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Nestled right on the border of St. Ann and Trelawny on Jamaica's North Coast, lies the scenic tiny seaside village of Rio Bueno. Running alongside this village is the Rio Bueno River. This river winds its way down, through various lush green plantations and tropical rainforests, to the Rio Bueno Harbor, which is the deepest… Read More
Dunn's River Falls in Ocho Rios is one of Jamaica's most popular attractions. It is definitely one of the most beautiful places you'll ever see. When you arrive, you'll be greeted by Dunn's River Fall's resident Donkey. His name is Bunny. Bunny is quite possibly the most famous donkey in the world. People from just… Read More
The Montego Bay Marine Park is an area of protected sea water on Jamaica's north coast. It is managed by the Montego Bay Marine Park Trust, which was set up to preserve the natural environment in the area and to educate the surrounding community of the importance of the marine resource. The Park includes the… Read More
If you're looking to have a peaceful day without leaving Kingston, Hope Botanical Gardens and Zoo is the place for you. The Hope Botanical Gardens, also known as the Royal Botanical Gardens, hail back to Jamaica's colonial past. In 1873, it was established by Major Richard Hope, who was a commander in the British Army… Read More
The Blue Mountain Peak is the highest point in Jamaica. It's 7,402ft high and famous for producing some of the world's best Coffee. Hiking the Blue Mountain, however, is less known as one of the premier activities on the Island. The peak is a natural paradise prized by nature lovers because of the wonderful scenery and… Read More
I went to Konoko Falls in Ocho Rios to celebrate my daughters 7th birthday. Two weeks before, I had never even heard of this place, which is surprising since I spent much of my childhood exploring the parish of St. Ann. In a conversation with a friend, they mentioned that a local business person had… Read More
There's more to Jamaica than excellent food, top-notch beaches, and vibrant nightlife. The island's lush mountainous interior and ample rainfall create the essential ingredients for our magnificent waterfalls. Most people know about the popular ones like Dunn's River Falls and Mayfield Falls. But there are others that are not as popular, but just as stunning… Read More
The Blue Hole in Ocho Rios Jamaica has become one of my favourite places. It's one of my more recent discoveries. I only found out about it because I've been spending lots of time in the countryside, trying to find those hidden gems in my country. Even my local Jamaican friends didn't know this place… Read More
This place is MAGICAL!!! Glistening Waters is a glowing lagoon on the north coast of Jamaica. This marvel is only found in 4 places in the world, however, this lagoon is the brightest of them all due to more consistent climate. Here, the fresh water from the Martha Brae River meets the salt-water ocean and… Read More
I finally made it to the Green Grotto Caves in Discovery Bay, Jamaica. I live about 5 minutes from this place. But for whatever reason, it took me all these years to finally make it inside. I had some family visiting from the UK so I figured we could all go and check it out… Read More
I recently got the opportunity to visit this firewater mineral healing spring in St. Ann's Bay. I've been hearing about it for some time now but it's not a popular attraction so it's difficult to find any information on it. I happened to be driving with a friend that knew where it was, so I… Read More
Mayfield Falls is an eco-tourism attraction centrally located in the parish of Westmoreland, Jamaica. It consists of 2 beautiful waterfalls, 21 natural pools, 52 types of ferns and lots of exotic flowers, plant species, birds, butterflies and wildlife native to Jamaica. Most parts of Mayfield Falls are pretty calm. You won't find some of the… Read More
Blue Lagoon is located in Port Antonio and is a wildly popular travel destination for families and honeymooners. With glistening, turquoise blue waters surrounded by lush greenery, Blue Lagoon is a vision of beauty. When you first arrive, you may notice that the lagoon looks familiar to you. If you've seen the Brooke Shields' movie… Read More
The Blue Mountains form the longest mountain range along the eastern edge of Jamaica and features majestic scenery. Many people hike or bike through the mountains to witness first-hand nature at its finest. This under-explored area is also the source of the world famous and fragrant Blue Mountain Coffee. The ride up to the Blue… Read More
YS Falls is located on the South end of Jamaica in a parish called St. Elizabeth. It's on a working cattle ranch and thoroughbred horse farm. Like much of Jamaica, the falls are surrounded by lush gardens and beautiful trees. Many people say YS Falls is one of the most beautiful sights they have ever… Read More
Reach Falls Jamaica is one of the most amazing natural waterfalls that the island has to offer. It is peacefully situated amongst exotic virgin rain forest. Located on the eastern end of Jamaica in Portland, it's approximately an hour and a half drive from Ocho Rios, Jamaica. This is truly beautiful setting. The water is… Read More


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Pablo Diego José Francisco de Paula Juan Nepomuceno María de los Remedios Cipriano de la Santísima Trinidad Ruiz y Picasso (1881-1973), best known as Pablo Picasso, was a Spanish-born painter and sculptor who lived most of his life in Paris and elsewhere in France. His accomplishments, flamboyant lifestyle, and genius for self-promotion combined to make him one of the best, most revolutionary, and certainly best-known of twentieth century artists.
"Cubism" was an artistic style pioneered by Picasso and Georges Braque, principally during the period 1907-1911, though copied and extended by others for many years thereafter. The painting here, dating from 1910 and titled "The Guitarist," is a prime example of Picasso's "analytical cubism."
Though somewhat influential in literature and music as well (and despite Picasso's brief membership in the French Communist Party in the mid 1940s), cubism and Picasso were judged too radical and unrealistic to be acceptable under Soviet socialist regimes, and therefore were labeled "decadent bourgeois art."
Thus, the narrator of Unbearable Lightness remarks, the approved regime of socialist realism ruled out Picasso and cubism as fit subjects for Sabina to study at the Academy of Fine Arts.

subset: commoncrawl


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SIP Trunking is the use of VoIP to facilitate a connection of a PBX telephone system to an IP telephony platform.
With Data Sense UK we can assure business continuity through our SIP trunking systems with centrally controlling the automatic call diverts. Eliminate cost of calls between sites to reduce call and line rental charges and operate your business without the limitations of a geographical number and have the opportunity work remotely where there is a data connection.
Through the internet ensure you have the essential disaster recovery routing for minimal disruption to your business.


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For time:
100 Overhead squats, 75#(55#)
*Each time the bar is put down, athlete must perform 15 bar facing burpees before picking it back up
Post times to comments and BTWB
Hold on to your butts folks. . . we got a lot of stuff on the calendar this summer.
*Verve will have one 7am WOD Saturday and Sunday this weekend. Verve will be closed the rest of the day for a seminar.
*Verve will have two off site swim WODs Saturday morning at 9:30am and 10:30am @ Glenarm Rec Center. Please sign up on MBO.
Glenarm Rec Center
2800 Glenarm Pl
Denver, CO 80205
Click here for a map
*CrossFit Cherry Creek is hosting the Triple Threat competition this weekend. Verve has 4 teams competing on Sunday only. Stop by and do some cheering!! Click here for directions and information.
*Verve will have it's first event of it's Summer Nutrition Series Saturday June 27th at 2pm @ Verve. It will be a lecture on the topic of your choice presented by Clara Wisner. Be sure to Vote for the topic you want to hear about. The voting box is at the front of Verve with your topic choices. The lecture is open to everyone at no cost.
*Verve will be hosting an i99 clinic Sunday June 28th at 12pm. More details will be posted next week along with a link to register.
*Verve is hosting the CrossFit Football Trainer Course Saturday and Sunday July 11th-12th. Spots are still available. For more information and to register, click here.
*The O2X Summit Challenge will be August 15th at Winter Park Resort. Verve trainers are signing up a team and looking to make some wagers against any other Verve teams that sign up!! For more info, click here.
*Front Range CrossFit is hosting the first annual Do More Charity Challenge October 3rd-4th. It's a 6 person team competition. Verve will put $600 towards registering a team. Click here for more info and rules.


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Article Side
Online Article Directory!
Tips for Stress Free Life
The process of underlying stressful life is extremely complex. To be aware how the mutual relations and the environment are interacting with the stressed life or how are they affecting a normal life of a person is to be laid emphasis today. The study suggests that the effects of stressful life events on the risk of depression are the essential points to be aware of. Genetic dissection of diseases such as cancer, asthma and heart disease is galloping ahead whereas development in other diseases like stress and depression is comparatively much slower. To be truth, evaluating the social, physical, mental and emotional impacts of the disease, in 1990 depression stood at the number 4th to be considered as the leading cause of global burden of illness and was predicted to be the number one in 2020.
So today stress has become the serious concern to be aware of as there is the rapid growth of the depressed and stressed patients we need to cast the close attention on this condition. Consequently, continue stress can lead you to be overloaded by the numbers of mental and physical diseases. Therefore before the cruel and evil shadow of stress come and clutch us we should tend to overlook the essential and most prominent remedies for it so that we may stay away from it. There are the following ways to have a stress free and fun life.
Of course, our healthy body for the stressed free life is as much concerned as an unhealthy for the death. So to have a depressed free life it is equally necessary to maintain our health. Eat timely do exercise regularly to keep you feel good and happy. You should be the one sided watcher that is don't look at your life on the negative point of view but be positive. Avoid your negative result and only stick to the positive one. Try to lessen your shoulder responsibilities by laboriously accomplishing them. Don't put off any work for tomorrow otherwise very soon you will be under the pile of heavy works. Also set specific time for your sleep. Getting sleep enough will bring a disastrous moment for you. Manage to have sound sleep for 6 to 8 hours daily. Fix time to retire to bed and wake up. Sometimes your being round the clock busy becomes the cause of your stressful life so spend sometimes for entertainment like watch movie, go for a trip, enjoy with your friends etc.
There are some other techniques to reduce your stressful life – according to the doctors; meditation keeps the person refreshed and healthy. It shortens your problems. It makes you more active and alert. It helps reduce your anxiety, tension, pain, frustration and tiresome. Like meditation exercise has also the significant role in lowering your stress. Regular exercise reduces the production stress hormones from the body such as ortisol, and neurotransmitters that are secreted during the stress response.
Ajay - About Author:
article has been written and posted by a health advisor working at bookmydoctor.com, who also provides free of cost consultancy to patients and advise on various topics like How To Reduce Stress By visiting the site, you can read articles on Reducing Stress and How To Reduce The Stress and For more information click the Hyperlink.
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catalog: DOSE128
artist: Wyrding
title: s/t
format: LP/CD
release date: March 1, 2016
There is something amiss in Wisconsin. Or perhaps the opposite forces are at work. More likely, it's a unique push and pull between the dark and light from which Wyrding pulls their inspiration. Founded by Troy Schafer and Bret Hartl, Wyrding began with a sound that took inspiration from the unique folk/neofolk leanings of Troy's long-standing project Kinit Her (With Nathaniel Ritter) and melded into a unique take on funeral doom metal. These initial experiments resulted in the two-track Agony in being EP. (Originally released in very limited quantities on Schafer's Shifting Sands Congregation imprint, and now available on cassette on Small Doses, and as bonus tracks on the CD of this debut album.)
As time passed, Wyrding evolved into a full, five-piece band, and their sound shifted with it. The music became more expansive and epic, and the vocals moved from the whispered growl on Agony in Being to a more crooned, almost operatic style. The results are still very much rooted in funeral doom, and the music is incredibly heavy without being heavy in the traditional metal sense – it relies on emotional mass and space more so than the typical huge distorted riffs.
The seven songs that make up Wyrding's debut album create the feel of a strange narrative – the plot isn't always easy to discern, but by the end, you know the band brought you along on a trip riddled with sadness, darkness, and perhaps even a bit of catharsis.
"Wyrding's hushed music flows over you like a warm blanket, Schafer's warm baritone voice soaring above fellow bandmates Brian Steele, Kyle Roessler, Bret Hartl and Jerry McDougal's smooth lugubre. Gone are the overt folk influences of Wyrding's demo, who have emerged from the cocoon with a sound akin to Benedictine monks performing funeral rites with modern electric instrumentation. Unlike their more conservative funeral doom brethren, Wyrding seeks out the dramatic not only in minimal execution, but minimal sound. They are quiet, unassuming, but are able to portray immensity with their subtle movements." – Jon Rosenthal, Invisible Oranges
"Wyrding is essential listening, not least because it serves as a forward and upward-looking counterpoint to the neo-traditional efforts of bands like Pallbearer and Bell Witch." – Evening of Light
The LP version of Wyrding's deput album is available on red vinyl with black swirl (limited to 100 copies) and standard black vinyl. Both versions are packaged in a matte sleeve with spot varnish with an insert. The CD version is packaged in a six-panel sleeve, and includes the two tracks from the Agony in Being EP.


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Orcel to switch from BofA Merrill to client VTBRiccardo Orcel, head of CEEMEA investment banking at Bank of America Merrill Lynch, is understood to be discussing a move to VTB, the Russian bank which is an important client of BofA Merrill. Because of the close ties between the ...
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IBM's 2013 Global Location Trends report ranks Denmark 2nd in the world measured by value and quality of inward investments. The top ranking confirms Denmark's position as an attractive destination for knowledge-intensive investments.
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International rankings show
No. 1 in the world on corporate values (IMD 2011)
No. 1 in the world on worker motivation (IMD 2011)
No. 2 in the world on quality of life (OECD factbook 2010)
No. 2 in the world on addressing Health, safety & environmental concerns (IMD 2010)
Top league on quality of life (IMD 2011)
Read more about living and working in Denmark
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Delonte Michael mccray
- 18 years old
- Date of birth: Jan 31, 1994
- Date of passing: Mar 3, 2012
|Let the memory of Delonte be with us forever
This memorial website was created in memory of our loved one, Delonte Mccray, 18, born on January 31, 1994 and passed away on March 3, 2012. We will remember him forever.
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1 /*2 * The contents of this file are subject to the terms of the Common Development3 * and Distribution License (the License). You may not use this file except in4 * compliance with the License.5 *6 * You can obtain a copy of the License at http://www.netbeans.org/cddl.html7 * or http://www.netbeans.org/cddl.txt.8 *9 * When distributing Covered Code, include this CDDL Header Notice in each file10 * and include the License file at http://www.netbeans.org/cddl.txt.11 * If applicable, add the following below the CDDL Header, with the fields12 * enclosed by brackets [] replaced by your own identifying information:13 * "Portions Copyrighted [year] [name of copyright owner]"14 *15 * The Original Software is NetBeans. The Initial Developer of the Original16 * Software is Sun Microsystems, Inc. Portions Copyright 2003 Sun17 * Microsystems, Inc. All Rights Reserved.18 */19 20 package org.netbeans.modules.openfile;21 22 import org.openide.filesystems.FileObject;23 24 /**25 * Interface for Open File implementations.26 *27 * @author Marian Petras28 */29 public interface OpenFileImpl {30 31 /**32 * Opens the specified FileObject.33 *34 * @param fileObject file to open35 * @param line line number to try to open to (starting at zero),36 * or -1 to ignore37 * @return true on success, false on failure38 */39 boolean open(FileObject fileObject, int line);40 41 }42
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Lest you think I am going soft, let me say right up front that I believe in honesty in relationships. It is one of the character qualities that those who seek husbands and wives most value. It is the foundation that trust is built upon.
Where I am taking exception is with the word "brutally". I know that people throw this word around to get across the idea of "the most honest version" of a story. I'm all over that. But there are people who are brutal in their honesty. And when they are they miss an important character quality: kindness. Rather than consider another person's feelings, they blurt out "truth" and wound them, often unnecessarily. This sounds more than a little bit selfish to me.
Sometimes the truth hurts, and the pain caused is unavoidable. Empathy helps to cushion the blow, and kind people think out their approach and try to be honest, but gentle. However, there are others who take grim pleasure in hurtful truth telling. They may think they are only being honest, but they are actually just acting self-righteous. This truth without grace is simply meanness – not a good character quality. I think of this well-known passage from the Bible that talks about loving others as the highest value. I suppose you could say that it is lovingly honest.
1 Cor 13 If I could speak all the languages of earth and of angels, but didn't love others, I would only be a noisy gong or a clanging cymbal. If I had the gift of prophecy, and if I understood all of God's secret plans and possessed all knowledge, and if I had such faith that I could move mountains, but didn't love others, I would be nothing. If I gave everything I have to the poor and even sacrificed my body, I could boast about it; but if I didn't love others, I would have gained nothing.
When searching for a mate, character (as in being honest, etc.) matters. Character decides what happens during crises, conflicts, temptations and trouble. Too often, certain character flaws are overlooked or minimized because our heart (feelings) overpowers our head (reality). One or two good character qualities do not make a person of good character. You have to look at the whole package. Of course, you have to be a person of good character to expect to attract a person with good values.
If you have been reading our blog for any length of time, you know that we bang on this theme of values and character a lot. Why? Many, if not most counseling issues stem from the breakdown of good values and character (The Bible calls this sin). When we improve character, we improve relationships, families and communities. Yes, it's a big deal.
I would encourage you to take an inventory of your character qualities (good as well as needs-to-improve). Are you kind, patient, honest, generous, God-loving, trustworthy, loyal, sober, diligent, responsible, forgiving, helpful, even-tempered, etc? Does this list describe the kind of person you would want to do life with?


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1. One of the biggest reasons to like Oklahoma this season is that the Sooners ranked seventh with 102 starts on the offensive line. Last week, the Sooners lost senior center Ben Habern (30 starts), who quit because of a chronic neck injury. Now guard Tyler Evans (29) has injured a knee and is expected to be out at least a few weeks. That would leave Oklahoma with 43 returning starts on the O-line, which would be ranked tied for eighth -- in the Big 12. Still think the Sooners are a top-five team?
2. Louisville coach Charlie Strong is concerned that his young players will hear they have been picked to win the Big East and think they earned something. "We are a young football team," Strong said. "We still have a lot of growing up to do." The Cardinals have nine seniors. Maybe four will start. "My major concern is the leadership, where guys don't get full of themselves." Last year, Louisville won at league champ West Virginia, and lost at home to Pittsburgh (6-7) the following week.
3. As of Monday evening, four Penn State trustees have joined to appeal the crippling NCAA sanctions levied against the university last month. That's four trustees out of 32. As a measure of solidarity or the lack thereof, it will be interesting to see how the other 28 trustees react. The attorney pursuing the appeal, Paul V. Kelly, knows his way around college athletics. He ran College Hockey, the promotional and marketing arm of the sport, until earlier this year.


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I use Olay regenerist as a night cream. I find it very moisturizing, and especially helpful around my eyes (I'm in my 30's)
A little goes a long way, one container lasts me approximately 2 months. On occasion I break out if I use it too generously in my problem areas (chin!)
Fantastic product, part of my daily routine
I have used this cream for many years and feel that it is keeping my skin younger looking. I use it twice a day every day on clean skin. It smells nice also.
I turned 40 and suddenly my skin is starting to sag/wrinkle (boo). Is that me in the mirror? I was looking tired and for years I've had mild adult acne....UNTIL NOW! I tried Olay Regenerist Sculpting Moisturizer and noticed a slight difference on day 1! 2 weeks later my skin feels plump, my pores are smaller and my breakouts are GONE. Yep...gone. I was always scared to try wrinkle creams before as my sensitive acne prone skin seems to explode with pimples when I use a new cream.
Not this one.
I can't wait to try more Olay Regenerist products.
Honestly, this is the worst product I've ever tried. I had a horrible allergic reaction, my face swelled up and I got chemical burns from it. I tried to get ahold of the company and ended up getting a "too bad" type message back. For anyone with any type of sensitive skin, I suggest you do not try this as it's just not worth the hospital trip. I had to end up taking antibiotics and put a skin cream on my face in order to get rid of it. It was horrifying.
I love this cream especially in winters. Great as everyday moisturizer, not oily, absorbs in skin faster and very light for combination skin. It does make my face feel refreshed and revitalized. Great as an anti aging moisturizer
I have somewhat sensitive skin and this moisturizer didn't effect my skin at all. Felt like silk going on and absorbed fully, no sticky residue left on skin. And it smells so good! I didn't see remarkable difference but I didn't use it for that long.
This anti aging micro sculpting cream smelt nice and when putting it on my face it felt thick. The formula felt like it was too thick and kind of oily. After about 5 minutes it actually doesn't feel as thick but it still feels like I have something on my skin. It would be nice if it soaked in better. Not for me #olay
These reviews are the subjective opinions of ChickAdvisor members and not of ChickAdvisor Inc.


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HELEN AND KYLE COMING BACK TO MARYLAND-OCTOBER 1st!
On October 1st, the MSWA presents its 4th annual Coaches Clinic featuring Olympic Gold Medalists from Maryland, Helen Maroulis and Kyle Snyder at 4pm (HS kids and under free to attend the 4pm session but must register).
Clinic will also feature Virginia Tech Head Coach Kevin Dresser and Brown University Head Coach and Maryland's own Todd Beckerman; plus Brad Howell, Rohan Murphy, Ronnie Wuest, and Gene Zannetti!
Coaches who attend the clinic will receive free gifts from vendor sponsors including Cliff Keen, WIN Magazine, MyHouse Apparel and MORE!
An awards banquet will conclude the clinic and will honor the Maryland All Americans, Triple Crown Winners, Eareckson Award Recipient Alan Meltzer, plus Helen and Kyle and the 4 Olympic wrestlers in Maryland Kerry Mccoy, Rob Eiter, Lloyd Keaser, and Ernie Fischer.
1972 Ford Mustang Sprint Edition Classic Muscle Car will be on hand for photo ops thanks to Rusty Smith
More info HERE


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Lucknow, Aug. 26: Mayavati today announced her decision to take her battle against the BJP beyond Uttar Pradesh.
Determined to hurt the BJPs poll prospects, the Bahujan Samaj Party leader said: My top priority is to finish off the BJP in the four states of Chhattisgarh, Madhya Pradesh, Delhi and Rajasthan where Assembly elections are due soon.
Mayavati was speaking at a hurriedly convened press conference after handing the resignation of her council of ministers to the governor. The pattern of her partys alliances and break-ups with partners shows that the BSP has gone on to add to its vote bank after every split.
I told the governor that I did not wish to head the caretaker government as I wanted to be free to campaign in the coming elections, she said.
This is bad news for the BJP, which had earlier hoped for the BSPs indirect support in the four states where it has major stakes. A senior party leader said: She had helped us by asking her Dalit supporters to vote for the BJP in Gujarat where the BSP had not fielded any candidate. We hoped she would do it again.
Sources close to Mayavati said some important Congress leaders, including chief ministers of two party-ruled states, had already approached her.
Mayavati believes the BJP central leadership let her down; so she is determined to pay them back.
My decision to part company with the BJP was not sudden and I have explained my stand in the 30-page chargesheet that I have sent to Prime Minister Vajpayee, she said.
A party leader said: The BSP has established significant presence in Madhya Pradesh and Chhattisgarh and is in a position to spoil the chances of any party in Delhi and Rajasthan.
Mayavati, clarifying that her split with the BJP was final and irrevocable, hinted that she was ready to do business with anti-BJP forces. I have already said at yesterdays rally that we can consider a tie-up with the Congress if the party is ready to give up its manuwadi practices.


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SAN BERNARDINO >> A 55-year-old San Bernardino man is behind bars accused of sexually abusing a girl under the age of 14, according to sheriff's officials.
Felipe Castro Flores was arrested Monday afternoon on suspicion of child sex abuse.
San Bernardino County sheriff's deputies were called to the 1000 block of Pacific Street, where the young victim told investigators she had been inappropriately touched by Flores, according to a sheriff's statement.
After interviewing Flores, authorities arrested the 55-year-old at the sheriff's headquarters, and he was booked into the Central Detention Center in San Bernardino in lieu of $250,000 bail.
The relationship between the victim and Flores was not released to protect her identity and privacy, officials said.
Anyone with information regarding this investigation can call the Sheriff's Detective Division at 909-387-3545. Callers wishing to remain anonymous can call the We-tip Hotline at 1-800-78-CRIME (27463) or leave information on the We-Tip Hotline at www.wetip.com.


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Carl Henry Louis Bocchini
This memorial website was created in the memory of our loved one, Carl Henry Louis Bocchini. We will remember him forever.
Memorial Tributes
Carl was a good friend me and many in our International Students, Inc. UC Santa Cruz program, a friendly greeter, Friendship Partner to grad students, idea-man, sponsor, kind host and friend to many, always eager to share Christ's love with everyone through humor and service. I could count on Carl to help me out in many a pinch and to always come up with a wisecrack to make us laugh-- a real man of God.
You are missed dear brother Carl!
I am so sorry to hear of the loss of Carl! The Bairds
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By: Dusty Langdon
"International Women's Day is a time to reflect on progress made, to call for change and to celebrate acts of courage and determination by ordinary women who have played an extraordinary role in the history of their countries and communities."
The official website for the United Nations states above the meaning and reasons behind International Women's Day, which takes place every year on March 8.
Because there are too many extraordinary women in history who have made a mark on our societies, this article will focus on only a few. However, there are so many, many more women deserving of recognition that you can check out on March 8, and throughout the whole year.
International Women's Day is all about the achievements of extraordinary women all over the world and history. It is for celebrating mothers, daughters, sisters, and friends who have helped shaped the world as you know it. Just a few of these incredible women are recognized in history books, and even fewer are known today for what they did for society.
- Elizabeth Blackwell, for instance, who according to the Scholastic Teacher's Activity Guide, was the first woman to earn a medical degree from a college. She was rejected by all the major medical schools in the nation because of her sex. She then founded a women's medical college to train other women to become physicians.
- Ella Fitzgerald, who is considered one of the greatest jazz singers of all time was the winner of 12 Grammy Awards and was awarded the Presidential Medal of Freedom.
- Shirley Jackson, who is the former head of the United States Nuclear Regulatory Commission. She is the first female African American to receive a doctorate from the Massachusetts Institute of Technology. Her outstanding leadership in education, science, and public policy demonstrates the capability of women to be leaders in the field of science and technology.
- Jackie Joyner-Kersee conquered the Olympic sport of heptathlon. She won the Olympic gold medal for the United States in 1988 and 1992, and set the world record. She was also the first American woman to win Olympic gold in the long jump.
- Ellen Ochoa is an astronaut and researcher of advanced optical information systems. She flew her first shuttle mission in 1993 as a Mission Specialist with the Discovery crew, conducting atmospheric and solar studied to better understand the effect of solar activity on the Earth's climate and environment. She is the first Hispanic woman to be named an astronaut, and has logged over 500 hours in space.
- Sandra Day O'Connor is the first woman appointed to the position of U.S. Supreme Court Justice and has carved a place for women at all levels of the legal profession.
Each of these women have set new standards for women and girls everywhere. They have become role models for young women to look up to and strive to be like. They have helped women realize their dreams and wishes can be achieved with hard work and perseverance, even when living in a male-dominated society.
These women, along with many more, are just a small number of incredible women who have advanced women's places in society over the years. However, being an incredible woman does not necessarily mean you must be an astronaut or a Supreme Court Justice. Women who are mothers, housewives, daughters, fast-food workers, mechanics, authors, and any other profession are all women who deserve to be celebrated on March 8. Every woman in your life has helped to shape you and the world you live in and those reasons alone are reasons to rejoice them every day of the year.


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Accession Number : AD0824042
Title : MICROWAVE N-PORT FERRITE CIRCULATORS.
Descriptive Note : Rept. no. 4 (Final) 1 Mar 66-30 Jun 67,
Corporate Author : SPERRY MICROWAVE ELECTRONICS CLEARWATER FL
Personal Author(s) : Hagler, Thomas A.
Report Date : NOV 1967
Pagination or Media Count : 118
Abstract : This report describes the development, construction, and final performance of 5-port single-junction ferrite circulators in C band stripline, X band stripline and X band waveguide configurations. The theory of N-port circulators, including postulated modes of operation for 5- and 6-port circulators, is discussed. Complete test data are given for the three exploratory development models in each of the three configurations. The C band stripline units, which incorporate true single-junction center conductors with reduced stripwidth impedance matching, have a 42.5% operating bandwidth. In a typical unit, maximum VSWR is 1.5:1, maximum insertion loss is 0.9 db, and minimum isolations between port 1 and ports 3, 4, and 5 respectively, are 18 db, 28 db, and 53 db. An operating bandwidth of 34% was achieved in the X band stripline circulators using pentagonal-junction center conductors and dielectric impedance matching transformers. Maximum VSWR over this bandwidth is 1.5:1, maximum insertion loss is 0.7 db and minimum isolation is 13 db. The X band waveguide circulators incorporate two circular ferrite pucks separated by a metal tuning post and concentric sleeve of dielectric material. VSWR in these units is less than 1.5:1, insertion loss is less than 1.0 db, and isolation is greater than 10 db over a 3.4% bandwidth. (Author)
Descriptors : (*WAVEGUIDE CIRCULATORS, FERRITES), ATTENUATION, C BAND, ELECTRICAL IMPEDANCE, STRIP TRANSMISSION LINES, STANDING WAVE RATIOS, X BAND, TRANSFORMERS, DIELECTRICS.
Subject Categories : Electrical and Electronic Equipment
Distribution Statement : APPROVED FOR PUBLIC RELEASE


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Google News's personalization features has received a couple of new features today that make Google News a little more personal. Starting today -- if you're logged in -- you may also find stories based on articles you've clicked on before.
First, for signed-in users -- "News for You" will now include stories based on your news-related web history. If you click on a lot of articles about a certain topic, this section will make sure that you get a chance to see more articles on that topic.
Second, "Recommended Sections" feature has beed added in the right side column that suggests topics you can add to your news page as custom sections, based on stories you've clicked on before.
If you don't want to see personalized news based on your Web History, you've a few options:
- Click "Standard U.S. Edition" link at the bottom of Google News. This willn't delete any of your News settings or Web History. It'll switch you to an unpersonalized version of Google News for the duration of your current session. (To switch back, click on "Personalized U.S. Edition".)
- Delete your web history. (Google News may take some time to update.)
- Log out of your Google Account.
[Source: Google News blog]


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Words by Alexander Norton
THE DAILY MENTION – Ross Rawlings
Ross Rawlings
When on the source, graduate photography page, I came across a picture as I did not know what to look for, and it was an image of two people, in an intimate relationship performing some kind of physical act, done in such a way that it makes me look and discover the whole series. I have no prior knowledge of Ross Rawlings work, but I felt compelled to write about it, as it is so intimate yet public.
I have always been a fan of photographs and relationships and how they work together, and what kind of implications about publicity of those chosen private moments, to become part of a stream of images online. Does the act of recording this action of lust, love and contempt for each other affect the scenario. Does the fact that a project is being made affect how they communicate, does he talk to her less, does she reveal less as she knows it will be shown to more than him. These are intimate moments, and what is beautiful is that we can see them, because these are moments when we are in the scenario we do not see them, and to take a step away and record it as it happens is both difficult, not just physically but emotionally also.
What is beautiful, and it is, is that you begin to feel for this woman on show, and when I say on show, I do not mean that negatively, it is a collaborative understanding that the issue of the heart will be taken seriously and treated with care. It could be talked about the visual qualities that enhance the sensation intended when you see the images, and the series finds a way of making us fall in love with this girl, but why should we fall in love with her, when the experience is someone elses. What I love about this kind of work, is that it raises many issues about photography itself, but ignores them and carries on, because the act on show is so powerful we just ignore the reason why we are seeing them, we just look.
Ross mentions in his synopsis, the use of the shutter release which enhances the almost elevated experience you gain from this, as the photographer himself is as much to do with the frame than the subject in question. I think this is to remember, as the love between two people is such a fragile thing, but its so strong that you want to hold onto that feeling permanently, because that feeling, is the best feeling we can feel as people. To borderline cheesy, we live to feel the love of another in many ways, from a lover, friend, family member and an obsession or activity.
Read Alex's blog here: http://alexandernortonblog.tumblr.com


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4037 Arbutus Avenue, Grove City, Ohio
$120,000Contract Pending
$120,000Contract Pending
4037 Arbutus Avenue, Grove City, Ohio 43123 - Commercial lot or rehab. Currently a single family home on corner lot next to police station. Great commercial potential or investment!
Listing Courtesy of Timothy E Beathard (740) 852-1181 - Timothy E. Beathard & Assoc. (740) 852-1181
"The Parrett Group was amazing to deal with. They always went above the call of duty and were helpful before we listed, durring the sales process and after closing to ensure that we were completely satisfied. Anyone buying or selling a home in The Parrett Group's territory that uses another agent is wasting their time and money."


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2005 Chevrolet TrailBlazer - Consumer Review
Build Quality
Reliability Value
Build Quality
Reliability Value
1 of 1 people found this review helpful
For the Money, Best SUV on the Road
By Mel Turner on
Vehicle2005 Chevrolet TrailBlazer LS 4WD 4dr SUV (4.2L 6cyl 4A)
ReviewWe've put over 134,000 miles and this baby still sounds brand new. Engine so quiet, I can't hear it idle at red lights. I only use Mobile 1 synthetic oil. Only two problems since ownership, fuel gauge and ignition switch. Warranty covered them. This is the only vehicle my wife drives. We have two Volvo s80s that's collecting dust in the garage. I'll probably have to sell them. Suv only requires maintenance every 50,000 miles besides oil change. One more thing, this suv is great on the road. The only suv we'll ever own. We love our trailblazer. For longevity, only use synthetic oils and lubricants. With my Continental tires, I get 18 city and 22 highway.
Best FeaturesLow maintenance, strong 6 cyl engine, comfort, price, on the fly 4wheel drive. 4wheel performs exceptional in Colorado mountains and snow. Chevy, please do not stop manufacturing Trailblazers.
Worst FeaturesDesign SUV so that rear seats fold flat.
Recommend (1) (0)
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Jacque Fresco Quotes About CrimeQuotes about: Crime
Crime is based upon need, making money. People sell drugs to make money. But if everybody is cared for, they don't sell drugs and if there's no money you can't sell drugs even if you wanted to. There'd be no such thing as gambling, prostitution, or selling out, or paying off a senator or a governor. There are no senators, there are no governors so you can't pay them off. If you take away the basis or the condition that generate abhorrent behavior, you don't have abhorrent behavior.
What will drive people if they don't have money or reward? The reward is the end of war, the end of poverty, most crime, and the end of begging for medical care. Everyone will be cared for and educated. There will be no taxation, and no advantage group. No technical elitism, or any other kind of elitism. If that isn't incentive enough, then I don't know what is.
People say that the monetary system produces incentive. This may be true in limited areas, but it also produces greed, embezzlement, corruption, pollution, jealousy, anger, crime, war, poverty, tremendous scarcity, and unnecessary human suffering. You have to look at the entire picture.
War is not the only form of violence imposed on people through inadequate social arrangements. There is also hunger, poverty and scarcity. The use of money and the creation of debt fosters economic insecurity, which perpetuates crime, lawlessness and resentment. Paper proclamations and treaties do not alter the facts of scarcity and insecurity, and nationalism tends only to propagate the separation of nations and the world's people.
If we are genuinely concerned about the environment and the fellow human beings, and want to end territorial disputes, war, crime, poverty, hunger, and the other problems that confront us today, the intelligent use of science and technology are the tools with which to achieve a new direction – one that will serve all people, and not just a select few.
A Resource-Based Economy is in the application of the methods of science with human concern and environmental concern. If we used the scientific method throughout the world, the probability of war drops to zero. The probability of human suffering disappears. Deprivation, poverty, crime - all those things tend to disappear because there's no basis. I'm strictly concerned with the environment that people are raised in and if that environment is altered, so will behaviors be altered.
Jacque Fresco
- Born: March 13, 1916
- Died: May 18, 2017
- Occupation: Futurist
- Cite this Page: Citation


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Facebook has reached a couple of milestones, with having passed about a billion active mobile users. This announcement was made during a recent conference call which was held in March, where CEO of Facebook, Mark Zuckerberg discussed the company's acquisition of Oculus VR.
While the active monthly app users of Facebook have shot up to 1 billion from 945 million in last December, the photo-sharing app Instagram has also seen a healthy growth. Its user base has doubled since Facebook acquired it in April 2012, now having more than 200 million users. Two years back, Instagram had only about a 100 million users. Also, Instagram's statistics till date shows that more than 20 billion photos have been shared and also there has been a steady growth of 50 million users over a period of the last six months.
With the huge growth in mobile advertisements, it is pretty much clear why Facebook is concentrating on mobile platforms. According to eMarketer, in 2012 only 11 percent of Facebook global ad revenues came from mobile, but last year, this figure jumped to 45 percent. Now this year, the figure is expected to rise to about 63 percent.
Earlier, in the press release announcing the Oculus deal, Zuckerberg said that the mobile is the platform of today, and they are planning ahead to get ready for the platforms of tomorrow. With this, we can definitely see that Facebook is a leading industry when compared to its counterparts.
We would love to know your comments about Facebook and how do you think it is rising towards a steep growth by defeating its industry rivals.


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Apple, Google Hiring Pacts Fixed Pay, Engineers Insist
In a brief supporting a renewed motion for class certification, a group of former software engineers for leading Silicon Valley companies argue they have proposed a reasonable method of demonstrating that the claims in their case had a wide impact on members of the proposed...
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This is a case study of a recent property I sold in Riverside, California. The purpose of this blog post is to familiarize you with the process of obtaining, listing, and selling an REO. This is a behind-the-scenes look into REO disposition.
The Property
The house I sold was a duplex in a Riverside neighborhood that was constructed in the 1970's. Each of the two units in this duplex home are 2 bedroom 1 bathroom, 906 sq. feet, with a one car garage.
The borrower's mortgage had been in default for more than 12 months, but the property was fully occupied and both tenants were still paying rent to the defaulting owner. We started legal eviction on both units, but since they were both occupied by tenants with written leases, we were limited to what we could do, and the eviction would take at least 90 days.
Evicting The Tenants
As the listing broker on this home, I went out to the house and met with both tenants. I offered Cash for Keys but both tenants were resistant to it, fearful that they wouldn't be able to find a similar place in the same rent range quickly enough to move. Since neither tenant wanted to do cash for keys, I made sure we got the written notices sent out from legal quickly because that shows them that the eviction is happening.
As the listing broker, my job is to sell this home which was now owned by a bank. Since the eviction is happening with or without me, I go out of my way to make the transition as easy as possible for tenants who are willing to cooperate. I told them that we were the owners and we will not collect rent anymore during the eviction process.
I waited until after I knew those notices had been delivered and went out there again to meet with them. I explained to them how detrimental an eviction would be on their credit and got one of the units to agree to move out in approximately 20 days from when I met with them. The other tenant was still resistant to move out, but I left him my information. Eventually the other tenant called me when he got more legal notices and he agreed to move out as well. This was the best decision for both sets of tenants because NOT having an eviction on their record made qualifying to rent a new home much easier.
Preparing The Home For Sale
After both sides were emptied, I did my physical inspection of the property and found the duplex to be in poor to fair condition. The house needed flooring replacement throughout, paint inside and out, water damage remediation in the bathrooms, appliance replacement, smoke detector and carbon monoxide detector installation, window replacement, water heater replacement, and fence replacement. Of that, we replaced the window and installed the smoke and carbon monoxide detectors. I replaced the window because it was a security issue and we were legally required to replace the smoke and carbon monoxide detectors. After we made those minor repairs, the property was cleaned and we performed basic landscaping.
Determining The List Price
Next we needed to set the list price of the home. This bank requires that two Broker Price Opinions (BPOs) be performed. Then a reconciled value is set for the list price.
When I did my BPO, I pulled 3 active listings and 3 competitive sales that were all within a half mile radius of the subject property and had sold within the last 90 days. This is the best way to understand the market's perception of value. Based on the condition and sales prices of the comps, I determined that the as-is value of this property should be about $165,000. I felt that due to the extremely distressed nature of this sub-market, repairing the property would not necessarily increase its value. I recommended the property be sold as-is for $165,000.
The second BPO came back and the broker agreed that in the duplex was worth $165,000 in its as-is state. This broker, however, thought that it was worth $170,000 repaired. As a result, the bank reconciled its value at requested the property be listed at $170,000.
Marketing The Home For Sale
We put up a For Sale sign in the front yard, installed lockboxes on the front door, created property fliers, listed the property in the MLS, and syndicated the listing to our network of websites including CAPropertyFinder.com, Trulia, Zillow, Realtor, and 40+ other real estate sites. We sent out feelers to our investor network and within the first 48 hours had received 9 offers.
We negotiated with everyone and ended up selling the property for almost $10,000 above list price. The escrow was opened 4 days after it was listed and the property closed exactly 30 days later. Pricing the property correctly and marketing the home properly allowed us to do that.
Home Sale Specs
This property was listed for 2 days until we accepted an offer. An average deal in our portfolio takes 39 days. We closed escrow in 30 days, whereas an average property in our portfolio takes 37 days. This property sold for 5% above list price, whereas an average property in our portfolio sells for 3% below list price.
In the last 150 days, 2 properties sold within a half mile. Those properties averaged 73 days on market and they sold for an average of 8% below list price.
List Your Home For Sale
Selling a home quickly and for above list price is a matter of experience, market knowledge, and connections in the industry. If you are seeking similar results and would like us to sell your home, call us at 1-800-287-1808.
You may also reach me directly, Scott Mehlman, at 310-430-6361. I am the managing broker of our office and I look forward to exceeding your real estate needs.
Search Southern California Homes for sale or view our REO Listings For Sale in California.
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The eternal struggle for anyone packing for a trip; do I bring a wide assortment of footwear because a sassy boot mood may strike me? Those extra shirts will fit in my weekend bag… maybe? Recently, the historical city of Victoria, BC, beckoned me to and I had to put all my MUA skills to work, planning and packing for a wonderful weekend away.
I am a high maintenance kind of woman sometimes; it really depends on what is happening. I can get down and dirty with the best of them outside, but I need my creature comforts, and for me that means makeup and hair products, what else?!
Here are some ideas on how to pack smarter so you keep your weekend away less about your significant other watching you get ready, and more about actually spending time out around the town together. These items can do double duty and will help you pack a little lighter for your next overnighter.
I struggle over what to pack all the time, but one way to approach this task is to see if there are any items that can do double duty, and conditioner answers the call.
You can use conditioner for your hair (obviously) but it can also be used to remove makeup (even waterproof), for shaving cream, and even to polish shoes in a pinch.
I tend to bring my own hair and body care products when I travel, as some hotels do not provide crueltyfree or good quality bath and body products. If you have allergies as well, it is always better to pack your own products to avoid a potential allergic reaction, especially when you are away on a weekend trip. Nothing kills the romance faster than a horrible rash all over your body!
Send Nudes
Makeup wise, depending on what your weekend looks like, you will need a few easy essentials to get your routine down to a minimal time frame. Whether you are planning a more casual weekend away in Whistler or a night on the town in Victoria, a nude makeup colour palette is always the answer.
For daytime, you can use a nice colour combo such as Elate Cosmetics Lithe shadow (a soft neutral gold) and Earthen (a matte neutral brown) shadow which you can also use to fill your brows in if you wish. This will set the stage for later, as you can enhance your existing makeup by building on top of it to darken it up into a neutral, smoky eye for that dinner out later on.
I would use Elate's Rise shadow (deep shimmery brown) to darken up the crease of your eye, and if you wanted to go even darker you could use Stillness, their matte black shadow. The beauty of Elate is that you can build your own palette, so everything you need; shadow, blush, bronzer and even powder foundation can be taken on the road in their ecofriendly, magnetic bamboo palette. Their shadows can also be used wet, so you can skip bringing eyeliner if you wanted to, just grab that angled brush for your brows and use it to create liner as well.
Elate also has a brilliant lipstick shade called Tantric, a romantic burgundy red, perfect for everyone. Dab a little on your lips using your fingertip to create a soft berry lip stain for daytime. You can also use it as a cream blush; it is perfect applied to apples of your cheeks with your fingers and blended out. And when you are heading out later on, you can use it full tilt on your lips for that gorgeous, kissable, classic bold lip. Paired with a neutral eye, it is a perfectly chic evening look that will go with anything.
Dry Shampoo
I cannot live without my dry shampoo, and I am not exaggerating. Dry shampoo can be used to remove the odours and oils that build up in your hair over the day, and can provide instant volume and lift those limp locks after a walk outside. It can also be used as texturizer; it will help your hair to have more grip if you want to put it up for dinner out later on. And, used sparingly, it can also be used as a deodorant. If you have lighter hair, it can also cover your roots up a little until your next colour. Added bonus: you are saving water and you are saving yourself time!
A great vegan crueltyfree dry shampoo is Tarte's Hair Goals dry shampoo. If you accidentally spill something on your outfit at dinner, dry shampoo can help lift grease stains from fabric, just be sure to clean the spot with soap and water as soon as possible. And as if that was not enough, dry shampoo can also deodorize your shoes; just spray and let dry overnight.
Before you go, pick your favourite scent to take along to use on your trip, many are available in a mini or roller ball format. Scent is tied to memory and emotions; long after your trip you can dab some of your scent on, and immediately you will have memories of your getaway flood back, and bring a smile to your face.
I hope that these tips and tricks will help you pack a little lighter and a lot smarter for a getaway or every day. We all have little time to waste when getting ready to face the day, so sometimes finding a smarter way to work is what we need.
Peace, Love, and Lipstick!
Next week: Beat the Heat – Tips and Tricks for Making Your Makeup Last in Heat Like a Pro


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Previous studies have shown that protease‐activated receptors (PARs) play an important role in various physiological processes. In the present investigation, we determined the expression of PARs on human lung fibroblasts (HLF‐1) and whether they were involved in cellular differentiation and pro‐inflammatory cytokine and prostaglandin (PGE2) secretion. PAR‐1, PAR‐2, PAR‐3, and PAR‐4 were detected in fibroblasts using RT‐PCR, immunocytochemistry, and flow cytometry. Increased expression of PAR‐4, but not other PARs, was observed in fibroblasts stimulated with phorbol myristate acetate. The archetypical activators of PARs, namely, thrombin and trypsin, as well as PAR‐1 and PAR‐2 agonist peptides, stimulated transient increases in intracellular Ca2+, and promoted increased α‐smooth muscle actin expression. The proteolytic and peptidic PAR activators also stimulated the release of IL‐6 and IL‐8, as well as PGE2, with a rank order of potency of PAR‐1 > PAR‐2. The combined stimulation of PAR‐1 and PAR‐2 resulted in an additive release of both IL‐6 and IL‐8. In contrast, PAR‐3 and PAR‐4 agonist peptides, as well as all the PAR control peptides examined, were inactive. These results suggest an important role for PARs associated with fibroblasts in the modulation of inflammation and remodeling in the airway.
- Cell differentiation
- cytokines
- inflammation
- lung
Bronchial asthma is a disease characterized by pulmonary inflammation and airway wall remodeling (Bousquet et al. 2000; Martinez and Vercelli 2013) and these features are major contributing factors toward the development of irreversible airway obstruction, bronchial hyperresponsiveness, and eventual decline of pulmonary function (Vignola et al. 2000; Chiappara et al. 2001; Fanta 2009). Although many cell types are involved in these processes, increasing evidence suggests that fibroblasts within the airway wall are primarily involved in airway wall remodeling and asthma. Increased numbers of subepithelial fibroblasts in the airways of asthmatic subjects have been reported (Holgate et al. 2000) and the number of fibroblasts has been positively correlated with the thickness of the basement reticular membrane (Holgate et al. 2000) as well as the severity of asthma (Benayoun et al. 2003). The increase in airway fibroblast proliferation may, in part, be due to increased concentrations of growth factors in the extracellular space. Transforming growth factor (TGF)‐β, fibroblast growth factor, endothelin‐1, and epidermal growth factor are predominantly derived from the airway epithelium (Chiappara et al. 2001) and have all been found to be elevated in asthmatic patients (Goldie and Henry 1999; Knight 2001). The role of these profibrotic mediators is to promote interstitial collagen deposition into the extracellular matrix, leading to thickening of the basement membrane (Holgate et al. 2000).
In addition, lung fibroblasts also proliferate, differentiate, and synthesize extracellular matrix components in response to Th2 cytokines such as IL‐4 and IL‐13 (James et al. 1989; Silvestri et al. 2001; Sabatini et al. 2002). These cytokines have also been shown to upregulate adhesion molecule expression on the surface of fibroblasts and promote inflammatory cell trafficking to the airway wall. Taken together, these findings suggest that lung fibroblasts may respond to various growth factors and profibrotic mediators in the extracellular space, stimulating cellular proliferation and augmenting the initial preinflammatory stimulus. Such structural changes are likely to increase airway wall thickening and stiffness, contributing to persistent airway obstruction and hyperresponsiveness observed in asthma (James et al. 1989; Saunders et al. 2009).
Interestingly, airway fibroblasts from asthmatic patients demonstrate an increased ability to differentiate into myofibroblasts (Michalik et al. 2009) which possess ultrastructural features intermediate between fibroblasts and smooth muscle cells. These cells are characterized by the presence of contractile proteins such as alpha‐smooth muscle actin (α‐SMA) (Darby et al. 1990; D'Andrea et al. 2001). Myofibroblasts are present in the asthmatic airways (Brewster et al. 1990; Holgate et al. 2000) and the number of these cells is increased upon bronchial allergen challenges in asthmatic subjects (Gizycki et al. 1997). In addition to matrix deposition, cytokine release, and expressing adhesion molecules, myofibroblasts are also likely to further contribute to luminal obstruction through contraction of the airway wall (Thannickal et al. 2004; Nihlberg et al. 2006).
In addition to profibrotic mediators and inflammatory cytokines, airway fibroblasts also respond to endogenous proteases such as thrombin and mast cell tryptase (Akers et al. 2000; Chambers et al. 2000). The effects of these proteases are, in part, mediated by a family of G‐protein‐coupled protease‐activated receptors (PAR‐1 to PAR‐4). The extracellular portions of each PAR are cleaved by proteases, resulting in a new amino terminus which then binds to the second extracellular domain, initiating intracellular signaling and activation of cellular processes. Stimulation of PARs is associated with multiple signals transduction pathways, including inositol trisphosphate hydrolysis, increased cytosolic free Ca2+ concentration, and NF‐κB expression (Macfarlane et al. 2001). In airway epithelial cells, activation of PARs stimulates the release of pro‐inflammatory cytokines, suggesting that PARs promote tissue inflammation and repair (Asokananthan et al. 2002; Lan et al. 2002).
The precise role of PARs in fibroblasts function in the lung remains unclear, but there is evidence to show that activation of PAR‐1 or PAR‐2 promotes fibroblast proliferation (Akers et al. 2000; Shimizu et al. 2000; Frungieri et al. 2002) which may be dependent on the autocrine release of connective tissue growth factor (CTGF) (Chambers et al. 2000; Shimizu et al. 2000). In addition, PAR‐1 or PAR‐2 agonist peptides also induce cytoskeletal changes in these cells (Bogatkevich et al. 2001; Ge et al. 2003). These studies suggest that PAR activation in fibroblasts may promote airway wall remodeling (Akers et al. 2000; Chambers et al. 2000).
Whether activation of PARs results in inflammatory cytokine and prostaglandin E2 (PGE2) release from fibroblasts and promotes myofibroblast differentiation has not been examined in great detail. To this end, we examined the activation pathway of all four PARs and assessed the functional significance of PAR activation in human lung fibroblasts. In particular, we have focused on changes in intracellular Ca2+, cytokine and PGE2 secretions, and α‐SMA expression.
Materials and Methods
Human lung fibroblast cells (HLF‐1; American Tissue Culture Collection, Manassas, VA) were kindly provided by Professor Darryl Knight (School of Biomedical Sciences and Pharmacy, Newcastle University). Synthetic PAR agonist peptides, as well as control peptides, were synthesized with amidated C‐termini (purity > 85 %) by Proteomics International Ltd. (Perth, WA, Australia). The sequences of the agonist and control peptides were, respectively: PAR‐1, TFLLRN‐NH2 and FTLLRN‐NH2; PAR‐2, SLIGKV‐NH2 and LSIGKV‐NH2; PAR‐3, TFRGAP‐NH2 and FTRGAP‐NH2; and PAR‐4, GYPGQV‐NH2 and GYPGVQ‐NH2. As it has previously been shown that the human PAR‐1 agonist peptide, SFLLRN, could cross‐activate PAR‐2, the more specific xenopus‐human hybrid PAR‐1 agonist peptide TFLLRN‐NH2 was used in this study (Hollenberg et al. 1997). Thrombin was purchased from CSL Limited (Melbourne, VIC, Australia) and Sigma Chemical Co. (St Louis, MO). Tissue culture and molecular biology reagents were purchased from Invitrogen Corp. (Carsbad, CA). Unless otherwise stated, other general chemicals were purchased from either BDH (Kilsyth, Vic., Australia) or from Sigma Chemical Co.
Protease activity
Trypsin activity was confirmed using the active site titrant, p‐nitrophenyl p'‐guanidino benzoate, as described previously (Chase and Shaw 1969). Thrombin activity was determined using the thrombin substrate N‐ benzoyl‐DL‐arginine p‐nitroanilide (King et al. 1996). Thrombin, obtained from CSL Limited (0.5 nmol L−1 p‐nitroaniline released min−1 mg−1), was approximately 50‐fold less active than that purchased from Sigma Chemical Co. (23.4 p‐nitroaniline released min−1 mg−1). Thrombin from the former source was used in cell culture experiments, whereas the latter was used in Ca2+ flux studies.
Human lung fibroblast cell culture and PAR stimulation
HLF‐1 fibroblasts were cultured in 75 cm2 tissue culture flasks (Nalge Nunc International, Naperville, IL), and grown in DMEM medium supplemented with 10 % (v/v) fetal calf serum (FCS), and antibiotics. Cells from the flasks were trypsinized and seeded into 24‐well tissue culture plates at a density of 2.5 × 105 per well for 24 h, and serum‐starved overnight. Serum‐deprived HLF‐1 cells were exposed to varying concentrations of thrombin (0.005–50 U mL−1) or trypsin (1.25 × 10−4–1.25 nmol L−1), as higher concentrations of these peptidases caused detachment and cellular death (data not shown). HLF‐1 cells were also stimulated with PAR peptides (100–500 μmol L−1) or phorbol myristate acetate (PMA, 10 ng mL−1). In some experiments, cells were incubated with indomethacin (10 μmol L−1) prior to stimulation to assess the role of cyclooxygenase in PGE2 release. Aliquots (100 μL) of cell culture supernatants were removed at various time points (0, 3, 6, 12, 18, and 24 h) after stimulation, centrifuged at 12,000 × g for 5 min at 4°C, and stored at −20°C until ready for cytokine quantitation. Cells stimulated in this way showed greater than 85 % viability at the conclusion of the experiments, as determined by trypan blue exclusion and lactate dehydrogenase assays (not shown).
Reverse transcription–polymerase chain reaction (RT‐PCR)
Total RNA was prepared from HLF‐1 fibroblasts using Tri‐Reagent according to the manufacturer's instructions (Molecular Research Center, Cincinnati, OH). Briefly, cells were lysed and the homogenates were transferred to 1.5 mL microcentrifuge tubes. Chloroform (200 μL) was added to each tube, shaken by hand, and left at room temperature (RT) for 10 min. Tubes were then centrifuged at 13,000 × g for 15 min at RT, after which the aqueous phases were transferred to new tubes containing 0.5 mL of isopropanol. The tubes were left overnight at −20°C, and then centrifuged at 13,000 × g for 15 min. The supernatants were discarded, and 1 mL 75% (v/v) ethanol was added to the pelleted RNA in each tube. The pellets were resuspended and tubes centrifuged at 13,000 × g for 8 min, after which the supernatants were discarded and the pellets air‐dried, and dissolved in 10 μL of diethyl pyrocarbonate (DEPC)‐containing water. For reverse transcription, RNA was primed with 250 ng of oligo‐dT12‐18 primer, and reversed transcribed in a mastermix totaling 50 μL, containing 7.5 mmol L−1 MgCl2, 0.4 mmol L−1 of each dNTP, 10U RNase inhibitor, and 2.5 U avian myeloblastosis virus reverse transcriptase (Promega, Madison, WI). The mixture was incubated at 42°C for 60 min, and terminated by heating at 90°C for 2 min. The synthesized first strand cDNA was stored at −20°C.
Forward and reverse primers used for amplifying human PARs were prepared commercially based on published sequence data (Wan et al. 2001); PAR‐1 sense 5′‐TGTGAACTGATCATGTTTATG‐3′, antisense 5′‐TTCGTAAGATAAGAGATATGT‐3′, (PCR product, 708 bp); PAR‐2 sense 5′‐ AGAAGCCTTATTGGTAAGGTT‐3′, antisense 5′‐AACATCATGACAGGTCGTGAT‐3′ (PCR product, 582 bp); PAR‐3 sense 5′‐CTGATACCTGCCATCTACCTCC‐3, antisense 5′‐ AGAAAACTGTTGCCCACACC‐3′, (PCR product, 382 bp); PAR‐4 sense 5′‐ ATTACTCGGACCCGAGCC‐3′, antisense 5′‐TGTAAGGCCCACCCTTCTC‐3′ (PCR product, 392 bp). Amplification of GAPDH, with the sense and antisense primer pair 5′‐ CCCATCACCATCTTCCAGGAGC‐3′ and 5′‐CCAGTGAGCTTCCCGTTCAGC‐3′ (PCR product, 471 bp), acted as an internal control (Prime et al. 2000). For the polymerase chain reaction, 1.5 μL cDNA was mixed in a reaction vial containing 2.5 pmol L−1 of each forward and reverse primer, 0.2 mmol L−1 of each dNTP, 2 mmol L−1 MgCl2, 0.5 U Platinum Taq DNA polymerase, 1.2 μL of 10 × Taq DNA polymerase buffer, and made up to 12 μL using DEPC water. The conditions for amplification were as follows: for PAR‐1 and PAR‐2, 94°C for 45 sec, 55°C for 45 sec, 72°C for 2 min and 30 sec, for 35 cycles; for PAR‐3, PAR‐4 and GAPDH, 94°C for 45 sec, 65°C for 45 sec, 72°C for 2 min and 30 sec, for 35 cycles. Electrophoresis was performed using 2% (w/v) analytical grade agarose gels which were subsequently stained with ethidium bromide, and visualized under a UV transilluminator. Densitometric analysis was performed using NIH Image software (National Institute of Health, Bethesda, MD).
Immunocytochemistry and fluorescence/confocal microscopy
Immunolocalization of PARs on fibroblasts was performed using cells cultured on glass slides. The cells were rinsed with PBS and fixed in 4 % (v/v) paraformaldehyde in PBS for 30 min at RT. The cells were permeabilized with 0.2 % Triton X (v/v) in PBS for 5 min at RT, and rinsed in PBS. After blocking the cells in 20 % normal horse serum, the fibroblasts were incubated with antibodies raised against individual PARs, including mouse monoclonal antibodies (Santa Cruz Biotechnology Inc., Santa Cruz, CA) raised against PAR‐1 (ATAP2 sc‐13503) and PAR‐2 (SAM11; sc‐13504), as well as rabbit antibodies (commercially prepared by Chiron, Clayton, Australia) raised against PAR‐3 (37TLPIKTFRGAPPNSFEEFP55) and PAR‐4 (28EDDSTPSLLPAPRGYPGQV39). Equivalent dilutions of isotype‐matched mouse and rabbit antibodies (Dako, Via Real Carpinteria, CV) were also used as appropriate controls. The primary antibodies were incubated overnight at 4°C. Following two washes in PBS, the cells were incubated with appropriate Alexa 488‐conjugated secondary antibodies (Molecular Probes, Eugene, OR) for 60 min at RT, rinsed twice in PBS, mounted in a nonfluorescent mounting medium, and kept in the dark until ready for viewing using fluorescence (TE2000‐U; Nikon Corp., Chiyoda‐ku, Tokyo, Japan) or confocal microscopy (Biomedical and Imaging Analysis Facility, UWA, Perth, WA, Australia).
Flow cytometry
Fibroblasts cultured in 24‐well tissue culture plates were removed by gentle pipetting the cells previously incubated in 8 mmol L−1 EDTA in PBS at 4°C for 5 min. The cells were centrifuged (500 × g, 6 min), fixed with 4 % (v/v) paraformaldehyde in PBS for 30 min at RT, and blocked in 10 % (v/v) normal horse serum (30 min, RT). Specific anti‐PAR antibodies were diluted in PBS or PBS with 0.1 % (w/v) saponin, to assess cell surface and intracellular PAR staining, respectively. The primary antibodies, as well as the isotype‐matched control antibodies, were incubated overnight at 4°C. Following two washes in PBS, the cells were incubated with appropriate biotin‐conjugated secondary antibodies for 2 h at RT, and washed twice in PBS. The cells were then incubated with phycoerythrin (PE)‐conjugated streptavidin (BD Bioscience Pharmingen, San Diego, CA) for 2 h at RT in the dark, washed twice in PBS, and stored in the dark at 4°C. Cells were assayed on a FACS Caliber (Biomedical and Imaging Analysis Facility, UWA), and files of 104 events were analyzed using FlowJo software (Tree Star Inc., Ashland, OR).
Detection of IL‐6 and IL‐8 by ELISA
Supernatants from stimulated fibroblasts were assayed for IL‐6 and IL‐8 using ELISA. Briefly, 96‐well plates were coated with 100 μL of the appropriate antibody (500 ng mL−1 in 0.1 M NaHCO3/NaCO3 buffer) and incubated overnight at 4°C. The plates were then washed three times with washing buffer (PBS containing 0.5 % (v/v) Tween‐20) and blocked with 100 μL of washing buffer containing 1 % (w/v) BSA at RT for 1 h. The plates were washed three times with washing buffer before 100 μL aliquots of each sample, or cytokine standards, were added to the wells. The plates were incubated overnight at 4°C and then washed three times. The appropriate biotinylated secondary antibodies were then added to each well at RT for 1 h and washed three times. Streptavidin‐horseradish peroxidase conjugates were added to each well, incubated at RT for 30 min, and the plates then washed three times. Finally, 100 μL of peroxidase substrate (K‐Blue ELISA substrate, Graphic Scientific, Brisbane, QLD, Australia) was added to each well, and reactions terminated by addition of 1 M phosphoric acid. Optical densities were determined using a microplate reader (Spectramax 250; Molecular Devices Co., Sunnyvale, CA) at 450 nm, and cytokine concentrations determined by interpolation from the standard curve using the SoftMax‐Pro software (Molecular Devices Co.), and expressed as pg per 2.5 × 105 viable cells.
Detection of PGE2 by ELISA
PGE2 was measured using a competitive PGE2 enzyme immunoassay (Cayman Chemical, Ann Arbor, MI) as described previously (Ge et al. 2003). PGE2 released was expressed as pg per 2.5 × 105 viable cells.
Measurement of cytosolic Ca2+
Changes in cytosolic Ca2+ concentration were measured using the membrane permeable fluorescent Ca2+ indicator, Indo‐1 (Teflabs, Austin, TX), as described previously (Asokananthan et al. 2002). In brief, cells were grown on coverslips to 80 % confluency and loaded with RPMI containing 6 μmol L−1 of Indo‐1 and 0.1 % (w/v) Pluronic F‐127 (Molecular Probes Inc., Eugene, OR) for 45 min at RT. After incubation, cells were washed with two changes of RPMI to remove the excess dye prior to challenges with PAR activators. The ratio of fluorescence emission at 405 and 490 nm was measured using a spectrophotometer (Cairn Research Ltd, Faversham, Kent, U.K.) attached to an inverted microscope (Nikon Corp.), configured for epifluorescence. The excitation wavelength of 340 nm was provided by a variable monochromator system (Cairn Research Ltd). The ratio of the emission intensities was used as a measure of changes in cytosolic Ca2+.
Statistical analyses
Unless stated otherwise, data were expressed as mean ± standard error of the mean (SEM). Statistically significant differences between means were determined using the unpaired Student's t‐test. P values less than 0.05 were considered statistically significant.
Expression of PARs in HLF‐1 human lung fibroblasts
mRNA for PAR‐1, PAR‐2, PAR‐3, and PAR‐4 was detected by RT‐PCR in HLF‐1 fibroblasts, and the sizes of each amplicon detected corresponded to those previously reported (Ge et al. 2003; Jin et al. 2003) (Fig. 1, upper panel). The intensity of the PAR‐4 amplicon was less than that observed for PAR‐1, PAR‐2, or PAR‐3, but treatment of cells with PMA (10 ng ml−1 for 24 h) upregulated PAR‐4 expression, but not other PARs (Fig. 1, lower panel). In addition, HLF‐1 cells were shown to express PAR proteins as judged by both immunocytochemistry and flow cytometry (Fig. 2). The intensity of immunoreactive PAR‐1, PAR‐2, and PAR‐3 was greater than that observed using with PAR‐4. Flow cytometry showed expression of PARs on the surface as well as in the cytosol.
Changes in intracellular Ca2+ induced by peptidases and PAR peptides in HLF‐1 cells
Both thrombin and trypsin induced changes in intracellular Ca2+ in HLF‐1 cells, as judged by increases in fluorescence ratio (Fig. 3). In addition, treatment of cells with either PAR‐1 or PAR‐2 agonist peptides also induced changes in intracellular Ca2+ (Fig. 3). Intracellular Ca2+ increased within a few seconds of exposure to proteases and PAR agonist peptides, and gradually returned to baseline within 120 sec. PAR‐3 and PAR‐4 agonist peptides, as well as the controls peptides for each of the four PARs, were inactive. In addition, PAR‐1 and PAR‐2 agonist peptide‐induced changes in Ca2+ were significantly attenuated when cells were previously challenged with thrombin, and trypsin, respectively. The effects of the PAR‐1 and PAR‐2 agonist peptide were slightly reduced in cells prestimulated with trypsin and thrombin, respectively (Fig. 4).
Induction of α‐SMA expression in HLF‐1 cells by proteases and PAR peptides
Stimulating HLF‐1 cells with thrombin, trypsin, and PAR‐1 and PAR‐2 agonist peptides for 24 h clearly induced the expression of α‐SMA in HLF‐1 cells. In contrast, cells stimulated with PAR‐3 and PAR‐4 agonist peptides, or PAR control peptides, expressed no detectable α‐SMA (Fig. 5).
Thrombin and trypsin induce cytokine release from HLF‐1 cells
Thrombin stimulated the release of IL‐6 and IL‐8 from HLF‐1 cells after 24 h in a concentration‐dependent manner (Fig. 6). The maximal release of cytokines occurred when the cells were stimulated with 50 U mL−1 thrombin. Similarly, trypsin concentration dependently stimulated the release of IL‐6 and IL‐8 from HLF‐1 cells after 24 h. Maximal release of IL‐6 and IL‐8 release was observed when the concentration of trypsin used was at 12.5 pmol L−1 and 1.25 pmol L−1, respectively. The maximal release of IL‐6 and IL‐8 induced by thrombin was, respectively, 7‐fold and 29‐fold greater than that induced by trypsin (Fig. 6).
PAR agonist peptides induce cytokine release from HLF‐1 cells
Exposure of HLF‐1 cells to 400 μmol L−1 PAR‐1 or PAR‐2 agonist peptides for 24 h also stimulated the secretion of IL‐6 and IL‐8 (Fig. 7). The release of IL‐6 and IL‐8 induced by the PAR‐1 agonist peptide was approximately 2‐fold greater than that induced by the PAR‐2 agonist peptide. The PAR‐3 and PAR‐4 agonist peptides, as well as the control peptides for each of the four PARs, did not significantly modulate basal cytokine secretion. The PAR‐1 and PAR‐2 agonist peptides concentration dependently induced the release of cytokines, with maximal cytokine release obtained when 300–500 μmol L−1 of agonist peptides were used (Fig. 8). The release of cytokines induced by 400 μmol L−1 of PAR agonist peptides was also time dependent (0–24 h), with maximal IL‐6 and IL‐8 release observed 6 and 18 h after peptide stimulation, respectively (Fig. 8).
HLF‐1 cells respond to additive combinations of PAR agonist peptides
Exposure of HLF‐1 cells to PAR‐2 agonist peptide in combination with PAR‐1 agonist peptides (each at 400 μmol L−1) resulted in the statistically significant additive release of both IL‐6 and IL‐8 (Fig. 9). Combinations of PAR‐3 or PAR‐4 agonist peptides with PAR‐1 or PAR‐2 agonist peptides did not cause any additive or synergistic cytokine release (Fig. 9).
PAR agonist peptides induce PGE2 release from HLF‐1 cells
Stimulation of HLF‐1 fibroblasts with PAR‐1 and PAR‐2 agonist peptides significantly increased the release of PGE2 by these cells. However, neither PAR‐3 or PAR‐4 agonist peptides nor the control peptides had any effect (Fig. 10). PAR agonist peptide‐induced PGE2 release was significantly inhibited by pretreating the cells with 10 μmol L−1 indomethacin (Fig. 10).
We have been able to demonstrate the expression of PAR‐1, PAR‐2, PAR‐3, and PAR‐4 using RT‐PCR, immunocytochemistry, and flow cytometry. Previous studies by Sokolova et al. (2005) have demonstrated detection of PAR expression using immunohistochemistry, while others have examined the expression of PAR mRNA levels using RT‐PCR (Ramachandran et al. 2006; Ortiz‐Stern et al. 2012). In line with previous observations (Ramachandran et al. 2007) PAR‐4 did not appear to be abundantly expressed on human lung fibroblasts unlike the other PAR receptors. We also showed that PMA, an activator of protein kinase C (Saitoh and Dobkins 1986) increased PAR‐4 mRNA transcripts in human lung fibroblasts. These findings, coupled with the lack of functional responses to PAR‐4 agonist peptides in naive HLF‐1 cells, suggest that PAR‐4 may not play a significant role in protease modulation of lung fibroblast function during tissue homeostasis. However, inflammatory trauma and pro‐inflammatory cytokines such as TNFα and IL‐1β may increase PAR expression in various cells including skeletal muscle, airway epithelium, and endothelial cells (Nystedt et al. 1996; Mbebi et al. 2001; Lan et al. 2004).
The activation of PARs by thrombin and trypsin, as well as the peptide activators of PAR‐1 and PAR‐2, was accompanied by changes in intracellular Ca2+ in lung fibroblasts which is consistent with PAR‐mediated signaling via Gq. As some PAR‐1 peptides have previously been shown to cross‐activate other PARs (Howell et al. 2001), cross‐desensitization experiments between PAR‐1 and PAR‐2 were conducted, and it was demonstrated that responses to the PAR‐1 (TFLLRN) and PAR‐2 (SLIGRL) agonist peptides were significantly attenuated when the cells were pretreated with thrombin and trypsin, respectively.
α‐SMA expression is an indicator of actively proliferating fibroblasts, and a distinctive marker of myofibroblasts differentiation. We now demonstrate that the agonist peptides to both PAR‐1 or PAR‐2, but not PAR‐3 nor PAR‐4, stimulate α−SMA expression in human lung fibroblasts. Similar results have been obtained by Bogatkevich (Bogatkevich et al. 2009) demonstrating that inhibition of thrombin leads to prevention of α‐SMA expression in human lung fibroblasts in a PAR‐1‐dependent manner while Borensztajn's work has indicated that PAR‐2 activation can also lead to α‐SMA expression in vivo (Borensztajn et al. 2010). Increased numbers of α‐SMA‐ expressing myofibroblasts have previously been reported in the airways of subjects with asthma and idiopathic pulmonary fibrosis (Brewster et al. 1990; Saunders et al. 2009). Although important in wound healing (Thannickal et al. 2004), myofibroblasts are the predominant pulmonary cells contributing toward increasing submucosal thickness and collagen deposition in the airway wall of asthmatic subjects (Roche et al. 1989; Brewster et al. 1990; Ward and Hunninghake 1998). In asthma, both the expression of PARs (Knight et al. 2001), as well as the concentrations of PAR activators such as thrombin (Gabazza et al. 1999), are increased in the airways, and thrombin inhibitors have been shown to attenuate airway wall remodeling in a murine model of bleomycin‐induced pulmonary fibrosis (Howell et al. 2001). Our findings suggest that PAR‐differentiated myofibroblasts are likely to promote tissue repair processes, but in chronic airway inflammatory disorders, myofibroblasts may adversely contribute to airway hyperresponsiveness and irreversible airway wall remodeling through increased collagen secretion and cellular proliferation (Darby et al. 1990; Phan 2002).
There is increasing evidence to suggest that fibroblasts play an important role in lung inflammatory processes. In this study, we showed that the activation of both PAR‐1 and PAR‐2 stimulated the release of the immunomodulatory mediators IL‐6, IL‐8, and PGE2, with the PAR‐1 agonist peptide exhibiting greater potency than PAR‐2. Mediator release induced by either PAR‐ 1 and PAR‐2 agonist peptides was shown to be additive, with the secreted levels of IL‐6 and IL‐8 being significantly higher when the cells were stimulated with both PAR‐1 and PAR‐2 agonist peptides in combination, compared to the singular effects of PAR‐1 or PAR‐2 agonist peptides.
These findings were similar to our previous observations in airway epithelial cells (Asokananthan et al. 2002), and along with the present data using cross‐desensitization Ca2+ assays, we confirmed that the PAR agonist peptides used in this study were acting through distinct receptors, PAR‐1 and PAR‐2. Similar results have been obtained in a study using thoracotomy samples, where PAR‐2 agonists upregulated granulocyte colony‐stimulating factor, IL‐8 and VCAM‐1 in bronchial fibroblasts (Ramachandran et al. 2006), while another group found thrombin stimulation of human lung fibroblasts via PAR‐1 promoted secretion of CCL2, a chemotactant which acts on monocytes, natural killer cells, T cells, and fibrocytes (Ortiz‐Stern et al. 2012). Sokolova and colleagues also found similar effects of trypsin and thrombin stimulation of lung fibroblasts on calcium flux and PGE2 (Sokolova et al. 2005).
Interestingly, both the PAR‐3 and PAR‐4 agonist peptides were inactive. Indeed, it was previously demonstrated that PAR‐3 is not functionally coupled to intracellular signal transduction pathway, but rather functions as a cofactor for the thrombin‐sensitive receptor, PAR‐4 (Nakanishi‐Matsui et al. 2000). However, as PAR‐4 is not abundantly expressed in human lung fibroblasts, the functional significance of PAR‐3 in these cells is currently not known. It is of interest to note that the PAR‐3 peptide may have other biological effects, such as stimulating intracellular Ca2+ changes in rat astrocytes and human vascular smooth muscle (Wang et al. 2002; Bretschneider et al. 2003) and there is recent evidence to suggest that it may cross‐activate PAR‐1 and PAR‐2 (Hansen et al. 2004).
Proteases such as thrombin and trypsin detected in the airway may be expressed by resident pulmonary cells. For example, the expression of immunoreactive thrombin in alveolar macrophages has been reported (Howell et al. 2002), while trypsin has been detected in the respiratory epithelium (Cocks et al. 1999; Danahay et al. 2001; Cederqvist et al. 2003; Jin et al. 2003; Miki et al. 2003). Furthermore, it is established that the concentrations of these proteases are elevated in the bronchoalveolar lavage fluid in diseases such as asthma (Gabazza et al. 1999; Terada et al. 2004). Based on these findings, PAR activation by secreted proteases on lung fibroblasts is likely to have profound implications on the pathogenesis of asthma. In addition to cellular proliferation (Trejo et al. 1996; Akers et al. 2000) and pro‐collagen secretion (Chambers et al. 1998), the simultaneous activation of PAR‐1 and PAR‐2 on lung fibroblasts is likely to have an additive effect on the inflammatory process in asthma. Furthermore, the differentiation of these cells into myofibroblasts via PAR activation is also likely to compound the disease process toward irreversible airway wall remodeling.
In conclusion, we report the presence of PAR‐1, PAR‐2, PAR‐3, and PAR‐4 in human airway fibroblasts. PAR‐1 and PAR‐2 expressed by these cells are functionally responsive to thrombin and trypsin, respectively. As PAR subtypes exhibit varying activation sensitivities to different proteases, the expression of multiple PARs within a single population of cells may discriminate different proteases in the extracellular space (Shapiro et al. 2000; Wang et al. 2002). Moreover, the activation of PAR‐1 or PAR‐2 in airway fibroblasts promotes myofibroblast differentiation (as indicated by increased expression of α‐SMA expression) and the release of immunomodulatory mediators such as IL‐6, IL‐8, and PGE2. The release of cytokines induced by PAR agonist peptides was both time and concentration dependent. Our findings are strongly suggestive that PAR activation is likely to perpetuate airway inflammation and tissue remodeling, two features which are present in airway disorders such as asthma.
Conflict of Interest
None of the authors have a financial relationship with a commercial entity that has an interest in the subject of this manuscript.
Funding Information
This work was supported by the National Health and Medical Research Council (NH&MRC) and the Healy Medical Research Foundation.
N. Asokananthan and R. S. Lan contributed equally to this work.
- Manuscript Received: November 7, 2014.
- Manuscript Revised: January 11, 2015.
- Manuscript Accepted: January 13, 2015.
- © 2015 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.


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The Los Angeles Clippers bowed out of the playoffs early once again this season, this time in a seven-game series against the Utah Jazz in the first round. Now, the team faces a big offseason in which stars Blake Griffin and Chris Paul can opt out of their contracts and join JJ Redick as free agents. The core of this Clippers recent run of success has come on the back of the Paul/Griffin/DeAndre Jordan combination, so there's some big questions looming.
Some folks in the NBA community have advocated breaking up that core of the team by "blowing it up" this offseason either via sign-and-trade or by simply letting those stars walk. Others, however, think the Clippers need to just keep their guys this offseason.
"I think the Clippers in the offseason, they don't really have a lot of work to do other than to keep their guys," said ESPN NBA insider Ramona Shelburne on CBS Sports Radio's After Hours with Amy Lawrence show. "They just have to do whatever they can to keep all of their free agents because the way that free agency is set up and also, the way their cap situation is set up, they can't replace anyone. If you lose them, you have no money and absolutely no flexibility to go and replace them, it's just a hole. As you can tell, they already don't have enough, so you don't want to dig another hole. You can kind of deal with the fallout later. I think we've all been asking the wrong question of should they blow it up? They don't have a choice, they have to keep them together. If even one of those guys leave, they're back to before Blake and Chris even got there. And, they're worse off because they don't have any draft picks.
They don't have any young guys, they don't have any draft picks. If they lose any of these guys, they can't replace them. In Oklahoma City, yeah that's tough they have to find people to put around Russ who can fit his game and be complementary to him. They started doing that already this year in the season with some of the trades they made, but at least you have flexibility, at least you have players that other teams want, at least you have options. The Clippers, the only option they have is to play defense to keep their guys."


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For sale by kobby 29 Dec 2:09 amSouth Dayi, Volta
Double door sharp refridgerator for quick sale.bought new and used slightly but still looks new and fresh and problem free.it is energy efficient and chills and freezes like shit.Am selling because I need money for fees.
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Got a few bucks to spend on your wardrobe? Save it. For the next few weeks, at least. Retailers citywide are moving up sales that would normally wait until December. A clerk at Jeffrey told us today to check back in two weeks — very early for the exclusive boutique.
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Desktop Client FeedDemon Will Support Google Reader Synchronization
FeedDemon — one of the best windows OS based rss desktop clients which synchronize rss feeds and lets you give you websites updates on your desktop, you can instantly read lots of websites changes/updates and recently they connecting this synchronization with Google reader. You already have experiences with bloglines, pageflakes and netvibes; all the web based service which synchronize data from your desired websites through rss feed and you can get access in web interface, not your desktop.
The latest version of FeedDemon 2.7 is available in download page but in FeedDemon 3.0 you might see this news functions and still in beta version. In before it has one options for sync, it only syncing in between of the software client and with newsgator.com server. Now, it will connect your account with Google reader, lets you options to connect and you may as well sync in between of feeddemon client plus google reader.
Before you go forward, in both site newsgator and on google reader you need accounts. As because you have to use google account to sync with that service and have to give the access permission to the desktop client from your PC. When you making any changes in your google reader account, it will automatically changed to your newsgator accounts too. Also when you're changing anything else to your desktop client "on feeddemon" it will updated to google reader. Thanks to Steve to let us know about the upcoming great features.


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Today's M3talinks for May 14th
- Pretty Link WordPress Plugin — Blair Williams – Shrink, track and share any URL on the Internet from your WordPress website. Unlike other link shrinking services like tinyurl, budurl, and bit.ly, this plugin allows you to create shortlinks coming from your own domain! Pretty Link tracks each hit on your URL and provides a full, detailed report of where the hit came from, the browser, os and host. Pretty Link is a killer plugin for people who want to mask their affiliate links, track clicks from emails, increase the reach of their website by spreading these links via Twitter, forums or comments on other blogs.


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At Pumptheory we love software so much we talk about it in our spare time and give it away for free!
Pumptheory is with pumptheory's Mark Aufflick heavily involved in the Australian iOS and Macintosh development community and is the main organiser of CocoaHeads and NSCoder's Sydney chapters where he has also been known to give the occasional presentation. Mark provides various pieces of opensource code to the Cocoa community, such as Net::APNS — a Perl module which manages communication with the Apple Push Notification Service.
Mark is also strongly involved in the global Perl community, developing and maintaining a number of CPAN modules and being a frequent contributor to Perlmonks.org.
In recent times Mark was a significant contributor to the OpenACS web framework community. OpenACS remains one of the richest online community frameworks available, opensource or commercial.
You can see some other opensource and community contributions by Mark on his personal website at http://mark.aufflick.com/software/.