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Product Description
Motorola 420-440 MHz UHF Super Stubby Antenna
Compatible with: EVX-S24 Radios
Color: Black
Warranty: 1 Year
Height: 2" (5.08 cm)
Stubby Antennas are great for belt-worn radios - the shorter antenna prevents snags; to keep the user moving freely. This super stubby is so small, the radio can be even be placed in the users pocket, for complete discretion.
Additional Information
|Part Number
|What's in the Box
|1 Antenna
|1 Year
|Compatible With
|Motorola EVX-S24


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BOSTON, Mass. — Three teams from the Vergennes Union High School rowing club each posted top-three finishes in their divisions at the Nov. 17 New England Youth Rowing Championships held in Boston, Mass., at which 28 teams in all competed in three divisions.
According to team coach Dennis West, in each division the teams raced three times, and the times from the three races were combined to determine the winners.
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Women's Basketball
11/20 Smith vs. Midd........................... 80-47
Men's Basketball
11/20 Midd. vs. Green Mt. ................ 117-81
Men's Hockey
11/30 Conn. College at Midd. ............. 7 p.m.
12/1 Tufts at Midd. .............................. 4 p.m.
When I look out across the economic horizon I feel concerned. The issues are so huge and advice so plentiful I hardly feel there is hope for change. Then I pull back and say "I'm not going to take in all the issue but what do I have control over today that in some small way has a positive impact?"
This week's writer is Mike Palmer, president of Ethics By Design, which is a Middlebury company that helps nonprofit organizations, commercial enterprises and government agencies set up effective ethics programs.
ADDISON COUNTY — Will Stevens — a state representative and co-proprietor, with his wife Judy, of Golden Russet Farm — stands in the kitchen of his Shoreham farmhouse, talking to a customer on his landline wall phone. He confirms the produce pick-up schedule, then stops mid-sentence, frowning.
"Hello?" he says, then hangs up with a resigned shrug.
FERRISBURGH — After more than a decade of planning and effort and a few setbacks along the way, the 13-year-old dreams for Ferrisburgh's Rokeby Museum to become a major Underground Railroad history site and dramatically expand its ability to handle visitors is a giant step closer to reality.
Brandon general contractor Naylor & Breen Builders recently finished a two-story, roughly $1.5 million visitor center on the Route 7 site that will go by the name the Rokeby Museum Underground Railroad Education Center.
MIDDLEBURY — Peter Padua hadn't thought much about testing his artistic abilities until he was a senior in high school in New Jersey. It was 1941, the same year as the Japanese attack on Pearl Harbor, and Padua — who enjoyed his wood shop class — thought he would try his hand at his first carving: A mountain deer, her head craned way back as if staring at the stars.
MIDDLEBURY — Carol Peddie was closing in on her 50th birthday in 2009 and decided it was time to make some changes in her life. One of the biggest changes was in how she would define the word "vacation." To Peddie, a vacation now means traveling throughout the world improving the lives of strangers less fortunate than she.
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New FEMA Base Elevation Requirements are Released
Last Updated on 19 December 2012
This past weekend, FEMA released the new
flood elevation maps
for those areas that were affected by Super Storm Sandy. Here you will be able to search by address and find out what the
Advisory Base Flood Elevation
(ABFE) is for that property. It is choked full of other information as well. After entering the address, and clicking "Get Details" you will not only get the new AFBE, it will also give you the old base elevation, as well as what Flood Zone the property is in. There are also numerous links to other reports and information regarding the elevation of the property.* While this is going to apply to any new homes that are built in flood zones, it will also apply to those homes that the are deemed to "substantially damaged" by the Township in which the property is located. On the
Long Beach Township
home page, they explain this a little more in the "Repair Your Flood Damaged Home" section about a quarter of the way down the page. They also explain that if the home is deemed to be substantially damage, you must now meet the the required flood elevation for that location. They also give some guidance as to what the Township will deemed as substantially damaged. Of course each town's requirements may be slightly different, but this is an example what homeowners need to take into consideration when they are deciding what to do with their damaged homes. *
Update as of December 19, 2012 – All owners should click on the "Link to AFBE Map" to view the actual map. It gives two different numbers as a base elevation, the 1% Elevation and the 2% Elevation. Here is a link to the
, but its essentially the percentage chance that the elevation will be breached by water in a given year. The general difference in feet between the two number are 4-5 feet. For example, our LBI office has a 1% advisory base elevation of 9 feet, and a 2% advisory base of 13 feet. These are the numbers to pay close attention to. We are hearing that Long Beach Township will adopt the 2% ABFE in their buiding ordinances, which means new homes and those deemed substantially damaged will have to raise their house to the 2% elevation. Regarding what is deemed substantially damaged, we are hearing that if the damage to your home is 49-50% of the buildings value, it will be deemed substantially damaged and be forced to comply with the flood regualtions. On Friday, Dec 21, 2012, Long Beach Township is expected to introduce its new FEMA ordinances, so stay tuned…
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Near Constantine
Locals sometimes refer, half-
The property offers six guest rooms in all, while there's also a recently built Garden Annexe around a hundred feet from the main building itself.
Being a free house, the Trengilly offers a regularly changing array of unusual ales as well as a wide selection of wines and malt whiskies – it's as popular with local residents as guests. Food is available both in the bar and the quieter bistro.
Once a modest farmstead (probably dating from the late eighteenth or early nineteenth centuries), its name means "settlement above the trees" in Cornish. Indeed, its surroundings have probably changed very little since it was first built.
The thriving village of Constantine is only a mile away, while the pretty waterside hamlet of Port Navas is a little further to the east. There are also several places to visit nearby, including the National Seal Sanctuary at Gweek and the world-
Well behaved dogs are welcome for a small supplementary charge.
Reservations can be made via Booking.com and LateRooms.com


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Table Mountain Veterinary Clinic is looking for a Full Time Associate Veterinarian for our very busy 5 doctor practice. Our practice has multiple opportunities to see a variety of cases. We have digital radiography, modern surgical and dental equipment, and low turnover in our staff. We utilize a radiologist for ultrasounds in house, and to read our films. We bring in a surgeon and a cardiologist regularly. We are growing and need to fill this position with someone who can jump in and begin seeing clients right away. We do have mentorship available for surgery and dentistry cases for a newer grad.
We are located in beautiful Golden, Colorado, a very desirable place to live. Biking, hiking, skiing, camping, all things outdoors in our spectacular mountains!
Our caseload offers a variety of general practice cases, medical and surgical, and includes the following:
Job Summary
In addition to the responsibilities outlined below, the Associate Veterinarian is an invaluable member of the hospital team who works directly with a the Hospital Manager and supporting medical staff. This individual advances the quality of medicine in the practice and provides exceptional client service to our clients and patients.
Job Responsibilities
Skills and Basic Qualifications
Equal Employment Opportunity
It is the policy of the company to afford equal opportunities to all applicants and employees regardless of race, color, religion, sex, national origin, age, non-disqualifying disability or status as a disabled or Vietnam era veteran.


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The browser wars are about to heat up again. This time, however, the battle will take place in the mobile space.
This week, ABI Research released a report that said, in essence, that the fun is about to start. The analysts say that people generally don't browse much on mobile devices. At the same time, smart phones have great browsing capabilities and an increasing number of sites are customized to serve mobile surfers. The release goes into good detail on the current status of mobile browsing. While the technology pieces all are not quite there yet, they are rapidly falling into place.
Signs of interest are all around. Opera Mini has been downloaded about 40 million times for use on smart and feature phones. Earlier this week, Opera offered developers access to a version of the Mini customized for Google's nascent Android platform. The story -- which links to the download site -- notes that Android phones are not yet available.
It's been a busy month for Opera. Last week the company rolled out its beta of version 4.1. A PDA Street story highlights some features, which include faster downloads, searching within Web pages and others.
As if to set the stage for the coming battle, StatCounter has released statistics on mobile browser usage. The iPhone/iPod Touch is the most-used mobile browser, according to a story at AfterDawn. The browser -- which is a version of Safari -- is ahead of Symbian in the U.S. but trails it worldwide. Windows Mobile is in third place in the U.S. and overseas.
The stats show much room for growth: iPhone/iPod accounts for only 0.23 percent of browsing in the U.S. The firm says that 0.19 percent of browsing worldwide is done by iPod/iPhone, while 0.25 percent of browsing is done on Symbian run devices. By all accounts, not much mobile browsing is happening.
If all that isn't enough to convince observers that a serious mobile browser war is upon us, how about this: At CTIA 2008 in Las Vegas earlier this month, Microsoft said it will offer "full Web browsing" on cell phones this year. This Reuters report says that the company will make Internet Explorer Mobile available to phone vendors during the third quarter. In a related development, the company announced the Windows Mobile 6.1 operating system.
Aside from the particulars raised in these reports, the focus on mobile browsers just makes sense: People love the Internet and are increasingly mobile. Right now, mobile browsing is at a relatively rudimentary stage. Look for that to change very soon.
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Weight loss has been a struggle among most people around the country. This is due to the unfruitful programs and medications that are numerous in the market on https://nutrimost.com/members/testimonials/. Luckily, there has been a program that has helped people lose weight successfully and maintain it henceforth. Nutrimost has been gaining popularity constantly due to its success.
Dr. Rob Vasquez, who is based in San Antonio, used Nutrimost to overcome various chronic medical conditions on nycfatloss.com he had been struggling with and lose weight. He used the technique to overcome headaches, asthma and allergies. Vasquez explained how he had struggled with weigh loss in his adult life by engaging in every imaginable weight-loss program on nutrimostrecipes.com with no success. He said that he only found a long lasting solution after engaging in Nutrimost.
Vasquez said, "I found Nutrimost, did it, and I lost 35 pounds in 40 days. The best part is that I lost over 60 pounds, and that was a year ago, and I've been able to keep it off." He sought to use his experience to help his clients live a healthy life. Vasquez has used Nutrimost to help the people in San Antonio. People who have enrolled in this program have reported higher energy levels, better control of chronic illnesses such as Diabetes Mellitus especially Type 2, hypertension and chronic pain.
Nutrimost Mechanism
Nutrimost has been successful because of its strategy in tackling the various causes of weight gain/ obesity. Each person has a different cause of weight gain and this system targets the individual cause to fight this condition. The goal of Nutrimost is to help its clients achieve a healthy lifestyle. Generally, it balances hormones on http://nutrimostrecipes.com, increases metabolism level of a person and balances neurotransmitter. More uniquely, it sets the weight point of a person such that they won't struggle with weight gain again.
Information on this article was published originally in KSAT12 news site (http://www.ksat.com/sa-live/sa-doctors-weight-loss-story-inspires-him-to-help-others-lose-weight) and The Grand Island Independent site (http://www.theindependent.com/news/business/nutrimost-nebraska-program-offers-customers-a-path-to-better-health/article_9d366bee-4947-11e5-af54-dfb77f991d1d.html)


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Thank you for visiting today. If this is your first visit, take your time and look around. I have plenty of information and resources available to you. If you are a return visitor, thank you. I would love to hear from you and tell you how I can serve all your real estate needs.
Sold for $395,000 on Mar 29, 2019
Super cute house on quiet street, close to parks. Fully renovated, 3 finished levels. New Kitchen, new bathrooms, new windows. Very Convenient.
The Key to Your Real Estate Success®
Damian Buckley | Office 301-907-7600 | Direct 301-215-4748 | Contact Us
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To start off, I'm an idiot. I did a very, very, very stupid thing. I converted my hardtail to SS and I love it. I was running a fixed tensioner... the Sette Chain Guide II. Worked nicely, did the job. Well I couldn't find my 6mm wrench to torque the hanger bolt down to secure the tensioner, so I just quick swapped it out with a hex bolt I had with matching threads. I tightened it down, went for a ride, came back, messed around with my debate between my spring tensioner vs my fixed tensioner, etc... couple screws/unscrews later I was torquing the hanger bolt down to secure the fixed tensioner and out came the threads. How nice. I guess I should have been more aware of the fact that a socket wrench could apply a bit more torque than a small handheld allen key.
Called a local Diamondback dealer for a derailleur hanger, said it'd be a 50/50 shot if it comes in today. They closed 3 minutes ago. I'm out of luck for the hanger. But me and a few buds were excited about riding tomorrow at a new trail we found. I was pretty bummed, considering it's the only working bike I have right now, so I thought Saturday would be a dull day for me.
But instead I busted out some wire ties from my toolbag from work and wire tied the tensioner in the up position. Went for a 5 min ride around the block. I had one skip which I can't seem to duplicate no matter how hard I ride it. The chain is tight so I have no idea how it happened! Other than that, it held up nicely.
Where's the trail at? I'm ready.


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You may be very upset with the fact that you do not have a good place to store all your wine bottles. You may have a nice home but do not have a wine cellar. How many wine bottles would you need to have stored? If you only need a dozen wine bottles stored in a nice, cold place for long periods of time, it could be very wise for you to check out the many wine coolers that are currently available on the market. To be honest, it should not be very difficult for you to find a very nice wine cooler. You should be able to find out which wine cooler brand might be the best for you to purchase. Are you willing to go online and find the ideal wine cooler for your home? If this is the case, it could be very wise for you to check out all Danby wine cooler reviews.
Yes, you may be able to figure out which of the many wine coolers could be exactly what you want. You will need to check out each wine cooler and its specifications. You can get a wine cooler that can hold a maximum of 12 wine bottles at once. Actually, you can find Danby wine coolers that can hold 34 bottles of wine. There is another wine cooler that can hold a maximum of 75 wine bottles. You will need to figure out which wine cooler would be exactly what you need to have in your wine. If you need to have many wine bottles stored in the cold over long periods of time, you should never check out the one that can hold 75 bottles of wine. I hope you do not have much trouble finding the ideal wine cooler that will look great in your home.


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Dear Friends,
we are happy to share with you the Menu we will serve for dinner "at Palazzo" in Venice, next Friday, 4th September 2015.
Last few slots are available…don't miss it!
If you want to make your reservation, contact us at firstname.lastname@example.com.
See you in Venice!
Metti una sera a cena SupperClub


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Looking over my story about Samira Soueidan, I realized I left out some vital information about why the Justice Ministry would appeal such a case. Do they not like Egyptians? Do they not like widowers?
Those who've followed my reporting on the issue will know that the appeal is actually more or less framed around the idea of the 400,000 or so Palestinian refugees being resettled in Lebanon.
Lebanon has always been paranoid that Palestinian refugees will be granted Lebanese citizenship and resettle here, upsetting the demographic balance in this hysterically confessional country. The argument put forward by officials in public is that they don't want the Palestinians to relinquish their right to return to the country they were expelled from. (This is coincidentally one of the main reasons Lebanon also does not grant the Palestinians the right to work in most professions).
But scrutinize the issue and you will find this policy is nothing but lies. Christian Palestinians were granted Lebanese citizenship a long time ago, simply because they would bolster Christian numbers. So it has nothing to do with the rights of Palestinians to return to their occupied country and everything to do with sectarianism. By granting Soueidan the right to nationality, the Justice Ministry saw a dangerous precedent being set that could pave the way for all stateless children and husbands to demand their right to citizenship.
It's especially difficult to comprehend the logic behind Lebanon's sexist nationality laws when looking at the facts on the ground: According to a UNDP-funded study, only 2% of all Lebanese women married to non-Lebanese are married to Palestinians. This shows the law for what it is:deeply flawed and only existing to serve the patriarchal sectarian system.
Nationality and the Palestinians8 Jun
Looking over my story about Samira Soueidan, I realized I left out some vital information about why the Justice Ministry would appeal such a case. Do they not like Egyptians? Do they not like widowers?
Rich old widows and manipulative foreign men3 Jun
NGO hits out at General Security over nationality laws
By Dalila Mahdawi and Carol Rizk
Daily Star staff
Friday, May 21, 2010
BEIRUT: A Lebanese non-governmental organization (NGO) lashed out on Thursday at recent comments by the director of General Security, Wafiq Jezzini, accusing him of "humiliating" racism and sexism.
The Collective for Research and Training on Development-Action (CRTD.A) also asked the Lebanese government to clarify what progress had been made in enacting a decree granting free of charge residency permits with up to three years validity to the non-Lebanese husbands and children of Lebanese women.
The decree, proposed by Interior Minister Ziyad Baroud, was approved by Cabinet on April 21, but has not yet come into effect, leading human rights activists to suspect it was being delayed on purpose.
Last week, Jezzini told the Cabinet Baroud's decree contravened Lebanon's labor laws and accused non-Lebanese husbands of Lebanese women of entering the country illegally and marrying much older "rich widows" to financially exploit them.
Jezzini, whose remarks were published by Al-Akhbar newspaper on May 14, also claimed that granting complementary residency permits to the non-Lebanese husbands and children of Lebanese women would lead to "social problems."
Lebanese law permits men to pass on their nationality to their non-Lebanese wives and children but bars women married to non-Lebanese from doing the same. Deprived of state protection and recognition, those without citizenship live in a precarious legal vacuum and cannot benefit from state education or health care, work in the formal economy or vote.
Non-Lebanese husbands and children must apply for costly residency permits on an annual basis or face imprisonment and deportation.
"Giving complementary residency permits would encourage these people to enter Lebanon on the pretext of tourism or work and then not leave," Jezzini said. "They marry Lebanese women to benefit from the provided facilities and nothing more, and this can lead to social problems and hurt society and the economy."
He added: "[General Security] has mentioned in previous correspondences that … Lebanon has become a target country for immigrants. This flow is either legal or clandestine … [and] has led to a relatively large number of foreigners living illegally in Lebanon, many of whom – notably Egyptians, Iraqis and Syrians – marry Lebanese women and have children even if they are already married in their native country.
"They do not take age differences into consideration and sometimes marry rich widows because they are looking for a refuge or a way out."
Roula Masri, gender program coordinator at CRTD.A, said Jezzini's tone was "humiliating" and "totally offensive." Jezzini was suggesting that foreign men come to Lebanon to find "old and unmarried women," she told The Daily Star. The security official also suggested that Baroud's decree "would give working class men the right to come and marry women who have passed the suitable marriage age and to exploit them," Masri said.
CRTD.A asked the government to elucidate what progress it had made toward ratifying Baroud's law. "It's been a month since the endorsement so it's unusual that it's not yet passed into effect," Masri said, adding that most laws only need two or three weeks to enter into force.
The NGO also issued a statement responding to Jezzini, saying his comments were "offensive to Lebanese women, their husbands, and to the working class."
It added: "The head of General Security should not have generalized but should rather have focused on determining clear and transparent standards. He should also not have interfered in the personal affairs of the right of Lebanese women to choose their husbands."
Jezzini's comments were especially offensive as "dozens of families live in constant fear of being deported," CRTD.A said. According to Masri, the Iraqi husband of a Lebanese woman was deported on Sunday even though his papers were in order.
(The Daily Star :: Lebanon News :: http://www.dailystar.com.lb)
Inside the mind of a Lebanese politician3 Jun
In all countries of the world, politicians are meant to serve their constituents. In Lebanon, politicians (except a minor handful) serve themselves and then, if they have enough energy to spare, their religious community. Samira Soueidan, a Lebanese mother of four, apparently committed a crime when she married an Egyptian man. Her husband is now dead but she continues to pay for this crime- the Lebanese government does not recognize women's right to pass on citizenship and so her children are stateless. They have never been to Egypt and do not have nor wish for Egyptian citizenship. They are Lebanese to everyone except the State.
Samira wanted recognition her children are Lebanese, and so she went to the courts. The first court said yes, her children should be granted citizenship. Then the Justice Minister decided to appeal. I repeat, the Justice Minister. He wins, Samira's children and by extension all Lebanese women lose. The irony is too much to bear.
If you want to read more about the case, here it is:
Wednesday, May 19, 2010
BEIRUT: A landmark ruling which granted citizenship to the children of a Lebanese mother was overturned by an appeals court Tuesday in a move that has left human rights activists reeling.
Samira Soueidan filed a lawsuit five years ago demanding her four children be granted citizenship rights following the death of her Egyptian husband. Lebanese law permits men to pass on their nationality to their non-Lebanese wives and children but bars women married to non-Lebanese from doing the same.
Leaving the courthouse in Jdidet al-Metn, Soueidan said she had "lost the battle but not the fight" and vowed to take her case to the Court of Cassation, Lebanon's highest appeals court. "I'm not going to stop here," she said, adding that her children had been born and raised in Lebanon and should be viewed as Lebanese.
In a breakthrough ruling last July, Judges John al-Azzi, Rana Habka and Lamis Kazma ruled in favor of granting Soueidan's two sons and two daughters citizenship. The judges said their decision was based on the fact there was no law prohibiting a Lebanese mother from passing on her nationality to her children after the death of her husband. They also noted that aspects of Lebanon's nationality law were "obscure" and that current legislation favored foreign women over their Lebanese counterparts.
The verdict was applauded by civil society organizations and suggested a breakthrough for thousands of other families suffering because of the sexist legislation. Ironically, however, it was appealed by Justice Minister Ibrahim Najjar.
The Court of Cassation, presided over by Judge Mary al-Maouchi and two other women consultants, accepted Najjar's appeal and overturned Azzi's ruling, saying it contravened Articles 3 and 537 of Lebanon's Civil Law code and the nationality law. "Judicial courts are not concerned with granting nationality rights [in cases where it was not granted at birth] as this is a right only enjoyed by the president," stated the 17-page ruling, a copy of which was obtained by The Daily Star. Soueidan will have to pay all the legal fees incurred in the case.
Representatives from the Collective for Research and Training on Development-Action [CRTD.A], a Lebanese non-governmental organization that has been supporting Soueidan in her fight, said they were disappointed but "unsurprised" by the legal setback. "The approach of the Lebanese government since Azzi's ruling has not been encouraging," said Roula Masri, CRTD.A gender program coordinator.
She claimed the judge had been subject to harassment and "humiliation" from leading officials in the Justice Ministry.
Azzi was also reportedly unofficially banned from talking to journalists, who were required to submit interview requests to the Justice Ministry. A request submitted by The Daily Star several months ago was never answered.
There are around 18,000 Lebanese women married to non-Lebanese in Lebanon, according to a study by the UN Development Program. Around 80,000 children and husbands are potentially made stateless by the current legislation.
Deprived of state protection and recognition, those without citizenship live in a precarious legal vacuum and cannot benefit from state education or health care, work in the formal economy or vote. They are vulnerable to arbitrary detention, have difficulty accessing the legal system and live under constant fear of deportation.
Lebanon's nationality law was formulated in 1925, at a time when Beirut was under French mandatory rule. "It's about time to amend such an outdated law," Masri said. That the law continues to be applied flies in the face of "claims that we live in a democratic country that can compete in the international arena."
A 1994 amendment of the nationality law gave the child of a Lebanese mother and foreign father the right to obtain citizenship. But in order for it to be granted, the child must marry a Lebanese citizen and live continuously in Lebanon for at least five years, including one year after marriage.
Tuesday's ruling, together with the discriminatory nationality legislation, contradicts Article 7 of the Lebanese Constitution, which guarantees men and women equality before the law, Masri said. "The only way women can win their rights is by amending the current nationality law," she said, adding that CRTD.A would respond to Soueidan's defeat with a major public event. Masri also decried the fact that Soueidan's hopes for nationality rights were thwarted by women judges. "It's a pity to find women not supporting other women's rights."
Combatting a memory for forgetfulness3 Jun
Sodeco's war-weary Barakat building to be renovated
Structure to house public memorial to civil conflict
By Dalila Mahdawi
Daily Star staff
Wednesday, June 02, 2010
BEIRUT: For years, architect Mona Hallak lugged a beautiful antique floor tile around Beirut, eager to show it to anyone who would spare her a few minutes of their time.
She had pinched the tile from a war-ravaged building in Beirut's Sodeco Square that she'd lost her heart to in the 1990s.
Even in its decrepit state today, it's not hard to see how it captivated her. The architecture of the Barakat building, an imposing yellow structure straddling Independence Avenue and the Damascus Road, is one of a kind in Lebanon.
Built by two different architects in the 1920s and 1930s, the Barakat building mixes elements of Art Deco with sweeping Islamic arches, stain glass windows and oriental balustrades. Although it is impossible to tell from outside, the four-story building actually consists of two distinct structures unified by freestanding balcony columns. "The building symbolized the whole of Beirut," says Hallak. "It symbolized how divided the country was and how it was camouflaged as being united."
When sectarian tensions spilled over into Civil War in 1975, the Barakat building was one of the conflict's many casualties. It was taken over as a militia headquarters, with snipers taking advantage of commanding views from every room in the building to kill those on the streets below in relative safety.
The fighters added their own layer of architecture to the structure – concrete buttresses to fortify the walls and ceilings, sand bags, and lots of graffiti. Within a few years, the Barakat building, once a sign of liberalism, progressive thinking and cross-cultural dialogue, had been transformed into a pock-marked symbol of bitter hatred, division and ruthless killing.
When the war ended, the Barakat family hoped to cash in on the post-war construction craze and sold the building to a development company. It would have been demolished if one day in 1997, Hallak, who was passing through the area, hadn't looked up at the building's façade and noticed the iron railings from the balconies were missing. Hallak rushed inside to find workers preparing for demolition. "The tiles were piled in a corner ready to go and the destruction permit was hanging on the wall," she recalls. "I never thought this building would go down … I went crazy."
Slipping one of the tiles into her handbag, Hallak hurried to her office, rallied her colleagues and began a concerted media campaign to preserve the building.
The architect visited officials from the Culture Ministry, the governor of Beirut and foreign ambassadors hoping to find a sympathetic ear. She would pull out the filched tile and tell her audience, "this is the tile- imagine how beautiful the house is!"
After years of tireless campaigning and with the support of the Italian and French embassies, Hallak finally achieved what many had thought was impossible. Beirut Municipality revoked the demolition order and in 2003 expropriated the building.
The war-weary structure is now being renovated and converted into Beit Beirut, a museum of memory, war and contemporary history. Prime Minister Saad Hariri inaugurated the project in early April and actual restoration and construction is due to begin in October at an estimated cost of $10 million.
Its restoration and modernization is being carried out by architect Youssef Haidar, with technical assistance from the Municipality of Paris.
Once opened, Beit Beirut will be the closest thing Lebanon has to a public war memorial. Traces of the war like the fighter-built fortified walls will be preserved and incorporated as part of the museum's permanent exhibition. "When you are there you feel the futility of war," says Hallak. "It is exactly what a war memorial should be."
Although Beit Beirut will chiefly be a museum, it will be "much more" than that, says Haidar. He hopes the building will help the Lebanese confront and reconcile their painful past.
The revamped Barakat building will connect to a new edifice built on an adjacent lot through a large spiral staircase, with both structures boasting state-of-the-art solar power systems. Just as the Lebanese themselves should be, says Haidar, the building will look firmly into the future while paying tribute to its past. "We are dealing with the building as if it is a war wounded that is starting to heal again," says Haidar. "These traces cannot be erased, they are like scars."
The museum will have an auditorium for lectures and workshops for young people on issues relating to memory, history and war- issues Haidar says have not been addressed at all in Lebanon.
"We went from amnesty to amnesia," he says. "It's important that at Beit Beirut, we can make a start in order to be able to say 'never again.'"
Hallak envisions Beit Beirut as a living museum where visitors can interact and contribute to building up knowledge about their city. She's put forth a proposal to have a "Beirut for Everybody" section on the ground floor, where locals can bring in and exhibit anything from their grandmother's traditional Beiruti recipes to old cinema tickets. "We want to create a relationship between the city and the museum," she says.
In addition to a permanent installation of personal items collected from the building, Beit Beirut will also host rotating exhibitions by artists, architects and urban planners on themes relating to the war, public space and contemporary history.
"It will be a place that will teach Beirutis to love their city," says Hallak. "We don't love our city because we don't know it."
Haidar and Hallak also hope the success of the project will encourage municipalities across Lebanon to preserve other traditional buildings as Beit Beiruts.
"There are other buildings that can be worked in this way," Haidar says. "We don't want to just reduce the idea to this one building."
Although pleased with the renovations, Hallak has one minor criticism: she wishes a large ficus tree outside the Barakat building hadn't been chopped back.
Only a few feet tall when the war broke out in 1975, the tree had grown several stories high by the 1990s.
"The tree was the memory of the war," says Hallak. "That would have been the most romantic way to remember the war – with life."


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- Research
- Open Access
A 3-dimensional in vitro model of epithelioid granulomas induced by high aspect ratio nanomaterials
© Sanchez et al; licensee BioMed Central Ltd. 2011
- Received: 10 December 2010
- Accepted: 18 May 2011
- Published: 18 May 2011
The most common causes of granulomatous inflammation are persistent pathogens and poorly-degradable irritating materials. A characteristic pathological reaction to intratracheal instillation, pharyngeal aspiration, or inhalation of carbon nanotubes is formation of epithelioid granulomas accompanied by interstitial fibrosis in the lungs. In the mesothelium, a similar response is induced by high aspect ratio nanomaterials, including asbestos fibers, following intraperitoneal injection. This asbestos-like behaviour of some engineered nanomaterials is a concern for their potential adverse health effects in the lungs and mesothelium. We hypothesize that high aspect ratio nanomaterials will induce epithelioid granulomas in nonadherent macrophages in 3D cultures.
Carbon black particles (Printex 90) and crocidolite asbestos fibers were used as well-characterized reference materials and compared with three commercial samples of multiwalled carbon nanotubes (MWCNTs). Doses were identified in 2D and 3D cultures in order to minimize acute toxicity and to reflect realistic occupational exposures in humans and in previous inhalation studies in rodents. Under serum-free conditions, exposure of nonadherent primary murine bone marrow-derived macrophages to 0.5 μg/ml (0.38 μg/cm2) of crocidolite asbestos fibers or MWCNTs, but not carbon black, induced macrophage differentiation into epithelioid cells and formation of stable aggregates with the characteristic morphology of granulomas. Formation of multinucleated giant cells was also induced by asbestos fibers or MWCNTs in this 3D in vitro model. After 7-14 days, macrophages exposed to high aspect ratio nanomaterials co-expressed proinflammatory (M1) as well as profibrotic (M2) phenotypic markers.
Induction of epithelioid granulomas appears to correlate with high aspect ratio and complex 3D structure of carbon nanotubes, not with their iron content or surface area. This model offers a time- and cost-effective platform to evaluate the potential of engineered high aspect ratio nanomaterials, including carbon nanotubes, nanofibers, nanorods and metallic nanowires, to induce granulomas following inhalation.
- Granuloma Formation
- Mannose Receptor
- Intratracheal Instillation
- Carbon Black Particle
- Asbestos Fiber
Carbon nanotubes (CNTs) are a commercial success of nanotechnology with an expanding global market and potential applications for biomedical devices, imaging, drug delivery, tumor targeting, electronics, composite materials and sensing applications [1]. The continued economic success of this industry is threatened by uncertainty and controversy surrounding potential adverse environmental and human health effects of engineered nanomaterials [2].
CNTs have been shown to induce significant toxicity using in vitro cellular assays and several in vivo rodent bioassays [3]. In rodent bioassays using intratracheal instillation [4] or pharyngeal aspiration [5], multiwalled carbon nanotubes (MWCNTs) produced significant lung injury, epithelioid granulomas, and persistent interstitial inflammation and fibrosis. Granulomas are aggregates of epithelioid macrophages elicited by biopersistent irritants or microorganisms that are not killed during the acute inflammatory response [6]. Two major types of granulomas are non-immune or foreign body granulomas (FBG) and antigenic or immunologic granulomas [6, 7]. Foreign body granulomas are characterized by aggregated epithelioid macrophages and multinucleated giant cells [8]. If the stimulus is biopersistent, granulomas may persist for months or years and lead to tissue destruction and fibrosis [7].
A major controversy in nanotoxicology is whether the observed induction of lung inflammation and fibrosis following the bolus delivery of MWCNTs by intratracheal instillation or pharyngeal aspiration is an artifact associated with agglomeration of the carbon nanotubes prior to administration [5, 9]. However, a recent inhalation study [10] and a pharyngeal aspiration study using commercial samples of well-dispersed MWCNTs also induced significant lung inflammation characterized by multifocal granulomas [11]. The specific physicochemical properties of carbon nanotubes responsible for lung toxicity are unknown. It is uncertain whether these pathologic responses are due to bioavailable metal catalyst residues, graphitic or amorphous carbon residues, length, geometry, high surface area, three-dimensional structure, agglomeration state, or some unique property of the carbon nanotube graphenic surface [12, 13].
There are numerous animal models of immune-mediated granulomas. In vitro models of immune-mediated granulomas have been induced by exposure of macrophages to pathogens or pathogen-coated beads. Macrophages exposed to heat-killed Candida albicans [14], Mycobacterium sp. [15], parasites such as Nippostrongylus brasiliensis [16], or antigens from Schistosoma sp. eggs bounded to 45 μm latex beads [17], produce granulomas in vitro.
Although non-degradable particles such as latex or dextran beads can induce foreign body or non-immune granulomas in monolayer cultures of macrophages [17], there is no in vitro model that recreates granuloma formation induced by biopersistent, high aspect ratio nanomaterials. To comply with a mandate issued by the U.S. National Academy of Sciences to replace animal testing with high-throughput in vitro cellular assays based on mechanistic pathways [18], we have developed a novel in vitro granuloma model. In this model, murine bone marrow-derived macrophages (BMDM) are cultured in a chemically-defined, serum-free medium as 3-dimensional spheroids in agarose wells. Cells maintain viability and macrophage phenotypic characteristics when exposed to sub-lethal doses of particulates up to14 days. Validation of this model for granuloma formation was done using carbon black particles (Printex 90) and crocidolite asbestos fibers as negative and positive reference materials, respectively, as recommended by an ILSI Working Group [19]. Three commercial types of MWCNTs were tested for their ability to induce granuloma formation using this in vitro 3D model. Induction of granuloma formation was evaluated by formation of stable cellular aggregates, analysis of morphological changes characteristic of epithelioid macrophages, and activation phenotype of macrophages in response to high aspect ratio nanomaterials.
Characterization of reference and sample materials
Summary of material characterization
Length (μm)
Surface area (m2/g)
Fe content
Carbon black
141 ± 0.9(1)
2.8 ± 2.6 (1)(2)
116 ± 112(1)(2)
6.1 ± 2.5(1)
163 ± 49(1)
< 0.1(5)
18.8 ± 7.4(1)
69 ± 30(1)
11.7 ± 3.6(1)(4)
80 ± 18(1)(4)
Determination of sub-lethal dose
Potential of high aspect ratio nanoparticles to induce granulomas
In 3D cultures, formation of cellular aggregates in response to asbestos fibers or MWCNT 1 and 2 was found to be dose-dependent. When treated at 0.05 μg/ml, macrophages form aggregates after 10 days (Additional file 1). When exposed to 0.5 μg/ml of asbestos fibers, cellular aggregates form after 3 days (Figure 4) and MWCNT 1 and 2 induced stable aggregates after 7 days (Additional file 1). Exposure to 0.05 or 0.5 μg/ml of MWCNT 3 induced aggregation after 3 days (Additional file 1). Exposure to carbon black did not induce cellular aggregation at any dose or time. In 2D cultures, untreated cells or cells exposed to 0.05 μg/ml (data not shown) or 0.5 μg/ml (Figure 4) of particulates showed formation of multinucleated giant cells. This effect may be due to prolonged attachment to a non-degradable surface in the presence of serum proteins as described previously [20, 21].
Morphological characteristics of in vitro granulomas
Untreated cells and cells exposed to carbon black particles show abundant cytoplasm, nuclei with an open chromatin pattern, prominent nucleoli, intact mitochondria, and short surface microvilli (Figure 5A and 5B). In contrast, asbestos fibers or MWCNTs induced well-formed, compact macrophage aggregates with ultrastructural features of epithelioid macrophages (Figure 5A). Most prominent is the extensive interdigitation of elongated, arborized surface microvilli. Particulates are contained in cytoplasmic membrane-bound vesicles with occasional free particulates observed in the cytoplasm (Figure 5B). Epithelioid macrophages contain abundant endoplasmic reticulum and large cytoplasmic vacuoles (Figure 5B) that are also characteristic of granulomas [22, 23].
Phagocytosis of high aspect ratio nanomaterials by macrophages in 3D cultures
Cellular activation in 3D cultures in response to particulates
Following exposure to microorganisms, macrophages undergo classical activation (M1 or pro-inflammatory phenotype) or differentiation towards alternatively activated macrophages (M2 or pro-fibrotic phenotype) [24–26]. Prototypical markers of the M1 phenotype include proinflammatory mediators including TNF-α, IL-1β, and IL-6 [27], and up-regulated iNOS expression [28]. The anti-inflammatory or profibrotic M2 phenotype is characterized by induction of arginase-1 (Arg-1) and increased synthesis and release of mannose receptor (MR/CD206) [26].
First, we determined the ability of 2D and 3D cultures of macrophages to express a pro- or anti-inflammatory response by exposure to 100 ng/ml of lipopolysaccharide (LPS) or 20 ng/ml of IL-4 and IL-13, respectively (Additional file 2). LPS induced TNF-α release, iNOS expression, and decreased MR expression (Additional file 2 Figure S1). IL-4 or IL-13 induced Arg-1 expression, as well as expression and release of both MR and YM-1 (Additional file 2 Figure S2).
Although the ability of macrophages to respond towards a pro- or anti-inflammatory stimulus under 2D or 3D conditions is not impaired, we were unable to detect significant differences in the levels of these inflammatory mediators using ELISA (TNF-α) or Western blot assays (iNOS and Arg-1) when macrophages were exposed to 0.5 μg/ml of particulates compared to untreated cells. Additionally, no significant differences in IL-1β release were detected following exposure to these reference and test particulate materials at the sublethal doses used in these experiments in the absence of priming by LPS or phorbol ester (Additional file 3).
Compared to untreated cells, exposure to asbestos or MWCNTs induced an early increase in TNF-α expression and decreased expression of MR between 1 and 3 days. After day 7, this activation phenotype changes with variable TNF-α expression and increased MR expression following exposure to all particulates (Figure 7).
Exposure to asbestos or MWCNTs in 3D cultures also induces early iNOS expression after 3 - 7 days, followed by a progressive increase in Arg-1 expression (Figure 8). Induction of both iNOS and Arg-1 in untreated cells or cells exposed to carbon black was also evident at later time points but to a lesser extent than asbestos or MWCNT-treated cells (Figure 8). Phenotypic heterogeneity is evident following all exposures. The kinetics of expression of different phenotypic markers is not identical for all markers evaluated: TNF-α expression is upregulated prior to increased MR expression and no co-localization of these markers is observed (Figure 7). In contrast, iNOS expression increases early and is prolonged with a gradual transition to increasing Arg-1 co-expression between 7-14 days. Untreated cells or cells exposed to carbon black showed low levels of Arg-1 expression (Figure 8). This phenotypic heterogeneity and co-expression of M1 and M2 markers by a subset of macrophages following exposure to asbestos or MWCNTs is similar to the complex phenotype shown by murine macrophages isolated from healing wounds [29].
Particulate agglomeration before and after exposure
This in vitro model of granuloma formation uses murine BMDM differentiated in vitro using macrophage colony stimulation factor (M-CSF) (Additional file 4 Table S1) The phenotype of these differentiated cells resembles mature circulating blood monocytes (Additional file 4 Figure S1). These cells are recruited into the lungs [30] or the peritoneal cavity [31] in response to particulate stimuli following an early influx of neutrophils. If the particulate stimulus is toxic and biopersistent, these elicited macrophages may further differentiate into epithelioid macrophages and form a granuloma [22, 23]. Activation of BMDM and formation of stable aggregates with the morphological features of a foreign body or non-immune granuloma are successfully reproduced in this serum-free, in vitro model only in response to high aspect ratio nanomaterials, not to spherical carbon black particles that have chemically similar surfaces, also composed of sp2-hybridized (graphenic) carbon.
The relative ranking of these materials for induction of granulomas in this 3D models is: MWCNT 3 > asbestos > MWCNT 1, MWCNT 2 > carbon back. Surface area is not a major determinant of granuloma formation in this data set because the MWCNT 2 with a surface area of 200 m2/g did induce granuloma formation while carbon black nanoparticles with a surface area of 300 m2/g did not.
During in vitro exposure in this 3D model, all of the nanomaterials underwent agglomeration (Figure 9), especially MWCNT 3 as illustrated in Figure 10. This phenomenon may be driven by several events leading to entangled fibrous agglomerates that persist after the cells are lysed. First, internalization of materials may lead to intracellular aggregation during phagocytosis. Second, over time, macrophages may undergo repeated cycles of phagocytosis and cell death, resulting in extracellular release and accumulation of agglomerated fibrous nanomaterials.
High aspect ratio nanoparticles also induce frustrated phagocytosis and formation of multinucleated giant cells similar to the response of macrophages to asbestos fibers or carbon nanotubes following pharyngeal aspiration [11] or intraperitoneal or pleural injection [35, 36]. Similar to the study by Poland et al. [35], this in vitro model used a range of commercial MWCNT samples with an iron content of < 0.1 - 1.8 wt-%; however, these samples induced granulomas to the same extent as crocidolite asbestos, the positive reference sample, with an iron content of 22%. Porter et al. [11] used the same MWCNT as our sample 3 and did not detect generation of hydroxyl radical using electron spin resonance (ESR). Although redox activity of commercial carbon nanotubes is lower than that of amphibole asbestos fibers [37], this does not rule out a role for secondary generation of ROS by macrophages during phagocytosis of high aspect ratio nanomaterials [13].
The phenotype of BMDM induced by exposure of 3D cultures to asbestos fibers or MWCNTs also recapitulates the proinflammatory response of macrophages following inhalation of asbestos fibers. For example, inhalation of chrysotile or crocidolite asbestos by rats increased formation of nitrotyrosine in alveolar macrophages and other target cells of the lungs and pleura [38] suggesting induction of iNOS similar to this in vitro model. Asbestos fibers delivered by inhalation to the lungs [39] or by intraperitoneal injection to the mesothelium [31] elicit an acute inflammatory response due to the proinflammatory mediators produced by resident macrophages [19]. Intratracheal instillation of asbestos fibers or MWCNTs in rats stimulated TNF-α release relative to ultrafine carbon black particles [4]. In this in vitro model, asbestos fibers and MWCNTs also upregulated TNF-α protein expression in comparison to carbon black three days after exposure (Figure 7).
The proinflammatory mediators TNF-α, IL-1β, and iNOS have also been observed during the formation of immune granulomas in vivo [40] and in vitro [41]. Less is known about the mediators involved in production and healing of non-immune granulomas. Non-immune or foreign body granulomas in the lung may progress to local fibrosis [42] similar to the reaction to well-dispersed MWCNTs delivered to the lungs of rodents by pharyngeal aspiration [5]. The phenotype of activated macrophages in non-immune granulomas has not been characterized; however, arginase-1, a marker of M2 macrophage activation, is induced after 10-14 days following exposure to asbestos or MWCNTs in this in vitro model (Figure 8). Induction of arginase-1 is associated with a profibrotic phenotype [43] and may be useful as a biomarker to predict potential fibrosis induced by high aspect ratio nanoparticles. Co-expression of proinflammatory as well as profibrotic phenotypic markers by epithelioid macrophages may be responsible for sustained inflammation and rapid development of fibrosis induced by MWCNTs in the lungs of rodents [4, 11].
The biological mechanisms responsible for formation of stable aggregates of macrophages elicited from circulating blood monocytes in non-immune granulomas are unknown [44]. Differentiation of monocytes into epithelioid macrophages is a morphological feature of granulomas. Epithelioid cells are large cells characterized by abundant endoplasmic reticulum, mitochondria, lysosomes, and cytoplasmic vesicles and vacuoles [22]. Interdigititation of long, surface microvilli may be responsible for stable adhesion of epithelioid cells and they are the ultrastructural hallmark of granulomas [23]; these morphological features are produced by exposure to asbestos fibers or MWCNTs, but not to carbon black (Figure 4). The complex 3D aggregate structure (Figure 1C) characteristic of these commercial carbon nanotube samples may promote macrophage aggregation and granuloma formation in vitro and in vivo.
A major caveat in in vitro nanotoxicology assays is the high dose delivered to target cells that may not reflect human exposure under realistic conditions [45]. In this in vitro model, 1.4 × 105 BMDM were exposed to 0.05-0.5 μg of test particulates. Elder et al. [46] established a subchronic NOAEL for Printex 90, the carbon black sample used in this in vitro study, and determined that approximately 0.1 mg was deposited in the lungs of rats after 13 weeks inhalation at a dose of 1 mg/m3. Assuming 3 × 107 alveolar macrophages per rat [47, 48], the dose used in this in vitro assay is close to the deposited dose of carbon black under conditions that do not induce impaired clearance or inflammation. For comparison, airborne dust levels of MWCNTs measured by Han et al. [49] in a research facility were 400 μg/m3 and this dose corresponded to the doses delivered to mice by pharyngeal aspiration that produced lung granulomas and diffuse interstitial fibrosis [11].
The distinctive morphological and phenotypic responses of BMDM to reference particulates and MWCNTs in serum-free 3D culture establish proof-of-principle for this in vitro model as a potential alternative to both traditional 2D monolayer cultures and to rodent bioassays for granuloma formation. This in vitro 3D model is a promising platform for testing of various commercial varieties of carbon nanotubes as well as other new high aspect ratio or one-dimensional nanomaterials including metallic nanowires [36], and ZnO or TiO2 nanorods and nanobelts [50]
Materials and methods
Isolation and culture of bone marrow-derived macrophages
This protocol was approved by the Brown University Institutional Animal Care and Use Committee in accordance with the Animal Welfare Act and the National Institute of Health Guide for the Care and Use of Laboratory Animals. Murine bone marrow-derived macrophages were differentiated from bone marrow cells isolated from 6- to 8- week-old C57BL/6J male mice (Jackson Laboratories, Bar Harbor, ME). Briefly, animals were sacrificed by CO2 asphyxiation, femurs were removed, and bone marrow was harvested by flushing with 3 ml of Dulbecco's Modified Eagle medium (DMEM; Invitrogen, Carlsbad, CA) supplemented with 20% fetal bovine serum (FBS; Atlanta Biologicals, Lawrenceville, GA), 10 units/ml penicillin, and 10 μg/ml of streptomycin (Invitrogen, Carlsbad, CA). Cells were collected in a 50 ml conical tube on ice, centrifuged at 500 g for 5 min at 4°C. Cells were resuspended in fresh media supplemented with 20 ng/ml of M-CSF (R&D Systems, Minneapolis, MA) and incubated overnight at 37°C in a humidified atmosphere of 5% CO2. Non-adherent cells were harvested and transferred to 10 cm non-tissue culture-treated polystyrene dishes. After 3 days the medium was replaced with fresh medium and, on day 7, non-adherent cells were removed. Adherent cells were washed 5 times with PBS, and 10 ml of serum-free medium (SFM) was added to each dish: X-Vivo 15 medium supplemented with 1X HL-1 (Lonza Rockland Inc., Rockland, ME), 20 ng/ml M-CSF and 10 units/ml penicillin and 10 μg/ml of streptomycin. On day 10, cells were detached with 5 mM of EDTA, centrifuged at 500 g for 5 min at 4°C, resuspended, and plated in SFM at 0.14 × 106 cells/ml. A 200 μl volume of cell suspension (35,000 cells) was added to each well of a 96-well microplate. For three dimensional cultures, wells were previously coated with 50 μl of 3% (wt/vol) low melting point agarose (SeamPlaque, Lonza Rockland Inc.) dissolved by boiling in endotoxin-free water (Sigma-Aldrich, St. Louis, MO). Coated wells were equilibrated with SFM for 24 hrs at 37°C and 5% CO2 in humidified chamber prior to use. In order to avoid endotoxin contamination, glassware and plastic containers were washed with PyroCLEAN™ detergent (AlerCHEK, Portland, ME) and autoclaved before use. Cells were kept up to 14 days and 2/5 of medium was replaced with fresh medium every other day. Light microscopy did not reveal any contamination with fibroblasts after 14 days.
Materials and cell treatments
Crocidolite asbestos fiber stocks ranging from 0.1 to 35 μm in length and 30 to 3000 nm in diameter, originally characterized by the International Union Against Cancer (UICC, Timbrell, 1971/72), were purchased from Duke Scientific (Palo Alto, CA). Carbon black particles (Printex-90, Evonik, Essen, Germany) with an average primary particle size of 14 nm and MWCNT 3 produced by chemical vapor deposition (CVD) with average dimension of 13 μm in length and 85 nm in diameter (MWCNT 3, MWCNT-7, Mitsui & Co. Ltd; Tsukuba, Ibaraki, Japan) were a gift of Dr. Günter Oberdörster (University of Rochester, Rochester, NY). MWCNT 1 (mean length 7 ± 2 μm, mean diameter 140 ± 30 nm) and MWCNT 2 (mean length 30 μm, mean diameter 35 ± 10 nm) produced by CVD were purchased from MER Corporation (Tucson, AZ). Size distribution of MWCNTs in serum-free medium was evaluated by SEM and TEM using NIH Image/ImageJ software (Table 1).
Asbestos fibers were baked in an oven at 250°C for 16 hrs to remove endotoxin. Stocks at 0.5 mg/ml in endotoxin free water (Sigma) were autoclaved and stored at 4°C. This sterilization protocol did not decrease the biological activity of asbestos fibers following intraperitoneal injection in mice [31] or redox activity as assessed by plasmid DNA break assay [37]. Removal of endotoxin and sterilization of carbon nanomaterials were done by heating at 400°C for 30 min in a furnace under continuous flow of nitrogen gas to prevent oxidation.
Stocks at 0.5 mg/ml in sterile endotoxin free water were stored at 4°C. Working solutions were prepared by dilution of stock suspensions in SFM. Briefly, stock suspensions were water-bath sonicated for a minimum of 30 min using a tabletop ultrasonic water bath (Branson Ultrasonic Corporation, Danbury, CT). After 30 min, the sample was mixed by pipeting up and down during sonication and 1 ml of the stock was transferred to a clean tube using wide bore, low retention tips (Axygen INC, Union City, CA), then brought to the desired concentration with SFM and sonicated for an additional 5 minutes. A volume of 50 μl of working solution was added to each well at a final volume of 250 μl. Dry materials and suspensions were handled under a Class IIB laminar flow hood equipped with Hepa filters with external exhaust. Endotoxin levels of the stock suspensions were less than 0.24 EU/ml as determined by gel-clot assay (Embryotch™ Laboratories, Wilmington, MA).
Determination of cell viability
Total DNA content was used as an indicator of total cell number. At the specified time, culture medium was removed and cells were digested with fungal proteinase K (Invitrogen) at a final concentration of 50 μg/ml (≥ 1 unit/ml) in TE buffer (200 mM Tris-Cl, 20 mM EDTA, pH 7.5) overnight at 37°C and stored at -20°C until use. Digested cell suspensions were brought to 300 μl of final volume with TE buffer and a 20 μl aliquot was used to determine DNA content. Double-stranded DNA was quantified using Quant-it Picogreen dsDNA kit (Molecular Probes, Eugene, OR) following the manufacturer's protocol for a 200 μl reaction assay. Briefly, samples were diluted with TE buffer to a final volume of 100 μl and 100 μl of the Quant-it Picogreen working solution was added to each sample, mixed, and incubated for 5 min at room temperature protected from light. Samples were excited at 480 nm and fluorescence emission intensity was measured at 520 nm using a fluorescence microplate reader (SpectraMax M2, Molecular Devices, Sunnyvale, CA). Under these conditions, the linear detection range of the assay was used for all treatments and no interference due to test particulates was observed.
Indirect immunofluorescence
For intracellular staining of TNF-α and mannose receptor (MR), cells were pre-incubated with Brefeldin A (BD Biosciences, Franklin Lakes, NJ) for 6 hrs prior to fixation. Cells were fixed in 4% paraformaldehyde in PBS for 30 min at room temperature, washed 3 times with 0.1% BSA/PBS, and permeabilized in 0.25% (w/v) saponin/blocking solution for 10 min. After washing, cells were incubated for 30 min in blocking solution. Following incubation overnight at 4°C with primary antibodies against arginase-1, iNOS, TNF-α (BD Biosciences) or MR (AbD Serotec, Oxford, UK) in blocking solution, cells were washed with PBS and incubated with secondary antibodies in blocking solution for 2 hrs at room temperature. After three washes with PBS, cells were mounted on glass slides using Vectashield (Vector Lab. Inc., Burlingame, CA) with DAPI as a nuclear counterstain. Samples were analyzed using SlideBook software (Intelligent Imagine Innovations, Denver, CO) and spinning disk confocal scanning microscopy (Olympus IX81, Olympus Corporation, Tokyo, Japan) using appropriate filters for dual wavelength immunofluorescence.
May-Grünwald-Giemsa stain
Bone marrow-derived macrophages in agarose-coated wells were transferred to glass LabTek wells (Nunc, Thermo Fisher Scientific Inc. Rochester, NY) and incubated for 6 to 24 hrs to allow cell attachment and spreading. Attached cells were fixed with methanol pre-chilled to -20°C. Slides were immersed for 3 min in filtered May-Grünwald (Sigma) and 2 min in Giemsa (Sigma) followed by 30 sec in acetone, 2:1 acetone: xylene, 1:2 acetone: xylene and xylene. Slides were air-dried and mounted with Permount™ Mounting Media (Fisher, Pittsburgh, PA). Light micrographs were taken using a Nikon Eclipse 501 (Nikon Instruments Inc, Melville, NY) microscope. Images were acquired and analyzed with SOPT Insight FireWire imaging software (SPOT Imaging Solutions, Sterling Heights, MI).
Toluidine blue stain
Macrophages in 3D cultures were fixed as pellets at room temperature for 30 min in 2% glutaraldehyde in 0.1 M sodium cacodylate buffer, pH 7.4, rinsed, and stored at 4°C in 0.1 M sodium cacodylate buffer with 8% sucrose for further processing. Cells were post-fixed with 1% osmium tetroxide in 0.1 M sodium cacodylate buffer for 30 min at room temperature and rinsed 3 times with buffer for 5 min. Following dehydration in graded ethanols (70%, 90% and 95%) for 10 min and 3 changes of 100% ethanol for 5 min, samples were infiltrated for 12-18 hours in a 1:1 mixture of 100% ethanol and embedding resin (Spurr Low Viscosity Embedding Medium, Electron Microscopy Sciences, Hatfield, PA). Samples were placed in 100% embedding media for 4 hours, and placed in a mold with fresh resin. Blocks were polymerized in an oven at 65°C overnight and sectioned on a Reichert Ultracut Ultramicrotome at a thickness of 0.5 μm using a diamond knife (Diatome US, Hatfield, PA). Following toluidine blue stain, slides were air dried and mounted in Permount™ Mounting Media (Fisher). Light micrographs were taken using a Nikon Eclipse 501 microscope (Nikon Instruments Inc, Melville, NY) and SOPT Insight FireWire software.
Transmission Electron Microscopy
90 nm sections from the same plastic-embedded block used for toluidine blue stain were stained with 1% uranyl acetate in 50% methanol for 10 min and lead citrate for 5 min, placed on copper grids, and viewed with a Phillips 410 Transmission Electron Microscope equipped with an Advantage HR CCD camera (Advanced Microscopy Techniques, Danvers, MA). Images were acquired and analyzed with AMT's imaging software.
Scanning electron microscopy (SEM)
Morphological analysis of materials prior to exposure
Materials were dispersed in serum-free medium at different concentrations, followed by water-bath (Branson Ultrasonic Corporation) sonication for 30 min and vacuum filtered through a PVCD membrane (Millipore, Billerica, MA).
Morphological analysis of materials post-exposure
After 14 days of exposure, cells from 3D cultures were collected, washed, and resuspended in lysis buffer (2% Triton X-100, 10 mM Tris-HCl, pH 8, 150 mM NaCl, 10 mM NaN3, 10 mM EDTA, 5 mM iodoacetamide, 2 mM phenylmethylsulfonyl fluoride, 1 g/ml pepstatin, and 1 M leupeptin). After incubation for 30 min on ice and vortexing every 10 min, cell lysates were centrifuged at 10,000 × g for 10 min at 4°C in a Sorvall Legend Mach 1.6 R centrifuge (Kendro Laboratory Products Inc, Asheville, NC). Supernatants were discarded, pellets were resuspended in 50 ml of methanol, followed by water-bath sonication for 1 hr and vacuum filtered through a PVCD membrane (Millipore).
Cellular phagocytosis and aggregation
After 0.5, 6, and 24 hrs of exposure to 1.25 μg/ml (1 μg/cm2) of asbestos fibers or MWCNTs, cells in 3D cultures were transferred to a glass coverslip, incubated for 15 min to allow attachment and fixed with 2.5% gluteraldehyde in 0.1 M cacodylate buffer.
All images were taken using a LEO 1530 Thermally-Assisted Field Emission (TFE) Scanning Electron Microscope (SEM) (LEO, Germany).
Statistical analysis
Data were analyzed using MatLab software (MathWorks, Natick, MA). All results are presented as the mean ± standard deviation (SD). Differences were compared by Scheffé's method after analysis of variance (ANOVA). A p value < 0.05 was considered as statistically significant.
This work was supported by the National Institute of Environmental Health Sciences at the National Institutes of Health (Superfund Research Program P42 ES013660 , R01 ES016178, and T32 E07272).
We would like to thank Professor Jeffrey Morgan for helpful discussions about 3D cell culture models.
Authors' Affiliations
- Madl AK, Pinkerton KE: Health effects of inhaled engineered and incidental nanoparticles. Crit Rev Toxicol 2009, 39: 629–658. 10.1080/10408440903133788PubMedView ArticleGoogle Scholar
- Stern ST, McNeil SE: Nanotechnology safety concerns revisited. Toxicol Sci 2008, 101: 4–21.PubMedView ArticleGoogle Scholar
- Donaldson K, Aitken R, Tran L, Stone V, Duffin R, Forrest G, Alexander A: Carbon nanotubes: a review of their properties in relation to pulmonary toxicology and workplace safety. Toxicol Sci 2006, 92: 5–22. 10.1093/toxsci/kfj130PubMedView ArticleGoogle Scholar
- Muller J, Huaux F, Moreau N, Misson P, Heilier JF, Delos M, Arras M, Fonseca A, Nagy JB, Lison D: Respiratory toxicity of multi-wall carbon nanotubes. Toxicol Appl Pharmacol 2005, 207: 221–231.PubMedView ArticleGoogle Scholar
- Porter AE, Gass M, Bendall JS, Muller K, Goode A, Skepper JN, Midgley PA, Welland M: Uptake of noncytotoxic acid-treated single-walled carbon nanotubes into the cytoplasm of human macrophage cells. ACS Nano 2009, 3: 1485–1492. 10.1021/nn900416zPubMedView ArticleGoogle Scholar
- Majno G, Joris I: Cells, tissues, and disease: principles of general pathology. 2nd edition. New York: Oxford University Press; 2004.Google Scholar
- Kunkel SL, Lukacs NW, Strieter RM, Chensue SW: Animal models of granulomatous inflammation. Semin Respir Infect 1998, 13: 221–228.PubMedGoogle Scholar
- Mukhopadhyay S, Gal AA: Granulomatous lung disease, an approach to differental diagnosis. Arch Pathol Lab Med 2010, 134: 667–690.PubMedGoogle Scholar
- Warheit DB: Long-term inhalation toxicity studies with multiwalled carbon nanotubes: closing the gaps or initiating the debate? Toxicol Sci 2009, 112: 273–275. 10.1093/toxsci/kfp237PubMedView ArticleGoogle Scholar
- Ma-Hock L, Treumann S, Strauss V, Brill S, Luizi F, Mertler M, Wiench K, Gamer AO, van Ravenzwaay B, Landsiedel R: Inhalation toxicity of multiwall carbon nanotubes in rats exposed for 3 months. Toxicol Sci 2009, 112: 468–481. 10.1093/toxsci/kfp146PubMedView ArticleGoogle Scholar
- Porter DW, Hubbs AF, Mercer RR, Wu N, Wolfarth MG, Sriram K, Leonard S, Battelli L, Schwegler-Berry D, Friend S, et al.: Mouse pulmonary dose- and time course-responses induced by exposure to multi-walled carbon nanotubes. Toxicology 2010, 269: 136–147. 10.1016/j.tox.2009.10.017PubMedView ArticleGoogle Scholar
- Donaldson K: Resolving the nanoparticles paradox. Nanomedicine (Lond) 2006, 1: 229–234. 10.2217/174358220.127.116.11View ArticleGoogle Scholar
- Sanchez VC, Pietruska JR, Miselis NR, Hurt RH, Kane AB: Biopersistence and potential adverse health impacts of fibrous nanomaterials: what have we learned from asbestos? Wiley Interdiscip Rev Nanomed Nanobiotechnol 2009, 1: 511–529. 10.1002/wnan.41PubMed CentralPubMedView ArticleGoogle Scholar
- Heinemann DE, Peters JH, Gahr M: A human in vitro granuloma model using heat killed Candida albicans cells immobilized on plastic culture wells. Scand J Immunol 1997, 45: 596–604. 10.1046/j.1365-3083.1997.d01-435.xPubMedView ArticleGoogle Scholar
- Puissegur MP, Botanch C, Duteyrat JL, Delsol G, Caratero C, Altare F: An in vitro dual model of mycobacterial granulomas to investigate the molecular interactions between mycobacteria and human host cells. Cell Microbiol 2004, 6: 423–433. 10.1111/j.1462-5822.2004.00371.xPubMedView ArticleGoogle Scholar
- Seitzer U, Haas H, Gerdes J: A human in vitro granuloma model for the investigation of multinucleated giant cell and granuloma formation. Histol Histopathol 2001, 16: 645–653.PubMedGoogle Scholar
- Shikama Y, Kobayashi K, Kasahara K, Kaga S, Hashimoto M, Yoneya I, Hosoda S, Soejima K, Ide H, Takahashi T: Granuloma formation by artificial microparticles in vitro. Macrophages and monokines play a critical role in granuloma formation. Am J Pathol 1989, 134: 1189–1199.PubMed CentralPubMedGoogle Scholar
- NRC: Toxicity Testing in the 21st Century: A vision and a strategy. Washington, DC: National Academy Press; 2007.Google Scholar
- Oberdörster G, Maynard A, Donaldson K, Castranova V, Fitzpatrick J, Ausman K, Carter J, Karn B, Kreyling W, Lai D, Olin S, Monteiro-Riviere N, Warheit D, Yang H: Principles for characterizing the potential human health effects from exposure to nanomaterials: elements of a screening strategy. Part Fibre Toxicol 2005, 2: 8. 10.1186/1743-8977-2-8PubMed CentralPubMedView ArticleGoogle Scholar
- Collier TO, Thomas CH, Anderson JM, Healy KE: Surface chemistry control of monocyte and macrophage adhesion, morphology, and fusion. J Biomed Mater Res 2000, 49: 141–145. 10.1002/(SICI)1097-4636(200001)49:1<141::AID-JBM18>3.0.CO;2-APubMedView ArticleGoogle Scholar
- Jenney CR, Anderson JM: Adsorbed serum proteins responsible for surface dependent human macrophage behavior. J Biomed Mater Res 2000, 49: 435–447. 10.1002/(SICI)1097-4636(20000315)49:4<435::AID-JBM2>3.0.CO;2-YPubMedView ArticleGoogle Scholar
- Turk JL, Narayanan RB: The origin, morphology, and function of epithelioid cells. Immunobiology 1982, 161: 274–282.PubMedView ArticleGoogle Scholar
- Wang NS: Electron microscopy in diagnostic pathology. In Electron Microscopy of the Lung. Edited by: DE S. Marcel Deker, Inc New York; 1990:429–490.Google Scholar
- Gordon S, Martinez FO: Alternative activation of macrophages: mechanism and functions. Immunity 2010, 32: 593–604. 10.1016/j.immuni.2010.05.007PubMedView ArticleGoogle Scholar
- Gordon SB, Read RC: Macrophage defences against respiratory tract infections. Br Med Bull 2002, 61: 45–61. 10.1093/bmb/61.1.45PubMedView ArticleGoogle Scholar
- Martinez FO, Helming L, Gordon S: Alternative activation of macrophages: an immunologic functional perspective. Annu Rev Immunol 2009, 27: 451–483. 10.1146/annurev.immunol.021908.132532PubMedView ArticleGoogle Scholar
- Mantovani A, Sica A, Sozzani S, Allavena P, Vecchi A, Locati M: The chemokine system in diverse forms of macrophage activation and polarization. Trends Immunol 2004, 25: 677–686. 10.1016/j.it.2004.09.015PubMedView ArticleGoogle Scholar
- Jacobs AT, Ignarro LJ: Lipopolysaccharide-induced expression of interferon-beta mediates the timing of inducible nitric-oxide synthase induction in RAW 264.7 macrophages. J Biol Chem 2001, 276: 47950–47957.PubMedGoogle Scholar
- Daley JM, Brancato SK, Thomay AA, Reichner JS, Albina JE: The phenotype of murine wound macrophages. J Leukoc Biol 2009.Google Scholar
- Lehnert BE: Defense mechanisms against inhaled particles and associated particle-cell interactions. Rev in Mineraology 1993, 28: 427–469.Google Scholar
- Moalli PA, MacDonald JL, Goodglick LA, Kane AB: Acute injury and regeneration of the mesothelium in response to asbestos fibers. Am J Pathol 1987, 128: 426–445.PubMed CentralPubMedGoogle Scholar
- Lam CW, James JT, McCluskey R, Hunter RL: Pulmonary toxicity of single-wall carbon nanotubes in mice 7 and 90 days after intratracheal instillation. Toxicol Sci 2004, 77: 126–134.PubMedView ArticleGoogle Scholar
- Shvedova AA, Kisin E, Murray AR, Johnson VJ, Gorelik O, Arepalli S, Hubbs AF, Mercer RR, Keohavong P, Sussman N, Jin J, Yin J, Stone S, Chen BT, Deye G, Maynard A, Castranova V, Baron PA, Kagan VE: Inhalation vs. aspiration of single-walled carbon nanotubes in C57BL/6 mice: inflammation, fibrosis, oxidative stress, and mutagenesis. Am J Physiol Lung Cell Mol Physiol 2008, 295: L552–565. 10.1152/ajplung.90287.2008PubMed CentralPubMedView ArticleGoogle Scholar
- Porter D SK, Wolfarth M, Jefferson A, Schwegler-Berry D, Andrew ME, Castranova V: A biocompatible medium for nanoparticle dispersion. Nanotoxicology 2008, 2: 144–152. 10.1080/17435390802318349View ArticleGoogle Scholar
- Poland CA, Duffin R, Kinloch I, Maynard A, Wallace WA, Seaton A, Stone V, Brown S, Macnee W, Donaldson K: Carbon nanotubes introduced into the abdominal cavity of mice show asbestos-like pathogenicity in a pilot study. Nat Nanotechnol 2008, 3: 423–428. 10.1038/nnano.2008.111PubMedView ArticleGoogle Scholar
- Donaldson K, Murphy FA, Duffin R, Poland CA: Asbestos, carbon nanotubes and the pleural mesothelium: a review of the hypothesis regarding the role of long fibre retention in the parietal pleura, inflammation and mesothelioma. Part Fibre Toxicol 2010, 7: 5. 10.1186/1743-8977-7-5PubMed CentralPubMedView ArticleGoogle Scholar
- Guo LMD, Lui X, Vaslet C, Hurt RH, Kane AB: Iron bioavailability and redos activity in diverse carbon nanotubes samples. Chem Mater 2007, 19: 3472–3478. 10.1021/cm062691pView ArticleGoogle Scholar
- Tanaka S, Choe N, Hemenway DR, Zhu S, Matalon S, Kagan E: Asbestos inhalation induces reactive nitrogen species and nitrotyrosine formation in the lungs and pleura of the rat. J Clin Invest 1998, 102: 445–454. 10.1172/JCI3169PubMed CentralPubMedView ArticleGoogle Scholar
- Kagan E, Oghiso Y, Hartmann DP: Enhanced release of a chemoattractant for alveolar macrophages after asbestos inhalation. Am Rev Respir Dis 1983, 128: 680–687.PubMedGoogle Scholar
- Lukacs NW, Chensue SW, Strieter RM, Warmington K, Kunkel SL: Inflammatory granuloma formation is mediated by TNF-alpha-inducible intercellular adhesion molecule-1. J Immunol 1994, 152: 5883–5889.PubMedGoogle Scholar
- Seitzer U, Scheel-Toellner D, Toellner KM, Reiling N, Haas H, Galle J, Flad HD, Gerdes J: Properties of multinucleated giant cells in a new in vitro model for human granuloma formation. J Pathol 1997, 182: 99–105. 10.1002/(SICI)1096-9896(199705)182:1<99::AID-PATH807>3.0.CO;2-XPubMedView ArticleGoogle Scholar
- El-Zammar OA, Katzenstein AL: Pathological diagnosis of granulomatous lung disease: a review. Histopathology 2007, 50: 289–310. 10.1111/j.1365-2559.2006.02546.xPubMedView ArticleGoogle Scholar
- Wynn TA: Fibrotic disease and the T(H)1/T(H)2 paradigm. Nat Rev Immunol 2004, 4: 583–594. 10.1038/nri1412PubMed CentralPubMedView ArticleGoogle Scholar
- Majno GaJI: Cells, tissues, and disease: principles of general pathology. Oxford University; 2004:446–476.Google Scholar
- Oberdörster G: Safety assessment for nanotechnology and nanomedicine: concepts of nanotoxicology. J Intern Med 2010, 267: 89–105. 10.1111/j.1365-2796.2009.02187.xPubMedView ArticleGoogle Scholar
- Elder A, Gelein R, Finkelstein JN, Driscoll KE, Harkema J, Oberdörster G: Effects of subchronically inhaled carbon black in three species. I. Retention kinetics, lung inflammation, and histopathology. Toxicol Sci 2005, 88: 614–629. 10.1093/toxsci/kfi327PubMedView ArticleGoogle Scholar
- Tao F, Gonzalez-Flecha B, Kobzik L: Reactive oxygen species in pulmonary inflammation by ambient particulates. Free Radic Biol Med 2003, 35: 327–340. 10.1016/S0891-5849(03)00280-6PubMedView ArticleGoogle Scholar
- Oberdörster G, Yu CP: Lung dosimetry--considerations for noninhalation studies. Exp Lung Res 1999, 25: 1–6. 10.1080/019021499270385PubMedView ArticleGoogle Scholar
- Han JH, Lee EJ, Lee JH, So KP, Lee YH, Bae GN, Lee SB, Ji JH, Cho MH, Yu IJ: Monitoring multiwalled carbon nanotube exposure in carbon nanotube research facility. Inhal Toxicol 2008, 20: 741–749. 10.1080/08958370801942238PubMedView ArticleGoogle Scholar
- Hamilton RF, Wu N, Porter D, Buford M, Wolfarth M, Holian A: Particle length-dependent titanium dioxide nanomaterials toxicity and bioactivity. Part Fibre Toxicol 2009, 6: 35. 10.1186/1743-8977-6-35PubMed CentralPubMedView ArticleGoogle Scholar
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text:
Defy volunteers are influential
business leaders who, like our
entrepreneurs-in-training (EITs),
have often defied many odds in
their lives to achieve success.
If you've profited from the wisdom of great mentors and have a passion for helping others do the same, then you'll have a powerful and rewarding volunteer experience with Defy.
Defy seeks successful business executives, entrepreneurs, investors and other skilled, passionate professionals (e.g., lawyers and accountants) to coach our EITs to launch their business ventures. At Defy, you'll use your core competencies to "pay it forward." If believing in underdogs, and seeing an idea grow from conception to completion is what you crave, then you'll love volunteering with Defy.
Tim Draper
(business plan competition judge)
Founder and Managing Director
Draper Fisher Jurvetson
Menlo Park, CA
From my experience, I think Defy Ventures is a very worthy cause—something I rarely say about nonprofits. The prison system really needs to be rethought. The U.S. puts more people in prison per capita and for longer periods of time than in any other country. It's time for a fresh look at crime and what happens to people who commit crimes.
Brian Korb
(mentor and Investment Committee Member)
New York, NY
Defy runs a tight, professional ship that pays attention to the details. It's a quality product and a winning model, and I'm excited to be part of it. Defy believes that the raw DNA of excellent entrepreneurship can be molded. If you've paid your dues and want to be a productive member of society, then you get a shot. That's what this country is about. I believe in the multiplier effect of Defy on EIT families and communities—Defy graduates are hiring people and adding to the tax base.
The U.S. prison system makes people feel like they're worthless. Defy graduates feel better about themselves when they realize that people care about them. Watching EITs create their business plans, pitch their companies, and grow as individuals is amazing. Many of them do it better than I do! However, Defy doesn't coddle its EITs. It isn't afraid to cut someone out of the program if they're not proving that they want to be a successful entrepreneur.
Before I started volunteering with Defy, I hadn't known anyone who spent time in prison and was even somewhat afraid to meet the EITs. Now I'm excited to see them! The whole experience humanizes the prison issue and strips down the human experience to its essentials. It has really humbled me—with a few different circumstances, my life could easily have been really different. I've realized just how much early circumstances play a big role in life and it's unfair.
Doug Beck
(mentor and business plan competition judge)
DWS Funds (a division of Deutsche Bank)
New York, NY
What really inspires me about Defy is having a hands-on impact on people's lives. I've been involved with several nonprofit organizations, but my experience with Defy is more engaging and real. It's miles ahead of anything I've ever done to give back. It's not just about writing a check. I put in real sweat equity and I'm proud to pass on the things I've learned in my life.
I believe that anyone with initiative, regardless of background and baggage, can control their future. However, I do realize that we live in a complex economy and its many hurdles can be very intimidating. Defy has so many dimensions—it's a virtuous cycle that benefits not just the EITs, but their families and communities.
I talk about Defy to my friends, colleagues, and family at least once per week, and even have photos of the EITs in my office. It's been a wonderful experience to help my mentee clarify and execute his vision. Seeing people refuse to quit helps put in perspective how fortunate many of us are in the corporate world and how important it is that all parts of society are looked after.
Malte Barnekow
Founder and CEO
The 86 Company
New York, NY
When I first came to a Defy event, I saw how different the EIT's lives were from my protected background. But what really struck me was how all the EITs came up to me to thank me for being at the event. I experienced a kindness from them that is very different from the picture that society paints about people who have criminal histories.
Ryan Evans
(business plan competition judge)
Founder and President
Bitesize PR and Lift Marketing
Chicago, IL
Defy is very personal to me because I spent time in prison, so I know what they're going through. Getting back on track was extremely difficult for me, even though I had a lot more connections, skills, and resources than Defy EITs have.
Our society has said that they've paid the price for their crimes, but the sentence should not continue for the rest of their lives. Even a short prison sentence is a life sentence. People don't realize how much help people need when they get out of prison—skills, confidence, and people who care about them. At Defy, I really feel like I'm helping to rebuild lives.
Charles Sims
Managing Partner
Empirica Strategic Management
New York, NY
Defy is all about looking forward and not being a prisoner of your past; your mistakes don't have to define you for the rest of your life.
The program is really rigorous. We say, "You can do it," and the EITs live up to those expectations. The level of understanding that the EITs display about business concepts like break-even and profit is impressive. With all the deadlines and performance expectations, Defy instills the importance of good execution and builds a set of behaviors that sets graduates up for success in other areas of their lives. Working with my mentee has been great because she's so coachable. In turn, the experience is also teaching me how to be a better communicator and listener.
Shavonne Correia
(business plan competition judge)
Vice President
Pantheon Partners
New York, NY
So many organizations help students at a young age before they make mistakes; fewer help people after they've made mistakes.
I walked into Defy not knowing what type of people I would meet, but I've been blown away by the caliber and dedication of the EITs. The energy they bring to the program is just phenomenal. It's obvious how serious they are and I'm so impressed with how thoughtful they've been in developing their businesses.
When I volunteer, I really feel like I'm doing something worthwhile. The EITs really listen and take my advice. It's been amazing to see them achieve more than many people who don't have the major disadvantage of having a criminal record.
Todd Young
(mentor and chairman of Young Professionals Board)
Portfolio Manager
Swiss Re Asset Management
New York, NY
I love working with Defy EITs because they are taking control of their futures. Their attitude, the quality of their work, and their ideas really inspire me. I've seen so much growth in the EITs since I first met them during the final executive interview session to get into the program – they're much more confident, disciplined, and polished. But it's not just about entrepreneurship. Defy also becomes a family for its graduates, and a source of love and hope.
Katie Sanford
(mentor, Young Professionals Board)
Chief Operating Officer
Chou, Chou LLC
New York, NY
Before Defy, I had never been involved with anyone whose life was as different from mine, no less someone who had been incarcerated. It didn't know what to expect, but on the first day, Defy really broke down barriers when they made us hug everyone and the EITs shared why they went to prison.
What's amazing about volunteering with Defy is how real the work is. The EITs really want to be there and are serious about turning their lives around. They also have no problem talking about how they messed up, have dealt with the consequences, and are living life differently now. There's no finger pointing or blaming others for their actions. They take responsibility.
When I first started to get to know my mentee, we didn't talk about business. We discovered that we like the same kind of books. We're both nerds in the same way, and it's so cool to have this in common with someone who's had such a different life than me. In NYC, there's a lot of emphasis on power, glamour, and lifestyle. Working with Defy strips all that away so that you're just looking at the person. We're both creatures of God who were created equally, despite our circumstances.
The EITs work just as hard, if not harder, than I do as an entrepreneur. They worked their butts off to prepare for the business plan competition. As a competition judge, it's not just about giving them a pat on the back and saying "good job." I felt like I could be tough on them because they worked hard and raised the bar for themselves. I'm tough on my mentee because he's smart and because Defy is tough on him, too.
In return, I feel like the experience has helped me focus on what's real and important in life. I've also honed my own business skills. Becoming entwined with souls who deserve so much and who work so hard to transform their lives is amazing.
As a member of the young professionals board, it's also been rewarding to share my experience with my friends. I'm really thrilled that it's been so infectious in my personal life.
Anna Akbari
Sociology of Style and Closet Catharsis
New York, NY
As an entrepreneur, I'm used to hustling and taking huge risks, but it's even more difficult for people who don't come from a background with resources like mine. I think Defy EITs have done more introspection and are committed to living a more positive life than the average citizen because of the obstacles they've overcome. I really like Defy's tough love approach. Too many programs take a handout approach, but I like working with an organization that has high expectations.


text:
David Harris on the Relationship Between Prosecutors and Police at Transnational Workshop on prosecutorial power
Published September 24, 2014
Professor David Harris participated in a transnational workshop on prosecutorial power held at Washington & Lee University School of Law on April 1 and 2, 2010. Participants from Sweden, the U.K., Italy, Germany, Bosnia and Herzegovina, the Netherlands, and other countries joined American scholars and prosecutors in examining trends in prosecutorial practices the world over. Professor Harris spoke about the relationship between prosecutors and police in the United States. The workshop was organized by faculty members of the Max Planck Institute for Foreign and International Criminal Law and Washington & Lee School of Law.
Referred Person:


text:
This is pretty much just a repository of my work at this point. Or at least the part of my work that's people-friendly. Still, as astronomically late as this update is, it's a pretty big one. Here's the finished version of a cover I did of Dimholt - a Bulgarian black metal band, who also happen to be good friends of mine. My version is different, though, to say the least. This song is very special for one other reason. I've started work on my first solo album and it's pretty much the prototype of what the album will sound like. I've barely started on it, but I'm really excited about it.
Also, the sound on this prototype wouldn't even be possible with the huge amount of help that a friend of mine, Lyubomir Stefanov, provided. The man's an absolute genius, that's all I can say.
Articorse - Blindead (Dimholt Cover)
And the other update is a collaboration I did with YouTube DooM Cover Master elguitarTom. Check him out, his covers are brilliant!
elguitarTom feat. Articorse - Them Barrels (DooM/Alice in Chains Cover)


text:
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Pharmacy Technician | Every Other Weekend | Day/Ev (661049)
Last Updated: 2019-06-17
Employer: Christiana Care Health SystemLocation: Newark, DE
The Christiana Pharmacy Department is currently recruiting for a part time Certified Pharmacy Technician to work every other weekend. This is a day/evening shift rotation position. Holidays as required.
The Christiana Hospital Pharmacy and the Wilmington Hospital Pharmacy operate 24 hours a day.
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Registered Nurse - Part Time - PACU - Wilmington (660474)
Last Updated: 2019-06-14
Employer: Christiana Care Health SystemLocation: Wilmington, DE
Christiana Care Health System, a Magnet health care organization with over 1,100 beds between its two hospitals (Christiana Hospital & Wilmington Hospital) and the only Level I trauma service on the East Coast corridor between Philadelphia & Baltimore, has been honored repeatedly as "One of America's Best Hospitals" by U.S…
Licensed Practical Nurse PD/80 - VNA (660196)
Last Updated: 2019-06-13
Employer: Christiana Care Health SystemLocation: Camden, DE
If you are a nurse, and you are compassionate about nursing, Christiana Care Visiting Nurse Association (VNA) is the place for you. Whether families need help during the day or night, this expert care is available for medically fragile children and complex adults. Our highly skilled staff can…
Clinical Pharmacist | Part Time | 24 Hours Every T (659949)
Last Updated: 2019-06-12
Employer: Christiana Care Health SystemLocation: Newark, DE
Christiana Care Health System is currently seeking a Clinical Pharmacist work part time, 24 Hours every two weeks, at the Christiana Hospital location.
- Evaluate drug efficacy, document drug use patterns, monitor therapeutic goals and outcomes, review for contraindications/precautions, drug -drug, drug-food, drug-laboratory, and drug-disease state potentials as well…
Administrative Assistant I (659734)
Last Updated: 2019-06-11
Employer: Christiana Care Health SystemLocation: Wilmington, DE
PLEASE NOTE: This position is a 40-hour a week position for 42 weeks and offers benefits at a part time rate. Please only apply if you are interested in this schedule/status.
To assure the everyday flow of office operations and programming for the SBHC as well…
Medical Laboratory Scientist | 28 Hours/Week | Day (659613)
Last Updated: 2019-06-10
Employer: Christiana Care Health SystemLocation: Newark, DE
Christiana Care Health System Blood Bank Services department is currently recruiting for a part time. 56 hours/every two weeks, Medical Laboratory Scientist. This is a day/evening shift rotation position. Every other weekend and holiday required.
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Ultrasound Technologist I | Part Time Christiana H (659297)
Last Updated: 2019-06-08
Employer: Christiana Care Health SystemLocation: Newark, DE
Christiana Care Health System is one of the largest health care providers in the Mid-Atlantic Region. Named one of "America's Best Hospitals" by U.S. News & World Report, we have an excess of 1,100 beds between our two hospitals and are committed to providing the best patient care in the…
Registered Nurse-Vascular Access Team-Part Time Mi (658789)
Last Updated: 2019-06-06
Employer: Christiana Care Health SystemLocation: Newark, DE
Christiana Care currently is currently recruiting for a Registered Nurse in the Vascular Access Nursing team at Christiana Hospital to be responsible and accountable for pediatric, adult, adolescent, and geriatric patients to provide Vascular Access Nursing care to patients through the nursing process in collaboration with other healthcare team members.
Phlebotomy Technician | 20 Hours/Week | Day Shift (658512)
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Employer: Christiana Care Health SystemLocation: Wilmington, DE
The Blood Collection Department at Wilmington Hospital is currently recruiting for a Phlebotomy Technician to work part time, 16 hours per week primarily in out patient phlebotomy. This position will work every Monday and every Friday from 8:30 am. to 5:00 pm. Occasional Saturdays will be required and hours on…
Surgical Technologist (658507)
Last Updated: 2019-06-05
Employer: Christiana Care Health SystemLocation: Newark, DE
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