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As a wedding photographer I am always on the hunt for that perfect second shooter. Over the last 4 years I have shot 120+ weddings and worked with over 70 different second shooters. Most of them have been fantastic but I have always thought it would be nice to have a list of second shooter tips I could send them before the wedding day. I'd love to hear what you think of the tips in the comments section below. Let me know what I left out and feel free to share stories (good or bad) about working with second shooters in the past.
So without further ado, here is my list of "26.5 Tips on Becoming A Perfect Second Shooter."
1. Plan to show up early. In other words don't be late. You never know what kind of traffic you will encounter on the way to the shoot location, so just plan on getting there 30 minutes early. Once you arrive on location, you are welcome to hangout in your car or even walk the grounds to scout out interesting locations. But don't embarrass the lead photographer by arriving late.
2. Leave your business cards at home. You have absolutely no need for them at the wedding. Instead of having your own cards with you, you should ask the primary shooter for 4 or 5 cards of theirs you can have on hand in case anyone asks for one. I often travel for weddings and hire local second shooters to assist me. On those occasions, if I really like the person I am working with I will introduce them to the local wedding planners with the hope they will be able to do some work together in the future, but don't ever walk up to the planner and introduce yourself and hand them your card.
3. Don't check your phone, update Facebook, or Instagram photos. If you really need to check your phone for whatever reason, it should only be when you are completely out of sight. Bathroom stalls make great hiding spots. If you are checking your phone in public where people can see you, you're giving off the perception you are not working.
4. Think like a dental assistant. One of my most pleasant second shooter experiences was with Ryan Reed. Ryan observed that in the first couple hours of shooting I had a pattern of switching between certain lenses. When I would grab one from my bag, she would grab the other and have it on her ready to hand me. All the while she was still shooting and making great shots. I wouldn't necessarily recommend this to everyone - as I would hate for someone to drop a lens for example - but Ryan pulled it off effortlessly and it really was quite helpful.
5. Don't add the Bride and Groom or anyone in their Bridal Party as Facebook friends. It's great to have good relationships with these people and of course as lead shooters we want our assistants to be friendly throughout the day. But leave it at that. There really is no need to add them as Facebook friends. If they hunt you down and try to add you, consider ignoring the request or consulting with the lead shooter if it's ok to accept them.
6. Speaking of Facebook, do not post any photos to your page before getting prior approval from the lead shooter. My personal rule is that my second shooters can post to Facebook only after I have delivered all the images to my clients. If they post to Facebook before I do that and someone recognizes the bride and groom and tags them in the image then now all the friends and family will believe that photographer was the lead photographer. Some lead shooters will even have rules against ever posting to Facebook. Make sure to ask them ahead of time so you know their rules.
7. Help the lead photographer stay hydrated. On the wedding day there are times when things are a bit hectic. Especially when I am working with families or large bridal parties I am constantly talking and directing people with no rest in between. It is always refreshing to have a second shooter that is aware of the situation and will grab some water for me to stay hydrated.
8. Don't be afraid to shoot lots of photos. Occasionally I will get a second shooter that seems to be afraid of giving me too many images at the end of the night. Don't be. Of course this doesn't mean to have your camera on high speed machine gun mode. But be always on the look out around you. During family formals capture some of the candid moments of people standing around, kids chasing each other and people enjoying their cocktails.
9. Don't delete images from your cards. No matter how bad or embarrassed you are because of an image, don't delete it. It might actually be something I could use and with the dynamic range in our cameras these days an overexposed or underexposed image can often be saved. But even more important is that when you delete images on your card and keep shooting on that card you are doing something called back-filling. I will write an article about the process in the next few weeks. In short though it makes it much harder to recover images from your memory card if for some reason it goes corrupt.
10. Don't drink alcohol. Yes most weddings will have free booze. But don't touch it. Practically every job out there does not allow you to drink while working, the same should apply to wedding photographers.
11. Don't cross shoot your lead photographer. Basically what this means is don't be in the background of my shot. If you have a great shot of the bride walking down the aisle as I shoot from the front of the church that is totally fine. But make yourself small and duck behind the bride so I cannot see you. If you can see me, I can see you. Stay out of the background.
12. Shoot a different lens than me. If I am shooting a wide angle, then shoot telephoto. Be aware of what I have on my camera and try to do something different.
13. Don't shoot over my shoulder unless I ask you to. I always prefer the second shooter to get shots from different angles than mine. Every so often though rather than popping on a different lens I might just ask my second shooter to fire off a shot using a different lens from mine in the same spot. By doing that we can get a few different variations of the photo and move on quickly.
14. Refrain from asking questions about how to use your camera in front of anyone. I love to help and have no issues if a second shooter asks me tips on how to use their camera, or what white balance I am dialed into etc. But do it quietly and do it when no one else is around. It looks highly unprofessional if you ask where others can hear you.
15. Don't pose the bride and groom unless given permission ahead of time. While you might have an amazing idea, others, including the bride and groom will often feel like you are just wasting everyone's time. It's better to let the lead shooter run the day. If you have a great idea mention it to them on the side. If it's an idea that I think will work I'll either run with it or even say something like, "My assistant here has a great idea that will look really cute. She is going to tell you guys all about it."
16. When you arrive, first thing, ask your lead if you would like to time sync your cameras. While this is something that is quite easy to do using Lightroom (I showed how in this article) it is always nice when a second shooter arrives on site and remembers to ask to do this. Also no matter how accurate your camera time is, sync it to whatever the lead shooter already has. Often they have already started shooting and if they change their time it will mess things up for later. Even if their camera time is entirely incorrect. The only time I would ask the lead about changing their camera time is if they have not started shooting any photos yet.
17. Pay attention to the small details that will make the photo better. Is the groom's tie crooked? Feel free to help fix it. Does the bride's dress need to be fluffed out? Go fluff it. Both the couple as well as the lead photographer will appreciate you when you see and fix those small details that will later show up in the photos.
18. Dress nicely. It's a wedding. Unless it's Honey Boo Boo's family getting married, don't show up in jeans and tennis shoes. Shoot your lead an email ahead of time and ask what they plan on wearing. That will give you an idea as to what you should wear as well.
19. Be ready to shoot the wedding as if you were the primary shooter. This means have a backup camera ready, have plenty of memory cards and batteries. Just because you are the second shooter doesn't mean you should take the responsibility any less seriously. If something were to happen to the lead shooter you will need to step up to the plate and take over. If your camera stops working in the middle of the wedding, you need to have a back up ready to go. If not, you make the lead shooter look terrible in front of their clients.
20. Make sure you are shooting RAW. This should be an obvious one, but I didn't want to fail to mention it. If you shoot JPEG images than the post processing has to be done entirely different. By shooting RAW files I can adjust all the white balances equally and have more dynamic range to work with.
21. Download your RAW files at the end of the night. Before my second shooters leave I have them download their memory cards of RAW files directly onto my laptop. That then gets backed up to my external hard drive before I travel home. If the lead shooter doesn't have a laptop with them, I would have the following available. Pull out your laptop from you car and download the photos onto a small external hard-drive. You should be able to find one for about $50. Then give the hard-drive along with a self-addressed pre-paid shipping envelope. Just wait till you see the smile on your lead shooter's face when they realize how easy you made the process for them.
22. Don't chit chat too much. You might be a bubbly full of personality kind of person ready to hand out hugs to everyone you see. Well don't. Keep chit chat to a minimum with others in the bridal party. Of course always be smiling and working hard, but don't spend your minutes in small talk. The relationship should be between those there and the primary shooter. You are there simply to assist.
23. Don't share images from the back of your camera. Even though you might have nailed the most epic photo in the history of weddings, don't share it. Remember you are working for someone. If you step up and share the photo with the bride and groom or even other people at the wedding you are basically saying, "Look at this shot that I got. Not the other photographer. I got this one." It is quite a slap in the face to your primary shooter.
24. Don't complain about the terms of second shooting later to others. If you worked your butt off on the wedding day and walked away with a $200 check but feel you deserve more, don't complain about it. Whatever the terms are that you decided on before accepting the job, be happy with it. Don't bad mouth the lead photographer before, during or after the wedding. You might get a few amens from those listening to you, but honestly it does nothing more than make you look bad.
25. Do give the lead photographer updates on time. While shooting I love when the second shooter chimes in quietly every so often, "Hey Trevor, it's 6:40pm." By knowing the time I can plan the shoot accordingly and know if we need to speed up a little to get all the shots in.
26. Don't leave early. Unless you arranged for someone to take your place, you need to absolutely be in for the long haul. If the lead photographer asked you to be there till 8pm and you cut out at 7pm it can put them in an ugly situation. They can even lose money over the situation if the client finds out about it and realizes they didn't get all the hours they paid for.
26.5 Grab a few photos of the lead photographer working. This is just a half tip. Definitely not required but always nice to have. As you are shooting, grab just a handful of shots of the primary working. They will appreciate the shots and be able to use them on their website or social media pages.
Alright, so what did I leave out? Let me know in the comments below some tips for second shooters. I would also love to hear your stories, good or bad, about working with second shooters in the past. Lastly, if you enjoyed the tips be sure to like the story, tweet and share it with others.
133 Comments
I Agree with many of these but 12 "donts" and 1 "do" is a little preachy.
Just because he didn't explicitly say "do" doesn't mean there weren't many other do's. For example: 1, 4, 7, 12, 18, 20, 21, and 26.5 could all easily have been prefaced with "do." And for that matter, aren't almost all #-of-blah-that-you-should-do blog post going to sound a little preachy? Whatever happened to looking for the positives rather than try to nitpick at small stuff? K I'm done ranting.
I agree with a lot of this but a few are iffy such as asking about settings. Most of the time when discussing settings with a photographer it's quick and to the point and we usually get the "I don't understand a word you guys just said" from whoever is around which just makes it look like us photogs speak some special language.
Ignoring the Bride and Groom on social media is rude if they go out of their way to add you. You are there to talk with them through the day and I truly think a small friendship is created. Ignoring them is not something I would agree with.
A few more points I was iffy about but I think 75% of this was spot on and the other 25% was a lot of personal opinion. Great article.
edit: wouldn't agree with**** for the second part of the above comment.
I agree with you, I dont think that would be nice to ignore the request if they add you..
Great post and I couldn't agree more! :)
This is awesome Trevor...So helpful. Thank you! And I especially love the lead photo at the top ;)
Definitely agree with not going out of your way to add them. That would make much more sense from that point of view :)
Wardrobe. Always ask the primary what to wear so you don't end up like this :)
Epic timing. Have a wedding Saturday and just forwarded this to my 2nd even though we did lunch today. But this was far more comprehensive of a list.
I love this list, thanks for sharing!!!
I agree with not going out of the way to add them but "If they hunt you down and try to add you, consider ignoring the request "? Nope, not going to ignore them and would NEVER ask my second to ignore them.
I agree with Trevor. If you follow all of these guidelines the point is that there's no reason why the wedding party would search for you online to add you anyways...i've had some seconds try this and they always come off as desperate...Trevor just put all my same feelings very nicely into words. I tend to believe that seconds should be grateful for the opportunity to work with the lead and leave it at that. There are special exceptions I'm sure. :) That's all.
If you haven't solicited the attention bridal party in anyway, and they still want to have contact with you, that tells me one thing: The primary dropped the ball and you picked it up to make their day special in some way.
You have to play it by ear. I would never try to "steal" or even solicit someone's client, but if they seek me out, they might actually be running away from the primary shooter.
Great tips. I dont agree with tip #10 - I always drink alcohol on the job :) Dont go crazy of course, but i think there is nothing wrong with one glass of wine. It really depends on the kind of event and type of people you're working for. You can always drink white wine instead of red... less noticeable
Awesome Trevor!! I swear I've done about 5 don'ts in the past! Ha ha... always learning as a primary and a second shooter (when I do have time to do this). Great post! Thanks for sharing!
Uh.... well most of this is OK, I think you really don't have to ever consider any of this if you actually just find one or two people to work with consistently, your return from them is better, and in the end you have a better product because of it. Consistency is so important I don't understand how anyone can hire such a large amount of different 2nd shooters, I mean, maybe, but it just makes so much more sense for many reasons to just hire the same one or two people for the majority of every season.
Plus, this way you can both perform almost as primaries, except the other shooter knows they have other duties that they must do based off of the situation and what you ask of them. This works far better than just relying on them to be only this or that. I find this approach much better than staying to rigid in the lines of 1st and 2nd.
Great post! Definitely have broken a few of these myself. ;) Some stuff on here I didn't even think about! Love it!
Great post! Definitely have broken a few of these myself. ;) Some stuff on here I didn't even think about! Love it!
True dat. Thinking back on our Orlando airplane shoot, you take your own rules seriously. If only I knew that a going rate for a second shooter was $200!
good list, but some of them would seem as common sense, but i guess not if you have to list them and it happened. best practices, i usually only work with people i know or friends, so i don't have to worry about them making my business look bad. besides the 2nd angle and shot, i like having someone to talk to beside the peeps at the wedding. 10-12hrs wedding day can feel like a longer day when im shooting alone.
I agree with all except for the last part of 21."Then give the hard-drive along with a self-addressed pre-paid shipping envelope." ARE YOU CRAZY. Sorry but this should never happen. If anything just bring your own laptop and burn off a copy of the photos.


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Re: "Command-line parameter in scripting is deprecated"
Simply want to open the winscp site using a script. The script contains:
open sftp://email@example.com/
Yet when the script run, the host is searched for and does not appear.
I assume you use wrong syntax of WinSCP command-line, making WinSCP use some random part of the syntax as a host name.
Please read scripting and command-line documentation properly:
If that does not help, please start a new thread and attach a full log file showing the problem (using the latest version of WinSCP).
To generate log file, use
/log=path_to_log_file command-line argument. Submit the log with your post as an attachment. Note that passwords and passphrases not stored in the log. You may want to remove other data you consider sensitive though, such as host names, IP addresses, account names or file names (unless they are relevant to the problem). If you do not want to post the log publicly, you can mark the attachment as private.

subset: commoncrawl


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The sun touches the horizon and will soon be sinking into the sea. So whats the time at home? If I call now, will I be waking the family? A glance at the new Portofino Dual Time, a smile. The watch, radiant and resplendent, vies for attention with the sunset. A never-ending source of pleasure, its elegant lines and perfect proportions are a feast for the eyes of its owner, who now realizes that he can make the call without any uncertainty.
The time at home,with your nearest and dearest, is shown unerringly by the new 64710 calibre, which is manufactured exclusively for IWC by MHVF (Manufacture Horlogère ValFleurier). The 24-hour display, discreetly positioned in the upper half of the dial, reliably displays the time back at home. A red hand completes a full revolution once a day, while the index markers and imprinted numerals show the time from 0 to 24 hours. To facilitate this, the centre of the 24-hour display is divided into two differently coloured zones. The lower half, which shows the time from 6 to18 hours is lighter in appearance. The upper half, from18 to 6, is darker. This effect was achieved by IWCs designers by using a special kind of finish on the dial. As you would expect in a watch from the Portofino family, the effect is not brash, but extremely understated.
Local time is displayed conventionally on the hour and minute hands. To set it preferably before landing the owner simply pulls out the crown from its basic winding position to position 1. With the crown in this position, the hour hand can be moved backwards or forwards in one-hour steps to show current local time whenever required. If, in the process, the hour hand passes the 24 mark, the date at the destination in question is likewise adjusted, either forwards or backwards. To set the hour hand, it is first necessary to disengage the drive from the watch movement. Together with the small seconds and minute hand, the movement continues to run uninterrupted and keep the time precisely. If the time in general requires adjustment, the crown is pulled out to position 2. A hacking seconds permits the Portofino Dual Time to be set with down-to-the second accuracy.
Behind all this ingenious functionality is the completely new 64710 calibre, which stands for relentless progress in terms of both construction and design. The bearings which are used to seat the moving parts,for example, are made not only of synthetic ruby but also of high-tech zirconium oxide. The state-of-the-art design principles are likewise reflected in the indexless adjustment system. Precision setting of the enormously accurate movement is achieved via four weight screws on the balance rim, which oscillates at a rate of 4 hertz or 28,800 beats per hour. Two barrels ensure a power reserve of 72 hours when the watch is fully wound. A bidirectional pawl-winding system transmits energy to the barrels every time the wearer of the Portofino Dual Time moves his arm. The rotor with the engraving Probus Scafusia Good, solid craftsmanship from Schaffhausen and the exquisitely finished movement can be viewed through the sapphire-glass back.
As befitting a watch of its class, the superb movement is housed in a typically elegant, Portofino-style case measuring 45 millimetres in diameter. The Portofino Dual Time is available in a fabulous solid red gold case with a black dial, or in simple stainless steel with a choice of either a black or a silver-plated dial. In the red gold version the characteristic indices and numerals are likewise made of solid gold. The arched-edge sapphire glass has an antireflective coating on both sides and sets off the lightness and elegance of the watch to perfection.
|Mechanical movement, 24-hour display showing home time, hour hand adjustable separately in one-hour steps via the crown, small hacking seconds, date display
|Calibre 64710
|28,800A/h / 4 Hz
|Power reserve
|Case in18-carat red gold, black dial, Santoni strap made of dark brown alligator leather, pin buckle in18-carat red gold.
Stainless-steel case, silver-plated dial, Santoni strap made of black alligator leather, pin buckle in stainless steel.
Stainless-steel case, black dial, Santoni strap made of black alligator leather, pin buckle in stainless steel.
|Sapphire, arched edge, antireflective coating on both sides
|see-through sapphire-glass back
|3 bar
|45 mm
|12 mm


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School Overview
Blue Tongue Helicopters offer you, the enthusiastic potential pilot, plenty of helpful hints to assist in making quality choices in this "not always so nice" industry. The school consists of a C.A.S.A. approved, air conditioned Office Building and Student Classrooms as well as a separate hangar for aircraft storage. The balconies are spacious with relaxing tropical gardens surrounding the main building.


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I admit it. I've indulged in the annual clerical ritual: I've used Christmas to lament America's spiritual decay and rampant materialism. Ecclesiastical moaning has become part of the Christmas tradition, bundled in the same package as the eggnog, tree-trimmings and Uncle Sal's weird presents ("Thanks, Sal. I love it. One question: What is it?"). Cue in the condemning sermon.
I'll break the ritual and ponder the full, personal brunt of the Christmas message. How has it touched me?
My thoughts turn to God, who chose to live as a human being, beginning as a baby. God did not merely appear human or become similar to a human. God was human, stripped of divinity's power. The baby howls at night. He needs food. Mary must change his clothes. Joseph stumbles out of bed, a bleary-eyed craftsman who wishes his son didn't wake him at 3 A.M. My thoughts turn to me: I too was a howling baby who woke up my parents. God empathized with me even then: The Cosmic Being had once been an infant.
My thoughts turn again to that Being: God grew up as a boy, experiencing a childhood so normal that the Gospel writers barely mention it. My thoughts turn to me once more and my own childhood memories, and then I remember myself as a quivering adolescent, confronted with the reality that I had been a ball of self-pity. But instead of finding God's frown, I found empathetic love: God had been an adolescent – not an adolescent like me, for Jesus was sinless – but he had been an adolescent nonetheless. Divine love enveloped me and prompted me to hand my life to Christ. Unlike many, I can pinpoint the date I was "born from above:" July 6, 1973.
I remember my subsequent spiritual growth; I remember becoming trapped in a nowhere career; I remember God releasing the trap. God was always present, nurturing me with the love of someone who had been there. He often didn't grant my desires because my desires were too small. His vision for me was far greater than my own.
I remember the joys of meeting my future wife. I remember confronting a potentially fatal illness and the possibility of leaving my new bride a widow. God was with me then; he knew the travails of deep suffering when Jesus was nailed to the cross. God's empathy has remained through the years. I felt it in my joy at the birth of our son; I felt it when we wept at the loss of a child through an ectopic pregnancy; I felt it through decades of nurturing broken churches. I feel it now as I delight in our grandson and struggle again with that illness. I've been tagged "incurable."
God laughs. God empathizes. God weeps.
We do not worship a God who is merely all-powerful, all-knowing, and all-loving. Our God is all those things and more: The Almighty has experienced human joy, human pain, human suffering, and human victory. Baffling as it seems, God not only possesses divinity; God possesses humanity. He knows power and weakness.
I'm sure Mary cooed and sung lullabies to Jesus. What were her intuitions as she suckled him? Were there moments when she looked into her baby's eyes and saw the eons staring back at her?
I'm sure she was too struck with awe to lament society's spiritual decay.


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"The warning bell for net neutrality rang Monday when President Donald Trump designated Ajit Pai, a Republican member of the FCC and a net-neutrality critic, to be the agency's new chairman," The Chicago Tribune reports.
"Current FCC rules essentially ensure that service providers act as gateways to a free flow of information and creativity rather than gatekeepers who reward the highest bidders. But Pai could swing the balance of power away from consumers and entrepreneurs and put it in the hands of corporate behemoths such as Verizon, Amazon and Netflix who can in essence buy faster and more efficient access at the expense of their less well-financed competitors."
"If he gets his wish, almost everyone who does business on the internet will feel an impact."


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Is Michael Ledeen Serious?
Here is his latest effort at The Corner:
Is Obama naive?
I don't think so. I think that he rather likes tyrants and dislikes America. I think he'd like to be more powerful, I think he is trying to get control over as much of our lives as he can, so that he can put an end to the annoying tumult of our public life. As when he said (about health care) to the Congress, "Okay, you've talked enough, now it's time to do the right thing (my thing)." And he's trying to end American power in the outside world. He's saying "I'm going to stop us, before we kill again."
There is nothing unusual about elitist hatred of freedom. Back in the 18th century, when book publishing really got going, British authors were infuriated that they had to submit to the judgment of a marketplace. They didn't want to be judged by people who were obviously inferior to them, and there was a great rage among the intelligentsia, including some very famous men. And in modern times, we can all name famous intellectuals who fawned all over Mussolini, Stalin, Fidel, and even Hitler.
American politics are very fractious, and always have been. Leaders are constantly frustrated, and some of them come to yearn for an end to our freedom. They think they know best, they just want to tell us what to do and have us shut up and do it. I think Obama is one of them. He's not naïve. It's different. He doesn't like the way things work here, he thinks he can do much better, and he's possessed of the belief that America has done a lot of terrible things in the world, and should be prevented from doing such things ever again. The two convictions mesh perfectly. It's The Best and the Brightest run amok.
Democratic leaders' envy of tyrants' power can be understood. But it can't be forgiven.
This is criticism from the Andy McCarthy and Victor Davis Hanson school. The conceit is that unlike less savvy observers, these men — due to their long experience, clarity of mind, and exceptional discernment — understand the true psychological motivations of President Obama.
So on health care, it may appear to the untrained observer that President Obama is pushing his agenda because he deems it wise to reform health care, and naturally enough judges his own policy preferences to be the correct course. Ah, but Michael Ledeen is knowledgeable about Mussolini, Stalin, Castro, and Hitler, who also thought their policy preferences to be correct, and implemented them, so he can see the true colors of our president. Why, if Hitler were alive today, he'd probably forgo setting buildings afire, instead marching right down to Congress to say, okay, enough talk about health care — now pass my bill!
And of course the neo-fascist Obama also wants to end his country's influence on the rest of the world, um, just like Mussolini, Stalin, Castro, and Hitler, because that's what Democrats who envy powerful tyrants do? That explains President Obama's choice of a radical dove as Secretary of State, his refusal to use force to free American hostages or kill terrorists, and the pacifist attitude he's taken toward Pakistan.
Yes, if only we could have a president like the one Mr. Ledeen, Mr. Hanson, and Mr. McCarthy last supported, one who doesn't bully Congress and try to radically increase the power of the executive. Perhaps some of you might object that President Bush asserted the ability to declare American citizens enemy combatants without offering any evidence for his designation, that he secretly wiretapped the conversations of American citizens, that his administration developed an assassination program that it kept secret from Congress — but leaders are not to be judged by their actions! Only a sophisticated analysis of their inner psychology can truly enlighten us as to their character and motivations.
UPDATE: Rich Lowry agrees that Mr. Ledeen's post is wrongheaded.


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One of only two Gamma Knife radiosurgery centers in the state offers brain tumor patients a safer treatment option
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Since its opening in 1998, the care team at the Palmetto Health Gamma Knife Center has treated thousands of patients with this non-invasive procedure that selectively destroys tumors and blood vessel abnormalities deep within the brain.
Conditions treated by Gamma Knife include:
Columbia native Elizabeth White is one of many patients who has benefitted from the technology. Recalling that frightening time she knew something was terribly wrong with her health, White said, "I started feeling a slight pain at the top of my head and my balance was off. If I was walking and I had to step up on a curb, I couldn't step up and I didn't know what was going on."
After a series of scans, it was discovered that White had a tumor pressing on her cerebellum, plus a number of "hot spots" on her lung, thyroid and in her brain.
"In about a four-month period I had the brain tumor taken out around my cerebellum, most of my thyroid removed, and I had a lung biopsy," said White. "They called me 'The Enigma' because it took the doctors about two months to figure out where all the spots were coming from."
Following her surgeries, White started ongoing treatments at Palmetto Health Gamma Knife Center, where Mangubat has been catching remaining metastatic lesions in her brain.
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Advantages of Gamma Knife radiosurgery at Palmetto Health include:
For more information, visit PalmettoHealth.org/GammaKnife or contact Palmetto Health Gamma Knife Center at 803-434-3497.
World-class neurological care and the most advanced brain surgery, spine and stroke treatments in the Midlands.


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Offer IP for lease
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Obstructionist Zimmer Propaganda
Zimmer is saying NO to open space in SW Hoboken
Don't be fooled by Dawn Zimmer's statement today that it's in progress and park land is coming and that she'll continue to buy more land after that.
She says: "the process is going on"… "at one acre, it's large enough."
If anything, the Zimmer administration has managed to halt progress, prevent others from bettering Hoboken, and to undermine the potential for a real 6-acre park. What we get left with instead is obstructionist Zimmer propaganda.
Conveniently, she left out some crucial information, that all of this is in the courts because it's being contested.
Zimmer says "it doesn't take years to get the park land," and yet, she has been an elected official for 6 years and Mayor for 4 years and she hasn't bought ONE INCH of land. Is she telling us how incompetent she is? That a good leader wouldn't take years to buy the land – but a unqualified, incompetent politician like Zimmer needs decades?
Obviously Zimmer's own spin machine is making her dizzy.
It has been discussed to death that you designate the area in need of redevelopment and then you buy the land all at once. Over the last six years, it has been brought up at numerous council meetings that the "piece-meal" approaches to buying land and building parks does not work. The politicians all agreed, and Candidate Zimmer was one of those people saying she would do it the right way. Unfortunately, Mayor Zimmer is now touting a piece-meal approach. Imagine that. But we already know that an undefined plan leads to failure, and that's why Zimmer hasn't succeeded in buying parkland of any kind.
So now Zimmer is saying the residents can wait for the SW Park, a park that she has decided will be less than 1-acre. Because we all know 1-acre is better than the 6-acres promised over the last 7 years (by her!)
The take away from Mayor Zimmer's letter:
- Zimmer is saying NO to open space in SW Hoboken.
- You want the Jackson St. lots to be turned into green public spaces? Sorry, Zimmer says NO.
- You want a SW park? You'll have to wait, maybe years – maybe forever.
In Zimmer's mind the park doesn't matter as much as getting her re-elected and her letter is another in a long line of her public relations bulls%#t.


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We run all over the place a lot of Texas Colorado Pennsylvania those are some big runs we do a lot do a lot of Wisconsin they go to Utah sometimes I steady get like 2500 to 3000 miles a week
Find Trucking Jobs
Climate Express - Union, MO
Discussion in 'Discuss Your Favorite Trucking Company Here' started by Efon, Mar 18, 2012.
Page 4 of 5
Scooter Jones Road Train Member
This thread is 6 years old. Efon left Climate Express a couple of mos after he started there.
Since this post shows up as first forums result I'll add some up to date info for the company.
First of all drove for them for a year from 2017 to 2018 and I'm back.
They have automatic Volvo 670s and 760s (phasing out the 670s)
Governed at 65
Pay is in the upper 40s with 9cpm of that being per diem.
They run mostly out of St Louis area and back.
Runs are normally to texas, Carolinas, Pennsylvania, and Denver but they do run all 48 except west coast and no north east.
They are a good company to drive for and easy enough to get along with.
Let me know if you have any questions.
P.s. I realize this is a Necro but this post still comes up as first forums result on Google so I'm sure there will be people by that are interested in this information.CarsNCameras Thanks this.
bryan21384 Road Train Member
- 7,901
- Sep 18, 2009
Trucking Jobs in 30 seconds
Every month 400 people find a job with the help of TruckersReport.
Page 4 of 5


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Dan Rank Testing
Dan Tests
Reading Time: < 1 minute The U.S. Soo Bahk Do Moo Duk Kwan Federation's Technical Advisory Committee provides 2 opportunities per year in each Federation region for qualified students to test for 1st, 2nd and 3rd Dan ranks. Higher rank candidates are tested at an annual, week long Masters exam called the Ko Dan Ja Shim Sa. Federation Regional Dan…Read More


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Before he gets to that Bumblebee spin-off, Michael Bay has to wrap up Transformers 5, also known as "the last one of these I'm making, I swear." The film, which is part of a planned Transformers trilogy that includes Bumblebee's lone outing, is scheduled for release on June 23, 2017, and it doesn't take the leader of the Autobots (or even Mark Wahlberg) to see that leaves all of 13 months to get this thing in the can. Make that 12 months—Paramount has just released a new promo for the film, which reveals its title and production start date.
So it looks like Transformers 5 has officially become Transformers: The Last Knight. Combined with the new, worse-for-wear logo, the revised title could be hinting at a more medieval setting. Maybe Optimus Prime pops through a wormhole on the way back from his robot-god-killing mission and ends up traveling back in time. Or maybe he just saves Queen Elizabeth II's life in the present and ends up knighted for his troubles. What are we saying? The last knight is obviously Mark Wahlberg.


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Share Your Photos!
Saskatoon, Saskatchewan
We would love if you shared your NYAW event pictures and stories with us!
Saskatoon, Saskatchewan
We would love if you shared your NYAW event pictures and stories with us!
Toronto, Ontario - Art Fusion Festival
Well done everyone! National Youth Arts Week 2013 was even better than we could have hoped for, with Launch Parties that took place in Halifax, NS, Saint-Jérôme, QB, Toronto, ON, Iqaluit, NV, Calgary, AB and Vancouver, BC.
Remix our logo! Download the template, jazz it up and use it to promote your own NYAW event! You can sell or give-away t-shirts, posters, buttons or use it on your website to make noise for NYAW. Share your remixed logos on our Tumblr site for Canada to see, and we'll feature them!
Looking for ideas of what to do for National Youth Arts Week? Love to dance? Check out our newest toolkit, How to run a Dance Flashmob!
Toronto, Ontario - Live Painting Day
If you are a young artist, local community organization, community group, youth arts organization or corporation or granting body, there are many ways for you to get involved!


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Christof Veillon : man of many passions
It is said that there are two kinds of people : those who are led by their heads, and those who are led by their hearts. Regarding Christof Veillon there is no question as to what kind of man he is, for it was out of love of acting that he pursued his career, and it was his love for a woman that brought him to Atlanta.
It was never in the plans for Christof Veillon to settle in the U.S., much less pursue an acting career. In fact, his interest in the craft developed relatively late as a teenager, who used to spend his free-time watching movies with his buddies. Raised in a family that did not place great importance on culture and the arts, it was his French teacher in high school who first introduced him to the world of acting beyond the cinema. In preparation for the French Baccalaureate, this exceptional and astute teacher took his class to the rehearsal and performance of L'Ile des esclaves by Pierre de Marivaux. Never before had Veillon been exposed to the fun and magic of the theatre, and from then on the decision was made.
He left his hometown near Lyon and headed to Paris to study acting at the Ecole Florent, where at 19, he performed for the first time on stage. He went on to perform at prestigious venues in France such as the Comédie Française and the Théâtre de la Ville de Paris as well as in Italy, Austria, and Slovakia. In addition, he played in several TV movies, independent films, and in a series of radio plays on the national radio, France Culture.
Thanks to a director with whom he had worked with on several projects, he was introduced to the new passion of his life— a young American student working on the translation of one of Koltès' plays in conjunction with the U.S. Koltès Project in Atlanta. Veillon wined and dined her, and as he proudly shared, his future wife fell in love with him over a plate of his homemade ratatouille. The two married, and in 2010, he moved to Atlanta where his bride, Jennifer was finishing her PhD at Emory University and taking on a new job at Georgia Tech.
What struck Veillon upon his initial arrival to Atlanta was the spectacular aerial view of this city in the middle of the forest—a sight "that doesn't exist in Europe". Avid reader, film-goer, and nature-lover, he was also very impressed by the diversity and creative energy. "Atlanta gives off the vibe of being a place to create, change, propose new things, and that's a very nice feeling. I lived in Paris for 20 years, and it was more difficult. We had the feeling—even if it was false—that everything had already been done," said Veillon.
As many expatriates who have settled in the U.S. for love rather than work know, it is not necessarily easy to build a career from the ground up on foreign soil, yet Veillon has wasted no time in building his network and getting work. The Théâtre du Rêve (TdR), Atlanta's own French-speaking professional theatre company with whom he had worked in Paris several years back, offered a wonderful opportunity for him to immediately jump in the water and make contacts in his field. He has played in several of the TdR productions and has also directed and performed a tribute to the famous French singer Serge Gainsbourg at the Alliance Française d'Atlanta. In addition, he's been the lead in two independent films and recently played in the last James Wan's feature film, "The Conjuring". One might assume that as an actor, Veillon would have no problem mastering the English language, and from what he demonstrates on the stage and the screen, that is certainly true, but little does one known about the effort, focus, and practice that goes into perfecting his American articulation and pronunciation.
"When I arrived, I hardly spoke English," Veillon confided. Although his wife is American, they only speak French at the house, especially since they became parents five months ago. In preparation for his audition with a booking agent, he arrived half an hour early in order to get the script, and then hid in his car to practice the pronunciation with his wife over the phone. Veillon's skills paid off, and he now has an agent to help him land roles in this city that is quickly becoming an important player in the entertainment industry.
"It's a bit precarious for artists [in the U.S]. In France, we have structures in place to protect them, but I really feel like I'm in the right place at the right time," shared Veillon.
As he strives for the stars, Veillon remains focused on the local community and creative opportunities available. He's very excited about a French-English acting class that he began teaching recently with the TdR for children—many of whom come from the local French community—as well as the upcoming class for adults this fall. In addition, Veillon is planning a series of small theatrical presentations within the framework of TdR based on and presented in conjunction with the High Museum's fall exhibition on the Jardin des Tuileries. He is also very conscious of his new role as a father, and with that the new dimension that the future takes on. His daughter, June Marguerite, has become Veillon's newest and most precious passion, and as he looks into the unknown future full of possibilities, her happiness is at the center, and everything else will, like always, fall into place.


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Kay Sibiya is one of South Africa's most sought-after bachelors. The media personality was thrust into the spotlight when he bagged a role on SABC 1's flagship show, Uzalo and since his appearance on the show, he has solidified himself as an actor and influencer.
After he left Uzalo, the actor had a short stint on Mzansi Magic's, The Queen and although we have not seen him on our television screens in a while, he is still a part of popular culture.
The actor set social media alight when he posted snaps of him and a girl who is believed to be his girlfriend, although they have not confirmed their relationship, social media users are convinced that this is the woman in Kay's life. Take a look at the snap below.
Despite his acting career, Kay is also known for his fashion sense and a couple of months ago, we wrote an article highlighting his style.


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Medic dies during Skype with wife
HOUSTON (AP) — The Texas-based wife of an Army medic serving in Afghanistan witnessed her husband's death as the two video chatted via Skype, his family said Friday.
The circumstances of Capt. Bruce Kevin Clark's death Monday were not immediately available.
"We are entrusting the military with investigating and with finding out what happened to Capt. Clark," Bradley Taber-Thomas, Clark's brother-in-law, told The Associated Press.
The Pentagon said the 43-year-old officer was formerly from Spencerport, N.Y., and that his death is under investigation.
"Bruce's wife tragically witnessed her husband's death during one of their regular Skype video-chats on Monday," Taber-Clark said in a prepared statement. "At the time of the incident, the family was hoping for a rescue and miracle, but later learned that it was not to be.
Clark, an Army chief nurse, was assigned to the William Beaumont Army Medical Center in El Paso and deployed in March. Taber-Thomas said Clark is survived by his wife and two daughters, age 3 and 9.
Taber-Thomas said the soldier's body was returned Thursday to Dover Air Force Base.
"Although the circumstances were unimaginable, Bruce's wife and extended family will be forever thankful that he and his wife were together in his last moments," he said.
A funeral is planned in Spencerport, N.Y., a suburb of Rochester and his wife's hometown, but details were not immediately available. Clark and his wife moved there in 2000 and he joined the Army six years later.
A memorial service also was planned for Addison, Mich., where Clark graduated from high school.
"While we understand the interest our family's tragedy will attract given the unique circumstances, we ask that our privacy be respected during our time of grief," Taber-Thomas said.


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Research Article | Open Access
Remco Franssen, Alinda W. M. Schimmel, Sander I. van Leuven, Simone C. S. Wolfkamp, Erik S. G. Stroes, Geesje M. Dallinga-Thie, "In Vivo Inflammation Does Not Impair ABCA1-Mediated Cholesterol Efflux Capacity of HDL", Cholesterol, vol. 2012, Article ID 610741, 8 pages, 2012. https://doi.org/10.1155/2012/610741
In Vivo Inflammation Does Not Impair ABCA1-Mediated Cholesterol Efflux Capacity of HDL
HDL provides atheroprotection by facilitating cholesterol efflex from lipid-laden macrophages in the vessel wall. In vitro studies have suggested impaired efflux capacity of HDL following inflammatory changes. We assessed the impact of acute severe sepsis and mild chronic inflammatory disease on the efflux capacity of HDL. We hypothesize that a more severe inflammatory state leads to stronger impaired cholesterol efflux capacity. Using lipid-laden THP1 cells and fibroblasts we were able to show that efflux capacity of HDL from both patients with severe sepsis or with Crohn's disease (active or in remission), either isolated using density gradient ultracentrifugation or using apoB precipitation, was not impaired. Yet plasma levels of HDL cholesterol and apoA-I were markedly lower in patients with sepsis. Based on the current observations we conclude that inflammatory disease does not interfere with the capacity of HDL to mediate cholesterol efflux. Our findings do not lend support to the biological relevance of HDL function changes in vitro.
1. Introduction
High-density lipoprotein (HDL) cholesterol levels exhibit strong inverse relations with cardiovascular disease risk [1–3]. In the light of this, raising HDL has been a prime target in current drug therapy. HDL exerts a variety of antiatherogenic effects [4]; it improves endothelial function, inhibits thrombosis, and has potent antioxidative properties as well as anti-inflammatory effects [5–7]. Most importantly, HDL is a key player in reverse cholesterol transport (RCT), a process that facilitates transport of cholesterol from peripheral tissues to the liver [8, 9]. In the course of this process, small HDL precursors, such as lipid-poor apoA-I and pre-β HDL, acquire free cholesterol and phospholipids through the action of ATP binding cassette transporter A1 (ABCA1) [10]. Maturation occurs through the action of lecithin cholesterol acyltransferase (LCAT), and further lipidation of these particles occurs by interaction with the ATP-binding cassette transporter G1 (ABCG1) resulting in the formation of α-HDL particles [11]. Finally, HDL lipoproteins are—as described for mice—directly taken up in the liver by scavenger-receptor-B1 (SR-B1) or HDL lipids are transferred to apo-B-containing lipoproteins via the action of cholesterol ester transfer protein (CETP) and will be subsequently cleared through hepatic low-density lipoprotein (LDL) receptors [5].
More recently, concerns have been raised that, beyond HDL quantity, HDL quality changes may also contribute to impaired cardiovascular disease (CVD) protection [12]. Inflammation is a central feature during all stages of atherosclerotic plaque formation with cytokines and chemokines orchestrating the influx of immune cells in disease vessels [12, 13]. At the same time, cellular activation status of local macrophages determines oxidation of subintimal cholesteryl esters and plaque stability [14]. Systemic inflammatory disease, such as rheumatoid arthritis, has been associated with an increased CVD risk [15]. Patients with active Crohn's disease (CD) also presented with accelerated atherogenesis [16], although a recent meta-analysis was unable to substantiate an increased CV mortality rate in CD [17].
Recent in vitro findings suggested that the propensity towards CVD risk in chronic inflammatory disease might in part be related to an impaired protective capacity of HDL particles. It has been shown that myeloperoxidase (MPO), a potent inflammatory mediator predominantly derived from polymorphonuclear cells, can modify apoA-I, resulting in the generation of "dysfunctional" HDL particles with attenuated capacity to mediate ABCA1-dependent cholesterol efflux [18–22]. Colocalization of MPO and apoA-I in the human plaque has fuelled the concept that MPO may also be involved in HDL modification in vivo [20, 23, 24].
In the present study we set out to analyse whether systemic inflammatory disease, dependent upon the severity of the disease, affects the capacity of HDL to mediate cholesterol efflux. HDL from patients in a mild chronic (Crohn's disease—CD) and acute severe inflammatory state (sepsis) was tested in in vitro cell-based efflux systems. A fibroblast cell line, expressing only ABCA1 was used to test the initial hypothesis that inflammation may specifically affect apoA-I-mediated cholesterol efflux. THP1 cells, a macrophage cell line expressing ABCG1, ABCA1, and SR-B1, were used to reflect total in vivo cholesterol efflux. These studies may imply that quantity of HDL may be more important for total cholesterol efflux than the quality of HDL.
2. Methods
2.1. Patients
Ten patients with Crohn's disease (CD) were recruited at the outpatient clinic (inflammatory bowel disease) at the AMC. During study visits, disease activity was assessed using the Harvey Bradshaw index (HBI, a research tool taking into account clinical parameters such as, general well-being, abdominal pain, number of liquid stools per day, abdominal mass and complications). Five patients with were considered to have active CD and five patients with were in remission. Additionally, we have selected 8 patients with severe sepsis from the ICU unit of the AMC. Sepsis was defined as the presence of a hemodynamic compromised state with temperature above 38°C and CRP levels above 50 mg/L. Additionally 5 healthy controls were recruited from the general population. The institutional review board at the AMC approved the study. All subjects gave written informed consent.
2.2. Blood Collection and Biochemical Measurements
Blood was collected after an overnight fast in EDTA-containing tubes and directly placed on ice. Blood of the patients with sepsis was collected during the admission to the hospital disregarding the fasting status of the patient. Plasma was isolated by centrifugation at 4°C, 3000 g for 15 minutes and stored at −80°C for further analyses. Cholesterol and triglycerides were analysed using a commercially available enzymatic method (Randox, Crumlin, UK) on the Cobas Mira autoanalyzer (Roche, Basel, Switzerland). HDLc and LDLc were analysed by a commercial assay (WAKO, Neuss, Germany). ApoB and apoA-I were analysed using a turbidimetric assay (WAKO, Neuss, Germany). Plasma C reactive protein (CRP) was analysed with a high-sensitivity nephelometric assay (Roche, Basel, Switzerland). Serum amyloid A (SAA) concentration was analysed with a commercial available ELISA (EL10015) from Anogen (Mississauga, Canada). HDL preparations of all patients were included in the efflux studies.
2.3. HDL Isolation
HDL (: 1.063-1.21 g/mL) was isolated by gradient ultracentrifugation in a TLR 100.4 rotor (Beckman Coulter, Fullerton, USA). In short, 0.9 mL human plasma was brought to density g/mL by addition of KBr. UC-tubes were filled with 4 ml 0.9% NaCL. 0.9 ml high density plasma was loaded underneath the NaCL using large injection needles. HDL fraction was harvested after centrifugation for 2 h, 12°C, 100,000 rpm in a Beckman Table top ultracentrifuge. HDL was subsequently dialysed against PBS (0.9% NaCL, 10.9 mmol/L Na2HPO4·2H2O, 1.8 mmol/L NaH2 PO4·2H2O, pH 7.4; Fresenius Kabi), and apoA-I concentration was measured and used directly in the efflux experiments. Then, HDL was obtained by precipitation of the apoB-containing lipoprotein fraction with polyethylene glycol 8000 (Sigma P-2139) as described [25].
2.4. Preparation of MPO-Modified HDL
HDL was purchased from Calbiochem (cat. nr. 178452) and used for the MPO-modification reaction exactly as described [19]. In short, HDL (1 mg/mL apoA-I) was added to the reaction mixture containing 50 mM phosphate buffer, pH 7.0, containing 100 μM DTPA, 100 μM NaCl, 140 μM hydroperoxide, 57 nM purified human MPO (BioDesign Int; cat and A63100H). Incubations were performed at 37°C for 1 h. MPO-modified HDL was dialyzed against PBS and used as a positive control in the efflux experiments.
2.5. HDL-Mediated Cholesterol Efflux from Fibroblast and THP1 Cells
Primary human fibroblasts (NHDF-Ad- dermal fibroblasts; Cambrex, CC-2511) expressing human ABCA1 were grown in Hanks Balanced Salt Solution (HBSS) medium without Ca and Mg, containing 10% FBS (GIBCO 14170-088) and 0.1% penicillin/streptomycin at 37°C under 5% CO2. To measure cellular cholesterol efflux, fibroblasts were plated and grown until 80% confluency; at this point cells were labelled for 24 h, with 0.5 μCi/mL 3H-cholesterol in labeling medium (D-MEM : F-12 (1 : 1), Glutamax, 0.1% penicillin/streptomycin, 0.2% BSA, and 30 μg/mL cholesterol in the presence or absence of 3 μM LXR agonist T0901317 (Cayman Chemicals Comp.; art 71810) for fibroblasts. After labeling, efflux was induced by the addition of 0.5 mL of efflux medium, consisting of D-MEM : F-12 (1 : 1), Glutamax, 0.1% P/S, 0.2% BSA, and the patients HDL, or HDL from control subjects, isolated by apoB precipitation or HDL isolated by ultracentrifugation at a concentration of 10 μg/mL apoA-I, or HDL isolated from plasma obtained from controls, for a period of 4 h. Efflux medium without apoA-I was used as a negative control, and commercially available apoA-1 at a concentration of 10 μg/mL was used as a positive control. At the end of the incubation period, medium was expired. Cells were extracted with 2-propanol. Both medium and cell extracts were transferred into a vial containing scintillation fluid and counts were assessed (Packard, Tri-Carb 2900TR Liquid scintillation analyzer). Fractional cholesterol efflux was obtained by measuring the release of radio-labeled cholesterol into the medium. The percentage efflux equals the counts in medium divided by total counts in medium and cells combined, subtracted by counts to medium without HDL (control).
THP1 cells (ATCC) were grown in RPMI 1640, glutamax, 25 mM HEPES, 10% FCS, and 0.1% Penn/Strep at 37°C under 5% CO2. One day prior to the experiment, cells were differentiated into macrophages by addition of phorbol 12-myristate 13-acetate (PMA: 50 ng/mL; Sigma p8138) to the cells. The efflux procedure was similar to that described above for fibroblasts. All experiments were performed in triplicate on the same day in different wells.
2.6. Statistical Analysis
All data are presented as mean (±SD) unless stated otherwise. Statistical analysis was performed in SPSS version 16. All differences in sepsis and controls were analyzed using unpaired Student's t-test statistics. The differences in CD-active, CD-remission, and controls were analyzed using two-way ANOVA and LSD post hoc analysis. A value <0.05 was considered as being statistical significant.
3. Results
3.1. Baseline Characteristics
The baseline characteristics of patients with Crohn's disease are shown in Table 1. Patients with CD-active, reflecting a mild inflammatory state, had 10-fold higher plasma CRP levels than those with Crohn's disease in remission (; Table 1). In line, patients with CD-active had a tendency to lower plasma HDL levels (CD-active: mmol/L versus controls: mmol/L; ; Figure 1). ApoA-I levels were similar to those in controls. As a consequence, the HDLc/apoA-I ratio in patients with active Crohn's disease was not significantly different (Figure 1).
|Data are presented as mean (±SD). TC: total cholesterol; LDLc: low density lipoprotein cholesterol; HDLc: high-density lipoprotein cholesterol; TG: triglycerides; apo: apolipoprotein. ; versus controls.
In patients with sepsis, plasma CRP levels were very high (>200 mg/L). Subsequently, plasma HDLc levels were strongly decreased ( mmol/L; controls: mmol/L; ; Figure 1). Plasma apoA-I levels were 64% lower than in controls (0.5 g/L versus 1.4 g/L; ); (Figure 1), reflecting a strong decrease in circulating HDL particles. The HDLc/apoA-I ratio was markedly reduced as compared to controls ( mol/mol versus mol/mol; ; Figure 1). Plasma SAA levels in patients with sepsis were significantly higher than in controls (sepsis: >400 mg/L and controls: 2.5 (range: 1.0–7.2) mg/L). Plasma SAA levels in CD-active and CD-remission were measured in a different, but comparable cohort of patients. Plasma SAA levels in CD-active was 23 mg/L (range 3–148) and in CD-remission 5.4 mg/L (range 1.2–69).
3.2. MPO-HDL Displays Impaired Cholesterol Efflux Capacity
We first tested whether in vitro modulation of commercially available human native HDL by MPO impaired the capacity of HDL to mediate cholesterol efflux, measured over a 4 h time span in LXR-stimulated fibroblasts to test ABCA1-mediated efflux. THP1 cells were used to test total efflux capacity mediated by ABG1, ABCA1 and SR-B1 and ABCA1-mediated efflux under LXR agonist stimulation of ABCA1. We have chosen to standardize the amount of HDL added to the incubation system based on apoA-I concentration in the HDL preparations, so that in every efflux experiment comparable amounts of apoA-I are present, reflecting an equal number of HDL particles. In line with expectations, cholesterol efflux capacity of MPO-HDL was significantly decreased by 62% () in fibroblasts and 32% () in THP1 cells (Figure 2) under both conditions, providing evidence that our experimental setup is viable.
3.3. Cholesterol Efflux Capacity of HDL from Patients with Crohn's Disease
HDL was isolated by ultracentrifugation as well as by precipitation to avoid bias through the presence of serum proteins in the PEG-precipitated HDL preparation, which may interfere with the cholesterol efflux measurement. Again equal amounts of apoA-I (10 μg/mL) were added to each well in the efflux experiment. Surprisingly, HDL isolated by ultracentrifugation (UC-HDL) showed similar ABCA1-mediated cholesterol efflux in fibroblasts as compared to controls; (Figure 3(a)). UC-HDL showed increased total cholesterol efflux potential in THP1 cells (CD-active: +28%, versus controls; CD-remission: +24%; versus controls (Figure 3(b)). PEG-HDL (10 μg/mL apoA-I), as a control, isolated from CD-active and CD-remission patient's plasma induced a similar ABCA1-mediated cholesterol efflux in fibroblasts as well as total cholesterol efflux potential in THP1 cells as compared to control PEG-HDL (Figure 3).
3.4. Cholesterol Efflux Capacity of HDL from Patients with Sepsis
UC-HDL from patients with sepsis, reflecting a severe inflammatory state, showed a similar ABCA1-mediated cholesterol efflux potential from cholesterol-laden fibroblast cells in the presence of a LXR agonist (Figure 4). Cholesterol efflux potential from THP1 cells to UC-HDL isolated from plasma of patients with sepsis was increased as compared to controls (+14%, ; (Figure 4). Cholesterol efflux potential of PEG-HDL in cholesterol-laden fibroblast cells was similar to controls (Figure 4), whereas cholesterol efflux capacity of PEG-HDL in THP1 cells was significantly increased (+58%; ; Figure 4).
4. Discussion
We demonstrate that the capacity of HDL (normalized for apoA-1 concentration) of patients with severe acute or mild chronic inflammatory state, to mediate cholesterol efflux from cultured cholesterol-loaded fibroblasts treated with a LXR agonist to stimulate ABCA1 expression and THP1 cells, was not decreased as compared to normolipidemic controls. Even in patients with sepsis, suffering from a severe inflammatory disease state, HDL efflux capacity is, if anything, increased compared to healthy controls. These data underline that in vitro findings regarding HDL "dysfunction" cannot be extrapolated to an in vivo situation.
In the present study we have used two different cell systems to determine HDL-mediated cholesterol efflux potential. Fibroblasts are known to express the ABCA-1 receptor abundantly which can be further increased by treating the cells with a LXR agonist, whereas THP1 cells (under lipid-laden conditions) express ABCA1, SR-B1 and ABCG-1 and thus model lipid-laden macrophages in human atherosclerotic plaques [26]. In line with previous reports, we observed that HDL when exposed to MPO in a test tube displays impaired capacity to mediate cholesterol efflux from both fibroblasts as well as THP1 cells, [20, 24] showing that ABCA1-mediated cholesterol efflux is highly impaired and thus provides validation of our efflux system. Two different disease models, one representing an acute, severe inflammatory state (sepsis) and the other representing a mild inflammatory state (Crohn's disease), have been used in the present study. Both disease states were characterized by increased plasma CRP and SAA levels, as well as decreased plasma HDLc levels, albeit not significant in CD-active, which is in line with HDL acting as an inverse acute phase reactant [5]. Our original hypothesis was based on the observation that in vitro inflammatory challenges modulate HDL response to induce normal cholesterol efflux from cell culture systems. We expected to see a gradual decrease in HDL efflux potential with increasing severity of inflammatory state. However, despite the inflammatory conditions of these patients we observed a modest increase in the efflux capacity of HDL, but we feel that more robust analysis are required to further investigate this.
Thus, our data clearly indicate that impaired efflux capacity of HDL in inflammatory disease states does not play a significant role in vivo. Obviously, these findings do not exclude a decreased total efflux capacity, since HDLc and apoA-I concentrations are profoundly lower in patients with sepsis albeit not in patients with a mild inflammatory state (active Crohn's disease) [15, 27]. The efflux data were obtained through controlling for apoA-I levels in the different HDL preparations. Plasma apoA-I levels in patients with sepsis were 45% reduced. This means in theory that, although HDL was more efficient in accepting cholesterol from cells, this cannot be compensated for by the strong decrease in number of circulating HDL particles and thus an overall decreased total efflux capacity.
Interestingly, we observed a slight increase in HDL efflux capacity in an inflammatory disease state. Van Der Westhuyzen et al. have previously shown that, besides apoA-I, SAA is also capable of promoting cholesterol efflux [28]. Since we have used similar apoA-I concentrations in all our assays and more importantly since HDL particles in the inflammatory state contain an increased number of SAA molecules, this may have been the reason for the increase in efflux capacity. However, plasma SAA levels in patients with sepsis are more than 200-fold increased as compared to controls whereas plasma SAA levels in active Crohn's disease were only increased 10-fold compared to controls. Therefore, we conclude that plasma SAA levels do not contribute to the increased cholesterol efflux observed in both CD-active and sepsis.
Previous studies have suggested that smaller lipid depleted HDL particles serve as better cholesterol acceptors via the ABCA1-mediated efflux pathway [29]. Indeed, the estimated HDL-c/apoA-I ratio between controls and patients in a severe inflammatory state was significantly reduced suggesting the presence of smaller-sized HDL particles. In line, the average number of particles was reduced in our study as reflected by the decrease in apoA-I levels. These data lend further support to the concept that HDL quantity is the main determinant of cholesterol efflux capacity under these conditions. In contrast to our findings, Mc Gillicuddy et al. reported that, following an acute endotoxin challenge, several steps in the RCT pathway were disrupted [30]. Thus, in vitro exposure of macrophages to LPS significantly reduced ABCA1 expression thereby attenuating cholesterol efflux capacity towards apoA-I. Cholesterol efflux acceptor capacity of HDL obtained from subjects subjected to the LPS challenge was decreased with a concomitant decrease in plasma apoA-I level. There are several reasons that may underlie the discrepancy with the current findings. First, a single endotoxin challenge is not comparable to a more sustained inflammation, since a wide variety of adaptation mechanisms evolve over time [5, 31, 32]. Most importantly, these authors failed to normalize for the decrease in apoA-I levels after the endotoxin challenge in the cholesterol efflux assay, which invariably contributes to differences in cholesterol efflux. In fact, we demonstrated in septic patients that correction for changes in apoA-I result in a normal or even increased efflux acceptor capacity of HDL independent of the HDL preparation (PEG isolated or ultracentrifugally isolated).
4.1. Study Limitations
Our study has several limitations. First, the patients in the present study were older compared to the controls. Berrougui et al. have reported that particularly ABCA1-mediated cholesterol efflux decreased with advancing age [33]. This implies that in patients we may have observed lower values compared to the younger controls. However, since we find, if anything, higher efflux values, the age difference cannot have contributed to the present observation. Second, we included only a limited number of subjects in our study. However, since we observe an increased efflux in patients with an extreme inflammatory disease, it is unlikely that inclusion of a larger number of subjects would alter these findings.
4.2. Conclusion
Whereas exposure of HDL to MPO ex vivo leads to an impaired cholesterol efflux, native HDL preparations isolated from patients in mild and severe inflammatory disease states retain its ability to stimulate efflux. These findings do not lend support to the biological relevance of HDL function changes observed in vitro.
The authors would like to thank Mirjam van Aalderen for her assistance in the efflux studies and Joram van Miert for technical assistance. This work was supported by a Grant from the Dutch Heart Foundation 2008B070 to (R. Franssen).
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Copyright © 2012 Remco Franssen et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


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Ductless Heating & Cooling Units
Ductless heating and cooling systems are a smart alternative to traditional electric options for homeowners looking to increase savings without the headache of a time-consuming installation process. Ductless systems are remote-controlled heating and air-conditioning units, allowing homeowners to remain comfortable year-round. Other benefits you'll notice over time with your ductless system include:
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The Qin/Ch'in Dynasty (221 BC - 206 BC) was preceded by the Zhou Dynasty and followed by the Han Dynasty in China. The unification of China 221 BC under the First Emperor (Qin Shi Huangdi) marked the beginning of imperial China.
The Qin government was totalitarian, which placed absolute power in the hands of the ruler, ruling by means of strict laws and harsh punishments. Qin Shihuangdi ruled exactly like this and took severe actions to eliminate all challenges to his authority. Those who criticized Qin politics were executed. To silence the criticism of imperial rule, the First emperor had the Confucian scholars who condemned the Qin for inhumanity and injustice were purged. It is said that around 400 scholars who criticized the Qin government were buried alive. All books and histories of the empire other than those having to do with agriculture or medicine were burned. Anyone who wished to be educated received basic education by government staff themselves.
This anti-intellectualism ran counter to the atmosphere of free learning that had prevailed in the Qin state in the years before. To reduce the threat of insurrection, descendants of the ruling houses of the six conquered states were forced either to relocate to the capital or be exiled. All weapons owned by private citizens were confiscated, and the metal either was stored in the capital, or the metal was used to help make the gigantic metal statues and bells that decorated the imperial palace.
Feudalism was abolished, and land was taken from landowners and distributed among peasants, who were then taxed by the central government. A new code of law was devised and applied to all citizens equally, regardless of rank.
Conscription was devised; Males between seventeen years old and sixty years old was to serve one year...


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My pirate name is:
Dirty Mary Cash
You're the pirate everyone else wants to throw in the ocean -- not to get rid of you, you understand; just to get rid of the smell. You're musical, and you've got a certain style if not flair. You'll do just fine. Arr!
Get your own pirate name from fidius.org.


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Now that the 2014 elections are behind us, we thought we'd take a look at 11 actions Congress can take to improve the Affordable Care Act. Several of our suggestions have bipartisan support and, even in the 2015 session of Congress, a vote to do away with some of the less popular aspects of the […]read more
Health insurance will undergo significant changes in 2014, and many American families are wondering how to plan for them. Whether you have health insurance through an employer or buy it on your own, here are ten questions you and your family should ask right now. Question Why is it important? 1. Will health insurance cover my […]read more
The Affordable Care Act promised to deliver significant changes in health care and health insurance. During the past two years, we've learned a great deal about what is included in this sweeping new law. As with most wide-ranging laws, it can be hard to get the real facts and all too easy for misinformation about the law to […]read more
Supporters of the Affordable Care Act told us the law would make medical care and health insurance less expensive. Advocates said there were several reasons. More people will have insurance, including a lot more healthy young people. Competition created by health insurance exchanges will bring prices down. More people would gain access to preventive care. However, other parts of the […]read more
The promise of the Affordable Care Act is to make health insurance less expensive for American families. The promise is based on premium assistance from the federal government for people who qualify. But there is a surprise in store for many families. If you didn't know you might have to repay the premium subsidy you already […]read more


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Season 7, Episode 706 : Aired on Oct. 26, 2013 Episode Preview
Simple construction takes center stage as the editors of Woodsmith magazine build a rustic, step-back cupboard.
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The Department of Lands and Surveys (DLS) has participated in the co-financed project of the Public Administration and Personnel Department (PAPD) called "Reorganizing and Enhancing the Administrative Capacity of Public Service Departments", with a duration of seven years (from October 2008 until May 2015), the main objectives of which were to strengthen the administrative capacity of the departments involved, improve the services offered and enhance their internal functioning.
In the context of this project, the Department participated in the study "Performance Comparison, simplification of procedures, modernization / codification of the institutional framework and preparation of the Department for the implementation of ISO ". The study carried out by an external evaluator, which was the "PWC", lasted two years, from October 2012 to October 2014.
Main Activities:
Preparation of the DLS for the implementation of ISO 9001: 2008:
Subsequently, after all the above were successfully completed, according to the time-lines of the co-funded project of the PAPD, work started on the "ISO 9001: 2008 Uniform Quality Management Systems Certification" , having the Cyprus Certification Company as the contractor company, which was of four months' duration and was completed early in June 2015.
During this project, the contractor company initially collected the required information in relation to the requirements of the standard, with visits to the Department and discussions with the quality team. Then, it proceeded to its first conclusions / findings on the state of the existing Quality System that had been created in the DLS. After the first stage was successfully completed, i.e. without problems in the documentation of the System as it was built, it was followed by a third party external audit by the Cyprus Certification Company. During the external audit, the basic procedures of the Quality Management System were evaluated, the procedures based on the operation manuals of the branches, both in the Central and the District Land Offices, were audited, and the results of the internal audits and the management review, as well as the audit of the quality objectives and the results of citizen satisfaction survey were evaluated. The result of this project was the successful certification of the DLS with the ISO 9001: 2008 certification, without any serious observations or non-conformities.
The above certification constitutes recognition that the Department of Lands and Surveys, has the ability to operate on the basis of documented procedures and applies the relevant legal and regulatory requirements to meet the demands both of the Department itself and of the citizens. Quality will contribute to the more effective functioning of the Department, to the consolidation of a citizen- oriented culture, to the improvement of the level of services offered and to the increase of the degree of customer satisfaction.
ISO 9001 CERTIFICATE.pdf
IQNet CERTIFICATE.pdf


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Friends of the Brolga Theatre Group Inc has been established as a community based organisation to support and assist the work of The Brolga Theatre and to promote artistic ventures in this region.
There are two options for membership:
You can elect to become a member of either group and take advantage of the benefits available.
Friends of the Brolga are appreciated as genuine supporters of their publicly funded regional theatre and they receive the following benefits:
Many of our volunteers develop longstanding friendships with their other colleagues.
The Brolga is very privileged to have such a hard working dedicated group of volunteer workers who provide great service to our many and varied customers. The main aim of volunteer working members is to provide excellent customer service and facilitate the enjoyment of patrons at all performances.
Volunteer working members choose to assist in the operation of the theatre in a number of different roles including working as ushers, door-keepers, front-of-house operations and assistance with the liquor and coffee bar.
Volunteers wear a uniform provided by the theatre and undertake training in each of the work areas nominated in operational procedures, as well as Workplace Health and Safety. They are rostered to work on a rotational basis giving all trained members an equal opportunity to assist at events and performances.
The support of our volunteers makes our centre one of the most successful and vibrant regional theatres in Australia
Call the Brolga Box Office on 4122 6060