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Evaluation of Alternative Coordination Systems Between Producers and Packers in the Pork Value Chain
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Volume (Year): 06 (2003)
Issue (Month): 02 ()
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Please report citation or reference errors to , or , if you are the registered author of the cited work, log in to your RePEc Author Service profile, click on "citations" and make appropriate adjustments.:
- Johnson, C. Scott & Foster, Kenneth A., 1994. "Risk Preferences And Contracting In The U.S. Hog Industry," Journal of Agricultural and Applied Economics, Southern Agricultural Economics Association, vol. 26(02), December.
- Hayri �nal & Laurian Unnevehr & Aleksandar Bekric, 2000. "Regional Shifts in Pork Production: Implications for Competition and Food Safety," American Journal of Agricultural Economics, Agricultural and Applied Economics Association, vol. 82(4), pages 968-978.
- Mark Drabenstott, 1999. "Consolidation in U.S agriculture : the new rural landscape and public policy," Economic Review, Federal Reserve Bank of Kansas City, issue Q I, pages 63-71.
- John D. Lawrence & V. James Rhodes & Glenn A. Grimes & Marvin L. Hayenga, 1997. "Vertical coordination in the US pork industry: Status, motivations, and expectations," Agribusiness, John Wiley & Sons, Ltd., vol. 13(1), pages 21-31.
- Hennessy, David A. & Lawrence, John D., 1999. "Contractual Relations, Control, and Quality in the Hog Sector," Staff General Research Papers 1706, Iowa State University, Department of Economics.
- Perry, Janet E. & Banker, David E. & Green, Robert C., 1999. "Broiler Farms' Organization, Management, and Performance," Agricultural Information Bulletins 33739, United States Department of Agriculture, Economic Research Service.
- James M. MacDonald & Michael E. Ollinger, 2000. "Scale Economies and Consolidation in Hog Slaughter," American Journal of Agricultural Economics, Agricultural and Applied Economics Association, vol. 82(2), pages 334-346.
- Melton, Brian & Huffman, Wallace, 1995. "Beef and Pork Packing Costs and Input Demands: Effects of Unionization and Technology," Staff General Research Papers 5043, Iowa State University, Department of Economics.
- Hayenga, Marvin L., 1998. "Cost Structures of Pork Slaughter and Processing Firms: Behavioral and Performance Implications," Staff General Research Papers 1254, Iowa State University, Department of Economics.
- Martin, Laura L., 1997. "Production Contracts, Risk Shifting, And Relative Performance Payments In The Pork Industry," Journal of Agricultural and Applied Economics, Southern Agricultural Economics Association, vol. 29(02), December.
- Jill E. Hobbs, 1997. "Measuring the Importance of Transaction Costs in Cattle Marketing," American Journal of Agricultural Economics, Agricultural and Applied Economics Association, vol. 79(4), pages 1083-1095.
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New UK Blog
I'm a bit late in noticing this but the Piddinghoe Gamers have just launched a blog on their website here. The latest entry about the price of poker chips and their downward spiral is pretty upsetting. It rubs in the fact that I bought my set 18 months too early at a very steep price when you could only get… Read more »


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|Reviews for Seizensetsu
| ClumsyWrighter chapter 17 . 12/14/2014
whahaha i love how hes mind games totly backfires because of ash's perenalety XD
| ClumsyWrighter chapter 16 . 12/14/2014
how the hell did you make this story sadisfing DX
| ClumsyWrighter chapter 5 . 12/14/2014
"Trip is not yandere"
he's not?
enyways... whaty is this, corps party?
| ClumsyWrighter chapter 1 . 12/13/2014
then thet mean thet Ash is muts older... oh god... how i love stories like this... okey okey he ise nt thet mutsh older but still i think this will be Nice X3
| violet -x3 stone chapter 20 . 8/1/2014
YES JUST YES! Did you know this is my first time reading Kodakshipping? So holy hell you were awesome! Best first time. (That sounded inapropriete.) but I cursed describing it, AKA it was so good, it was worth cursing. I want more TRIP! Nom, nom, nom. I feel like writing some Kodakshipping now, if I do, I want to dedicate it to you since you inspired me. Haha, anyways, this crazy fangirl is out!
Lots of hearts,
| violet -x3 stone chapter 12 . 7/31/2014
I cried so hard at the end. You don't even know. I must keep this short though because of wifi should be asleep and sneaking around the house to better wifi spots. But I love you an this story. I cried. And the feels. Gotta love Kodakshipping. Lots of love your way.
| caird56 chapter 20 . 6/19/2014
OMG I FRICKEN LOVED THIS STORY BEST STORY EVER EVEN BETTER THAT EVERY COMASHIPPING STORY I'VE READ (yeah I'm a comashipper to me it's the best shipping ever)
| AbiWritesThings chapter 12 . 1/25/2013
Oh... My... God. I've read rape stories or chapters before but this is the only one that's actually ever made me cry. While I was reading it I just wanted to go kill the guys screaming that they just raped a fucking TEN YEAR OLD! You are one sick fuck and I kind of admire that
| KrazyKat12 chapter 17 . 11/12/2012
I'm certianly a fan of this story. I've loved it so much so far. The thing about Cilan, Chili, and Cress being the shadow triad, I actually took that seriously. Like they put that in the pokemon anime(that would be F-ing awesome!). At first I thought it was just you who thought that, but recent searching proved me wrong. Appearently a lot of people think so. I wonder why? Like, who connected the dots there, and thought of it first? Oh well, anyway, awesome story and I LOVE IT.
| Won-Chan108 chapter 1 . 9/24/2012
Before I read this, PLEASE answer these questions:
1. Who is Blair?
2. Is Ash uke, or seme, in the relationship with Trip?
| weet-woo chapter 1 . 7/18/2012
woot hint of ashXcilan :3
| lemonlime77 chapter 20 . 7/5/2012
Need... Sequel...
Need.. More... KODAK!
But seriously this story was AMAZING! It's kind of like if the anime had a plausible plot. And then you know, they added rape and stuff like that. Again, AMAZING!
| AmeNeko chapter 20 . 5/16/2012
I read this long before I got an account of FF.N, and I am seriously not a fan of WishfulShipping.
But I LOVE THIS STORY! Trip is my favorite character in Pokemon, and thanks to you, I'm watching Umineko and Higurashi and much much more!
I spotted a few errors here and there, but it's really well written. I applaud to it all!
The interactions between Trip and Ash were adorable. Might be one if my favorite Ships now. And Cilan being a part of the Shadow Triad? Great! Keep up the good work, as I am looking forward to a sequel!
| MewMewExorcist chapter 3 . 4/24/2012
*sweatdrops* Cilan won that battle... Nice story.
This is just... well, to say awesome or epic would be underestimating it. Seriously, this is just incredible, and I love it so much... I was actually so absorbed in the lemons that I didn't even notice all the description! And you're right, there does need to be more Kodakshipping out there... a LOT more.
Well, great job. It's been a few months, but hopefully you'll write that sequel. I would love to know what happens next, so hopefully it'll be out soon! :)


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Animal Shooting Games
This is the place to play free animal shooting games games in popular categories such as animal adventure, animal hunting, animal racing, animal arcade, animal action, animal fighting, animal killing. I would recommend animal shooting free online games. Some teach new skills with games like animal, adventure, shooting, action, killing games.
You can find the newly animal shooting games games here.


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Purpose of United Methodist Women
United Methodist Women shall be a community of women whose PURPOSE is to know God and to experience freedom as whole persons through Jesus Christ; to develop a creative, supportive fellowship; and to expand concepts of mission through participation in the global ministries of the church.
Special Events
Save the Date!
Annual Day - Dorrenceton United Methodist Church, Kingston, Pa - October 17, 2015 (Flyer and Registration Form)
Northeast Jurisdiction Quadrennial Meeting, June 16-18, 2016 - Syracuse, NY - "A Fresh Wind Blowing"
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August Newsletter
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Support the United Methodist Women Legacy Fund:
Purchase items from Equal Exchange and not only will you be supporting fair trade, but in addition, 25% of your purchase goes directly to the Legacy Fund! Browse their website and make purchases.
Contact Us
Susquehanna Conference United Methodist Women Officers for 2015:


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People just haven't been going out - most of the places around here aren't booking bands - or are booking really cheap ones. Our winter schedule hasn't been this thin for a very long time. How are you guys making out compared to last year?
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How is your winter schedule?* * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * *
My cover band
HARD WORK BEATS TALENT WHEN TALENT DOESN'T WORK HARDTags: None
We actually have a packed schedule for OH. 7 gigs between now and end of the year. The crowds have been up and down though... last Friday we had a thin (but fun) crowd but Thanksgiving weekend (Saturday) was awesome. I have a feeling it will be thin as we get closer to Christmas. Hope I'm wrong though.
Everything went to **************** in October.. we had a great run through 2012.. but crowds lately are just thin. We're playing a gig this weekend at another regular hotspot, and if we don't really bring it, I think we might be in jeopardy of not getting booked there in 2013..
Nearly every place we play has told us they are holding off on 2013 bookings, either to decide whether to have them, or decide on slashing pay and bringing in bands willing to work for nothing (I'm guessing). Notably, one place we finally got into booked us on a rotation, based on how well we did there in the summer on fill-in gigs.. so there's that.
We are going to weather this storm.. but it's looking pessimistic for the near future. A lot of places can't even sell out for new years.. and are starting to panic. We're not booked new years.. only offer we got was for peanuts (less than our minimum), so we declined.Sig Fail
Back in April or so when we were booking fall/winter, we purposely didn't book dates from Thanksgiving until second week of Feb. We did this knowing that the crowds are thinner throughout that time period, plus to accomodate band member schedules for vacations and family time, as well as to retool the setlists.
We're going on a little hiatus while one of the band members works through some personal stuff. Too bad, the last few gigs have been great and it felt like a little momentum might be building for us. I think we only have one January gig left on the calendar and we're not booking anything new until we see how things shake out.>>>Click here for my list of smooooov deals!
We have three rooms that we can depend on to book us in the winter - and one of them is in trouble, so I don't know how long they will last. We've dropped two rooms since last year ($$$$) and another room isn't booking until 2013 and then wants us for their new beach location, so we don't know how that will work out. Two other rooms are seasonal, so we need to wait for them. There is one other room where we have one more show to play and they haven't been doing much in the booking department other than DJs.
It's not like we're over-saturating the market, but people aren't going out like they did just a year ago. We haven't maxed out our best-drawing room since last year. Our venues tell us that our crowds are still better than most/all of the other bands that they have, but if that is the case, then they have got to be feeling the crunch.
This is not the best thing to happen after purchasing a major PA upgrade and my own investment in a keyboard rig.* * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * *
My cover band
* * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * *
My cover band
we are slow for the off property stuff. Got a regular rotation mkt days show in jan. The new cd is charting in the top 10 on the cashbox pop country chart so , we may see some insteresting stuff happen after the first of the year. Its always feast or famine with us it seems lol."you mess with him and you mess with the whole trailer park"
We've mostly been off since our singer had her baby. Played a couple gigs in Oct with her sub that went well crowd wise. November we had 1 bar gig that was packed and then a private gig last weekend. We booked nothing for December by design. January we have 4 gigs booked, which is about the limit we'll do in a month. 2 restaurant/bars, a casino, and a private. Both bar/restaurants are excited to have us back after the layoff and I'm sure they will be crowded since we haven't been there for a while with our usual singer.My Live Gear: Roland FA-08, Hammond SK1-73, Moog LP
My Band: http://www.bksband.com
Slow, slow, slow for us so far. Winter's usually are, but this one seems even slower than usual. After NYE, I don't think we have anything yet on the books until May. Stuff always seems to pop up, but I think we had at least one gig a month booked at this time last year._________________________________________________
band websites: http://www.JumpStartYourParty.com
It's been slow for us after a busy spring and somewhat busy summer. One of the venues that was booking us monthly cancelled their Friday night music series (including dates we had booked in both Nov. and Dec.), another place is keeping us on for dates this month and in January, Feb, and March, but they've reduced their pay-scale substantially.
Ah, well. Onward.....
We were recently in self imposed slow period while we changed drummers. This Saturday - we'll resume our regular schedule of about 1.5 gigs per month spread out over 5 venues. That is the speed at which we like to move.
I am glad we dodged college football season if we had to have any down time. This being the weekend after the SEC championship game - I am hoping that the focus will be back on music at the places that we play.nine8central
It's slow for me. I was doing 18-20 a month in the summer, I predicted it would drop down to about 12-15, and it's actually looking like about 8-10 if I'm lucky. Considering I play full-time I was getting a little nervous. I'm so burned out at this point between gigs and school I simply don't care and I'll live off student loans for a couple of months if I have to.
I will say that when I'm done with school in a year I won't be doing more than 75 dates a year. Playing full-time at 43 is not even close to being fun after awhile.


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"Convenient Selcuk" Selcuk by eugini2001
Selcuk Travel Guide: 339 reviews and 945 photos
Selcuk is a nice place: Ephesus, St. John Basilica, bus station in a walking distance from every hotel, many places to dine out,so it's small and convenient. Beaches are in 15 min ride on a mini-van distance. Also every Saturday and Wednesday it a shopping paradise, you can buy everything very cheap if you know how to bargain. I bought very cool tablecloth (cotton 100%) for 30$, the sale person asked for 50$. My child bought a lot of stuff also. You can buy a lot of tasteful fruit and vegetables also, since Saturday is a farm market day. All this stuff is organic and taste so gooooooooooood! We stayed 4 nights there and enjoyed it very much!
Our guide said this mosaic is the oldest in Ephesus.Our guide said this mosaic is the oldest in Ephesus. It's next to the sarcophagus of Cleopatra's sister. This cruel woman killed all her folks in order to became a queen , her life still was not perfect ,but l like her, the little ugly Jewish conqueror.She built some Egyptian buildings in Ephesus too, devoted to her Gods of course. That's way some columns looks so much different from Greek or Roman ones.
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Just wanted to post a couple of beanies I finished over the last few days. I couldn't figure out how to post two images so if one of them doesn't show up just go to my site (www.earthtodavid.com). I posted them there also. By the way I hope all you guys had a happy halloween. I spent the whole evening at home in front of the tv watching the Ghost Hunters Live showing on Sci-Fi. It was kinda boring but a little creepy and fun at the same time.
Williamsburg Print And Autumn Stripes
Submitted by madhatter7zero on Sat, 2008-11-01 20:53
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Visit Crafty Andy's Blog
Permalink Submitted by Crafty Andy on Sat, 2008-11-01 22:26
Beautiful hats, I love the
Permalink Submitted by crmartin on Sun, 2008-11-02 07:41
Beautiful hats, I love the color combinations. I would think they will be very popular.
I love your new hat, the
Permalink Submitted by bedauph on Sun, 2008-11-02 20:17
I love your new hat, the colors are very cheery and happy. Nice.
Nothing wrong with staying
Permalink Submitted by pjmma on Mon, 2008-11-03 10:38
Nothing wrong with staying home and watching the tube, especially when you've got a knitting project to go along with it! The hats look great!


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|Showing 1-20
Sanjay Bangar
Most India batsmen have done justice to their talent - Bangar
Sep 6, 2015: A year into his role as India batting coach, Sanjay Bangar talks about his overall experience, what the India batsmen have been working on, the Rohit v Pujara debate and Virat Kohli's technical adjustments
India news
Critics need to be less harsh on Rohit - Bangar
Sep 6, 2015: Sanjay Bangar, the India batting coach, has hit back at Rohit Sharma's critics, saying that the batsman has "definitely contributed to the team's cause"
Why is Pujara given short shrift?
Aug 31, 2015: He averages better than Rohit Sharma but still has to fight for a place in the Test side, mostly because he doesn't play ODIs
Sri Lanka v India, 3rd Test, SSC, 2nd day
Resolute Pujara shines in stop-gap role
Aug 29, 2015: Cheteshwar Pujara's century was proof that at times in Test match play, survival need not mean mere tentativeness but the ability to wait for simpler things, like the loose ball
Sri Lanka v India, 3rd Test, SSC, 2nd day
Pujara's Asia average, and Prasad's home run
Aug 29, 2015: Stats highlights from an engrossing day at the SSC
Sri Lanka v India, 3rd Test, SSC, 2nd day
Pujara's fighting ton holds India together
Aug 29, 2015: Cheteshwar Pujara scored his first Test century since the last month of 2013 to deny Sri Lanka clear advantage in the rain-hit SSC Test
India tour of Sri Lanka 2015
Pujara's chance to beat his blues
Aug 25, 2015: After a ten-month free-fall, Cheteshwar Pujara will turn out for India once again at the traditional batting paradise that is the SSC. Can he make it count?
India in Sri Lanka 2015
Bullish Shastri backs India's chances
Aug 10, 2015: After arriving late in Sri Lanka, India team director Ravi Shastri made the team's situation around Cheteshwar Pujara more than evident, saying the best five batsmen would play
India A v Australia A, 2nd unofficial Test, Chennai
I will open the batting - Cheteshwar Pujara
Jul 28, 2015: Cheteshwar Pujara has said that KL Rahul, Umesh Yadav and Amit Mishra will be rested for the second unofficial Test against Australia A in Chennai and that he would open the innings for the side
India A v Australia A, Chennai, 4th day
Don't think there's anything too wrong with Pujara's game - Dravid
Jul 25, 2015: Rahul Dravid is confident Cheteshwar Pujara's Test career will soon be back on track, and believes his recent run of poor form isn't due to any technical issue in his game
India A v Australia A, Chennai
Fifties for Rahul and Pujara, others struggle
Jul 22, 2015: A sluggish surface at the MA Chidambaram Stadium produced an attritional first day on which India A took the early initiative through a century stand between KL Rahul and Cheteshwar Pujara before losing five wickets for 94 runs to Australia A's constricti
'My father made me a Test cricketer'
Jun 21, 2015: The India batsman on the advantages of having his father coach him in cricket, and life
Bangladesh v India, only Test, Fatullah, 2nd day
Pujara the loser in 6-5 roulette
Jun 11, 2015: If India continue playing five bowlers in their upcoming Tests, it would leave four batsmen fighting out for two spots, and at the moment, Cheteshwar Pujara isn't in favour
India news
Have come back a better player - Pujara
May 19, 2015: India batsman Cheteshwar Pujara believes he has come back a "better player" after his stint in English county cricket with Yorkshire
Yorkshire v Hampshire, LV= County Championship Div One, Headingley, 3rd day
Pujara cuts Hampshire down to size
May 12, 2015: All the chairs with a good view of the play at Heagingley on day three were filled with people ready to resume watching Cheteshwar Pujara and Jack Leaning butcher James Vince's attack during an unbroken fifth-wicket stand that gave Yorkshire control.
India news
'A test of character, technique and toughness' - Pujara
Apr 6, 2015: Cheteshwar Pujara says he expects to be a "different player" when he returns from his stint with defending English county champions Yorkshire
County news
Yorkshire sign Pujara after Younis pulls out
Apr 1, 2015: Younis Khan, the Pakistan batsman, has pulled out of his deal to play for Yorkshire this season but the club have moved swiftly to sign Indian Test No.3 Cheteshwar Pujara.
Kings XI release Pujara, Balaji and Kartik
Dec 15, 2014: Kings XI Punjab have released Cheteshwar Pujara, Murali Kartik and L Balaji from their squad for the next season of IPL
Australia v India, 1st Test, Adelaide, 3rd day
'Can't score double-centuries all the time' - Pujara
Dec 11, 2014: Cheteshwar Pujara got out for 73 when he was looking good for a lot more, but he isn't too disappointed that he missed out on a hundred
Australia v India, 1st Test, Adelaide, 3rd day
Kohli century leads India's reply
Dec 11, 2014: On Test captaincy debut, on the ground where he had scored his maiden Test century, Virat Kohli made his seventh hundred to lead India's reply to Australia's 7 for 517 declared
|Showing 1-20
'Show some respect to your players'
The cricket world reacts to the incidents of bottle-throwing during the second India-South Africa T20 in Cuttack that caused two interruptions in play
We need Pakistan as much as it needs us
The country produces captivating cricketers, and the global game cannot afford to rest until Pakistan is fully restored to the fold
What you need to know about the Cairns perjury trial
Background to the Chris Cairns perjury case and what to expect over the coming weeks
Who is the better bowler in challenging conditions?
Sehwag's debut in controversial game
After making his maiden Ranji Trophy appearance for Haryana, Virender Sehwag spent a day at the Sehwag International School. While offering tips to youngsters, he recalled some memories of his first Ranji match
We need Pakistan as much as it needs us (186)
The country produces captivating cricketers, and the global game cannot afford to rest until Pakistan is fully restored to the fold
Steyn v Anderson in Asia (107)
Who is the better bowler in challenging conditions?
Bhuvneshwar needs to stick to swing (54)
It looks like he has lost his main weapon in an attempt to get his bowling speeds up
Yasir threat looms for England (44)
The fastest Pakistan bowler to 50 Test wickets is reviving the art of legspin and is expected to bamboozle batsmen, just like Ajmal and Rehman did three years ago in UAE
'Show some respect to your players' (42)
The cricket world reacts to the incidents of bottle-throwing during the second India-South Africa T20 in Cuttack that caused two interruptions in play


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Liver Biology/Pathobiology
An essential role for monocyte chemoattractant protein-1 in alcoholic liver injury: Regulation of proinflammatory cytokines and hepatic steatosis in mice†‡
Article first published online: 12 OCT 2011
DOI: 10.1002/hep.24599
Copyright © 2011 American Association for the Study of Liver Diseases
Volume 54, Issue 6, pages 2185–2197, December 2011
Additional Information
How to Cite
Mandrekar, P., Ambade, A., Lim, A., Szabo, G. and Catalano, D. (2011), An essential role for monocyte chemoattractant protein-1 in alcoholic liver injury: Regulation of proinflammatory cytokines and hepatic steatosis in mice. Hepatology, 54: 2185–2197. doi: 10.1002/hep.24599
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Potential conflict of interest: Nothing to report.
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This work was supported by PHS grant #AA017545 (to P.M.) and AA017986 (to P.M.) from the National Institute of Alcohol Abuse and Alcoholism (NIAAA) and by the resources of University of Massachusetts Medical School CFAR (grant 5P30 AI42845), and its contents are solely the responsibility of the authors and do not necessarily represent the views of the NIAAA.
Publication History
- Issue published online: 30 NOV 2011
- Article first published online: 12 OCT 2011
- Accepted manuscript online: 8 AUG 2011 10:25AM EST
- Manuscript Accepted: 26 JUL 2011
- Manuscript Received: 17 FEB 2011
- Top of page
- Abstract
- Materials and Methods
- Results
- Discussion
- Acknowledgements
- References
- Supporting Information
The importance of chemokines in alcoholic liver injury has been implicated. The role of the chemokine, monocyte chemoattractant protein-1 (MCP-1), elevated in patients with alcoholic liver disease is not yet understood. Here, we evaluated the pathophysiological significance of MCP-1 and its receptor, chemokine (C-C motif) receptor 2 (CCR2), in alcoholic liver injury. The Leiber-DeCarli diet containing alcohol or isocaloric control diets were fed to wild-type (WT) and MCP-1-deficient knockout (KO) mice for 6 weeks. In vivo and in vitro assays were performed to study the role of MCP-1 in alcoholic liver injury. MCP-1 was increased in Kupffer cells (KCs) as well as hepatocytes of alcohol-fed mice. Alcohol feeding increased serum alanine aminotransferase in WT and CCR2KO, but not MCP-1KO, mice. Alcohol-induced liver steatosis and triglyceride were attenuated in alcohol-fed MCP-1KO, but high in CCR2KO mice, compared to WT, whereas serum endotoxin was high in alcohol-fed WT and MCP-1KO mice. Expression of liver proinflammatory cytokines tumor necrosis factor alpha, interleukin (IL)-1β, IL-6, KC/IL-8, intercellular adhesion molecule 1, and cluster of differentiation 68 was induced in alcohol-fed WT, but inhibited in MCP-1KO, mice independent of nuclear factor kappa light-chain enhancer of activated B cell activation in KCs. Oxidative stress, but not cytochrome P450 2E1, was prevented in chronic alcohol-fed MCP-1KO mice, compared to WT. Increased expression of peroxisome proliferator-activated receptor (PPAR)α and PPARγ was accompanied by nuclear translocation, DNA binding, and induction of fatty acid metabolism genes acyl coenzyme A oxidase and carnitine palmitoyltransferase 1A in livers of alcohol-fed MCP-1KO mice, compared to WT controls. In vitro assays uncovered an inhibitory effect of recombinant MCP-1 on PPARα messenger RNA and peroxisome proliferator response element binding in hepatocytes independent of CCR2. Conclusion: Deficiency of MCP-1 protects mice against alcoholic liver injury, independent of CCR2, by inhibition of proinflammatory cytokines and induction of genes related to fatty acid oxidation, linking chemokines to hepatic lipid metabolism. (HEPATOLOGY 2011)
Alcoholic liver disease (ALD) is a major health concern, and approximately 90% of heavy drinkers develop fatty liver disease or steatosis. Fatty liver is occasionally accompanied by, or progresses to, inflammation in human ALD. The essential role of innate immune cell activation and circulating endotoxin/lipopolysaccharide (LPS) in ALD has been proposed.1, 2 Circulating endotoxin activates liver macrophages and leads to the induction of cytokines, chemokines, and reactive oxygen species.3 Though proinflammatory cytokine production in the alcoholic liver is extensively investigated, the importance of chemokines is still unknown. Elevation of chemokines, such as interleukin (IL)-8, monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein 1 (MIP-1), in alcoholic hepatitis and cirrhotic patients and the correlation with the recruitment of polymorphonuclear leukocytes is reported.4, 5 However, the pathophysiological mechanisms affected by chemokines in ALD are yet to be determined. CC-chemokines induce the recruitment and activation of mononuclear cells, such as monocytes/macrophages, T cells and natural killer T cells,6, 7 and these cells play an important role in the development and propagation of alcoholic liver injury.8
MCP-1 or chemokine (C-C motif) ligand 2 (CCL2), an important CC-chemokine, recruits and activates monocytes/macrophages to the site of tissue injury and regulates adhesion molecules and proinflammatory cytokines tumor necrosis factor alpha (TNFα), IL-1β, and IL-6.9, 10 The pivotal role of MCP-1 in alcoholic liver injury was first recognized by studies showing higher amounts of MCP-1, as compared to other CC-chemokines, such as MIP-1α and MIP-1β, in the liver and mononuclear cells of patients with alcoholic hepatitis.4, 5 Subsequently, the pathogenic role of MCP-1 expressed by liver macrophages and endothelial cells was demonstrated in rodent models of alcoholic hepatitis.11 Besides macrophage activation, MCP-1 appears to play a significant role in hepatic steatosis or early liver injury. Recently, transgenic mice overexpressing MCP-1 in adipose tissue exhibited insulin resistance and increased hepatic triglyceride content.12 These studies were based on the observations that mice fed a high-fat diet led to MCP-1 induction in adipose tissue, but not liver.12In vitro studies also demonstrated that MCP-1 can induce lipid accumulation in hepatocyte cultures.13 In general, MCP-1 seems to play an important role in hepatic inflammatory responses and steatosis during tissue injury.
Previous studies from our laboratory and others have shown the pathophysiological importance of proinflammatory cytokines in ALD.1, 2, 14 However, the pathophysiological role of chemokines, such as MCP-1, in alcoholic liver injury is still uncertain. Based on the preferential elevation of MCP-1 among other CC-chemokines in alcoholic hepatitis patients4, 5 and its importance in the modulation of proinflammatory cytokines,9, 10 we hypothesized that MCP-1 contributes to chronic alcoholic liver injury and steatosis via the modulation of inflammatory cytokines. Using MCP-1-deficient mice, we sought to investigate whether MCP-1 and its receptor, chemokine (C-C motif) receptor 2 (CCR2), play a causative role in alcoholic liver injury.
Materials and Methods
- Top of page
- Abstract
- Materials and Methods
- Results
- Discussion
- Acknowledgements
- References
- Supporting Information
Additional descriptions of methods are available in the Supporting Information.
Animal Studies.
All animals received proper care in agreement with animal protocols approved by the Institutional Animal Use and Care Committee of the University of Massachusetts Medical School (Worcester, MA). Six- to eight-week-old female wild-type (WT) (C57BL/6) and MCP-1-deficient and CCR2-deficient mice (all generated on a C57BL/6 background; The Jackson Laboratory, Bar Harbor, ME) received Lieber-DeCarli diet (Bio-Serv, Frenchtown, NJ) with 5% (v/v) ethanol (36% ethanol-derived calories) for 6 weeks; pair-fed control mice received an equal amount of calories as their alcohol-fed counterparts with the alcohol-derived calories substituted with dextrin maltose. All strains of mice consumed comparable daily calories. In some cases, mice from both the alcohol-fed and pair-fed groups were administered an intraperitoneal (IP) injection of either 0.2 mL of 0.9% saline (phosphate-buffered, pH 7.4) alone as a vehicle control or 0.2 mL 0.9% saline containing 0.5 mg/kg of nonpurified lipopolysaccharide (LPS; from Escherichia coli 0111:B4; Sigma-Aldrich, St. Louis, MO) and sacrificed 2 hours later. There were 9-12 mice in each experimental group. Serum was separated from whole blood and frozen at −80°C. Liver tissue was rapidly excised and a portion was snap-frozen in liquid nitrogen and stored at −80°C. Additional portions of the livers were stored in RNA stabilization reagent (RNAlater; Qiagen, Valencia, CA) for RNA extraction or fixed in 10% neutral-buffered formalin for histopathological analysis.
Statistical Analysis.
Statistical significance was determined by analysis of variance (in vivo) and t-test (in vitro) using the GraphPad Prism 5.01 software (GraphPad Software, Inc., La Jolla, CA). Data are shown as mean ± standard error of the mean (SEM) and were considered statistically significant at P < 0.05.
- Top of page
- Abstract
- Materials and Methods
- Results
- Discussion
- Acknowledgements
- References
- Supporting Information
Chronic Alcohol Consumption Induces MCP-1 in Kupffer Cells and Hepatocytes.
MCP-1 is increased during ALD; however, its cellular source in the liver is not yet identified. Here, C57Bl/6 mice were fed the Leiber-Decarli alcohol diet or its isocaloric control (pair-fed) diet to determine the expression of MCP-1 in the liver. Chronic alcohol feeding for 6 weeks induced MCP-1 messenger RNA (mRNA) (Fig. 1A) and protein (Fig. 1B) in whole livers, compared to pair-fed controls. Next, to identify the cell types expressing MCP-1, we isolated hepatocytes and Kupffer cells (KCs) and estimated MCP-1 mRNA. Figure 1C shows that isolated hepatocytes as well as KCs express high amounts of MCP-1 mRNA in chronic alcohol-fed mice, compared to isocaloric pair-fed controls, with similar expression levels of baseline MCP-1 in hepatocytes relative to KCs (Supporting Fig. 1). Expression analysis of the CC-chemokine gene family revealed a significant increase in CCL4/MIP-1β and KC/IL-8/chemokine (C-X-C motif) ligand 1, with a maximal elevation in MCP-1 in livers of chronic alcohol-fed mice, compared to pair-fed controls (Fig. 1D).
MCP-1 Deficiency Protects Against Alcoholic Liver Injury.
To investigate the role of MCP-1 in ALD, WT and MCP-1 knockout (MCP-1KO) mice were fed the Leiber-DeCarli diet with 5% ethanol or isocaloric control diet for 6 weeks to induce ALD. Prolonged alcohol feeding resulted in liver injury, as assessed by significantly increased serum alanine aminotransferase (ALT) levels (Fig. 2A) and higher liver/body-weight ratio (Supporting Fig. 2A) in alcohol-fed WT mice, compared to pair-fed controls and MCP-1KO mice. Despite no liver damage, serum alcohol levels in MCP-1KO were comparable to alcohol-fed WT mice (Supporting Fig. 2B). Histological analysis showed micro- and macrosteatosis in chronic alcohol-fed WT mice, whereas fat deposition was not detectable in pair-fed controls and MCP-1KO mice (Fig. 2B). In agreement with the histological data, liver triglyceride levels were significantly higher in alcohol-fed WT mice, compared to pair-fed controls and MCP-1KO mice (Fig. 2C). Furthermore, chronic alcohol-fed WT and MCP-1KO mice showed significantly increased serum endotoxin, compared to pair-fed controls (Fig. 2D). Collectively, these data show that chronic alcohol feeding induces liver damage in WT mice, and regardless of high blood alcohol levels and elevated endotoxin, MCP-1KO mice are protected from alcoholic liver injury.
|Target Genes
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|Reverse Primer (5′ 3′)
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|tcg cac ttt ggt att ctt gga g
Chronic Alcohol Induced Up-regulation of Proinflammatory Cytokines Is Prevented by MCP-1 Deficiency Independent of Nuclear Factor Kappa Light-Chain Enhancer of Activated B Cell Activation.
MCP-1 plays an important role in the induction of proinflammatory cytokines at the site of tissue injury.10 Here, we investigated the effect of MCP-1 deficiency on alcohol-induced expression of cytokines in the liver. We elucidated the expression of circulating endotoxin (baseline)-mediated induction of proinflammatory cytokines TNFα, IL-1β, and IL-6, as well as CC-chemokine mRNA levels in liver of alcohol-fed WT and MCP-1KO mice. Here, we show that TNFα, IL-1β, and IL-6 mRNA was increased significantly in alcohol-fed WT mice, compared to pair-fed WT controls, whereas alcohol-fed MCP-1KO mice were unable to induce proinflammatory cytokine mRNA in the liver (Fig. 3A). MCP-1 deficiency also prevented chronic alcohol-induced liver tissue TNFα, as compared to WT mice (Fig. 3B). Interestingly, among CC-chemokine genes, KC/IL-8 mRNA was significantly decreased, but CCL4/MIP-1β and CCL5/RANTES mRNA was high in alcohol-fed MCP-1KO mice, compared to pair-fed controls (Fig. 3C). Furthermore, investigation of MCP-1-mediated adhesion molecules and macrophage markers demonstrated a significant induction of intercellular adhesion molecule 1 (ICAM-1) and cluster of differentiation (CD)68, but unchanged vascular cell adhesion molecule 1 (VCAM-1) and F4/80 in livers of alcohol-fed WT, but not MCP-1KO, mice (Fig. 3D). Because nuclear factor kappa light-chain enhancer of activated B cells (NF-κB) is important in chronic alcohol-mediated proinflammatory cytokine production and macrophage activation,15 we next determined whether the inhibition of inflammatory cytokines was regulated by the lack of NF-κB activation in MCP-1-deficient mice. Interestingly, our results show that NF-κB binding activity in whole livers was significantly increased in alcohol-fed MCP-1-deficient mice (Fig. 3E), compared to alcohol-fed WT and pair-fed MCP-1KO mice. Furthermore, increased NF-κB activation was observed in isolated KCs of alcohol-fed MCP-1KO and WT mice, compared to pair-fed controls (Fig. 3F). Immunohistochemical analysis revealed NF-κB p65 staining in nonparenchymal cells of alcohol-fed WT and MCP-1KO mice (Supporting Fig. 3). On the other hand, isolated hepatocytes showed decreased NF-κB activation in alcohol-fed WT mice, compared to pair-fed controls, and this inhibition was prevented in alcohol-fed MCP-1KO mice (Fig. 3F), likely contributing to NF-κB-mediated hepatocyte survival in alcohol-fed MCP-1KO mice. These results indicate that liver proinflammatory cytokine mRNA, ICAM-1, and CD68 are significantly decreased in chronic alcohol-fed MCP-1KO mice, compared to their WT counterparts, in an NFκB-independent manner.
Oxidative Stress and Sensitization to LPS-Induced Proinflammatory Cytokine Production Is Inhibited by MCP-1 Deficiency.
The classical feature of alcoholic liver injury is alcohol-mediated oxidative stress and increased sensitization to LPS, resulting in enhanced proinflammatory cytokine expression in the liver.1, 16 To further test the effect of sensitization to LPS in chronic alcohol-fed MCP-1-deficient mice, an in vivo LPS challenge (0.5 mg/kg body weight; IP) was administered at the end of the chronic alcohol feeding. Our results show that in vivo LPS challenge increased proinflammatory cytokine, TNFα, IL-1β, and IL-6 mRNA in liver of WT mice, as compared to pair-fed controls, and this induction was prevented in chronic alcohol-fed MCP-1KO mice (Fig. 4A). Interestingly, no changes were observed in Toll-like receptor 4 (TLR4) expression, a receptor for LPS (Supporting Fig. 4). We next determined whether MCP-1 deficiency would affect alcohol-induced oxidative stress and alcohol-metabolizing enzyme cytochrome P450 2E1 (CYP2E1) in the liver. Chronic alcohol-induced oxidative stress, as illustrated by increased thiobarbituric acid-reactive substances (TBARS) in WT mice, was significantly blunted in alcohol-fed MCP-1KO mice (Fig. 4B). However, CYP2E1 levels estimated in liver microsomal preparations from alcohol-fed WT and MCP-1KO mice remained similar (Fig. 4C). These results suggest that MCP-1 contributes to chronic alcohol-induced oxidative stress in a CYP2E1-independent fashion and sensitizes the liver to LPS, resulting in enhanced proinflammatory cytokine production.
Chronic Alcohol Feeding Induces Liver Injury in CCR2-Deficient Mice.
MCP-1 is known to mediate inflammatory cell activation in the liver via its receptor, CCR2.17 The importance of CCR2, predominantly expressed in monocyte/macrophage cells, is shown in liver diseases, such as fibrosis.18 To investigate the role of CCR2 in alcoholic liver injury, we fed CCR2-deficient mice (CCR2KO) with the Leiber-DeCarli diet containing 5% ethanol for 6 weeks. Similarly to WT mice, alcohol feeding increases serum ALT in CCR2KO mice, indicating liver damage in the absence of CCR2 (Fig. 5A). Furthermore, histological examination showed that micro- and macrosteatosis were observed in alcohol-fed WT and CCR2KO mice, compared to pair-fed controls (Fig. 5B). Quantitation of liver triglycerides exhibited significantly high levels in alcohol-fed WT and CCR2KO mice, compared to pair-fed mice (Fig. 5C), supporting histological findings. Thus, it is evident that chronic alcohol feeding induces liver injury irrespective of the absence of CCR2, and suggests that MCP-1-mediated protection from alcoholic liver injury is independent of CCR2.
PPARα and PPARγ mRNA Expression, DNA Binding, and Target Genes Are Increased in Chronic-Alcohol Fed MCP-1KO Mice.
Having observed inhibitory effects on inflammatory responses in the liver, we next wanted to determine whether the decrease in hepatic steatosis in alcohol-fed MCP-1KO mice (in Fig. 2D,E) was related to the regulation of fatty acid metabolism genes. We analyzed peroxisome proliferator-activated receptor alpha (PPARα) and peroxisome proliferator-activated receptor gamma (PPARγ), important transcription factors in metabolism as well as inflammatory responses.19 Though chronic alcohol feeding decreased PPARα mRNA in WT, alcohol-fed MCP-1KO mice showed comparable levels to pair-fed controls, indicating the prevention of PPARα down-regulation by alcohol (Fig. 6A). On the other hand, PPARγ mRNA was not affected in alcohol-fed WT mice, but was significantly increased in alcohol-fed MCP-1KO mice, as compared to controls (Fig. 6A). PPARα and PPARγ mRNA were not altered between genotypes, but showed a significant up-regulation in alcohol-fed MCP-1KO, compared to WT counterparts (Fig. 6A). Upon activation, PPARs translocate to the nucleus and bind to promoter elements of the target gene involved in fatty acid metabolism.20, 21 Nuclear PPARα and PPARγ levels were increased in alcohol-fed MCP-1KO mice, compared to pair-fed controls (Fig. 6B). Using electrophoretic mobility shift analysis (EMSA), we next analyzed the DNA-binding activity of PPARs in livers of alcohol-fed WT and MCP-1KO mice. Our results show that peroxisome proliferator response element (PPRE)-binding activity was significantly reduced in alcohol-fed WT mice, compared to pair-fed controls, whereas down-regulation of PPRE-binding activity was prevented in alcohol-fed livers of MCP-1KO mice (Fig. 6C). Similar to PPRE activation in whole livers, PPRE binding activity in isolated hepatocytes was significantly reduced in alcohol-fed WT mice, whereas this down-regulation was prevented in alcohol-fed MCP-1KO mice, compared to pair-fed controls (Fig. 6D). It is worthy to note that PPRE binding was significantly higher in alcohol-exposed hepatocytes (Fig. 6D) and whole livers (Fig. 6C) of MCP-1KO, compared to alcohol-fed, WT mice. Supershift analysis in whole livers of alcohol-fed MCP-1KO and WT mice revealed the presence of PPARα and retinoid X receptor alpha in the PPAR-binding complex (Fig. 6E). Next, to further evaluate whether increased PPRE-binding activity in MCP-1KO mice would result in target-gene induction related to fatty acid metabolism,20 we estimated mRNA levels of acyl coenzyme A (CoA) oxidase (ACOX), carnitine palmitolyltransferase (CPT-1), long-chain acyl CoA dehydrogenase (LCAD), and medium-chain acyl CoA dehydrogenase (MCAD). Our results show that ACOX (Fig. 7A) and CPT-1 (Fig. 7B) mRNA levels are significantly decreased in alcohol-fed WT mice, and this down-regulation was prevented in MCP-1KO mice. Furthermore, LCAD (Fig. 7C) and MCAD (Fig. 7D) mRNA did not show significant changes in alcohol-fed MCP-1KO and WT mice. These results indicate that MCP-1 regulates PPAR mRNA expression, nuclear translocation, DNA binding, and downstream target-gene expression related to fatty acid metabolism in alcoholic liver injury.
MCP-1 Negatively Interferes With PPARα mRNA and PPRE Binding in Cultured Hepatocytes.
Because the lack of MCP-1 correlates with PPRE binding and expression of fatty acid oxidation genes, we wanted to next evaluate whether MCP-1 would directly affect PPARα expression and DNA-binding activity in hepatocytes. To this end, we performed in vitro experiments using recombinant MCP-1 and determined its effect on PPAR agonist WY-14,643-induced PPARα mRNA and PPRE-binding activity in human hepatocyte Huh7 cells. In accord with previous studies showing a lack of CCR2 expression in hepatocytes18 and Huh7 cells,13 our results show an absence of CCR2 expression in hepatocytes and Huh7 cells, compared to a high expression in monocyte/macrophages (Supporting Fig. 5A). Recombinant MCP-1 significantly decreases baseline and WY-induced PPARα mRNA, compared to WY treatment alone, in Huh7 hepatocytes (Fig. 8A). Further, MCP-1 treatment of Huh7 cells significantly reduced baseline and WY-induced PPRE activation (Fig. 8B; shown in triplicates in Supporting Fig. 5B). Collectively, our results suggest that MCP-1 can directly inhibit PPARα induction and activation impeding fatty acid oxidation in hepatocytes.
- Top of page
- Abstract
- Materials and Methods
- Results
- Discussion
- Acknowledgements
- References
- Supporting Information
The significance of MCP-1 as a master regulator of monocyte/macrophage function has been proposed in various chronic inflammatory diseases.22 MCP-1 was previously identified to direct the trafficking of immune cells to the site of tissue injury.6, 7 However, recent studies have suggested a role for MCP-1 in metabolic diseases, such as diabetes and obesity-related insulin resistance and hepatic steatosis.12, 23 Here, we report on novel data that MCP-1 contributes to alcohol-induced fatty liver, likely via the down-regulation of PPARα and its target fatty acid metabolism genes, independent of its receptor, CCR2. These results, for the first time, indicate a link between inflammatory chemokines and lipid metabolism in alcoholic liver injury. We show that chronic alcohol consumption increases MCP-1 in KCs and hepatocytes in the liver. Deficiency of MCP-1 protects against chronic alcohol-induced liver injury by reducing the expression of proinflammatory cytokines and the macrophage activation markers, ICAM-1 and CD68, and increasing PPARα expression and DNA binding, leading to the induction of fatty acid metabolism genes.
Chronic alcohol-induced liver injury is characterized by steatosis, inflammatory cell activation, and hepatocyte damage.1, 2, 24 Inflammatory cell mediators produced in the liver during chronic alcohol exposure contribute to liver injury. For instance, TNFα induces hepatocyte apoptosis in the liver,25 whereas IL-6 can generate hepatoprotective or damaging effects, based on the target cells.26 Human studies and animal models of alcoholic hepatitis show that MCP-1 is up-regulated in KCs.11 Our novel observations show that chronic alcohol feeding induces MCP-1 in KCs and hepatocytes, suggesting a functional role in inflammatory responses and steatosis. Previous studies in genetically obese and high-fat-diet–fed mice showed that MCP-1 significantly increased in adipose tissue and plasma, but not in the liver, contributing to insulin resistance and hepatic steatosis.12 Recent studies by Obstfeld et al.23 demonstrate that leptin-deficient ob/ob mice exhibit increased MCP-1 in hepatocytes only. Contrary to obesity and diabetes, chronic alcohol induces MCP-1 in KCs as well as hepatocytes, ascribing a pathogenic role for MCP-1 in the alcoholic liver. Activating signals, including LPS/TLR4 and proinflammatory cytokines, are potent inducers of MCP-1.27, 28 Interestingly, recent studies show that homocysteine, increased in alcoholic liver injury,29 also induces MCP-1 expression in hepatocytes.30 The investigation of the cell-specific regulation of MCP-1 during alcoholic liver injury will provide further insights into its functional significance.
The contribution of MCP-1 in various models of liver injury has been under investigation. Though in some cases of liver injury, such as hepatic granuloma formation and obesity-induced fatty liver, the lack of MCP-1 is protective,12, 23, 31 in other instances, such as concanavalin A–induced liver injury and lethal endotoxemia, the absence of MCP-1 worsens disease.32, 33 Here, we show that MCP-1 deficiency is protective against chronic alcohol-induced liver injury, as indicated by decreased serum ALT and reduced steatosis. Patients with severe alcoholic hepatitis and cirrhosis displayed the highest elevation of MCP-1 in liver and plasma, compared to other CC-chemokines.4, 5 Previous studies indicated that CC-chemokines, including MCP-1, played a major role in late-stage alcoholic hepatitis directing the migration of inflammatory cells and leading to fibrosis and cirrhosis.8 Studies from Seki et al.18 indicated the significance of the MCP-1/CCR2 axis in liver fibrosis. Our studies provide novel direct evidence for the importance of MCP-1 in the pathogenesis of early alcoholic liver injury.
Chronic alcohol feeding induces gut permeability and increases serum endotoxin levels, which, in turn, upregulate proinflammatory cytokine production in the liver.2, 3 Our results show that similar to alcohol-fed wild-type, MCP-1KO animals also demonstrate an elevation in serum endotoxin, suggesting that chronic alcohol does not affect mechanisms related to gut permeability in MCP-1-deficient mice. MCP-1 regulates the production of proinflammatory cytokines and adhesion molecules in monocytes/macrophages.9, 10 Despite increased endotoxin, we observed a significant reduction in mRNA expression of proinflammatory cytokines TNFα, IL-1β, IL-6, and KC/IL-8 in the liver of alcohol-fed MCP-1KO mice, compared to WT controls. In addition, we also observed a significant decrease in adhesion moelcule, ICAM-1, and the macrophage activation marker, CD68, in alcohol-fed MCP-1KO mice. Furthermore, our data indicate that the down-regulation of proinflammatory cytokines, adhesion molecule, and macrophage activation marker is independent of NF-κB activation in KCs in alcohol-fed MCP-1KO mice. Noteworthy is the lack of reduction in NF-κB DNA-binding activity in isolated hepatocytes from alcohol-fed MCP-1KO, compared to the inhibition of NF-κB activation in hepatocytes of alcohol-fed WT mice, which indicates a role for NF-κB in hepatocyte survival. Future studies will delineate the mechanism of reduction in proinflammatory responses in alcohol-fed MCP-1-deficient mice.
Oxidative stress and sensitization to LPS are hallmarks of molecular mechanisms of alcoholic liver injury.1, 2, 16 Interestingly, our results show that MCP-1 deficiency prevents the induction of chronic alcohol-induced oxidative stress, compared to WT mice. A similar correlation between MCP-1 and induction of oxidative stress in a toxic model of acute liver injury, using carbon tetrachloride, was previously observed.34 How MCP-1 modulates oxidative stress pathways or reduces antioxidants will be investigated in the future. Similar up-regulation of microsomal CYP2E135 in alcohol-fed WT and MCP-1KO mice indicate the induction of oxidative stress independent of alcohol-metabolizing CYP2E1. Besides cellular mechanisms, such as TLR expression, oxidative stress contributes to LPS sensitization in ALD and enhancement of pro inflammatory cytokine gene expression.36, 37 An in vivo LPS challenge given at the end of chronic alcohol feeding led to an augmentation of proinflammatory cytokines TNFα, IL-1β, and IL-6 in WT mice, and this was prevented in MCP-1KO mice. Our results suggest that MCP-1 deficiency inhibits oxidative stress and also impedes sensitization to LPS, independent of TLR4 expression, during alcoholic liver injury. Studies to unravel the mechanisms of LPS sensitization affected by MCP-1 during chronic alcohol exposure will be examined in the future.
Among the various mechanistic studies for alcoholic steatosis, alterations in transcription factors, such as PPARα and PPARγ, controlling lipid metabolism have been recognized.24 Previous studies showed that alcohol feeding in rats decreased PPARα activation and downstream target genes important in fatty acid oxidation.19 Here, we show that alcohol-fed MCP-1KO mice exhibit increased PPARα and PPARγ mRNA expression, enhanced nuclear PPARα and PPARγ, PPRE activation in whole liver and isolated hepatocytes, presence of PPARα/RXRα in the DNA-binding complex, and induction of target genes, such as CPT-1 and ACOX—both enzymes critical in fatty acid oxidation. Previous studies have shown that a secretory product of adipose tissue, likely MCP-1, can induce lipid accumulation in hepatoyctes.13 Our in vitro findings demonstrate that recombinant MCP-1 down-regulates PPARα mRNA expression and DNA-binding activity in hepatocytes, likely contributing to increased triglyceride accumulation in ALD. These results suggest a direct effect of MCP-1 on PPARα and its target genes and thus steatosis.
MCP-1 mediates its action via receptor CCR238 or independent of CCR2.39 Our results show that CCR2KO mice induce alcoholic liver injury similar to alcohol-fed WT mice, indicating CCR2-independent effects of MCP-1. Furthermore, because hepatocytes do not express CCR2,18 as reported here (Supporting Fig. 5A), we predict that MCP-1 mediates its effects in the liver independent of CCR2. Another lipid-modulating transcription factor with anti-inflammatory properties, PPARγ, was up-regulated in alcohol-fed MCP-1KO livers. It is likely that PPARγ inhibits proinflammatory cytokine production in chronic alcohol-exposed MCP-1KO mice. Our results here show that MCP-1 expression is directly up-regulated in hepatocytes during chronic alcohol exposure and likely regulates fatty acid oxidation, resulting in steatosis. LPS can modulate fatty acid oxidation genes via TLR4/IRAK-1 and PPARα reduction.40 We predict that alcohol-mediated increase in circulating endotoxin (i.e., LPS) induces MCP-1 in hepatocytes and macrophages to regulate fatty acid oxidation pathways in an autocrine or paracrine fashion in the liver. Future studies, using MCP-1-targeting strategies, will provide mechanistic insights into the pathophysiological mechanisms affected by MCP-1 in alcoholic liver injury.
Overall, our studies show, for the first time, that MCP-1 in the liver regulates macrophage activation, proinflammatory responses, and hepatic steatosis in alcoholic liver disease. These studies provide a link between inflammatory cell activation and pathways of fatty acid metabolism during alcoholic liver injury likely involved in the amplification and progression of disease. Therefore, it appears plausible that pharmacological approaches to block MCP-1 in the alcoholic liver may be beneficial to early alcoholic fatty liver injury and also abrogate inflammatory pathways contributing to propagation in ALD.
- Top of page
- Abstract
- Materials and Methods
- Results
- Discussion
- Acknowledgements
- References
- Supporting Information
The authors thank Karen Kodys for labeling the oligonucleotides for the EMSA analysis.
- Top of page
- Abstract
- Materials and Methods
- Results
- Discussion
- Acknowledgements
- References
- Supporting Information
- 1Recent insights into the role of the innate immune system in the development of alcoholic liver disease. Exp Biol Med (Maywood) 2003; 228: 882-890..
- 2Signalling pathways in alcohol-induced liver inflammation. J Hepatol 2009; 50: 1258-1266., .
- 3Role of Kupffer cells and gut-derived endotoxins in alcoholic liver injury. J Gastroenterol Hepatol 2000; 15( Suppl): D20-D25., , , , , , et al.
- 4Distinct patterns of chemokine expression are associated with leukocyte recruitment in alcoholic hepatitis and alcoholic cirrhosis. J Pathol 1998; 186: 82-89., , , , , , .
- 5Serum concentrations and peripheral secretion of the beta chemokines monocyte chemoattractant protein 1 and macrophage inflammatory protein 1alpha in alcoholic liver disease. Gut 1999; 45: 416-420., , , .
- 6Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell 1994; 76: 301-314..
- 7Chemokines: leucocyte recruitment and activation cytokines. Lancet 1997; 349: 490-495., .
- 8Impact of alcohol on the ability of Kupffer cells to produce chemokines and its role in alcoholic liver disease. J Gastroenterol Hepatol 2000; 15: 349-356..
- 9Abnormalities in monocyte recruitment and cytokine expression in monocyte chemoattractant protein 1-deficient mice. J Exp Med 1998; 187: 601-608., , , , , , et al.
- 10Monocyte chemoattractant protein-1 regulates adhesion molecule expression and cytokine production in human monocytes. J Immunol 1992; 148: 2423-2428., , , .
- 11Chronic alcohol intoxication primes Kupffer cells and endothelial cells for enhanced CC-chemokine production and concomitantly suppresses phagocytosis and chemotaxis. Front Biosci 2002; 7: a117-a125..
- 12MCP-1 contributes to macrophage infiltration into adipose tissue, insulin resistance, and hepatic steatosis in obesity. J Clin Invest 2006; 116: 1494-1505., , , , , , et al.
- 13Monocyte chemoattractant protein-1 secreted by adipose tissue induces direct lipid accumulation in hepatocytes. Hepatology 2008; 48: 799-807., , , , , , et al.
- 14Signaling mechanisms in alcoholic liver injury: role of transcription factors, kinases and heat shock proteins. World J Gastroenterol 2007; 13: 4979-4985..
- 15The opposite effects of acute and chronic alcohol on lipopolysaccharide-induced inflammation are linked to IRAK-M in human monocytes. J Immunol 2009; 183: 1320-1327., , , , .
- 16Oxidative stress and alcoholic liver disease. Semin Liver Dis 2009; 29: 141-154., .
- 17Impaired monocyte migration and reduced type 1 (Th1) cytokine responses in C-C chemokine receptor 2 knockout mice. J Clin Invest 1997; 100: 2552-2561., , , , , , .
- 18CCR2 promotes hepatic fibrosis in mice. Hepatology 2009; 50: 185-197., , , , , , et al.
- 19Integration of metabolism and inflammation by lipid-activated nuclear receptors. Nature 2008; 454: 470-477., .
- 20Molecular regulation of adipogenesis. Annu Rev Cell Dev Biol 2000; 16: 145-171., .
- 21Convergence of 9-cis retinoic acid and peroxisome proliferator signalling pathways through heterodimer formation of their receptors. Nature 1992; 358: 771-774., , , , .
- 22Monocyte chemoattractant protein-1 (MCP-1): an overview. J Interferon Cytokine Res 2009; 29: 313-326., , , .
- 23C-C chemokine receptor 2 (CCR2) regulates the hepatic recruitment of myeloid cells that promote obesity-induced hepatic steatosis. Diabetes 2010; 59: 916-925., , , , , , et al.
- 24Alcohol and lipid metabolism. Am J Physiol Endocrinol Metab 2008; 295: E10-E16., .
- 25Ethanol potentiates tumor necrosis factor-alpha cytotoxicity in hepatoma cells and primary rat hepatocytes by promoting induction of the mitochondrial permeability transition. Hepatology 2000; 31: 1141-1152., .
- 26Cell type-dependent pro- and anti-inflammatory role of signal transducer and activator of transcription 3 in alcoholic liver injury. Gastroenterology 2008; 134: 1148-1158., , , , , , et al.
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- 32MCP-1 protects mice in lethal endotoxemia. J Clin Invest 1997; 99: 2832-2836., , , , , , .
- 33C-C chemokine ligand 2/monocyte chemoattractant protein-1 directly inhibits NKT cell IL-4 production and is hepatoprotective in T cell-mediated hepatitis in the mouse. J Immunol 2003; 170: 5252-5259., , , .
- 34Prevention of severe toxic liver injury and oxidative stress in MCP-1-deficient mice. J Hepatol 2007; 46: 230-238., , , , , , et al.
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Supporting Information
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- Materials and Methods
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- Supporting Information
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100 years of the T-shirt
Seems hard to believe something as ubiquitous as the T-shirt was ever . . . new. But that's what it was back in 1913, when the U.S. Navy ordered them for sailors to wear under their uniforms. To celebrate the fashion staple's 100th birthday, we've gathered a quick look at its history and some classic and not-so-classic tees to keep you cool the rest of the summer.
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Explore NY State: 10 spots to take your breath away 29 chains Long Island wants now NYC Wine & Food Festival, more NYC weekend picks 13 best Halloween costumes for 2015 20 best places to see fall foliage in the Northeast Peek inside Long Island's most lavish homes 10 best tours at LI wineries 20 crazy-cute pet costumes Spooky cocktails for Halloween 25 weekend getaways every New Yorker should take 29 festive fall cocktail recipes 10 off-the-beaten-path wineries on LI


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Hotel Style
Propio del lugar, rústico
Contemporaneo, diseño
Resort, vacacional
Hotel Category
Lujo exclusivo
Lujo informal
Chic, con encanto
Delser, Verona
|Strada dei Monti, 14/B, 37124, Verona, Verona, Véneto, ITALIA.
Descripción: DelserEnjoying a fine location on a hillside overlooking the plains of Verona and the river Adige, Delser is situated in a magnificently converted "castellum aquae", the water cistern of an 18th century noble villa. This luxury boutique property provides exclusive accommodation in a tranquil rural atmosphere.
Just over six kilometres from the historic centre of Verona, with all of its splendid Roman, Medieval and Renaissance attractions, Delser is perfect for visits to this magnificent region, away from the bustle and crowds but close enough to enjoy the sights at leisure.
Standing in a fine garden and surrounded by its own olive groves and vineyards, Delser offers splendid accommodation in its exclusively designed rooms and suites. Using fine local stone, the ancient foundations of the cistern, and noble timbers, the rooms and suites are decorated in an airy, minimalist style, combining modern furniture with carefully chosen antiques.
Descripción de habitaciones: DelserStandard Double Room:
These double rooms have oak-wood floors, marmorino stucco walls and are all decorated with XVI century antique furniture. Extra beds are available on request. Spacious private bathrooms have bath tubs and shower or shower only. Room size approximately 32 sq m.
Superior Double Room:
These double or twin bedded rooms feature oak floors, marmorino stucco walls and are all decorated with XVI century antique furniture. Extra beds are available on request. The spacious private bathrooms have bath tubs with shower. Room size approximately 38 sq m.
Exclusive Superior Double Room:
These large rooms feature oak-wooden floors, marmorino stucco walls are decorated with XVI century antique furniture. One large double or two twin beds, extra beds are available on request. The spacious en-suite bathrooms are equipped with bath tubs and a shower. Room size about 40 sq m.
Aclaraciones e información adicional: Delser
Pets allowed on request.
Airport shuttle services available on prior notice. Charges apply.
Laundry, baby-sitting and car rental are also available on request.
Información Básica:
Número de habitaciones: 8
Categoría oficial: Bed and Breakfast.Régimen de fumadores: There are non-smoking rooms available.
Tarjetas de crédito aceptadas: Visa, Euro/Mastercard, CartaSi.
Coordenadas GPS: 45° 28' 30.40'' N 10° 57' 30.13'' E
Aeropuerto más cercano: Verona Villafranca Airport - 15 km.
Instalaciones y servicios:
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Hornady Custom Lite Ammunition 12 Gauge 2-3/4" 300 Grain Flex Tip eXpanding Sabot Slug Box of 5
Product #: 921236 Hornady #: 86230 UPC #: 090255862300
You must be 21 years or older to order ammunition. Ammo must ship UPS ground. Due to Department of Transportation regulations, we cannot accept returns on ammo. Please check local laws before ordering.
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Hornady is known as an industry leader in creating innovative products that work. The SST Lite Shotgun Slugs are no different. Hunters know how effective a 12 gauge slug is for harvesting whitetail deer, however, that effectiveness comes at the price of heavy recoil to get maximum performance. Hunters now have a great alternative with the SST Lite Shotgun Slug. This load is ideal for youth, women or anyone looking to tame the recoil of their rifled barrel shotgun. This ammunition delivers up to 30% less felt recoil when compared to standard 12 gauge slug loadings. This load is loaded with a 300 grain FTX slug that leaves the barrel at 1575 FPS giving a capable range of 150 yards. This mild recoil of this load is perfect for gaining confidence in your slug gun by being able to practice at the range and not developing a flinch from heavy recoil.
Technical Information
Gauge/Bore: 12 Gauge
Shell Length: 2-3/4"
Slug Type: SST
Slug Weight: 300 Grain
Ballistics Information:
Muzzle Velocity: 1575 fps
Muzzle Energy: 1652 ft. lbs.
Technical Information
Ballistics Information:
Delivery Information
The total weight of the product, including the product's packaging as it sits on our shelf. Since some products ship directly from the Manufacturer, their weight may be unavailable. Because of variations in the manufacturing process, similar products may have differing weights. Unless the product is designated as "Ships Alone", Shipping Weight does not include the shipping box or packing materials associated with your order.
Product ships from MidwayUSA, separate from any other products on your order due to its size. Products that are Shipped Alone incur separate "Per Order" and "Per Pound" shipping charges.
Product is regulated by the US Department of Transportation (DOT) and cannot be shipped via USPS or air shipping methods. DOT-Regulated products must ship UPS Ground.
Product is restricted by shipping carriers and cannot be shipped to certain locations or via certain shipping methods.
- Cannot ship via air
- Cannot ship via USPS
- Cannot ship to PO Box
Additional shipping charge for HazMat products. Carriers charge extra to ship packages with products they consider hazardous, such as reloading powder and primers.
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Martin Hampton
Does anyone know how to add the fast forward and rewind buttons into the .swf file? I can only get the play/pause/stop ones.
Ken Clarke
While I've never done it myself, I'd try this. In the Flash Production options, in the General Tab, in the behavior section, you can check "Produce raw movie for further editing in Flash MX editor. Theoretically you could then just edit the RAW file to include those buttons and have Flash MX export the .swf file.
If you try that, please post back and let us know how it works out.


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MAC Feeling Flush Mineralize Blush ($23.00 for 0.10 oz.) is described as a "melange of pink, deep blue-pink, and soft brown." It swirls together and creates a plum-tinged pink with blue undertones and a satin sheen. The mineralize blushes from this collection have a very different texture than past mineralize blushes. These have the texture of the In the Groove mineralize eyeshadow trios–very silky smooth, soft, and really no frost or grittiness. The overall feel is reminiscent of Chanel's Joue Contrastes (not quite as silky). These feel even softer and have less of a sheen the the less frosted side of the In the Groove mineralize blush duos.Read Full Review
MAC Giggly Mineralize Blush Swatches, Photos, Review
MAC Giggly Mineralize Blush ($23.00 for 0.14 oz.) is a new and limited edition duo-toned blusher from MAC's Quite Cute Collection, which will launch in-stores on April 7th. It's described as a "light pink with pearly plum heart." The outer color is a darkened bubblegum pink with subtle blue undertones, while the heart itself is a frosted berry plum. When swirled together, the result is a heavily pinked plum with a soft, frosted sheen.Read Full Review
MAC Sakura Mineralize Blush Swatches, Photos, Review
MAC Sakura Mineralize Blush ($23.00 for 0.14 oz.) is a new and limited edition shade from MAC's Quite Cute collection, due in-stores on April 7th. MAC describes this shade as "lavender with pearly magenta heart." The outer edge is a pinked lavender with a pearled finish, while the heart is a darker pink, not quite magenta, and more of a frost finish. When swirled together, it creates a muted, medium-dark magenta pink with a soft frost finish and cool undertones.Read Full Review
MAC Miss Behave Mineralize Blush Swatches, Photos, Review
MAC Miss Behave Mineralize Blush ($23.00 for 0.14 oz.) is new and limited edition shade from MAC's Quite Cute collection, which launches in-stores on April 7th. It is one of three shades (the others being Giggly and Sakura). MAC describes it as "light beige with pearly mint green heart."Read Full Review
MAC Too Fabulous: Mineralize Blush Duo
On March 4th, Too Fabulous will launch launches in-stores, but you can expect it to pop-up online on either Monday (the 1st) or Tuesday (the 2nd) as well. Included in the collection are six shades of Mineralize Blush Duos ($23.00), nine shades of Cremesheen Glasses ($18.00), and six shades of Lip Pencils ($13.00). I'm missing one mineralize blush (Two Virtues) and one Cremesheen Glass (Loud & Lovely), as well as all of the lip pencils–I will have the missing pieces reviewed on the launch date once I get to the store and purchase them, so please be patient! Update (3/4/10): Everything is now reviewed and all that!Read Full Review


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Diamondbacks' Grant Heyman Great at First Impressions
With a major emphasis on hitting for power in professional baseball; organizations can never have too many power hitters being developed throughout their minor league systems. Taking this concept to heart, the Arizona Diamondbacks selected Grant Heyman in this past June's MLB draft with hopes of him becoming a major impact for the big league club in the near future. Known for his...
Via Baseball Essential
January 12, 2015 | Discuss
16 car big one notes and quotes from Daytona
RICKY STENHOUSE JR., No. 17 Cargill/Winn-Dixie Ford Fusion (INOVLVED IN WRECK ON LAP 19) – WHAT HAPPENED OUT THERE? "I am not really sure. I haven't seen anything yet. Our Cargill Ford was really fast. We had the outside lane working there and it seemed like some of the guys were struggling on the bottom and the middle and we got a little loose on the top. I save it and everything...
Via Speedway Digest - Home for NASCAR News
July 06, 2014 | Discuss
D-backs release minor league pitcher Placido for suspected theft
Some guys never learn. The Diamondbacks this week released pitcher Anderson Placido after he was accused of stealing from at least one teammate on the short-season Hillsboro Hops. Placido also was released by the Indians two years ago over similar accusations. According to a report on OregonLive.com, Placido snuck into the Hops clubhouse during the team's road game Saturday in Boise...
Via Fox Sports Arizona
July 03, 2014 | Discuss
18 drivers to battle it out on the tri-oval in the "Memphis 100 with Ricky Stenhouse Jr."
The USAC Silver Crown Series returns to Memphis International Raceway (MIR) on Sunday after a 10-year hiatus. There will be 18 drivers, including NASCAR driver and hometown favorite Ricky Stenhouse Jr., competing for the coveted win at the .75-mile race track. USAC Silver Crown Series regulars Tracy Hines and Kody Swanson are heading into MIR tied for first-place in the point...
Via Speedway Digest - Home for NASCAR News
June 27, 2014 | Discuss
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Peter Dunfield congratulates Elizabeth Manley after a skate. He coached Manley when she earned a silver medal at the Calgary Games. CBC
Peter Dunfield, a former Olympic skating coach who helped Elizabeth Manley to a silver medal in 1988, has died at the age of 82.
Dunfield and his wife Sonya helped coach Manley to three Canadian national titles and silver medals at the 1988 Olympics and Worlds. The couple had been living in Seattle and Peter Dunfield had recently developed heart and lung problems.
Manley announced news of his death through Twitter, writing: "My heart is empty tonight with the passing of my coach Peter Dunfield. You will be in my heart. Thank you for everything you taught me in life."
My heart is empty tonight with the passing of my coach Peter Dunfield . You will be in my heart… http://t.co/zQlzDu0EUB— elizabethmanley (@Lizmanley88)
May 27, 2014
While coaching in Ottawa, Dunfield was based out of the Gloucester Skating Club.
Skate Canada's Emery Leger coached alongside Dunfield in the early 90s and said he was a perfectionist.
"Detailed. Focused. Very Disciplined. Strict. He knew exactly what he wanted," said Leger.
Both Peter and Sonya Dunfield were elected to Skate Canada's Hall of Fame as builders in 2001.
Prior to coaching, Dunfield was a competitive skater. He was the 1951 junior national champion and the senior bronze medallist in 1952 and 1953, and also represented Canada on two World and North American championship teams.
His wife Sonya was a medallist at two World Championships and represented the United States at the 1952 Olympic Games.
"Their whole life was figure skating," said Leger. "They were really, what we call 'royals' in the skating family."
Dunfield is survived by his wife and two sons.


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Come join us this weekend at the Tonic Lounge to celebrate the release of our debut CD, "Trapped Again." Doors open at 9 pm with special guest the Scrubjays. Hope to see you and rock on!!
If you happened to be listening to KUFO or KNRK lately, you may have heard the title cut from our debut CD, "Trapped Again." If you want your fix of Mental Hygiene, call the radio stations and request to hear us!! Thanks to all of you that have pre purchased the CD or downloaded our music from ITunes. We love you and appreciate your support!!


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Get ready for a flavor EXPLOSION! There are 7 (SEVEN!!) new recipes on this week's meal plan -- the most we've ever had in one week!
You'll find it hard NOT to fall in love with the Thai PB&J Sandwich (NEW!), Corn & Black Bean Cakes (NEW!), and Roasted Butternut & Red Pepper Penne (NEW!).
Also featured on this week's menu: Pad Thai Soup (NEW!), Miso Maple Sweet Potato Tacos (NEW!), Creamy Wild Rice Soup (NEW!) and Banana ...
Mardi Gras is usually all about eating and celebrating in excess -- but it doesn't have to be!
Let the good times roll and enjoy a not-so-fat Tuesday with these 40+ plant-based recipes!
P.S. there will be a like batch cooking tutorial event on Saturday, February 21st -- see Friday's post for more info.
Cajun Tempeh Meatloaf from Holidays
Mardi Gras recipes from the blog:
Meet our Herbie of the Week: Beth Ann!
Beth Ann first shared her plant-based journey in a comment she left on my nutrition elitism post (you can the entire blog post here):
I thought what Beth Ann said was a great example of why "Progress, Not Perfection" is needed -- not to mention I couldn't stop starring at her amazing glow!
But there is SO much more to Beth Ann's story than what was mentioned in that brief comment ...
Celery in foil... what?
Some of these tips will surprise you!
But many more will help you save money, flavor, and freshness.
Keeping your ingredients at the top of their game is KEY if you want to be at the top of yours!
Thus, storing your produce RIGHT is part 3 of the Cook Like a Pro mini-series :)
Plus nothing makes me more sad than limp vegetables...
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Here's ...
With Mardi Gras right around the corner, this week's meal plan celebrates the taste of N'awlins with some all time favorites!
Enjoy two NEW! ragin' Cajun dishes like the Cajun Tofu & Quinoa (NEW!) and Sweet Potato Gumbo (NEW!). Also making their this week: Spaghetti Squash Stir-Fry (NEW!), Vegetable Paella (NEW!), and Maple Oat Crumble (NEW!).
And of course we can't forget these meal plan classic: "Beef" Stroganoff, Chili Fries (or Chili Cheeze Fries), Chocolate Strawberry Oatmeal (perfect for Valentine's Day!), and much more!
Cajun Tofu & Quinoa
NEW! Individual Recipes this week:
Plus these favorites are back: