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World News: 18:11 GMT Monday 17th July 2017. [Lundin Law PC via Globe Newswire via SPi World News]
LOS ANGELES July 17 2017 GLOBE NEWSWIRE -- a shareholder rights firm announces a class action lawsuit against Neurotrope Inc Neurotrope or the Company NasdaqNTRP for possible violations of federal securities laws from Jan uary 7,...
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The goof was on the 26th section of the Doctor Who Scarf. I knit it as yellow. It's supposed to be tan. I didn't notice until my boyfriend pointed it out to me today just as I was finishing the 37th section.
I just couldn't leave it. I couldn't. I know that not many people will notice a mistake like that, but I'll know it's there and I can't have that.
Yup. My scarf is in two sections. I pulled it apart at the join between the 25th and 26th sections. I'm going to knit the tan as I was supposed to then join with the gray section that's supposed to be next. Guess it's time to figure out how to graft two pieces of garter stitch together!
Also: It's a really good thing that my color joins aren't super knotted and short. I was actually able to pull it out without cutting anything, though what I'm going to do with that length of yellow I have no idea at this point. I'll figure something out.
And in the baking department…
I made cookies! I'm going to frost them in the morning (I'm too tired to do it tonight) but, cookies! Anise cookies! A holiday classic in my mom's family courtesy of my great grandmother (maternal grandfather's mother – if you can follow that).
I'm planning to bring some to knit night at my LYS tomorrow as a treat. I don't bake/cook often, but I shared the cake I made for my birthday and I figured I'd bring some of these. I wanted them anyways, and I knew my mom wouldn't object to my baking them, so now the house smells nicely of anise. It's a great smell this time of year. And the flavoring isn't even really that strong, it tastes nothing like licorice (or absinthe!), it's much more subtle.
I hope everyone had a good Christmas yesterday and though I'll have another post before New Years, Happy New Years anyways!
PS: Have a penguin pinata full of nips. We swung at it with a sword. I didn't end up drinking any of the nips, but a ton of fun anyways. Who doesn't like a pinata?

subset: commoncrawl


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Here's Patrick on his banjo and his father, Pat, on a tenor guitar. This video is about having a mellow jam session. They have the songs posted as jpg's so that you can print and play along as you wish. The tab is in "Banjo" tuning G-DGBD and might be "tweaked" to bari if you're quite diligent. The video is over 38 minutes and is fun to just listen along to as well.
Patrick's comments...
Recorded in 2001, Pat & Patrick Costello present the skills needed to learn frailing banjo (the same style Grandpa Jones played) in a series of lighthearted, easy to follow workshops that show you just how easy it really is to play a few tunes on the front porch. Chapter 8: Play-along jam session. Tab files for this workshop series can be downloaded from:


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As he walked by the Sea of Galilee, he saw two brothers, Simon, who is called Peter, and Andrew his brother, casting a net into the sea for they were fishermen. And he said to them, "Follow me, and I will make you fish for people." Immediately they left their nets and followed him. As he went from there, he saw two other brothers, James son of Zebedee and his brother John, in the boat with their father Zebedee, mending their nets, and he called them. Immediately they left the boat and their father, and followed him. (Matthew 4:18-22)
Today we hear the familiar scripture of Jesus calling men to be fishers of men. Jesus first asked Andrew and Simon Peter; then He called James and John to join his disciples.
There are many stories within the Bible that are known as 'call stories'. A call story is about someone being invited by God to become something new and unexpected. God calls this person to begin, and not only begin, but persist so that this new thing can take place.
One day Andrew, Simon Peter, James and John got up before the sun came up, walked down to the sea and hurled nets into the water, anticipating a catch of fish. It was a day just like every other day; it was dark and probably cool and the nets were smelly and heavy. They were doing what they did every day. There were fishermen.
Jesus came down to the seaside, amid the water and nets and fresh fish, roughly hewn boats of wood, the rhythm of the waves. He stood on the bank watching these men throw out the nets and then haul them back in, loaded with fish. He looked at these men, and in a very commanding voice, announced:
Follow me and I will make you fish for people. (Matthew 4:19)
I imagine they looked at this man on shore as if you he was a little crazy. Obviously, he knew nothing about fishing – the very idea that they could fish for humans!
Who is the crazy man, this itinerant preacher who calls them to fish for people? And where did he come from? The Sea of Galilee was in the middle of nowhere!
What was this man doing there? And why had he come all this way from his home in Nazareth?
And why did he choose these men?
The command / invitation to Simon Peter, Andrew, James and John to
Come, follow me, (Matthew 4:19)
was not:
- "I wonder if you would tag along with me,
if you have the time,
and it's not too inconvenient."
It is so difficult for most of us to imagine getting up and leaving everyone and everything they knew to follow Jesus. We surmise that the disciples were extraordinary, first-century super heroes of the faith that we can admire but not identify with.
But we know that Jesus did not read resumes before he called these men to be his disciples. He didn't care what their history was. And as we now know, sometimes he didn't always make the best decision on whom He called. Simon, who became known as Peter, denied Jesus three times during his trials. James and John, often called the 'Sons of Thunder', thought this calling was going to allow them to defeat the Romans and enthrone them in glory, quite the opposite of what Jesus taught. But Jesus called them, and in spite of their own personal deficiencies, He still made them his disciples. And they left their old life, its security, and even their families. They may have been afraid, but not so afraid that their faith in Jesus did not lead them to answer His call.
And why did they follow Jesus?
When they were called by Jesus, they must have felt the joy of the new world that Jesus was preaching. They were about to see miracles performed and illnesses cured. Jesus was going to show them a wonderful new world, touch everyone who heard Him, and then, make the ultimate sacrifice to bring about the new world.
If Jesus called this group of imperfect humans to be His fishers of men, then why wouldn't He call each of us to follow Him? Our own calling means the same kind of new beginning; each of us are called to go to that edge of safety so we can bring people to Christ. Jesus comes to us and chooses us, and sends us out to do something new.
We are called to be evangelists. . . to look for and bring people to Christ. We are called to say to others 'Come and See'. But we can't be fishers of people until we have been caught by Jesus. We need to fish for others using our own personal experience as bait.
Jesus issues the same call to us — to be in genuine and real relationships with the people around us, the way Jesus was and is in relationship with us. We need to bear each other's burdens, care for each other – especially the vulnerable – hold onto each other through thick and thin, always with the promise of God's abundant grace.
I remember giving a testimony at a church as part of the stewardship campaign about how the church and God had gotten me through a very rough time when my partner of 27 years was dying of cancer. After the service, a young man came up to me and thanked me for my testimony. It seems that he was so depressed and sure that God hated him that he had been planning to go home and commit suicide.
Imagine how I felt at that moment. I certainly had not given the testimony with the intent of fishing for people. But with God's help and direction, my little testimony was the bait that brought a young man back to God and salvation.
We are called to bring others to the kingdom of Christ. . . where we are all one in His love.
But, how do we do that?
Think of one person you have a relationship with: it could be someone you love or someone that irritates or frustrates you. Now, take a moment to pray for them; believe that God is using you to make a difference in their lives.
You are now a 'fisher of people' – you have joined the ranks of those that Jesus first called on the banks of the Galilee! You are caring for those whom God loves?
Let us pray:
We are lazy sometimes, God. Sitting comfortably here, it's easy to ignore the suffering of others, easy to forget the challenge to see your image in friend and stranger, easy to expect others to work for justice. We are lazy sometimes, God, and that diminishes us. Make us uncomfortable, make us alert, make us responsive to the call of Jesus. Above all, make us love, in your name. Amen.
Delivered at In The Garden, Trinity Episcopal Church on Capitol Square, Columbus, OH 26 January 2014


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Does anyone know the outcome of this court action? It presents an interesting question
Addicts, operator of BC safe-injection site go to court to keep facility open
918 words
17 August 2007
The Canadian Press
(c) 2007 The Canadian Press. All rights reserved.
VANCOUVER (CP) _ Two drug addicts and a group that runs Canada's only safe-injection site have launched court action, saying any effort by the federal government to close the place would violate addicts' Charter rights.
Shelly Tomic and Dean Wilson, along with the Portland Hotel Society which helps operate the facility, filed a statement of claim in B.C. Supreme Court on Friday.
They want the court to declare the operation of Insite, as the facility is known, the exclusive jurisdication of the province and that the federal government shouldn't play any role in its future.
Closing it would be a violation of Tomic's and Wilson's right to ``security of the person,'' the court documents say.
Insite opened four years ago as a pilot project in the Downtown Eastside for IV drug users to inject their own heroin and cocaine with clean needles under the supervision of a nurse.
Addicts who get their fix at the site -- instead of in back alleys and the area's single-room hotels -- can also access referrals to detox and rehabilitation services.
But the future of Insite is currently in Ottawa's hands because in order for the facility to operate, it needs the federal government to grant it an exemption from the Controlled Drugs and Substances Act.
The exemption is set to expire Dec. 31 and federal Health Minister Tony Clement has been noncommital about whether it will be renewed.
``Should Insite close due to the revocation of the exemption. . . Wilson, Tomic and other users will face risk of death or serious harm to their physical and mental health,'' the court documents say.
``In particular, Wilson and Tomic will face increased risks of overdose, infection, decline in their mental and psychological well-being and other health-related complications from drug use.''
The documents go on to say the services provided at Insite are health-care matters and are therefore ``within the exclusive jurisdiction of the province of British Columbia.''
Mark Townsend, director of the Portland Hotel Society, said the ongoing uncertainty is no way to operate a health service for vulnerable people.
``It's not even funded by the federal government, it's 100-per-cent funded by the province, yet they get to give us an extension here and there to create this endless kind of worry,'' Townsend said.
``We'd rather not be in court on this issue but what we really need is just to get the clarification that, really, this is about health care,'' he said.
The site, known as Insite, is the only facility of its kind in North America.
Tomic, a 39-year-old addict with a long history of addiction issues, said she got involved in the legal action because she's seen its positive effects first-hand
``A lot of people will end up dying if they shut it down because with the fear of the police picking them up, they start (injecting) too much just to get it in them before they're caught,'' said Tomic, who was one of the first to use the facility.
Tomic said she's been taking methadone for about 10 months as part of her treatment to withdraw from heroin. And despite staying clean so far, she said she still visits Insite to talk to the staff on days when she feels like relapsing.
Dean Wilson, the other addict named in the statement of claim, credited Insite for keeping him and his two sons alive.
``This is gold-standard medicine,'' Wilson said. ``Addiction is a health issue.
``This is a life-saving measure down here. It may not be a life-saving measure in (Toronto's) Rosedale or Mount Royal in Montreal but in the Downtown Eastside, it is a life-saving measure.''
The court documents note Wilson has been addicted to herion for about 38 years, contracted hepatitis C in 1969 and has had several close calls with overdoses.
The suit says before Tomic and Wilson began getting treatment for their addictions, they stopped in at Insite to inject about six times a day for Tomic and 15 times a day for Wilson.
Dr. Evan Wood, an epidemiologist at the B.C. Centre for Excellence in HIV/AIDS, said 24 studies have pointed to Insite's benefits.
Since 2003, research published in top journals has stated the site reduces public injections, overdose fatalities and the transmission of blood-borne infections like HIV and hepatitis C, Wood said, adding crime hasn't increased in the area despite initial fears by some in the community.
``From a scientific perspective, I believe this is a health issue,'' Wood said, adding he agrees with the court action.
``(Prime Minister) Stephen Harper has failed to show any leadership on this issue and the fact that it's going to the courts now is not surprising,'' he said.
``If the facility closes and somebody contracts HIV or dies of an overdose and has a compelling argument that this wouldn't have happened to them had the federal government not removed this service, then I think that raises legal issues as well.''
Monique Pongracic-Speier, one of three lawyers working on the case for free, said her clients are merely asserting their constitutional rights regarding the use of a facility that's helped them stay alive.
``After all, Insite is about saving lives. That's the bottom line.''
Wednesday, October 17, 2007
The question of charter rights
Does anyone know the outcome of this court action? It presents an interesting question
Posted by Jen at 10:44 AM


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Hi, I'm Ciar, a writer, gardener and mum of three under-10s aka the Little Weeds.
We moved into our urban garden a year and a half ago and it is very much a work in progress. In the sunny, south-facing front garden we are in the process of replacing overgrown shrubs with raised veg beds.
Our back garden is north-facing but on a slope, so gets plenty of sun in the spring and summer.
It starts off with a shaded patio and overgrown toolshed, then you go up curved steps to a lawn which has been destroyed this year by our trampoline and resident Guinea pigs, Nibbles and Gracie.
More steps lead up to more terraces – we have a lot of crumbling terracing; a jungle of fruit trees including a huge fig we have nicknamed The Whomping Willow after Harry Potter; a sandpit which is popular with the local cats and another large shed.
I have a clear vision but it is going to take some time to achieve. In the meantime, I am putting to use my RHS Level 2 Certificate in the Principles of Horticulture by planting a glorious jumble of flowers and vegetables (hence the name of the blog).
I am also lucky enough to volunteer in the garden at nearby Charleston farmhouse one day a week, where I am learning lots of skills and gaining ideas for my own garden.
This blog also talks about inspiring gardens we have visited, some of my favourite gardening books and gardening projects I've tried with my kids (when I can persuade them away from screens and outdoors) or as my husband aka Mr Carrot likes to call it "Forcing kids to dig dirt."
You can also find me on Instagram and Twitter as @wildecarrot


text:
Today was just beautiful! I love spring. I love the clouds that accompany storms...
and I love flowering trees. You can just see the touch of pink by the bridge. My poor redbud has been so slow to grow. It is just about to bloom. I heard that it takes a redbud about 7 years to really get rooted and to take off. We planted it 5 years ago, so hopefully soon it will begin to do something!
My flowering crab tree is on the verge of flowering too. I think tomorrow that will happen although I have seen it elsewhere in town already.
I had an errand to run yesterday. There was such a crowd at South Dillons and all along Washington Street people were lined up. I had no idea what was going on. So I asked Jack at Ruzens.
Evidently the eight-horse Clydesdales (as in Budweiser) along with the red beer wagon were marching down our main street in town.
Each wagon travels with a Dalmatian. I read that in the early days of brewing, Dalmations were bred and trained to protect the horses and guard the wagon when the driver went inside to make deliveries.
Progressive Dinner
Our ladies ministry planned a progressive dinner last evening. We started at the church with appetizers. Then the women were divided into 3 groups.
One of the groups came to my house for salad. I served both a Caesar salad and a tossed salad sprinkled with Wal-marts' Great Value Good Health Energy mix - a blend of soy nuts, cranberries, almonds and pumpkin kernels. We LOVE this! I served home-made honey mustard dressing. It is Lisa Prier's recipe (my daughter-in-law's mother).
Mix together in whatever portions tastes good to you!
To drink, we had the options of ice water, raspberry/mango tea and Mary Ellen Tippin's almond tea. Here is the recipe for those who asked:
10 cups water
2/3 cups lemonade mix
1 T instant tea
1 t. almond flavour
1 t. vanilla flavour
3/4 - 1 cup sugar
Mix well.
After an hour at my home, I joined the group and carpooled to Helen Wall's house for lasagna, green beans and French bread.
I left my camera at Helen's so didn't get pictures of the next stop - dessert at Julie Garrett's house. She fixed peanut butter pie!
I love Sunday mornings. The babies at Grace are so very precious as are the workers who faithfully come! Katelin comes to help with her daddy in the infant, or Bunny class. Isn't this sweet?
And Andi is so dedicated and has helped me for several years already. Wonderful with the children!
This week we will be parenting again! While Adam and Jill attend a pastor's conference in Chicago, we get to take care of these sweet girls!
Positively Perfect!
Sunday, April 10, 2011
Posted by Allen and Debby Graber at 6:17 PM
2 comments:
- Marc, Sarah, and Luke said...
The progressive dinner was fun! I didn't know if you saw me or not during the appetizers (I didn't get a chance to come over and say "Hi")....But, we had a great time at Marjean's, at Sandra's, and at Marilyn's....I definitely want to sign up again next year!
- April 10, 2011 at 8:40 PM
- Rich and Carolyn Dewey said...
Loved your "P's". Progressive dinner was "Pfun"!
- April 11, 2011 at 12:57 PM
Subscribe to: Post Comments (Atom)


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Find UK Tuition
Classical & Jazz Piano Lessons with Cadenza Music - Tooting, Balham, Streatham, Wandsworth Common
Cadenza Music Tuition
- Tooting (SW17) View on map
- Address: Tooting Broadway, SW17 0SU
- Telephone: 07432487947
This Study Centre is temporarily unavailable for contact
- Founded in 2017
- All tutors qualified to Masters degree level in music
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Subjects we cover
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- Personal Message
- Jazz and Classical piano lessons that bring together the skills and expertise of a team of tutors to deliver lessons to your home that are creative, inspirational, progressive and holistic.
- Classes
- One of the most distinctive features of lessons with Cadenza Music is the versatility and breadth of teaching available. All tutors are highly experienced and qualified in delivering piano and keyboard lessons across Jazz, Classical, Contemporary and Popular music. Each subject area is not isolated however. The benefits of multi-disciplined tutors is the huge creative resource that can be drawn from. Various facets of Classical piano can be used in Jazz tuition and vice versa. From this comprehensive approach to lessons each pupil benefits from a truly holistic experience whilst also having the opportunities to be exposed to approaches and styles that they may not have previously considered. Lessons are also available in Music Theory, Composition, SIbelius and Improvisation.
Group lessons are conducted by founder Daniel Nicholson every term to offer collaborative opportunities, themed workshops, mock exams and performance sessions.
These are held in Wimbledon at a Piano Showroom.
Classical Piano
Jazz Piano
Popular Piano
Electric Keyboard
Music Theory
ABRSM and Trinity exams (100% pass rate)
These areas are currently covered:
Wandsworth Common, Balham, Tooting, Earlsfield, Streatham, Southfields, Putney, Wimbledon
- Ages and Levels Covered
- No lower age limited but recommendation is to begin from 5yo.
No upper age limit. Adult learners are welcome!
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- Fees are maintained at a fair level so that the opportunities are available to all.
Profits from Cadenza Music Tuition largely get put back into the business to offer further enhancements to lessons.
30mins - £22.00
45mins - £28.00
60mins - £35.00
These fees include travel to your home!
- Centre Director(s)
- Daniel Nicholson BMus MMus
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- For profiles and more general info please follow the link below:
A major benefit of taking lessons with a music school such as Cadenza Music is the fact that your tutor will have the full support of knowledge and resource of the whole team. Issues and new ideas are regularly communicated and developed to ensure that there is sustained motivation, progression and enjoyment.
Cadenza Music operates across two sites. In Cardiff, founder Danie Nicholsonl teaches from his home teaching studio in Llandaff whilst a team of associate tutors provide home visit tuition to pupils across South West London. The whole team regularly meets and discusses teaching matters and experiences whilst Daniel provides half termly reviews and observations of lessons to ensure that the standards of tuition are being kept to the highest level. The needs of each pupil are at the forefront of lesson delivery and the independent feedback given by Daniel is another way to maintain this.
Supplementing the teaching are dedicated resources including a branded Practice Notebook, online resources from the Pupil's Portal, and Cadenza Music's own book series called Missing Bars which focuses on aspects of composition and improvisation.
As a whole, Cadenza Music tutors thrive in a creative and progressive environment and will teach in a way that is motivating, inspiring, enjoyable and challenging. The piano is one of the most versatile instruments there is and it is the aim of all Cadenza Music tutors to match that in their teaching
- Special Offers
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Key Staff
Daniel NicholsonDaniel is a the Founder of Cadenza Music Tuition.
He graduated from the Royal Welsh College of Music and Drama with Bachelors and Masters degrees in 2013 and began his teaching career in 2011. He studied with internationally acclaimed musicians and educators during this time and maintains some professional connections to the college. Today Daniel teaches one to one and group lessons from his home studio in Cardiff
Benjamin CookBen teaches in SW London with Cadenza Music and offers home tuition in Jazz and Classical piano. He holds Bachelors and Masters degrees and draws upon a huge creative resource formed from his multi-disciplinary background which also includes Composing for Moving Image. Ben has an empathetic approach and will always interesting ways to bring vitality to learning the piano.
Christina KarpodiniChristina started teaching with Cadenza Music in 2017. She began her musical studies at 7 years old in Classical piano and vocal ensemble participation. She took her first music Diploma in Music Harmony from the Conservatory of Music in 2011 and a second diploma in Counterpoint Music in 2013. In 2015 she took her Piano Teaching Diploma (equivalent to LRAM) and her Bachelor Degree in Music Studies, pathway Music Technology and Acoustics, with first-class Honors by the National University of Athens.
Joshua McConvilleJoshua holds a BMus (Hons) from one of the country's finest music departments during which he had the opportunity to study piano at the Royal Academy of Music under Professor Colin Stone. Josh also gained a PGCE in Primary Education from Goldsmiths College which he passed at Master's level. With over 5 years' experience of teaching piano, Josh is a highly experienced teacher and has taken pupils from the earliest stages of learning piano to successfully completing grades 4 and 5.
Liam HarmanLiam began playing and studying piano from the age of 10 alongside learning the Clarinet and Viola. His teaching career began in Spain during his Undergraduate studies where he taught privately and in music schools. He taught a wide range of pupils from age 5 to adults across a variety of styles and abilities. Liam was awarded a scholarship from RCM which presented him with world class opportunities including performing in the Royal Festival Hall and Buckingham Palace.
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Student Testimonials
"Once again, thank you so much for great teaching and encouragement for the girls. They have really benefited from your various methods of instruction and hopefully some seeds of musical enjoyment have been planted and nurtured."
- Kate (Parent)
"The lessons were interesting and stimulating and you managed to incorporate a fund of useful teaching tips into both the scale playing and the pieces. You even succeeded in making note learning an interesting exercise for Grade l. Any problems were broken down into smaller components and sorted out".
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"Dear Mr.Nicholson, I would like to start by thanking you very for that lovely lesson. I learnt so many new things in that session. I really like the way you put sight-reading and theory together because you know that will really help me."
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"You are a talented piano teacher and performer, but even more so your lovely caring nature really was there for us to see."
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I am an adult pupil based in London and have lessons with Ben. Ben is an engaging and inspiring teacher and I would definitely recommend his lessons to others. The whole business is run very efficiently and professionally and the annual showcase that Dan puts on each year is great!
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Couldn't be happier. Both my daughter and I have lessons with Dan. We're both making good progress and enjoy the lessons, which culminate in a showcase each year. Very happy to recommend!
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Dan has been providing me both piano and music theory/composition tuition for a couple of years now and I highly recommend him. I'm an adult trying to learn around a hectic schedule, but he's very patient and supportive even when progress slows during those busy periods. He's also able to effortlessly work with material he may not be familiar with, to help get you playing what you enjoy. Friendly. Professional. Great stuff!
- David (adult pupil)
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Bill Sussman Replaces Collective Bias Co-Founder John Andrews As CEO, President
by Chris Bahn
Posted 6/11/2013 04:05 pm
Updated 5 years ago
Bill Sussman has been named president and CEO of Collective Bias, the global social media marketing firm headquartered in Bentonville. Sussman replaces co-founder John Andrews, who has left the company after helping it secure $10.5 million in investment funding.
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2 Corinthians 2:1-5 English Standard Version (ESV)
2 For I made up my mind not to make another painful visit to you. 2 For if I cause you pain, who is there to make me glad but the one whom I have pained? 3 And I wrote as I did, so that when I came I might not suffer pain from those who should have made me rejoice, for I felt sure of all of you, that my joy would be the joy of you all. 4 For I wrote to you out of much affliction and anguish of heart and with many tears, not to cause you pain but to let you know the abundant love that I have for you.
Forgive the Sinner
5 Now if anyone has caused pain, he has caused it not to me, but in some measure—not to put it too severely—to all of you.
Cross references:
English Standard Version (ESV)
The Holy Bible, English Standard Version. ESV® Text Edition: 2016. Copyright © 2001 by Crossway Bibles, a publishing ministry of Good News Publishers.
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Congrats Maddy! I know she'll be great there, she'll be with just as many friends from home as I had. Meagan + Chris, Kailee, Jess + Stephen, Laura, Jena (?), and all the awesome people that are there already. I'm really so excited for her, she's gonna love college as much as I did I think. It's the bomb.
I think the talk from Elder Bednar was given at the MTC, so maybe it's not available anywhere else. It's about how to be truly converted (and how that's different than merely having a testimony) and how having a character of Christ is always looking and reaching outward instead of inward. Putting off the natural man and becoming more converted everyday through the Atonement. So eloquent and powerful though, I wish you all could listen to the real thing.
Yes, having a breakdown is a normal thing. It's a combination of frustration from learning (or not learning) the language, stress from teaching our investigators (especially when they don't keep their commitments!), and the rigorous study schedule we have. The pressure to do well is big too. That's why I feel so particularly blessed with peace and confidence because I haven't felt that way. This might be weird to say, but my spirit just feels so prepared for this kind of stress. It's not getting me down like others and I can be there to help them through their moments of "I can't do this!!". We've had some "Sister inventories" at our residence at night where we've had to talk through why we shouldn't go home, but at the same time it's okay to think about our families and home without it getting in the way of fulfilling our purpose. I haven't had a moment where I wanted to go home or when I didn't feel the strengthening and assurance from the Spirit that I'm going to be fine. I do have lessons where I just can't get the Spanish out and that's frustrating, but I usually always feel the Spirit in our lessons no matter how bad the Spanish and that's really all that matters. My companion this last week has been on an emotional roller coaster though 🙂 One hour she is freaking out because she can't Spanish then the next she's hugging me and telling me how appreciative she is. I'm definitely learning how to comfort and help others when I can't exactly relate to their feelings.
I got a haircut today! They have a barbershop here where we get them for free. I got it about shoulder length so I can still pull it up. Not sure how I like it yet because she didn't "do" it afterward, it was still kinda wet and weirdly wavy but it'll be good for a hot climate. I've never cared less about my hair anyway.
Haha, I was afraid of the facebook moms being crazy. No, that's not bad 🙂 I wouldn't want to either. Was that the general one or the one just for my mission?
I do remember Aunt Inabo. I definitely remember Aunt Rita; Uncle Dan has told me more funny stories about her. It must be weird to have almost everyone you know for so long pass away.
Geez, it never ends. I sometimes just don't get why people make their lives so much harder for themselves.
Last Tuesday night Elder Rasband and his wife came and spoke to us! I never realized how similar the jobs of missionaries and apostles are; they feel a real kinship with us. It was pretty much a big question and answer session. He talked about how the apostles assign missionaries to missions. He said it's like a picture of a globe and a section of it is lit up in neon. They just look into our eyes and it's like we're there. He says assigning missionaries is the most spiritually exhausting and demanding thing he does as an apostle. Cool stuff. In the choir that night we sang an arrangement of "Nearer, My God, To Thee" and I learned the background of the song. It was originally a poem about Jacob, being in the desert after getting the birthright from Esau and Esau wanting to kill him and being warned by Rebekah. He was just wandering, trying to find the family of his mother and thinking that his life was pretty rough when he laid his head on a stone and went to sleep. He then had a dream about a ladder, which modern prophets have identified as his endowment. He woke up praising God instead of pitying himself. He built an alter of stone and named the place "Bethel". "Beth" means "house of" and "El" is short for the name of Heavenly Father = "House of the Lord". Go read that hymn with that context and it takes on whole new meanings. Love it!
We had a couple of really spiritual moments during our lessons this week. We have been teaching Andrea and she wants to be baptized but she lives with her boyfriend who is a member but in-active and their daughter. I felt prompted to ask her to share her testimony to him and her simple, new faith brought so much into the lesson! She wants an eternal family so her support won't only help Sebastian but it will cement it into her own faith too.
Have I told you guys about Elder Steimle? It's so hard to describe him without actually meeting him. This week he said so many hilarious things. He mentioned that when he was around dogs he starts behaving like one. We obviously asked for a demonstration, so he showed us his pant, bark, and growl. Frighteningly accurate. He also says that female animals are attracted to him. He shampoos his leg hairs (is that normal?). He also said, "I could never enjoy a woman who was so fat that she couldn't run." Keep in mind that these comments had very little to do with the context of the conversation. Oh Steimtime.
My next story will demonstrate how the MTC has brought us to the brink of desperation in our forms of entertainment. We decided to start telling people that Adam Sandler had died, that we'd heard it from people at the temple. We very quickly saw the fruits of our labor. We'd exclaim the news and the missionaries in front of us would turn around and say, "Oh ya, from a heart attack, right?" Haha score.
Elder Nasilai says, "hello, you're amazing. P.S. Go Michigan P.S.S your cookies are even more amazing". We're gonna have fun in El Salvador. We were talking about how dangerous our area is and he was joking about how he's gonna buy guns when he's down there and eventually become one of the leaders of MS13 (a really scary gang down there). He goes, "Down there it becomes kill or be killed, and I ain't dyin'!". Then he goes, "No, I'm gonna come around a corner someplace and I'm gonna see Hermana Morse just curb-stomping someone. I'm gonna go see if she needs help and I'll just stand there and watch. She'll leave a Book of Mormon on the guy with a note saying 'Read this book, it's about repentance. P.S. I'm sorry' and she'll just walk away". Haha I'm actually worried about him getting into trouble down there though. He's gonna be with a little white companion and when someone tries something he's gonna defend him. Honestly though, if I was in a situation and I saw him coming I'd breathe a sigh of relief. Not that any of that is going to happen or that we'll even see each other there. We also decided that we're getting donkeys to ride around. Much more of an entrance that way.
Anyway, long email this week but it was a good one. I'll be forwarding more emails with pictures of everyone. The pictures in this email are of the record board and me with my certificate the MTC gave me for breaking it. Love you all so much!
Hermana Morse


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Christy Bishop has taught at Rocky Point Elementary for 15 years
Why teach? Pender County's Teacher of the Year Christy Bishop said her motivation is simply her love of children.
"I love being a part of their academic, their social, their emotional and just their overall well-being and development," Bishop said in a video posted by Pender County Schools.
She began her career as a first-grade teacher at Rocky Point Elementary, where she remained for 11 years. For the past four years, Bishop has been the Title 1 coordinator as well as reading interventionist at the school. She said her responsibility as Title 1 lead works with her role as literacy facilitator, allowing her to develop learning strategies specific to her students' needs.
With nearly 15 years under her belt, Bishop said she is proud to represent Pender County.
The "ah-ha" light bulb moments that her students have are especially rewarding, Bishop said.
"I am blessed to impart knowledge, and to be an influential factor in helping a child learn to read, a skill they will forever use for the rest of their life," she said in the video.
Bishop received a bachelor's of science in elementary education from the University of North Carolina Wilmington, where she later returned to get her master's in education.
She joins Tosha Diggs, principal of Trask High School, to represent Pender County Schools in the regional teacher and principal of the year competitions.
Reporter Courtney Sullivan can be reached at 910-343-2023 or firstname.lastname@example.com.


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Movie The Andy Griffith Show Goodbye, Dolly () download free
Book Title: The Andy Griffith Show Goodbye, Dolly
Directors: Lee Philips
IMDb Rating: 6.2
Country: USA
Language: English
Genre: Comedy, Family
Release Date: 1967-03-06
Runtime: 30 min
Stars: Andy Griffith,Ron Howard,Frances Bavier
Full movie description "The Andy Griffith Show Goodbye, Dolly":It's the end of an era in Mayberry when Walt Simpson, the milkman, needs to retire his horse-drawn milk wagon and start using a modern truck. He plans a nice home for his horse however and hires Opie to take care of it while he is away for a week. Andy isn't too keen on Opie taking on that responsibility remembering what had happened when Opie had agreed to deliver someone's morning papers only to have Andy delivering them himself. Opie is conscientious however, traveling every day to Simpson's farm to feed the horse. The problem is that it simply won't eat and becoming very concerned, gets Andy's help. The vet can't quite figure out what is going on but they soon find the root of the problem.
Reviews of the The Andy Griffith Show Goodbye, DollyI rate this episode a "10" just for the sake of "Dolly"-the horse. The horse that appeared in 'Goodbye, Dolly' (3/6/1967) shortly appeared one month later in the 'Star Trek' episode 'City On the Edge of Forever' (4/6/1967) with the same cart from 'The Andy Griffith Show.' 'Star Trek's' "City" had exterior scenes filmed on the Forty Acres back lot-where Dolly previously appeared on TAGS. Dolly resurfaced one season later in the 'Gomer Pyle USMC' episode 'Goodbye, Dolly' (2/23/1968). She worked both in the confines of Desilu and outside at Forty Acres. So, one must deduce that it was brought in a horse trailer three times to Forty Acres. Most actors rode on the bus or drove themselves. That horse needs its own page on IMDb!
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Paul's 365 Photo Blog
Random Daily Pictures.
Wednesday, September 24, 2008
Crystal Springs Dam
This is the dam that holds back the reservoir that was in yesterday's picture.
This is a great old dam, built in 1890. It survived both the 1906 and 1989 quakes, both of which hit the area pretty hard (remember, the reservoir sits on top of the San Andreas).
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Did You Know?
We provide international education to over 1,100 students ages 1-18. We are at the heart of our community, a hub for many expat families in living Beijing.
Through the Nord Anglia University we focus our teachers professional development to ensure that your child receives high quality teaching experience.
You will immediately recognise one of our students when you meet them by their evident respect for others, their intellectual and social confidence.
We are a high achieving school with excellent academic success. We believe in bringing out the best in every student of all abilities.
We have a dedicated team waiting to hear from you and ready to put you in touch with others whom you may wish or need to speak to.
Through our News & Insights section you can find out the latest from our school and from other schools in the Nord Anglia Education global family.
Did you know?
We teach the English National Curriculum, offer a German curriculum at primary level and IB Diploma for Years 12 & 13 students.
29 October 2016
French Y9
Year 7 German Native Cake
German Y9
Spanish Y9
Spanish Y11
Spanish macarena Y9
All the MFL teachers helped celebrating International Day. We had a variety of activities taking place in German, French and Spanish.
In German, Mr Mende's students tasted some tasty German cakes. Mr Müller shared his passion for Geography. The students were encouraged to spot and locate various German landmarks and cities.
In Spanish, the Year 11 students put their cooking skills to good use and explored Spanish cuisine by recreating certain dishes. The Year 9 students created flags from various Spanish speaking countries. In Ms Châtillon's Spanish class, students talked about Spanish music. They had a blast dancing the Macarena following instructions in Spanish.
In French, creativity was the word of the day! Our Year 9 students looked at the work of Henri Matisse, a famous French artist. After sharing information about the painter-sculptor, they were encouraged to create their own "chef d'oeuvre" inspired by the work of Matisse using collage.
We hope our students had a fantastic time learning about the cultural aspects of Germany, France and Spain.
Update from: Mrs. Linda Worsley, Head of Modern Foreign Languages


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Enhancement of Monocyte Procoagulant Activity by Adhesion on Vascular Smooth Muscle Cells and Intercellular Adhesion Molecule-1–Transfected Chinese Hamster Ovary Cells
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Background—Plaque erosion is a frequent finding in sudden death due to coronary thrombosis. The present study sought to investigate whether monocyte adhesion to human aortic vascular smooth muscle cells (VSMCs) induces procoagulant activity (PCA) and whether this could be mediated by intercellular adhesion molecule-1 (ICAM-1).
Methods and Results—We incubated mononuclear cells (MNCs) with VSMCs and ICAM-1–transfected Chinese hamster ovary (CHO) cells, investigated monocyte tissue factor (TF) mRNA expression by Northern blot analysis and TF protein expression by ELISA, and measured PCA. Incubation of MNCs with VSMCs for 6 hours increased PCA from 0.7±0.1 to 166.0±37.9 mU/105 cells (P=0.007), which could be inhibited in a dose-dependent manner by the addition of blocking anti–ICAM-1 monoclonal antibodies. Prestimulation of VSMCs with interleukin-1β enhanced surface ICAM-1 expression significantly but did not induce PCA in VSMCs. Incubation of MNCs with prestimulated VSMCs led to a further increase in PCA to 239.9±27.9 mU/105 cells (P=0.02 compared with incubation with unstimulated VSMCs). Incubation of MNCs with VSMCs enhanced TF mRNA after 2 hours and significantly increased TF protein content after 6 hours. Incubation of purified monocytes with ICAM-1–transfected CHO cells increased PCA from 1.2±0.2 to 81.9±3.3 mU/105 cells (P<0.001 compared with incubation with untransfected CHO cells) after 6 hours. This effect could be inhibited significantly by the addition of blocking anti-CD18, anti-CD11b, or anti-CD11c monoclonal antibodies. Similar results were obtained for MNCs.
Conclusions—Monocyte adhesion to VSMCs induces TF mRNA and protein expression and monocyte PCA, which is regulated by β2-integrin–mediated monocyte adhesion to ICAM-1 on VSMCs.
Thrombosis in acute coronary syndromes is caused by 2 different processes: either plaque rupture or plaque erosion.1 Plaque rupture of a lipid-rich plaque exposes the highly thrombogenic core to blood in the arterial lumen, thus initiating the coagulation cascade and thrombus formation. In plaque erosion, the overlying endothelium undergoes denudation, and thrombus formation occurs on the surface of the plaque.2 A recent study showed that erosion of these proteoglycan- and smooth muscle cell–rich plaques, which lack a superficial lipid core, is a frequent finding in sudden death due to coronary thrombosis.3 This form of endothelial loss and erosion shows marked surface accumulation of macrophages and lymphocytes.4 Furthermore, tissue factor (TF) content in coronary artery sections of patients with unstable angina correlates with areas of macrophages and smooth muscle cells.5 Nevertheless, the mechanisms that cause thrombosis after plaque erosion are as yet incompletely understood.
TF is an integral membrane protein, which binds to factor VII/VIIa and initiates the coagulation protease cascade.6 7 Several stimuli, such as lipopolysaccharide (LPS),8 cytokines,9 10 immune complexes,11 and lymphokines,12 13 14 induce TF expression in monocytes, and it has recently been shown that monocyte interaction with endothelium increases monocyte procoagulant activity (PCA).15 16 We speculated that monocyte adhesion to vascular smooth muscle cells (VSMCs) might induce monocyte TF expression, thus contributing to thrombosis after plaque erosion.
Monocyte adhesion to VSMCs is mediated primarily through intercellular adhesion molecule-1 (ICAM-1),17 which is increased in the surface of VSMCs in human atheroma.18 19 ICAM-1 tethers β2-integrins on the leukocyte surface, such as lymphocyte function–associated integrin-1 (LFA-1),20 Mac-1,21 and p150,95.22
The present study sought to investigate whether monocyte adhesion to VSMCs induces monocyte PCA and whether this could be mediated by ICAM-1.
Isolation of Mononuclear Cells
Peripheral blood was drawn from healthy volunteers, and the mononuclear cell (MNC) fraction was prepared by the Ficoll-Hypaque gradient technique (Pharmacia).9 The recovered cells were washed 3 times in PBS. Neutrophil contamination was consistently <5%.
Monocyte Isolation
Human peripheral monocytes were isolated from freshly drawn blood of healthy volunteers by sequential Ficoll-Hypaque and Percoll (Nycoprep, Nycomed) gradient centrifugation. The purity of monocytes was >80%.
VSMC Culture
VSMCs, cell line AOSMC (Cellsystem, Clonetics), were cultured in smooth muscle cell basal medium (SMBM, Cellsystem) containing 10% FCS (Gibco BRL). At passage 5, cells were subcultured in 24-well plates in SMBM with 1% FCS, grown to confluence, and used for incubation experiments. In some experiments, VSMCs were stimulated with interleukin-1β (IL-1β) (R&D Systems) at 100 ng/L for 24 hours, washed 3 times, and then processed as indicated.
Chinese Hamster Ovary Cell Culture
ICAM-1–transfected and untransfected Chinese hamster ovary (CHO) cells, purchased from American Type Culture Collection, were maintained in RPMI 1640 medium with 1% glutamine, 1% penicillin-streptomycin (all from Sigma), and 10% FCS. Cells were subcultured in 24-well plates, grown to confluence, and used for incubation experiments.
Incubation of MNCs/Monocytes and VSMCs or CHO Cells
Suspensions of washed MNCs or purified monocytes (cell count, 109/L; >95% viable cells; Trypan blue exclusion) were added to confluent VSMCs or CHO cells and incubated at 37°C with 5% CO2 in RPMI 1640 tissue culture medium containing 1 mmol/L CaCl2 and polymyxine (Sigma) at 10 g/L. In some experiments, VSMCs or CHO cells had been preincubated for 1 hour with anti–ICAM-1 MAbs (clone 84H10; reacts with extracellular part and blocks adhesion; Immunotech) or blocking anti–VCAM-1 MAbs (clone BBJG-V1; blocks adhesion; R&D Systems; and clone 1G11; blocks adhesion and signaling through VLA-4; Immunotech). In other experiments, purified monocytes or MNCs had been preincubated with blocking anti-CD11a MAbs (clone 25.3; recognizes the α-L-chain of LFA-1 and blocks adhesion, T-cell cytotoxicity, and natural killer cell activity; Immunotech), anti-CD11b MAbs (clone 44; recognizes the α-M-chain of Mac-1 and blocks adhesion; Cymbus Biotechnology), anti-CD11c MAbs (clone 3.9; recognizes the α-X-chain of p150,95 and blocks adhesion; Cymbus Biotechnology) or anti-CD18 MAbs (clone 7E4; recognizes a group I epitope on β2 and blocks adhesion; Immunotech) at 10 mg/L. Some incubation experiments were performed in the presence of fibrinogen (Sigma) at 0.44 μmol/L. MNCs alone, nonstimulated or stimulated with 1 mg/L LPS, served as control cells. At times indicated, cells were washed twice with PBS (Gibco BRL) to eliminate nonadherent cells. The adherent monocytes were harvested after incubation for 2 minutes with cold 0.2% EDTA/PBS, pH 7.4, at 4°C (lymphocyte contamination <5%). As shown by light microscopy, the residual number of adherent cells was negligible. For Northern blot analysis, cells were sampled after 2 hours, and for ELISA and determination of PCA, we harvested cells after 6 hours. In some experiments, we measured PCA after incubation for 2, 4, 6, 12, and 24 hours.
Flow Cytometry
Immunofluorescence staining and flow cytometry were performed as described previously.23 In brief, VSMCs were harvested and resuspended in fresh medium, and 40 μL of cell suspension was incubated with saturating concentrations of FITC-conjugated anti–ICAM-1 MAbs (Immunotech) for 15 minutes at 4°C. Cell suspensions were washed 3 times and stored in 1% paraformaldehyde at 4°C until flow cytometric analysis was performed within 12 hours.
MAb binding was assessed by flow cytometry using FACS-Calibur (Becton-Dickinson) equipped with a 488-nm argon laser at 500 mW.
PCA Assay
PCA of intact cells was assayed by the 1-step recalcification clotting time as described.9 24 In brief, cell suspensions (0.1 mL, 105 cells) were added to 0.1 mL of citrate-anticoagulated pooled plasma or factor VII–deficient plasma (Behring Diagnostika) and 0.1 mL of 50-mmol/L CaCl2 at 37°C. The time required for production of a fibrin clot was measured with a fibrometer (KC 2, Amelung). Each sample was run in quadruplicate. Units of TF were calculated from the plot of log(clotting time) versus log(units TF activity) derived from dilutions of the TF standard Thromborel S (Behring Diagnostika) as described previously.9 24
RNA Preparation and Northern Blot Analysis
Total RNA of 107 cells was isolated by the single-step method of Chomczynski and Sacchi.25
Total RNA (5 μg) of each sample was subjected to electrophoresis on a 1.2% agarose gel that contained 100 mmol/L MOPS (Boehringer), 40 mmol/L sodium acetate (Boehringer), 5 mmol/L EDTA, and 6% formaldehyde (Boehringer). The RNA was transferred to nylon membrane (Hybond-N) in 20×SSC (Merck) by capillary blotting overnight. Blots were baked and prehybridized at 42°C in 50% formamide (Merck), 5×Denhardt's solution, 5×SSC, 0.5% SDS (Merck), and 20 mmol/L salmon sperm DNA (Gibco BRL). Blots were probed with the 1.2-kb SAL-I fragment of pHTF8 (ATCC) and reprobed with the 0.95-kb PST-I fragment of pUC18-GAPDH (gift from Dr Roger Becker, Toxikologisches Institut, University of Mainz, Germany) to ensure integrity of total RNA and comparable RNA loading in each lane.
The cDNA probes were radiolabeled by random priming with [α-32P]dCTP (>6000 Ci/mmol) (Amersham). The blots were washed at 60°C in 1% SDS/2×SSC and autoradiographed with Kodak X-OMAT film at −70°C with an intensifying screen.
For the detection of cellular TF, a sandwich-type ELISA with 2 monoclonal antibodies (American Diagnostica) was used as described previously.9 Washed cells (105 cells) were homogenized by Ultraturax (Jahnke & Kunkel) in buffer containing 20 mmol/L HEPES, 1% Triton X-100, 1 mmol/L dithiothreitol, 5 mmol/L benzamidine, 10% glycerin (Sigma), 2 mmol/L EDTA, 100 mmol/L NaCl, and 300 000 IU/L aprotinin (Bayer). Supernatants of the homogenates were used for ELISA.
Endotoxin Assay
To eliminate endotoxin contamination, all crystalloid solutions were ultrafiltered (U2000TM, Gambro), and stock solutions of protein were decontaminated by polymyxine columns (Pierce). We performed all incubation experiments in the presence of polymyxine at 10 g/L. All solutions and cell suspensions were checked before and at the end of each experiment by Limulus amoebocyte lysate assay (Schulz) as described previously.26 The detection limit of the assay was 10 ng/L.
Statistical Analysis
Results of the experimental studies are reported as mean±SEM unless otherwise indicated. Differences were analyzed by ANOVA followed by Scheffé's test or by paired t test, as appropriate. A value of P<0.05 in the 2-tailed test was regarded as significant.
ICAM-1 Expression and PCA in VSMCs
In nonstimulated VSMCs, we detected a baseline ICAM-1 expression of 4.3±1.5 mean fluorescence channel, which increased after stimulation with 100 pg/mL IL-1β to 11.3±2.7 (P=0.04). Baseline PCA in VSMCs was 0.8 mU/105 cells and did not increase after stimulation with IL-1β (data not shown).
Induction of PCA in Monocyte/VSMC Coculture
Suspensions of MNCs alone and VSMCs alone showed a baseline expression of PCA <1.0 mU/105 cells, which did not change over 24 hours. Stimulation of MNCs with LPS (1 μg/mL) resulted in a PCA of 1053.4±123.7 mU/105 cells. Coculture of MNCs and VSMCs enhanced the monocyte PCA in a time-dependent manner: After 6 hours, we observed a peak PCA of 166.0±37.9 mU/105 cells (P=0.007 compared with control cells), which returned to baseline within 24 hours (Figures 1⇓ and 2⇓). Similar results were obtained for purified monocytes: Baseline PCA of purified monocytes was 1.9±0.3 mU/105 cells, and it increased after coincubation of 6 hours to 170.6±46.4 mU/105 cells (P=0.008) (Figure 2⇓). The PCA of VSMCs after 6 hours of incubation with MNCs was <1.0 mU/105 cells (Figure 2⇓). Incubation of MNCs with VSMC supernatant did not enhance TF activity of MNCs (data not shown).
Compared with incubation with unstimulated VSMCs, incubation of MNCs with IL-1β–prestimulated VSMCs led to a further increase in PCA to 239.9±27.9 mU/105 cells (P=0.02). The use of factor VII–deficient plasma or the addition of anti-TF MAbs at 40 mg/L abolished monocyte PCA after incubation with VSMCs, confirming that the observed effects were dependent on monocyte TF surface expression (Figure 2⇑).
Addition of blocking anti–ICAM-1 antibodies to coincubation inhibited monocyte PCA in a dose-dependent manner, whereas addition of blocking anti–VCAM-1 antibodies had no effect (Figure 3⇓).
TF mRNA and Protein Expression After MNC/VSMC Coculture
In pure, nonstimulated MNCs, no TF transcripts were detectable, but exposure to LPS was associated with marked induction of TF mRNA expression. After 2 hours of coculture with VSMCs, Northern blot analysis showed a substantial increase of TF mRNA in MNCs (Figure 4a⇓).
Nonstimulated MNCs incubated alone contained 17.2±2.8 pg of TF protein/105 cells, as shown in Figure 4b⇑. Stimulation with LPS produced a marked increase in TF protein, and culture with VSMCs increased cellular TF protein of MNCs significantly, to 29.9±3.9 pg/105 cells (P=0.01). VSMCs incubated alone contained only small amounts of TF protein (5.5±0.3 pg/105 cells) (Figure 4b⇑).
Induction of PCA in Monocytes by Coculture With ICAM-1–Transfected CHO Cells
Purified monocytes incubated with untransfected CHO cells showed little PCA (1.2±0.2 mU/105 cells), and coculture with ICAM-1–transfected CHO cells enhanced monocyte PCA significantly, to 81.9±3.3 mU/105 cells (P<0.001) (Figure 5⇓). Addition of blocking anti-CD18, anti-CD11b, or anti-CD11c MAbs almost completely abolished the increase in PCA after coincubation with ICAM-1–transfected CHO cells, whereas addition of anti-CD11a MAbs had no such effect (Figure 5⇓). The increase in PCA after coculture of ICAM-1–transfected CHO cells with MNCs was similar to that obtained after coculture with purified monocytes (from 1.0±0.1 to 75.5±1.4 mU/105 cells, P<0.001) and was also inhibited by anti-CD18 MAbs (Figure 5⇓). The presence of fibrinogen in incubation experiments of MNCs and ICAM-1–transfected CHO cells had no influence on the enhancement of monocyte PCA (Figure 5⇓). Addition of fibrinogen to suspensions of MNCs incubated alone had no effect on monocyte PCA (data not shown).
In the present study, we examined the effect of monocyte adhesion to VSMCs on PCA of peripheral blood monocytes and the potential role of cellular adhesion molecules for PCA induction. After coculture with VSMCs, we found an enhancement of monocyte PCA, which is regulated by β2-integrin–mediated monocyte adhesion to ICAM-1 on VSMCs.
After incubation of purified monocytes with VSMCs or ICAM-1–transfected CHO cells, the PCA of the adherent monocytes was significantly increased. Similar results were obtained after incubation of MNCs with VSMCs or ICAM-1–transfected CHO cells. Differential cell count of the adherent cells in experiments with MNCs revealed monocytes as the predominant cell type and a lymphocyte contamination in this population <5%. Hence, cell distribution of the adherent cells was comparable in the two experiments, with purified monocytes and with MNCs. Thus, to minimize the effect of separation-induced monocyte stimulation, most of the experiments were performed with MNCs. Our findings suggest that PCA is enhanced by adhesion of monocytes. Nevertheless, we were concerned about the possibility that lymphocyte-derived mediators could contribute to the observed monocyte response. However, because incubation of purified monocytes or MNCs with VSMCs or ICAM-1–transfected CHO cells increased monocyte PCA to almost the same degree, a major contribution of lymphocytes to PCA enhancement can be excluded.
The increase in monocyte PCA has to be attributed to TF activity, because the observed effects were absent in factor VII–deficient plasma and could be inhibited by the addition of blocking anti-TF MAbs. In support, we found an increase in TF mRNA after incubation for 2 hours and an increase of TF protein in cellular homogenates after 6 hours of incubation.
Induction of monocyte PCA occurred in a time-dependent manner, with a peak after 6 hours and a normalization to baseline after 24 hours. This time course of TF generation by adhesion to VSMCs is different from the E-selectin–mediated TF generation after monocyte adhesion to endothelial cells16 : Monocyte adhesion to endothelial cells leads to a rapid induction of functional TF activity after only 30 minutes.16 In our experiments, we observed an increase in TF mRNA after 2 hours of coculture, suggesting a transcriptional induction of TF expression. Thus, in contrast to the adhesion to endothelial cells, our data suggest a different regulatory mechanism that involves de novo protein synthesis rather than alteration of the functional expression of preformed TF.16
Consistent with previous studies, we showed by flow cytometry analysis that stimulation of VSMCs with IL-1β increases surface ICAM-1 expression.27 Incubation of MNCs with prestimulated VSMCs led to a further increase of monocyte PCA compared with incubation with unstimulated VSMCs. To elucidate the potential role of cellular adhesion molecules for PCA induction, we performed incubation experiments with blocking anti–ICAM-1 MAbs and blocking anti–VCAM-1 MAbs. We found that the induction of PCA could be inhibited by anti–ICAM-1 MAbs in a dose-dependent manner, whereas we could not find an inhibitory effect of anti–VCAM-1 MAbs in our model. Hence, ICAM-1 seems to have an important role in the observed effects.
To investigate whether adhesion of monocytes to ICAM-1 is sufficient to induce monocyte PCA, we performed further incubation experiments with ICAM-1–transfected CHO cells. Under these conditions, we also found a marked increase in monocyte PCA after 6 hours of coculture, although less pronounced than that observed after incubation with VSMCs. Therefore, we concluded that adhesion to ICAM-1 is an appropriate stimulus for the enhancement of monocyte PCA. ICAM-1 interacts with leukocytes by binding to the surface-expressed β2-integrins LFA-1 (CD11a/CD18),20 Mac-1 (CD11b/CD18),21 and p150,95 (CD11c/CD18).22 Addition of blocking antibodies against the β-chain (CD18) of these β2-integrins or against the α-chain of Mac-1 (CD11b) or p150,95 (CD11c), which share 63% homology in their amino acid sequence,28 inhibited the induction of monocyte PCA after adhesion to ICAM-1–transfected CHO cells completely, whereas blocking of anti-CD11a MAbs had no such effect. These data indicate that the β2-integrins Mac-1 (CD11b/CD18) and p150,95 (CD11c/CD18) play an important role in signaling TF expression after monocyte adhesion.
Fan et al showed by cross-linking experiments that priming of the cells by other agonists was needed to initiate TF transcription by β2-integrin signaling.29 In our system, monocyte adhesion to other VSMC surface adhesion molecules or contribution of VSMC-derived mediators could represent such a mild stimulus for TF induction. In experiments with CHO cells, monocyte adhesion to untransfected CHO cells itself increased PCA slightly, but adhesion to ICAM-1–transfected CHO cells enhanced monocyte PCA substantially. Adhesion of monocytes to VSMCs or CHO cells in itself may therefore act as a mild priming stimulus, but the marked increase in TF expression is mediated by cellular interaction via the β2-integrins Mac-1 or p150,95 and ICAM-1.
In addition to β2-integrins, ICAM-1 can bind fibrinogen.30 Fibrinogen is also a ligand for Mac-1 on monocytes and has been shown to serve as a bridging molecule between monocytes and endothelial cells.30 Fibrinogen may interfere with PCA induction through monocyte adhesion to VSMCs under physiological conditions, if it blocks the ICAM-1 moiety involved in signaling TF expression. Addition of fibrinogen to cocultures of MNCs and ICAM-1–transfected CHO cells, however, had no effect on the enhancement of monocyte PCA, and fibrinogen added to pure MNC suspension could not induce PCA. Thus, binding of fibrinogen to ICAM-1 does not appear to affect the ICAM-1 domain that induces TF expression in monocytes. Our experiments indicate that the induction of monocyte PCA by binding of VSMCs is independent of fibrinogen and thus can occur under physiological conditions.
Others have shown that appropriate engagement of the β1-integrin VLA-4 initiates TF biosynthesis.31,32 Nevertheless, MAbs against VCAM-1, the ligand for VLA-4 on VSMCs, did not suppress the PCA response after coincubation of monocytes and VSMCs in our experiments. Despite the use of 2 different MAbs known to block VCAM-1–supported adhesion and VLA-4 signaling, this finding does not fully exclude a role for VCAM-1–mediated interaction in our model, because the MAbs used may not block the bioactive site for the PCA response.
We were concerned about LPS contamination in our experiments and checked all suspension media by the Limulus amoebocyte lysate assay, thus ensuring that LPS contamination was <10 ng/L. Furthermore, we performed all incubation experiments in the presence of polymyxine at 10 g/L. Nevertheless, even traces of LPS as low as 10 pg/L have been shown to induce PCA in monocytes.12 In our study, PCA in MNCs incubated alone was <1.0 mU/105 cells, which precludes substantial LPS contamination. Moreover, the effect of PCA induction by adhesion to ICAM-1–transfected CHO cells could be inhibited completely by blocking antibodies against β2-integrins or ICAM-1, which confirms a specific effect of cell adhesion.
The finding that monocyte adhesion to VSMCs induces TF expression may be of major pathophysiological relevance. Recently, it was shown that plaque erosion is a frequent finding in sudden death due to coronary thrombosis, found in up to one third of the cases.3 Histological analysis of these lesions revealed smooth muscle cells and adherent monocytes/macrophages and lymphocytes as the major cell types.4 Furthermore, Moreno et al,5 in a recent study, showed an increased TF content in coronary artery sections of patients with unstable angina and a correlation with areas of macrophages and smooth muscle cells, suggesting a cell-mediated thrombogenicity in patients with acute coronary syndromes. The underlying mechanism for these findings remained unclear. We now present data that adhesion of monocytes to VSMCs, as can occur after endothelial denudation, induces monocyte PCA. Tethering of monocyte β2-integrins to ICAM-1 on VSMCs is a major mechanism of this effect. ICAM-1 expression in VSMCs is regulated by proinflammatory cytokines like IL-1β and TNF-α, increases monocyte adhesion, and in concert with VCAM-1, enhances the invasion of monocytes into the arterial intima.33 Our study showed that enhancement of monocyte PCA is another effect of β2-integrin/ICAM-1 binding. In addition, our findings suggest that this mechanism could play a major role in monocyte/VSMC interaction occurring in atherosclerotic plaques. VSMC-induced monocyte PCA may thus contribute substantially to the thrombogenicity of eroded plaques. Our findings could assist in the design of therapeutic strategies to reduce the thrombogenic potential of eroded plaques.
This study was supported by a grant from the Deutsche Forschungsgemeinschaft (Ne 540/1–2), Bonn–Bad Godesberg, Germany. We gratefully thank Tanja Breustedt and Ulrike Pfeifer for excellent technical assistance.
- Received January 22, 1998.
- Revision received March 20, 1998.
- Accepted March 26, 1998.
- Copyright © 1998 by American Heart Association
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- Enhancement of Monocyte Procoagulant Activity by Adhesion on Vascular Smooth Muscle Cells and Intercellular Adhesion Molecule-1–Transfected Chinese Hamster Ovary CellsNikolaus Marx, Franz-Josef Neumann, Dietlind Zohlnhöfer, Timm Dickfeld, Almuth Fischer, Susanne Heimerl and Albert SchömigCirculation. 1998;98:906-911, originally published September 1, 1998https://doi.org/10.1161/01.CIR.98.9.906
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- Enhancement of Monocyte Procoagulant Activity by Adhesion on Vascular Smooth Muscle Cells and Intercellular Adhesion Molecule-1–Transfected Chinese Hamster Ovary CellsNikolaus Marx, Franz-Josef Neumann, Dietlind Zohlnhöfer, Timm Dickfeld, Almuth Fischer, Susanne Heimerl and Albert SchömigCirculation. 1998;98:906-911, originally published September 1, 1998https://doi.org/10.1161/01.CIR.98.9.906


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