confirm the role of PlGF in CRC cancer cells, we generated the PlGF-over-expression stable clones in LoVo, SW480, HT29 and HCT116 cells as well as their empty vector control cells. The over-expression of PlGF in these stable clones have been validated and monitored periodically by quantitative PCR, which we got hundreds of fold increased expression of PlGF than the control cell lines. Migration assay was performed to compare the stable clones with and without PlGF and normalized to the empty vector control. Migration ability increased in LoVo and SW480 with stable expression of PlGF, but no change in HT29 cells and even decreased in HCT116 cells (Figure 1D). This data suggests that Flt-1 receptor may be critical for PlGF induced tumor cell invasion in CRC cells.
Stable clones have been validated for the presence of the transgene and for overexpression of PlGF protein by quantitative RT-PCR, Western blot and ELISA (Figure 1E, F and G). According to the datasheet, it showed PlGF (C-20) can be used for detecting variants of PlGF of human origin. The overexpressed PlGF was double confirmed by immunoblotting with Flag and PlGF. As increased invasion/migration was shown most prominently with the LoVo stable clone for PlGF, LoVo-PlGF and its control LoVo-pcDNA were used for the following experiments. The migration and invasion abilities increased 4.9 and 3.9 fold in LoVo-PlGF cells as compared to LoVo-pcDNA, respectively (Figure 1H, I). There was minor apoptosis difference by PI and annexin V staining between LoVo-pcDNA and LoVo-PlGF cells, 3.2% and 1.3%, respectively, and no difference in the proliferation status by MTS assay between these two cell lines (data not shown).
In LoVo cells with overexpression of PlGF, we found that phosphorylation of p38 increased but there was no increase in the phosphorylation levels of ERK and JNK MAP kinases (Figure 2A). It might be due to the exposure time which showing the phospho-ERK at both pcDNA and PlGF group. From the results, we could say there was no significant difference between the pcDNA and PlGF group for the expression of ERK (both phospho and total form). Inhibition of p38, either by using p38 chemical inhibitors (Figure 2B) or siRNA (Figure 2C), decreased the ability of LoVo-PlGF cells to migrate. All experiments had been performed at least 3 times.
Increased migration of LoVo-PlGF cells was due to increasing p38 MAPK activation. (A) Phosphorylation levels of p38, JNK and ERK are shown along with total protein levels of p38, JNK and ERK in LoVo-PlGF and LoVo pcDNA control cells. (B) Inhibition of p38 by chemical inhibitor, SB203580 (20 μM) as well as by siRNA of p38 (30 pmole/ml) (C), decreased the migratory ability of LoVo-PlGF. * indicated as P < 0.05.
Several lines of evidence have shown that p38 activation is required for MMP9 expression, which has been linked to tumor migration and invasion [22, 23]. We therefore checked MMP9 expression in LoVo-PlGF cells. Indeed, MMP9 expression was significantly increased in LoVo-PlGF cells compared to control at the message level (Figure 3A), and inhibition of p38 by both siRNA and chemical inhibition both decreased the expression of MMP9 at the protein level (Figure 3B and data not shown). The migration ability was inhibited by using the chemical inhibitor of MMP9 (Figure 3C).
PlGF induced MMP9 expression via p38 MAPK activation. (A) MMP9 expression was significantly increased in LoVo-PlGF cells compared to LoVo-pcDNA cells as assessed by quantitative PCR. (B) Inhibition of p38 by siRNA (30 pmole/ml) decreased the expression of MMP9 in LoVo-PlGF cells as detected by Western blotting. (C) By using the chemical inhibitor of MMP9 (30 μM), the migration ability was inhibited in LoVo-PlGF cells. In human colorectal cancer tissue, PlGF (D), MMP9 (E) and Flt-1 (F) expression were significantly higher in the advanced than localized disease group. There was a strongly positive correlation between the expression of PlGF and MMP9 (G), as well as PlGF and Flt-1 (H). * indicated as P < 0.05.
To investigate the clinical significance of this in vitro finding, we checked PlGF, Flt-1, and MMP9 expression in 80 human colorectal cancer tissues at the message level. We found, indeed, PlGF, MMP9, and Flt-1 expression were significantly higher in the advanced CRC group than the localized CRC group (Figure 3D, E and F). Moreover, the expression of MMP9 in human CRC tumor samples significantly correlated with the PlGF expression levels in the samples (Figure 3G), as well as the PlGF expression and Flt-1 expression (Figure 3H).
Next, we asked whether Flt-1 expression is required for PlGF-induced tumor invasion since only the Flt-1 expressing CRC cell line responded to exogenous PlGF. To address this question we used a siRNA approach to inhibit Flt-1 expression in LoVo-PlGF cells (Figure 4A). We have tried the Western blot for Flt-1, however, we could only detect the over-expressed one but not the endogenous one. This may be due to the low expression level of endogenous Flt-1. Therefore, we could only show the data by quantitative PCR. The migration ability decreased when Flt-1 levels were reduced by siRNA (Figure 4B). This data indicates Flt-1 is required for PlGF-induced CRC cell migration. To further confirm this result, we checked the effect of siFlt-1 on p38 phosphorylation in LoVo-PlGF cells. Indeed, downregulation of Flt-1 significantly attenuated the phosphorylation of p38 in LoVo-PlGF cells (Figure 4C). We also found that both migration and invasion ability decreased in LoVo-PlGF cells when the PlGF was knocked down (validated in Figure 4D) by using the siRNA approach (Figure 4E).
Flt-1 is required for PlGF-induced p38 phosphorylation, PlGF and Flt-1 were both important in promoting CRC cancer cell migration/invasion. siRNA (30 pmole/ml) inhibited Flt-1 expression and migratory ability in LoVo-PlGF cells as monitored by quantitative PCR (A) and migration assay (B). (C) Knockdown Flt-1 significantly attenuated p38 phopsphorylation in LoVo-PlGF. (D, E). The invasion ability of CRC cells decreased when PlGF was inhibited by siRNA. * indicated as P < 0.05.
To confirm the role of PlGF in CRC ex vivo, tumor xenograft assays were performed. During the observation period, three of the four LoVo-PlGF cells implanted mice had palpable nodules as early as the 3rd week, and all of them had measurable tumors by the end of the 14 week experimental period. In contrast, only two of the four mice in the LoVo-pcDNA group had palpable nodules, detectable only in the 10th week (Figure 5A). The LoVo-PlGF group also gained weight slower than the control group (Figure 5B, group effect, P = 0.0137). The body weight difference became even more significant during follow-up (interactive effect between time and group; P < 0.0001). Mice were sacrificed following week 14. Both the tumor radius and tumor volumes were larger in the LoVo-PlGF group (Figure 5C and D). PlGF expression was indeed significantly higher in the tumor tissue induced by LoVo-PlGF cell implantation (Figure 5E). LoVo-PlGF induced tumors had higher vascularity (Figure 5F), higher microvessel density (Figure 5G), and less caspase 3 staining (Figure 5H) than the control group.
Tumors grew faster and larger in animals receiving LoVo-PlGF xenografts. (A) At the 10th week after injection, tumors were visible in the LoVo-PlGF group but not in the LoVo-pcDNA group. (B) The LoVo-PlGF group lagged behind in body weight gain. Both the tumor radius (C) and tumor volumes (D) were larger in the LoVo-PlGF group than the control group. (E) PlGF expression was significantly higher in the tumor tissue induced by LoVo-PlGF cells. (F) Angiogenesis was also more advanced in the LoVo-PlGF group, with the microvessel density shown in (G). (H) Less apoptosis in LoVo-PlGF group than the control group. * indicated as P < 0.05.
We further analyzed a publicly available gene expression dataset from the GEO database. In this cohort, higher PlGF and Flt-1 mRNA expressions were observed in stage III-IV diseases (advanced disease) compared to stage I-II disease (localized disease) (Figure 6A and B). Patients that had high Flt-1 and high PlGF expression had shorter survival (p = 0.016) (Figure 6C). Flt-1 expression was correlated with PlGF (p = 0.003, correlation coefficient: 0.225) (Figure 6D). This result supports the in vitro study results, validating the results from NTUH cohorts, and strongly implies that high PlGF levels combined with high Flt-1 expression increase cancer invasion and lead to shorter survival.
PlGF correlated with Flt-1 expression in CRC tissue and patients with higher expression of PlGF and Flt-1 had worse prognosis in another CRC cohort. (A, B) higher Flt-1 and PlGF mRNA expressions were observed in stage III-IV disease (advanced disease) compared to stage I-II disease (localized disease). (C) Patients with high Flt-1 and high PlGF expression had shorter survival. (D) Flt-1 expression correlated well with PlGF levels.
In this study we demonstrated that CRC cells express PlGF and Flt-1 have higher invasion/migration ability. PlGF increased the invasion/migration ability of colorectal cancer cells by increasing the phosphorylation of p38 MAPK and upregulating MMP9 expression. Overexpression of PlGF decreased the apoptosis mildly, but did not affect the cell proliferation status. These in vitro results had been validated by using two independent CRC cohorts and showed that patients with high PlGF and high Flt-1 expression in CRC tissue had a poorer prognosis. These results revealed that, in addition to previously recognized effects on angiogenesis, PlGF also plays a hitherto unappreciated role in CRC carcinogenesis.
How the PlGF regulates CRC carcinogenesis? PlGF has been shown to increase tumor cell migration in lung cancer, leukemic and melanoma cells [24–26]. It also has been shown that the increased migration of leukemic cells was via the p38/ERK pathway resulting in Rho GTPases activation and caveolae formation . In addition, Loesch et al. demonstrated that the p38gamma MAPK cooperated with transcription factor c-Jun in trans-activating MMP9 which resulted in cell invasion . In line with these findings, our data linked the over-expression of PlGF with the upregulation of MMP9 expression by increasing phosphorylation of p38 MAPK in colorectal cancer cells. In addition, knockdown of p38 MAPK, either by chemical inhibition or siRNA, decreased the expression of MMP9 and the ability of tumor cells to migrate. Furthermore, PlGF expression levels in human CRC tissues correlated well with their MMP9 expression. Taken together, these data suggest that in colorectal cancer cells, PlGF induces tumor cell invasion and migration through upregulation of MMP9 expression by increasing p38 phosphorylation.
Flt-1, one of the well studied receptors of both PlGF and VEGF, plays an important role in regulating vasculogenesis and angiogenesis [28, 29]. In addition to angiogenesis, the VEGF-Flt-1 connection also plays an important role in the inflammatory process by activating monocytes/macrophages and inducing their migration [16, 17, 30, 31]. Previously, Xu et al. reported that overexpression of PlGF in HCT116 cells decreased tumor growth, cancer cell invasion and angiogenesis . This result appears to contradict our findings and the clinical observations of others that higher expression of PlGF correlates with poor prognosis [2, 6, 11–15, 33, 34]. To elucidate this discrepancy, we checked the PlGF major receptor-Flt-1 status of four different colorectal cancer cell lines, and found that Flt-1 was barely detectable in both HCT116 and HT29 cell lines. In fact, we found exogenous PlGF recombinant protein or stable overexpression of PlGF led to increased invasive ability only in cells with Flt-1 expression. These results suggest PlGF may play different roles in CRC cells depending upon whether the Flt-1 receptor is present, though the role of PlGF in tumor angiogenesis remains controversial [35, 36].
Flt-1 protein contains two isoforms, one is the trans-membrane receptor (mFlt-1) and the other is the soluble isoform (sFlt-1). sFlt1 is antiangiogenic as it can function as a decoy that traps the VEGF and PlGF, then prevents binding to VEGFR; whereas mFlt1 is proangiogenic [37, 38]. Our results demonstrated that high Flt-1 expression in colorectal cancer cells increased their invasive ability, and is associated with poor prognosis. Recently, Yao et al. also found this similar condition and reported that the role of PlGF in tumorigenesis largely consists of promoting autocrine/paracrine growth of tumor cells expressing a functional Flt-1 rather than stimulation of angiogenesis .
In addition to the well known effect on angiogenesis, PlGF/Flt-1 signaling plays a previously unappreciated important role in colorectal carcinogenesis by increasing the phosphorylation of p38 MAPK, thereby upregulating MMP9 expression; resulting in increasing cellular migration/invasion. Blocking PlGF-Flt-1signaling may be an alternative therapy for treating CRC.
We thank the second Core Laboratory of the Department of Medical Research of the National Taiwan University Hospital for technical assistance, Dr. Hwai-I Yang for his kind help in statistical analysis, Prof. Wen-Fang Cheng and Min-Hung Hsu for technical support in tumor xenograft experiment. We thank the critical review and comments from Dr. Ji Luo (Medical Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA). The in vitro work was supported partially by the National Science Council of Taiwan (100-2314-B-002-071-), and others by the Liver Disease Prevention & Treatment Research Foundation.
No conflict of interest to be declared by all authors.
SCW: study concept and design; acquisition of data; analysis and interpretation of data; drafting of the manuscript; obtained funding; approval of the final version of the manuscript. PNT: study concept and design; statistical analysis; analysis and interpretation of data; drafting of the manuscript; critical revision of the manuscript for important intellectual content. MTW: study concept and design; statistical analysis; analysis and interpretation of data; drafting of the manuscript. ZC: technical and material support; drafting of the manuscript. JMW: samples collection; critical revision of the manuscript for important intellectual content. All authors read and approved the final manuscript.


the example. i have just changed the register name like U0LSR to U1LSR.
a newbie in this line. thanks in advance?
> the example. i have just changed the register name like U0LSR to U1LSR.
> a newbie in this line. thanks in advance?
interrupt and then call one by one for uart0 uart1 respectively.
> > a newbie in this line. thanks in advance?
> interrupt and then call one by one for uart0 uart1 respectively.


Since reading Sherrill Sellman's book Hormone Heresy: What women must know about their hormones I've decided that I have to find a natural alternative to Hormone Replacement Therapy. I'm already not keen on medication, and the idea of pumping my body with a cocktail of synthetic hormones yet again (since I stopped the Pill) does not appeal.
He prescribed estriol (a form of oestrogen) for down below, and, to his credit, instead of imposing HRT on me, suggested I ask my buddies about HRT, and to do some research on the internet. So I did. Well I did the internet bit because I don't have many friends who have hit menopause yet.
I discovered that HRT is not all it's made out to be, and that some women have disastrous reactions to it. Instead of taking it and risk becoming oestrogen dominant which is not a good thing, I wanted to balance out the hormonal imbalance between oestrogen and progesterone in my menopausal body as naturally and side-effect-free as possible.
Dr Sellman suggests seeing a naturopathic doctor, and as luck would have it, the Pages Jaunes for Montpellier threw up two: a man and a woman. I looked at the woman's web page and noted she has 25 years of experience, and, from her photo saw that she has very probably gone through menopause. It seemed an obvious choice to select her rather than the man who did not have a web site and thus was just a name on a page.
Today was the day when I had my appointment. I took along my blood test analyses (usual stuff plus FSH which determines whether you're in menopause or not because it's a reproductive hormone - I am) and found myself in a little outhouse in the doc's garden.
She started by looking at my eyes. Apparently you can tell a lot from eyes. Before she looked at my blood tests, she flashed a torch at me and asked me whether I had some intestinal issues, and some joint problems, and said I have an excellent 'hygiene de vie' which means I'm in roaring good health.
I was pleased to hear that because I have made a lot of effort in the past year, and it's obviously paid off, not that I was abusing my health before, but I'm taking more care now (organic fruit and veg, less meat, dairy and bread, sport).
Rhadiola Rosea and Magnesium Marin to improve the quality of my sleep and help me relax.
I was in there for an hour and it cost me 65 Eur which should be reimbursed by my mutuelle. I have to get most of the supplements on the internet, and she indicated where I could go because there's such a variety out there.
I'll be chucking out the estriol which had revolting consequences.
In the same vein, I started doing a local yoga course this evening. It was a trial session, and although there was some noise from the other activities going on in the communal centre, I found it relaxing and pretty promising.
Got to keep my strength up for all this healthy living!
The other thing about HRT is that you really don't want to be pumping oestrogen into your body, when 1 in 10 women (or whatever the figure is) will develop breast cancer in their lives. Breast cancer medication is all about removing or reducing oestrogen, because it feeds the cancer.
I like the sound of your naturopath, and your approach. We can't be young forever, and surely some of the journey is allowing our minds and souls to catch up with the fact that our bodies are ageing.
Thanks, Iota. The naturopathe looks terrific. She's 63 but certainly doesn't look it.
These days we have to combat the toxic effects of our environment and food on our bodies that didn't exist in the past.
Houblon? That means beer. Menopause, I'm ready.
Interesting. I have been taking HRT for a while since having early onset menopause. It's been great for me and I was happy to take them as, from my own research, felt it was replacing what would normally be present before the menopause. The decision regarding when to come off them is now arising - for me the risks up to now have been worth taking but carrying on with HRT past the normal age for menopause is something I don't want to do.
Absolutely, I read that it's not a good idea to take it long term.
I'm glad you've been satisfied with it, no doubt you would have stopped it if you hadn't been!
We are not told about hormone imbalances enough (at all) and yet they have a major effect on our health and sense of well-being!
I'll say this, with respect to hot flashes.....my late grandmother suffered from far too many. Poor lady.
It sounds like you have a very competent doctor. Do listen to her advice.
I'm so glad I've seen this post (I'm so far behind on everyone's posts!) because you have just described me! The flushes which I've been coping with quite well are now so out of hand it's not true! I look forward to seeing how you get on with all those recommended supplements.
It's a fascinating subject. You must have enjoyed studying it, and I'm sure you put it all to good use! I'll look out for Kenton's books.


Faster perforating with less distortion. Cluster tooling from Wilson Tool can be manufactured with as few as 2 to as many as 120+ holes in any pattern or shape. In addition to speeding up perforating operations, these punches help control sheet distortion, and with the addition of a back bending die can also prevent oil canning or warping. Custom cluster tools are available in all tool styles provided by Wilson Tool.


Did You Know a Mortgage is a Pledge to the DEAD?
On December 3, 2015, I wrote and published an article titled The Occult Definition of Mortgage: Why You Should Never Get a Mortgage. On December 15, 2015, Youtuber Chua Chye Koon turned this article into a video. If you have not read this article and are interested in the forbidden secrets of the mortgage industry, I encourage you to watch the following short video. If you would rather read the article, visit this page.
To connect the dots, when you take out a mortgage, you unknowingly make a pledge (promise) to the dead. This is the general occult definition of the word mortgage. So, what is the dead? The dead represents the Dark Forces (groups of demons) and their corporations (corpses or dead bodies).


Dissinger Reed is proud to provide insurance and risk management for the NFHS Officials Association. With over 34 years of insurance experience for governing organizations and high school associations, Dissinger Reed understands the unique special risks involved with the NFHS and its members. The NFHS and Dissinger Reed have specifically designed an insurance plan to cover Officials and all registered member Associations.
All officials currently registered with a member state association, like the OSAA.
State and local officials' associations who have 100% membership in the NFHS Officials Association, which includes all Oregon chartered local officials association.
Assignors, instructors, and rule interpreters who are currently registered with the NFOA are covered under the Liability Policy for claims involving bodily injury and property damage. There is no coverage under the Liability Policy for any other type of claim.
Officiating duties include chain crews and attending officiating camps, clinics, and meetings.
Coverage for officials extends, not only to high school activities, but also to college, independent youth, adult, and recreational leagues. However, coverage is still limited only to sports recognized by the state high school association.


text:
Pandemic Impact
2019 National Coach of the Year: COVID 'threat is still out there'
By Dan Guttenplan, FNF Coaches Editor
Dutch Fork High (S.C.) football coach Tom Knotts couldn't have had a better 2019.
He led Dutch Fork to its fifth state championship — bringing his total state championship count to 12 between coaching jobs in North and South Carolina. At the conclusion of the season, he was named the 2019 National High School Football Coaches Association (NHSCA) National Football Coach of the Year.
Like so many coaches, Knotts hasn't been as enamored with 2020 so far. His team finally opened the season last Friday night after a difficult summer and delayed start to the fall season. He hosted Senior Night for his players on the first game of the season to make sure his upperclassmen didn't miss out on the ceremony if the season gets cancelled before its scheduled completion.
Knotts recently joined FNF Coaches for a Q&A.
What made you decide to host Senior Night so early in the season?
"I guess the fear that COVID would eliminate, pause or postpone the season. We went ahead with it since we knew we'd play on the first night. We recognized the seniors exactly the same way we always do."
Did you take any extra precautions in terms of social distancing or mask-wearing?
"We attempted to. I don't think we pulled it off very well. Most of the parents pulled their masks down for the pictures. The seniors come on to the field. We announced what they play, what their dreams and aspirations are, if they're attending college, scholarship offers, those types of things. Then they take pictures. We attempted to have social distancing in the stands, but the students didn't really adhere to it."
Why do you think it's so hard to adhere to those safety recommendations?
"Kids are ready to get out. Americans are ready to get out. But the threat is still there. We're playing a dangerous game, but hopefully life goes on and football goes on."
What is the status of school at Dutch Fork? Is it remote on in-person?
"We have a hybrid schedule, and a lot of people are not taking this seriously. On Friday, they announced we'll transition to a full-time schedule. My estimation is everyone is acting like this is coming to an end. I'm not sure that's the case. I'm hoping it doesn't have a negative effect. We talk about a Dutch Fork bubble. I think the kids have done a good job, but we're hoping that putting another 1,000 kids in the school won't result in an outbreak."
What's driving the conversation in your locker room? Email Managing Editor Dan Guttenplan or Tweet us @fnfcoaches. Don't forget to use that hashtag #FNFCoachesTalk
Q&A with Georgia state champion coach Tim McFarlin, whose team is in quarantine period
How Louisiana state champion coach creates pipeline to LSU, NFL
California Coach: A Playbook for Dealing with No Season
The Blueprint for a Winter Football Season
Masaki Matsumoto: 'It's important our kids...
California coach explains how he coached Alaska team...
7 Things to Consider If the Season Is Postponed Until...
Highland Park (Texas) coach Randy Allen on the impact...
FNF Coaches Talk — Culture Week continues
Six-time Kansas state champion coach: Culture...
FNF Coaches Talk — Coach Regalado's back...


Q: How many firms are currently outsourcing CAD services?
A: The CAD outsourcing marketplace is changing rapidly, especially over the last few years. Until recently, CAD outsourcing was not being openly discussed in the user meetings and conferences, however, that is changing quickly. Until a couple of years ago, only large Architectural and engineering firms were seriously pursuing CAD outsourcing and smaller firms were feeling left out. CTS Drafting and Design is changing that marketplace by offering even smaller firms an opportunity to leverage the competitive advantages of outsourcing and empowering them with CAD outsourcing solutions that really work. The number of firms that are currently outsourcing is still small (maybe 5% or less), however, within a short period of time (5-10 years), the number is expected to go up drastically and the companies that jump on the CAD outsourcing boat today, will have a strong competitive advantage, tomorrow.
Q: How small or large a project can you undertake?
A: We work with smaller shops to larger firms, however, the size of the project we work on is not important, it's the regularity of the work. For most of the clients, we have setup standards within our office and that has resulted in shorter turn around times for each of our clients. It takes a little time for our team to understand each specific clients way of working and develop an understanding that is critical for long-term CAD outsourcing. In general, we have clients who provide us work either continuously or regularly. Having said that, we also work on one time projects.
Q: Can you please explain the software capability of CTS Drafting and Design?
Architecture/Engineering: Autocad, Architectural Desktop (ADT), Chief Architect.
Q: Why CTS Drafting and Design? What differentiates you from the herd?
A: First, we are not brokers – the ones who accept projects from clients and then in-turn outsource to another company in Argentina, Philippines, China or India. We have our own production office and we control the quality of the people we hire, the processes and every small detail that matters. Secondly, we are experienced working on hundreds of projects and we have developed time-tested work processes to make outsourcing really work for you. Lastly, most firms work with large firms only – At CTS Drafting and Design, we work with a wide range of sizes of companies (1 Person Company to companies with hundreds of employees) – we bring the prowess of CAD outsourcing to the mainstream, not just to the big players.
Q: Can I mail, E-mail or courier the originals?
A: Yes, you can send your paper originals/copies directly to our office by what ever means are most convenient for you.
Q: Do you have a representative who can meet with us?
A:Yes. Our representative would be more than happy to meet you at your office to assist with your CAD outsourcing venture.
Q: Would you be willing to sign a Non-Disclosure Agreement?
A: Yes. We can sign a Non-Disclosure agreement upon request.
Q: Do you provide up front pricing?
A: Yes. our focus is in providing ongoing CAD support (long-term) and we do provide an upfront price to help you evaluate the services before committing.
A: The office is located in Effort, Pa.
A: CTS Drafting and Design has been in business since 2004 and over the years, we have worked with a number of clients across Pennsylvania and New York.
Q: I have more questions not answered here.


text:
Amy Mogensen
Can The Internet Overcome Its Chaos and Turn into a 21st-century Library?
Digital Library shelf. (2021). [Photograph]
The idea of the library is changing in the 21st century. The library as a collection of knowledge is no longer limited to a physical space; with the internet, information can exist anywhere and everywhere. How we access information is changing too; social media allows everyone to be creators of knowledge and sharers of information. Addressed here is how the internet and social media are changing how libraries and librarians operate in a digital society.
There are two important elements that constitute the library as an institution of information and knowledge:
the organization of a collection of records, and
the sense of community created by that collection of records.
Mittler (2016, p. 227) identifies that "cataloging is the main prerequisite for library usage" in modern libraries. When Internet Library Portals were first introduced in the late 1990s, Davidsen (2005, p. 14) recognized that their purpose was to "bring order out of [the] chaos" of the information readily available on the internet.
Be it a physical library or an Internet Library Portal, the structuring of information is recognized as a central idea to what defines a library. Indeed, Jacob (2002, p. 49) identifies the library as a space "both structured and structuring". This statement can be understood by recognizing that libraries order the information within them and are responsible for structuring how the community responds to and uses the information included in the collection.
Pipe, B., Cultura Travel, and Getty Images. (n.d.). New York Public Library [Photograph].
Fisher (2011, p. 263) notes that in the context of the Western world, the library profession was introduced in the Victorian Age, when "notions of public service were being formed". There is a relationship between the idea of public service and the accessibility of the information contained in libraries for the community.
Indeed, Jacob (2002, p. 41) believes that we should understand libraries "in terms of relationships" between the collection and the people who organize and use them. Jacob (2002, p. 42) expands on their definition to say that libraries are "ways of externalizing memories…[they are] socially active". Libraries are therefore not only spaces where information is ordered, but "relationships" where information is "socially active" in ideas of public service, community, and culture. To understand what a library is and how they are changing in the 21st century, we must understand libraries as spaces of both knowledge and culture.
Following the definition of the library established thus far, the internet cannot be considered a library as it does not organize the information it contains; as Davidsen has recognized, Internet Library Portals had to be introduced to achieve this structuring of information.
Where the internet does suit the definition of a library, though, is when we articulate the culture of the internet. Jacob (2002, p. 47) makes an interesting definition of libraries in the ancient world as "autarchic collections of memory". This definition can be adopted by the internet, where the internet is an autarchic collection of disorganized memories rather than the structured knowledge of a physical library. The greatest challenge to libraries in the 21st century is how the internet displays disorganized information. To overcome this challenge, we can develop our understanding of the library as "socially active", recognizing the relationship between the information and the people who use it.
To understand the library as "socially active", Jacob (2002, p. 56) argues that we must look at "the relationship between storage technologies and the circulation of knowledge in a particular society at a particular time". The internet is a "storage technology", and thus according to Jacob, we must look at how the internet is changing how we interact with knowledge. Fisher (2011, p. 274) identifies how "the post-modern influence that social media currently exert indicates that they will emerge as the major channel used to convey…ideas and responses, as well as in initiating discussion and setting agendas".
To see how libraries will change in the 21st century, we must look at how the relationship between individuals, the internet, and social media will change how information is shared in society.
Vladgrin and Thinkstock. (2014). Social media and information [Graphic].
When looking at how information is shared in society, we are required to look at how the role of librarians and information services professionals is changing. If the internet and social media give rise to "autarchic collections of memory", what is the need for the services of a professional librarian? Jacob (2002, p. 48) recognizes that librarians "manifest…power [of information]…through control": the question for our digital age, where social media democratizes who owns and controls information, is how a loss of control will change how information is shared in society.
Fisher (2011, p. 275) argues that social media has given rise to "information workers" and that these have replaced the services of professional librarians. Fisher decides this is not the way forward and that there is still a strong need for information services professionals. Even if the trend of "information workers" is not desirable for libraries, it is a responsible and accurate representation of how information is shared in our digital society, and therefore, we must continue to discuss how "information workers" are changing how we access and use information.
It is important to discuss how libraries are changing in the 21st century, as it helps us assess how the access and the use of information are changing in a digital world. Traditional libraries are physical spaces of ordered knowledge, with information being controlled by professional librarians. The internet and social media are changing how information is being created, used, and accessed by individuals, with less order and control.
In the 21st century, the library has become less of a physical space and more of a relationship between the people who access, use, and share information.
The critical question for us now, is how to train individuals to navigate this new reality. Are traditional education systems out of touch in doing this? How do we verify knowledge when everyone has an authority over the information? These are the exiting challenges we get to meet as we continue to understand our position as "information workers" in a digital society.
Davidsen, S. L. (2005). The Internet Public Library and the History of Library Portals. Journal of Library Administration, 43(1–2), 5–18. https://doi.org/10.1300/j111v43n01_02.
Fisher, B. (2011). Redefining librarianship. In D. Baker & W. Evans (Eds.), Libraries and Society: Role, Responsibility and Future in an Age of Change (1st ed., pp. 263–278). Chandos Publishing.
Jacob, C. (2002). Gathering Memory: Thoughts on the History of Libraries. Diogenes, 49(196), 41–57. https://doi.org/10.1177/039219210204919605.
Mittler, E. (2016). The Library as History: Library History Research after the Cultural Turn. Quærendo, 46, 222–240. https://doi.org/10.1163/15700690-12341352.
Digital Library shelf. (2021). [Photograph]. ZBW Mediatalk. https://www.zbw-mediatalk.eu/2021/08/why-modern-libraries-have-to-be-active-on-social-media-seven-glorious-reasons/.
Pipe, B., Cultura Travel, & Getty Images. (n.d.). New York Public Library [Photograph]. Lonely Planet. https://www.lonelyplanet.com/usa/new-york-city/attractions/new-york-public-library/a/poi-sig/382117/362079.
Vladgrin & Thinkstock. (2014). Social media and information [Graphic]. School Library Journal. https://www.slj.com/?detailStory=slj-social-media-resources.


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You are here: Home / comments / Panthers at Kings pregame primer – with Mike Richards
Panthers at Kings pregame primer – with Mike Richards
December 1, 2011 By John Hoven
When the Florida Panthers and Los Angeles Kings last played each other it certainly featured an exciting finish. Tied 2-2 with less than three minutes remaining in the third period, Anze Kopitar scored to put LA on top and they went on to win the game 3-2.
That was only Kopitar's fourth career game vs. Florida, largely because they're an Eastern Conference team – so the Kings don't see them very often.
Mike Richards, on the other hand, has faced them 23 times and picked up 13 points (three goals, 10 assists) in the process.
"They're always a good team," said Richards, who has scored nine goals in the the last 10 games for LA. "I know they were extremely well coached the last couple of years with Pete DeBeor there [since replaced by Kevin Dineen, more on that earlier today]. It was usually a tough place to play too, going down to Florida. But, they've made a lot of changes. They have a lot of skill in their line-up now – including Kris Versteeg scoring well, I played with him. They have skill and they ruffle things up a bit too. They're playing well right now and we have to play well too."
– Earlier today, Kings captain Dustin Brown and former King defenseman (now with the Panthers) Mike Weaver exchanged comments about each other here. Also, former King Marco Sturm paid a visit to MayorsManor here, talking about his return to Staples Center tonight and how he's happy to leave the lows of LA behind him.
– From the random trivia files – Kings defenseman Rob Scuderi played his first NHL game against the Florida Panthers back in 2004. He shared a few memories from that game and sized up tonight's contest here.
– As usual, we'll be running our first goal contest tonight too. If you'd like a chance at winning a free t-shirt from Glass Bangers, simply reply below with the name of the Kings player you think will score the first goal for LA tonight. You can even use your facebook profile to enter!
Finally, remember to follow along on the twitter or facebook links below too, as we'll have post-game notes and quotes live from the Kings locker room and Terry Murray's presser immediately following the game.
www.twitter.com/MayorNHL
www.facebook.com/MayorsManor
Filed Under: comments, hockey, Interview, Kings, Los Angeles, Mayor, MayorsManor, NHL, Panthers, preview Tagged With: comments, Florida, hockey, Kopitar, Los Angeles, Mayor, MayorsManor, preview, quote, Richards
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Judith Rodin to be Chubb Fellow at Yale
Judith Rodin, president of the University of Pennsylvania and former Yale provost, faculty member and dean, will speak at Yale as a Chubb Fellow on Tuesday, Oct. 12, at 4:30 p.m. in the Levinson Auditorium of Yale Law School, 127 Wall St. Rodin will...


Resort 2016 – Tenerife Outfits of the week!
It may not seem like it since I am posting on the same level as always, but for me it seems like a long time ago since I have written something on my blog. That's because I have been away to Tenerife to relax for a week and I hadn't brought my laptop with me. I created blogposts beforehand so I could stay up-to-date with you guys. So now, back in Belgium for a few hours, I am writing again to you guys!
Today I am bringing to you my Tenerife outfits of the week, inspired by the Resort 2016 collection. If you don't know what resort is, click here.
I had one of the best holidays I had in my entire life so far on Tenerife. It is such a beautiful island, with lots of breathtaking nature. I feel very inspired again, so be prepared for a shitload of posts coming, haha!
For those who don't know Tenerife, it is an island lying in the Atlantic ocean just before the African coast. It is the biggest of the Canarian island group.
Besides Tenerife, there is Fuertaventura, Lanzarote, Gran Canaria and a bunch of little islands in the Canarian islandgroup. They belong to Spain, and Spanish is the official language as well, but they feel much more Canarian than they feel Spanish thou.
I hope I got your enthusiasm to take a look at what I wore during this week!
Since the Canarian islands carry the name of the eternal spring, they always have a nice climate of 25 degrees celsius going on, even in winter! So my outfits were also very light, appropriate for the resort collection.
The first one is the most summer-like outfit. The flower shorts are by Hollister and the shirt is by Bershka. Mirror selfie alert, because I had to be quick. We were going out!
I love this outfit! On this day we were going for a hike near the beach. The shorts are by Bershka and the top is by Zara. The veste is by H&M and the scarf by Veritas.
This was by the port!
One of the colder days. Please excuse the hair, because we were on a boat so it was very windy! The leggings are by L&L and the top is by Zara. The veste is from Vero Moda. The bag is by a store called I Am. On this day we were discovering a very green nearby island so it was a little bit colder and there was more adventure, so that's why I chose this outfit!
This picture was taken in our resort. I am wearing a lace shorts by Forever 21 and a knotted shirt by Pull and Bear.
I chose only four outfits, because otherwise this post would be too long, I hope you don't mind and like the ones I chose!
That was it for today guys, I hope you enjoyed reading this and let me know your thoughts in the comments down below!
PS: Do you already follow me on Bloglovin'?
Disclaimer: Not sponsored. I bought everything on my own and these are my own opinions. Pictures are mine.


In the United States, an Alien is an individual who is not a citizen or national of the United States. Government use of the term Alien dates back to 1798, when John Adams signed the Alien and Sedition Acts into law. In general, immigrants are considered a subset of Aliens by the U.S. Government. There are several different ways that the term Alien is used, all denoting different immigration statuses: Deportable Alien, Inadmissible Alien, Nonresident Alien, Resident Alien and Undocumented Alien (also referred to as an Illegal Alien).
There is some discussion on whether using the term Alien is politically correct. For example, in 2009, a Halloween costume that came with a space alien mask and an orange jumpsuit with "Illegal Alien" stamped across the chest caused public uproar and was ultimately banned from Target stores.


Green Vale is a coed, nonsectarian elementary school for Pre-Nursery through 8th Grade, drawing families from all over Long Island. Since 1923, a program of exceptional academics and small class sizes is combined with emphasis on athletics, visual and performing arts, and character education at every grade level. After a culminating experience in the middle school years (6th-8th grades), GVS graduates go on to excel at independent, public, and boarding secondary schools.
"Beginnings" at Green Vale is a "Mommy & Me" experience. Taught by Cara Hershkowitz, Beginnings offers the very young child (9-23 months) the opportunity for exploratory, interactive and parallel play with their caregiver as well as their peers. We welcome all adult caregivers and have been fortunate to have mothers, fathers, grandparents, and nannies participate. The grown-ups enjoy connecting with other adults as the children start to experience what school is all about. Everyone loves starting their day at Green Vale.


boutique in Ithaca, New York.
and Joseph's coat of many colors.
she could give to her children.
when they saw the land being stolen.
wings for a white man.
can't see what's funny in that.
O to fly away from this loss.
Susan Deer Cloud is a Métis mountain Indian (mostly Mohawk/Blackfoot) from the Catskills. She has received a National Endowment for the Arts Literature Fellowship and two New York State Foundation for the Arts Poetry Fellowships. Her most recent books are Braiding Starlight, Car Stealer and The Last Ceremony. She is dedicated to getting out the voices of indigenous people through her editing work and by serving on the board of YANAN (You Are Not Alone) non-profit Native organization for stopping suicide among young Native people. Her website: sites.google.com/site/susandeercloud/.


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Q: Webservice Method generic list fill and keep for 1 day I have a webmethod inside a webservice that calls another webservice to get data and fills a generic list then it returns it, what i want to do is to save the list in memory, so the next time the webmethod is invoked it does not hit the other webservice but just returns the list, i have tried but when i invoke the web method for the second time the list count shows as 0, looks like garbage collection is cleaning all. any suggestions ?

A: It sounds to me like your list might either not be static, or it might constantly be new'd within a non-static constructor. There are three possible fixes for this:

*

*Make sure that your generic list is a static property which only get initialised within a static constructor.

*Seeing your time requirements I would also suggest potentially looking into MemoryCache or Cache.

*Use the WebMethod attribute and set a CacheDuration (i.e: [WebMethod(CacheDuration=86400)])

A: Store it in the ASP.NET cache.  Setting an absolute expiration of midnight should assure that you only get it once per day (unless it gets tossed from the cache due to space issues).
 [Web Method]
 public List<Foo> GetFoos()
 {
      var foos = Cache["FooList"] as List<Foo>;
      if (foos == null)
      {
          ... get foos from remote web service ...
          var expiration = DateTime.Today.AddHours(7);
          if (DateTime.Now >= expiration)
          {
              expiration = expiration.AddDays(1);
          }
          Cache.Insert( "FooList", foos, null, expiration, Cache.NoSlidingExpiration );
      }

      return foos;
 }

Note: you could also use output caching as well, but you're limited to a sliding expiration.  That is, it will be cached for a duration based on when the request occurs.  It's not clear that's what you want.  For example, what if the first request occurs at 11pm with a 24 hour duration, you wouldn't check again until 11pm the next day.  If you have data changing on a daily basis, you're better off using the ASP.NET cache in conjunction with output caching on a shorter duration to ensure that you get the latest, daily data in a timely fashion.
Updated example based on comments.

A: I have not tried this on a webservice, but I think output cashing would work.
[WebMethod(CacheDuration=86400)]
public string FunctionName(string Name)
{
    ...code...
    return(sb.ToString());
}

Read: How to perform output caching with Web services in Visual C# .NET


This highly polished contemporary wall sconce with its crisp, clean lines and cut-out frame, will make a stunning wall feature.
Its oval shape makes it wider than it is deep, with the light spilling down the wall and through the horizontal laser-cut slits.
But this silver wall light doesnt need to be switched on to shine; even in the day it reflects light from its gleaming surface.
Made from nickel-plated steel, this modern metal wall light will have a sophisticated elegance.


"Always the Bridesmaid, never the Bride"
The Just Add Ink challenge this week is from Nikki, 'Just Add Bows' and as you know, I always love adding bows to my cards, but this time, for me, it had to be an 'amazing' bow - a stand out bow!
This is a tri-fold card, so it needed to be on an easle!
The card was so thick that I made a box for it!
I am loving the new Modern Medley Designer Series Paper on page 154.
I have seen so many beautiful cards made using this paper, great for Christmas and male cards too! I used it >HERE< a few months ago.
Another favoutite stamp - Hello, Doily on page 122.
Simply Scored was used to create the box.
OMG! that is just AWESOME!!!!
Wow Wow Wow this is soooooooo stunning L! Love every square inch of this masterpiece, you are a superstar!
Love how elegant this card is. There is just something about this colour scheme that just makes me go "Oooh".
This is a gorgeous set! WOW and I love black and white.
Thank you so much for leaving a nice comment on my blog. Always appreciated.
Lou this card is just beautiful. I always love black & white together. You are soooo talented. Just love everything you do. Thanks for sharing!
How do you get the colored doilies? I'd really like to try this.