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where are all the good animes gone?
Where have all the good animes gone too?
I mean the movies or one ep. shows, not the lenghty ones. Such as Ninja Scrolls or Fist of the North Star, or Vampire Hunter D. These types of animes. If there are out there, can someone give me some names so I get them.
Or did animes just go in a totally different way that I did not notice?
All of those have been licensed..so they wouldn't be on here...your best bet to get them is to buy them...
on Animesuki they do however have New Fist of the North Star...but there's only 1 ep released
First of all, the entire anime industry isn't going to take some drastic turn towards exclusively 100+ episode series overnight and you just missed it. The works you're talking about spanned a decent amount of time, for one, and also, you've got to put a little effort into an anime to get anything out of it. If you want a decent storyline, you're usually going to need to see 9+ episodes before you feel like you've seen something "good." Exceptions to that include Ah! My Goddess (5 episodes), a few other shorts, and many full length movies. The point is, watch a series and you'll feel like you're watching something worthwhile. Anime generally doesn't deliver cheap, one-time, never-look-back entertainment like Hollywood here.
There are still several nice movies comming each year. Next year for example we will have Howl's Moving Castle from Studio Ghibli and Ghost the Shell Innocence from Production I.G
Eh? What you talking about? Dude, there's tons of great stuff this season!! :)
i am a big fan of Cat Soup (click the link and then the camera for screenshots) . 1 episode OVA, INCREDIBLY abstract... really messed up. heh. 30 minutes long ;)
I don't think that many OVAs and movies are being licesend, I think mainly they're just being passed over for long running series by the fansub community. There's really no shortage of stuff out there.
As for what is out there...
Macross Zero is pretty amazing so far. 5 episode OVA, up to the third episode now. I reccomend the Infusion or ANBU/AonE versions.
Strangely enough, the new Captain Herlock OVA went pretty unnoticed, only Anime Junkies picked it up as far as I know, but it's licensed now.
Wonderful Days is worth seeing. Not a classic that will be remembered for all time, but something a little different, worth checking out.
The BLAME logs are some of the best stuff out there right now. Six logs and one 'Extra', all about 6 minutes a piece. A promo for a series that's coming out later, based on an incredible looking manga.
I'm sure ther's more, but it really does seem like fansub groups lean towards long running series over a movie or short OVA.
I agree with Yaggy i miss things like ninja scroll and princess mononoke well done films :)
I thought that Spirited Away was excellently done too. :)
Strangly enough though, I actually thought that they did a better job with the english dubbed version than the original japanese version. Not sure why, but it just seems to me like the voice actors did a better job, and that's rare for me to say that. 0_o
what do you mean that where are the good anime gone????
full metal alchemist, chrno crusade are pretty good... i've been watching them.. anime movie such as grave of the fireflies, vampire hunter d are licensed or not on bittorrent. it's best if you download it on kazaa or mirc.
They aren't gone, they are just replaced with new stuff.
Ranma, Ah! My Goddess, You're Under Arrest, El-Hazard, Fushigi Yuugi, Marmalade Boy, Maze, Rurouni Kenshin, etc...
The list could continue forever. They were all amazing mid/late-90's animes, now all animes are CG coloured, have somewhat of a similar plot and animation style as animes from the mid/late-90's.
I doubt they are dead. Some are really dead (I'd be in heaven if they made a new Ranma series or COMPLETED the Ah! My Goddess series in anime!), while others are getting new episodes!
It's amazing how many people are simply not reading the posts in this thread, not even the original post.
i dont think he means downlaoding i think he means why arent there movies as good as the ones he said guys. also spirited away is good. and dont anime companies license movies more cause most people would buy that and not feel like they need to keep buying volumes for a 100 ep seires and so on. EH whatever
I read it as "I'm seeing lots of 100+ episode series here on animesuki...but where are all the movies and short OVAs?"
And it's true, there's tons and tons of series coming out, multiple times in some cases, and yet it seems that most fansub groups aren't going after hardly any movies...except those tied in with a series.
It'd be nice to see more movies get the fansub treatment, and don't try to give me no bull about all the movies getting licensed right away because that's simply not the case, and even if it where, there's an untold number of movies from just a few years ago, on back down the ages, that have never been subbed in any form.
Still, I believe there's a lot of much newer movies that are just getting overlooked. Strange, things used to be completely opposite on the fansub scene. Tons of movies, very few long running shows, and most of those had huge gaps or just dropped off after a while.
OK, What I meant is that it does not seems like the fansubbers are doing any movies or short ovas. That they are only doing long running series. If there are some Movies or Short OVAs, that fansubbers did what are they. Plus I do buy Videos when they come out. Also I import them if I really like it.
Have you tried End of Evangelion or Hoshi no Koe ? The latter is not a movie but a 1 ep OVA done entirely on a single old-school Mac and it looks damn good and the story is quite touching as well :)
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A COUNCIL tenant has been evicted from her home after it was revealed she was illegally subletting it to a friend.
Nicola Chin, aged 37, moved out of her house in Pawsons Road, Selhurst, in 2009 and is now living in Cornish Grove, Penge.
She gave her rent card and keys to a friend, who then moved in his family.
Croydon Council was alerted to the subletting when the friend tried to take over the tenancy.
Ms Chin was ordered to pay £10,000 in rent arrears to Croydon Council as well as £3,000 court costs after a hearing at Croydon County Court on January 17.

subset: commoncrawl


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Hey! I am 15 years old dancer. My height is 1,78. I'm looking for an ambitious and nice dancepartner to dance 10 dances or only LA. I have three years experience in dancing in open class both LA and ST. I live in Lithuania's capital city, but can relocate for promising partner. I speak Lithuanian, Russian and English.
You can contact me by email (firstname.lastname@example.com).
Dance is not hobby for me, it's my life.
Tuesday, 07 January 2014 19:21


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The Office of Residential Life works to build strong communities within each residence hall.
Our mission is to share and model a commitment to the education of students through our roles as administrators, teachers, and mentors.
Residence Halls
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5 Dining Halls (with vegetarian and Kosher options). 4 Cafes (2 of them run by students). 1 student-run pub. 1 s'more pit. yum.
Wellesley Fresh will keep you up-to-date on the weekly menu.
Res Life Staff
Meet the Res Life Staff.
As administrators and mentors for Wellesley students, the Res Life Staff works to build a strong community within the Residence Halls.


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For our children who have grown up over the last decade and never known an America not at war;
For the men and women, military and civilian alike, who have sacrificed on fields of battle from the hellhole of Mogadishu, to our embassies in Africa, to the USS Cole, to the skies over Pennsylvania, to the towers of Manhattan, to the Pentagon, to the barren wastes of Afghanistan, to the bloody streets of Iraq and to the countless hidden places where evil lurks and must be rooted out;
For all who love freedom and crave justice;
And for all who have prayed that evil would not go unpunished,
Today we give thanks to our Creator and to the brave Americans of the CIA and special forces who have fulfilled their mission by ridding the Earth of a great blight.
66 years to the day after the death of another enemy of mankind, a powerful victory over darkness has again been achieved.
Today we also give thanks to our leaders, Presidents Barrack Obama and George W. Bush, who did not waiver in their determination to defeat the forces of hatred.
Admittedly, they each made some poor choices, both in the execution of the fight against terror and in other areas of policy. Because we are free and because we do not fear brutal reprisal, we can and should speak out when our leaders come up short. It is in part for the right to question our commanders in chief and have a government of our own choosing that we have endured this fight.
Our political system is at times divisive, but our unity is greater than our divisions.
In our celebration over this triumph, let us be sober. The death of Osama bin Laden will not by itself bring peace to the world. We would like to think it will make the world safer, but even of this we cannot be certain. It may be that for a time his fanatical followers will hate us even more and be driven to further acts of aggression.
The death of Osama bin Laden will, however, give notice that justice is served and that one less thug inhabits our world.
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JL Audio W3 12" Enclosure with Subwoofer
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The biggest story to come out of the Miss USA pageant was not who got crowned but Miss Indiana's body.
Twitter went crazy with comments about her "normal" body. There were tweets thanking her for not being a bag of bones.
Here's the thing…her body is NOT normal. She is 5'8″ and wears a size 4. The average woman wears a size 12.
Take a look for yourself…you be the judge.


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The race towards March is on. Check out all of Wednesday's Top 25 betting action:
DePaul Blue Demons at Georgetown Hoyas (-15, 135)
It wasn't long ago that No. 11 Georgetown was an unranked team that began Big East play with a 2-3 record. However, the Hoyas will put the conference's longest winning streak on the line on Wednesday when they host DePaul. On Jan. 19, Georgetown provided South Florida with its only league win, but since that setback, the Hoyas have trumped three top-25 opponents and won seven straight overall.
The win streak has allowed Georgetown to move from the bottom half of the conference and into a three-way tie for first with Marquette and Syracuse, the latter of which it will meet Saturday. In the meantime, the Hoyas stand a good chance at remaining atop the Big East with a home game against the Blue Demons. DePaul snapped a nine-game slide with Saturday's victory over Rutgers, but has lost 11 straight against Georgetown.
Providence Friars at Syracuse Orange (-12.5, 130)
Providence has suddenly put itself in major contention for a postseason bid after a four-game win streak, its longest in Big East play since 2004. The healthier Friars attempt to beat their third ranked team of the month when they travel to Syracuse on Wednesday. Providence was without guards Vincent Council and Kris Dunn for 10 and nine games, respectively, with injuries and leading scorer Bryce Cotton missed two close Friars' losses. Providence must continue its strong play on the defensive end to win at the Carrier Dome for the first time since 1999.
Eighth-ranked Syracuse is only 3-3 its last six games, but is 15-0 at the Dome this season – 5-0 in league play. The contest will start a critical three-game stretch for the Orange, who host Georgetown on Saturday and play at Marquette on Monday. Syracuse won at Providence 72-66 on Jan. 9.
Minnesota Golden Gophers at Ohio State Buckeyes (-6, 121)
Both Minnesota and Ohio State have 18 victories this season, but neither is playing well entering Wednesday's matchup at Columbus. The visiting Golden Gophers are far removed from a 15-1 start, losing seven of their past 10 and falling into a three-way tie for sixth in the Big Ten standings. Minnesota is last in conference games in defending the 3-point shot, which could prove problematic against the Buckeyes.
Ohio State is third in the conference in 3-point shooting (36.5 percent), and junior Lenzelle Smith Jr. (39.1 percent) and Deshaun Thomas (38.5 percent) rank in the top 12 in the Big Ten in shooting from beyond the arc. Ohio State is fifth in the conference standings, three games out of first.
Wisconsin Badgers at Northwestern Wildcats (+7, 112)
No. 17 Wisconsin will be fighting to keep its Big Ten title hopes alive when it travels to slumping Northwestern on Wednesday night. The Badgers are two games behind conference co-leaders No. 1 Indiana and No. 5 Michigan State, but are flying high after a routine 71-49 win over No. 18 Ohio State.
The Wildcats enter on a three-game losing streak, having posted a season-low 41 points against in-state rival Illinois on their home floor on Sunday. Its the longest skid of the season for Northwestern, and even a matchup against a Badger squad that's been less than adequate on the road would seem a tough place to stop it. Wisconsin hasn't lost to the Wildcats since Jan. 7, 2009, in Evanston.
Kansas Jayhawks at Oklahoma State Cowboys (-1.5, 138)
Oklahoma State puts its seven-game win streak on the line Wednesday against No. 9 Kansas in a critical Big 12 contest, with both teams trailing first-place Kansas State by a half-game. The 14th-ranked Cowboys are trending upward behind the play of freshman point guard Marcus Smart, who scored a career-high 28 points in an 84-79 overtime win over Oklahoma on Saturday.
Kansas bounced back after suffering its first three-game losing streak since 2005 with dominating wins over Kansas State and Texas last week. Oklahoma State ended Kansas' 33-game home win streak with an 85-80 victory Feb. 2, but the Jayhawks have defeated the Cowboys in five of the last seven games. Kansas freshman guard Ben McLemore, who leads the Big 12 in scoring in conference play at 17.8 points per game, had 23 points against the Cowboys in the first matchup.
Houston Cougars at Memphis Tigers (-19, 158)
Memphis staved off a second-half rally from Marshall last time out to win its 16th straight game, the second longest streak in the nation. The 19th-ranked Tigers host Houston Wednesday in a Conference USA game needing two more wins to clinch their second-straight conference title. Memphis has try to avoid looking past the Cougars with Southern Mississippi, its lone threat in the conference, on deck.
Houston is coming off a heart-breaking loss to Tulsa in triple overtime. The Cougars scored five points in the final five seconds of regulation to force the extra sessions but ran out of gas in the final period. TaShawn Thomas was named the conference's Player of the Week averaging 27.5 points and 13 rebounds over a pair of games last week.
Colorado State Rams at UNLV Runnin' Rebels (-3, 138)
Colorado State has a clear route to sole possession of first place in the Mountain West. It will be far from easy though. To reach a showdown for the top spot in the league on Saturday with New Mexico, the 21st-ranked Rams must first end UNLV's 13-game home winning streak on Wednesday night. Colorado State has won six straight contests since losing at New Mexico on Jan.23. That came one game after beating UNLV 66-61 at home. The Rams have five players averaging at least 9.9 points and led the nation in rebounding through Monday.
UNLV has lost three of its last five games, but the two victories were against No. 16 New Mexico and talented San Diego State at home where the Runnin' Rebels are 15-1 overall and 7-0 in the league. UNLV, led by freshman Anthony Bennett, could use another key victory to seal an NCAA bid.
Washington Huskies at Arizona Wildcats (-12, 137)
A Pac-12 championship is still within reach for No. 12 Arizona heading into its game Wednesday night against visiting Washington. The Wildcats ended a two-game losing streak with a win Sunday against Utah, leaving them a game behind first-place Oregon with five remaining. Washington ended a three-game losing streak with Saturday's victory against Oregon State. The Huskies, the defending conference champions, were picked to finish fifth in the Pac-12 but are tied for eighth with Stanford.
Washington has one of the premier scorers in the Pac-12 in junior guard C.J. Wilcox but he was stymied by Nick Johnson when these teams met Jan. 31, scoring 11 points on 4-for-16 shooting in the 57-53 win by the visiting Wildcats. That was part of an overall decline by Wilcox, in which he shot 34.9 percent from the floor during an eight-game stretch that resulted in seven losses. He regained his touch against Oregon State, shooting 7-for-14 and scoring 24 points.
Santa Clara Broncos at Gonzaga Bulldogs (-15, 142)
No. 3 Gonzaga appeared to have cleared the last major hurdle on the way to an undefeated season in the West Coast Conference when it eased past Saint Mary's to cap a sweep against its biggest conference rival. The Zags host dangerous Santa Clara needing one more win to match the most its ever had in regular-season play. Kelly Olynyk continues to state his claim for national Player of the Year winning the conference's weekly honor for the second time last week.
Rising Santa Clara looks to end the Zags' eight-game winning streak in the one-sided series. Gonzaga has won 26 of the last 27 meetings. Santa Clara coach Kerry Keating has beaten Gonzaga just once, that coming in 2011. The Broncos beat Pepperdine last game 70-60 as senior Raymond Cowels III scored the 1,000th point in his career.


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The following are some of the incidents reported by the Wright County Sheriff's Office for the period ending Jan. 21:
On Jan. 16, Travis Lee Kruck, 25, no permanent address, was arrested in ROCKFORD on a Wright County warrant for flee police in a motor vehicle.
On Jan. 17, Randy Lee Peterson, 45, of Spring Lake Park was arrested in ALBERTVILLE on the charges of theft and flee police on foot.
On Jan. 18, Crystal Lynn Jones, 33, of DELANO was arrested in Delano on the charge of criminal damage to property.
On Jan. 19, Steven Dennis Sturman, 36, of Independence was arrested in DELANO on the charge of third degree DWI.
On Jan. 21, Brian Charles Grote, 26, of Maple Grove was arrested in ST. MICHAEL on the charge of second degree DWI.
There was one property damage accident, no personal injury accidents, no hit and run accidents and one car/deer accident.
There were six arrests for DWI, one underage consumption arrest, no school bus stop arm violations and no tickets for miscellaneous traffic violations reported this week.


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Lisa won't let Sofie bring Hallie to work, and Hallie gets a fever. Alison runs into Sofie at the hospital and learns Hallie needs a blood transfusion and Sofie is not a match. Sofie leaves a message for Cole. Alison suggests Gwen might be a match for Sofie. Gwen agrees to be tested and discovers she's a match. Gwen, Will and Sofie are shocked when Cole arrives.
Gray tells Matt he's going to use Henry and Vienna as bait to set their plan in motion. Alison convinces Casey to ask Matt about his dealings with Gray. Vienna avoids Gray's advances. Matt informs Gray that Alison and Casey are growing suspicious. Casey questions Matt's relationship with Gray. Matt denies knowing Gray and promises Casey he'll have nothing to do with him. Vienna tells Gray she's still in love with Henry and he covers his anger.
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This property has been removed by the agent. It may be let or temporarily removed from the market
Property Description
Letting information:
|Added on Rightmove:
|03 February 2012 (2056 days ago)
Key features
- Southsea
- House
- Part Furnished
- Patio
- 18 Bedrooms
- 1 Reception
- Gas central heating
- Part-double glazing
Full description
Overlooking the common very convenient for college, pubs and nightlife, partially dg, totally centrally heated, wash hand basins in some rooms, some rooms with included shower cubicles and WC's, four communal bathrooms, large comprehensively equipped kitchen, medium size common room/lounge, rear access yard for bike storage etc., some of the front facing bedrooms have panoramic views toward the harbour mouth and the Isle of Wight, some rear rooms have a pretty good view of adjacent fire escapes, there are generally five rooms per level so a group could reserve all of the rooms on one level at a variety of sizes and prices. The communal areas will be cleaned and serviced included in the rent, this will be like a mini hall of residence but without the restrictions. The smallest rooms are priced at £250 pcm and the large front rooms with views range in price up to £335 pcm, We will take single and groups of any number please ask for further details
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Tag archives for headache
Our patients don't practice evidence-based medicine? That doesn't justify doctors embracing quackery!
One of the most annoying phenomena when it comes to "complementary and alternative medicine" (CAM), which its advocates are more and more insistent on calling "integrative medicine" is how little the average doctor cares that pseudoscience is infiltrating medicine. The reason, of course, is that CAM advocates don't like the "alternative" part of the term…


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The Legal Clinic enables students to work with real clients, the community and the court system. The Clinical Programs consist of six clinics, three clinical projects, a Public Service Externship Program, and a Part-Time Student Practicum. Each of the clinics offers distinct experiences while emphasizing a similar approach to clinical legal education.
Clinic courses bring the theory of the classroom to life. Many of our graduates refer to their Clinic experience as the best course they took in law school. We are one of the few law schools to offer both day and evening students the opportunity to participate in a clinical course. In addition to providing law students with extraordinary educational experiences, our clinical programs also fill many unmet legal needs of the community.
Contact Information
UALR William H. Bowen School of Law
Legal Clinic, Room 105
1201 McMath Avenue
Little Rock, AR 72202-5142
Office: 501.324.9441
Fax: 501.324.9911
Kelly Browe Olson, Director of Clinical Programs
Terry Harrison, Higher Education Institution Coordinator
Tiffany Tackett Kell, Mediation Programs Coordinator


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We will drink your blut
Welcome once more to a new edition of Shonen Showdown, the weekly battle recap for whom the term "Paper Tiger" takes on a new and far more terrifying meaning!
This week's pages are backed up with true strength, courtesy of sweet, sweet fighting anime and manga recaps from myself, Pedro, Chris and Kristina, for Naruto, Bleach, One Piece, Hunter x Hunter, and Fairy Tail!
Get on below like you've been grabbed by a magic giant cat at read up on the week in hot combat, then tell us all about your favorite fight shows!
Chris Walden
Hunter x Hunter Episode 52
Gon and Killua finally get hold of Kurapika, and it seems he's less than pleased that they are going after the Phantom Troupe. It makes me wonder as to his reasons why, though. Sure, it seems that he's worried because of how strong they are, but is it also because he wants to deal with them himself? Regardless, he agrees to help the two out with their nen at a later date.
Meanwhile, the Phantom Troupe are running rampant still, and the yakuza inside the building are getting riled and being all yakuza-like. If wearing a suit and yelling at people is all it takes to be a member, I think I could do a pretty good job myself. Might need to lose the hair. They are soon shut up when the top dogs from the Zoldyck family show up.
Which leads to the crux of the episode, and what a great one it was. A two-on-one fight with grandpa and daddy Zoldyck fighting the leader of the Troupe? It's just as good as it sounds, and the fighting is top-notch, if a little short. We do get to see the ins and outs of Mr Troupe's ability, learning that he needs his book open to use the various techniques he has stolen.
Aside from the fighting itself, don't you just love the minimalistic approach with the audio during the battles? It's such an effective way to drive home the seriousness of a fight, and I wouldn't be the least bit surprised if we see this replicated in other shows in the future.
[Hunt for this show on Crunchyroll!]
Pedro Cortes
One Piece Episode 568-570
The Seakings drop a bit of science on Shirahoshi and let her know that she's their queen, the first one in quite a while. They also reveal that Noah was designed to be moved by them for MYSTERIOUS REASONS. Maybe one day we'll find out. For now, we discover that the only other human to hear the Seakings was Gol D. Roger. The Seakings drop off Noah and head out into the ocean while the Ryugu guards find the kidnapped mermaids in the treasure room. Seems that Caribou (remember him?) has been busy. Being the end of all the battles, the royal family apologizes for blaming the Strawhats, forcing the populace to reevaluate their prejudices.
That's immediately tested when Luffy comes back in dire condition. His wound has been bleeding and he's low on blood. On top of having a rare blood-type, the citizens of Fishman Island fear violating the old laws banning giving blood to humans. Being the boss that he is, Jimbei steps in and volunteers. When Luffy regains consciousness, he offers Jimbei a spot in his crew.
Elsewhere, Neptune frees Hordy's human slaves and sentences the remainder of the pirates to hard labor. He also closes the Fishman District. In all of the hullabaloo, Jimbei and the Strawhats escape. Jimbei rejects the offer due to personal responsibilities, but says he will join after he meets his responsibilities. One of Neptune's soliders catches up and offers them a banquet for their troubles. The Strawhats can never refuse food and a party, so they come back to the palace for a good time. After the party, Robin and Neptune step out and talk about what she read on the poneglyph. Neptune talks about Joy Boy, a guy who lived on the surface 800 years prior to the start of the series and how he left Noah behind for somebody to finish his work. Robin reveals that Noah is one of the ancient weapons, revealed by the poneglyph she read back in Skypiea. The other ancient weapon? Shirahoshi herself.
Jimbei pulls the Strawhats aside and gives them the run down on what's happened the last couple of years. For one, Aokiji and Akainu had a falling out involving the succession to Fleet Admiral. They fought for 10 days, leading to Akainu barely winning the fight and the new rank. Second is that Blackbeard has taken over Whitebeard's territory. While the Strawhats are talking Caribou tries to kidnap Shirahoshi, but gets kicked out by Luffy. The episode ends with the representatives of Big Mam, one of the Four Emperors, arriving for their monthly candy tithe.
Josh Tolentino
Naruto Shippuden episode 284-285
This is the sort of story I like to see in mid-arc filler. Rather than making up adventures that can never be acknowledged or simply contradicted once the show gets back into line with the manga, Naruto's using its padding-out time to add substance to engagements that the manga could only give a few panels' to otherwise.
In this case it's Kakashi's group's engagement with the Seven Swordsmen of the Mist and other assorted undead ninjas. Episode 284 covers Sai fighting that ninja with the hammer-and-chisel sword, and 285 covers a bunch of no-name ninjas (plus Omoi) fighting another group of no-names who, between all of them, got a total of one page's worth of space when they first arrived.
Now, that's not necessarily a problem, but in a setting that is stuffed up the bum with major supporting characters whose exploits are still virtually unknown, there's a lot of more significant material that could have used expanding. It's not like we're scraping the barrel bottom here.
Kristina Pino
Fairy Tail Chapters 304-305
It's probably a good thing that I couldn't do last week's chapter since it wasn't too loaded.
It's been revealed that Mavis is some kind of demon tactician that has almost everything figured out for our strongest Fairy Tail team in the games. It was pretty awesome to see Phase 1 of her plan in motion, and I'm excited for next week's dog fight between Grey and Rufus. What I'm looking forward to even more is the fight versus that horrible woman! Who'll it be -- Erza? The entire team versus just her?
As for Jura... I really think that it won't come to a battle with them. At least... I hope not. Jura is so strong that it would kind of take away all his prestige to get his butt whooped by anyone, wouldn't it? He's had some difficulty in past fights, but there shouldn't be anyone in the games that are a match for him one on one. Maybe if all of one guild takes him on at once or something... yeah, I guess I could understand that. Though, what I forsee is that someone's going to play dirty and he'll end up teaming up with Fairy Tail again. Is that a little too predictable? Then again, I may totally be off the mark.
As for the other team, they made it to Lucy's cell but then all were caught by the mysterious Princess of Fiore. The manga suggests that her intentions are unclear, and her expression after "capturing" the crew looked kind of... regretful? Is she against Eclipse? Probably. Will she do something to help them? Also a big question mark. I'm interested in knowing what it is they're going to see in that prison; if perhaps a dragon will come for them, or anything else.
More than anything, what kind of bothers me is the inconsistency with Lucy's powers and her spirits. When it's convenient, they come out and help her on their own. She's also capable of summoning a few at a time if she wanted to. Yet, she's in great trouble and has been separated from her keys -- and they do nothing to assist her. Couldn't Virgo have dug her out of the castle? And more importantly - couldn't the Exceeds and Mirajane have lifted everyone clear of the trap they all fell in? These things bug me. I'll never understand these random plot holes and convenient fixes.
Pedro Cortes
One Piece Chapter 684-686
Smoker and Vergo begin their battle in earnest while Sanji, Tashigi and the G5 marines try to escape the flames in the room. Unfortunately, an explosion causes some of Shinokuni to seep into the room and petrify some marines. Caesar cackles and sets up the floor of the lab to be flooded with gas. We also find out that the accident four years prior was caused by an overly ambitious Caesar, not Vegapunk.
Back in the garbage area, Luffy wakes up and bumps into a tiny dragon. Oh, THIS is Momonosuke. The little punk is snooty, just like his father. Being such a snoot, the kid refused to eat anything after being kidnapped, saving him from the candy addiction the other kids are suffering. This also lead to him sneaking into an open lab and eating an experimental artificial Devil Fruit, which turned him into the little dragon. He also overheard Caesar and his plans, so Momonosuke swallows his distaste of pirates and escapes with Luffy out of the top of the trash chute. There's a quick montage showing what everybody is up to, ending with a 20 minute countdown until Shinokuni enters the black everybody is in. Oh boy, I wonder how long these 20 minutes will last..
While Mocha runs away with the candy, she remembers how the kids looked before their transformations and addictions. Monet proves to be more vicious than originally thought, as she repeatedly attacks Nami, Chopper and Robin instead of fighting Zoro. The girls and reindeer escape, leaving Zoro to fight Monet. The harpy points out that Zoro's only been defending, which probably means he doesn't want to kill a woman or he doesn't know enough about Haki to deal damage. Good thing Tashigi and the Marines arrive. Tashigi knows how to use Haki AND has no qualms about slicing up another woman.
Josh Tolentino
Bleach chapter 513
What?!...what?!. This is pretty much the second time I've done this, and I can't help but to simultaneously facepalm and rejoice.
Last week I noted that it would be supremely unconvincing for Ichigo to be able to engage Juha Bach on anything resembling an equal level without some kind of new twist in the formula. There's simply been no buildup or action to imply that he could have even taken on the sort of people Bach wiped the floor with just minutes before. Unlike with Aizen, Ichigo didn't spend months in time-dilated nega-space duing brain training. He was stuck in a box listening to his friends die.
That's why I'm rejoicing. I am rejoicing that my prediction was correct. Ichigo does have a new twist, one that plausibly puts him in striking distance of big boss power-level. And yet I facepalm because of this new twist, one that plausibly puts him in striking distance of big boss power-level. I know it had to happen, and to be frank I'm at a loss for better ideas of how to do it, but my goodness is it dumb.
The twist? Ichigo's a Quincy.
Or, at least, can now manifest the same "Blut" power as these bad dude Quncies. Yep. I'm pretty sure your palm is on your face as well.
Basically, he's like the Avatar of Bleach, some kind of ubermensch capable of mastering every major power everyone else has, from Zanpakuto to Bankai to Visored mask to Fullbring to Blut. He's Hope Summers or Rogue on steroids and injected with an unhealthy dose of Mary Sue formula. Except he's not a Mary Sue in the traditional sense, because he's the officially sanctioned protagonist.
Well, at least it's not a variation on Getsuga Tensho.
Naruto chapter 607
Obito's set of flashbacks end, finally, with us knowing nothing that we didn't know already. Obito's pissed, has taken Madara's name to lay the foundations for Akatsuki, and summoned the Nine-Tails to Leaf Village. We still don't know why Rin was killed, and doubtless we'll find out once he and Kakashi begin their final battle.
Meanwhile, real Madara's pitting his hard wooden dragon against Naruto's flaming fox (that sounds vaguely obscene) and we'll see how that goes, but my interest will be seeing Kakashi dealing with the sheer power disparity between himself and Obito. They have the same basic Sharingan trick (Kamui), but it's been years since Kakashi's been able to put his whole "Copy Ninja" moniker to use. He'll have to come up with something new, or get some outside assistance (maybe from Guy or the Kages?) to take Obito down.
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AUTO RACING; Nascar's Drive for Diversity Is Producing Mixed Signals
By VIV BERNSTEIN
Published: November 8, 2006
Nascar officials have promoted their diversity efforts for years, hoping to erase a long-held image of stock-car racing as an all-white sport. But the Drive for Diversity program, which began in 2004 and was billed as an attempt to integrate drivers, has yet to produce a member of a minority group, or a woman, who has moved into the upper echelons of the sport.
And now the father of an African-American stock-car driver once called a top racing prospect said the highly publicized Nascar-supported program was a failure. Joe Henderson Jr. said his son, Joe III, was under contract to MB2 Motorsports from 2005 to 2006 and was used for publicity but was given poor equipment in 2005 and not even provided a racecar in 2006.
''It's a sham,'' Joe Henderson Jr. said in a telephone interview Friday. ''The program is not designed to be successful because, No. 1, it's not properly funded.
''They claim that it's a pipeline. Well, nobody came out the pipe.''
It is not the only issue affecting the handful of minority drivers who have dreams of rising to Nascar's top level, the Nextel Cup. At least three African-Americans, including Joe Henderson III, have been the targets of racial incidents at tracks in the South. Most recently, there was an incident in October at Hickory Motor Speedway in Newton, N.C., where a witness said that more than 100 fans crowded a fence next to the track to yell racial epithets at Marc Davis, a 16-year-old African-American driver.
The notions that racial incidents are widespread and that minority drivers are placed in hostile environments were disputed by Malcolm Calhoun, vice president and general manager of Calhoun Enterprises, the parent company for Access Marketing & Communications, which runs the Drive for Diversity program.
''I don't want to do anything to put anyone in harm's way,'' said Calhoun, who said the Drive for Diversity program was a success and pointed out that several drivers had won races with the equipment they have received. Calhoun also said some drivers in the program would be promoted next year to the lower-level Nascar Busch East and West series, which is still a level below the third-tier Craftsman Truck Series and far below Nextel Cup competition.
Not everyone agrees that the program is a success.
''Nascar's not trying to develop nobody,'' said Henderson Jr., who is considering a lawsuit against MB2, Nascar and Access Marketing. ''They have nobody ready to move up. It's a smoke-and-mirror program.''
Joe Henderson III of Franklin, Tenn., was one of the five original Drive for Diversity drivers selected for the program in 2004. None have moved into the top three series.
Joe Henderson III signed a two-year deal with MB2 Motorsports at the start of the 2005 season. He raced in Late Model competition at small tracks in the South in 2005, but his father said that the equipment was not competitive and that the team was understaffed.
At the same time, Joe Henderson Jr. said his son was called upon to make numerous publicity appearances for the Drive for Diversity program. He also was highlighted in a documentary called ''Black Wheels,'' in which Henderson III was called ''the most prominent name mentioned for future stardom'' in Nascar's diversity efforts. But he said his son was ostracized after complaining about equipment. Joe Henderson III, 21, is no longer competing.
''MB2 is very committed to the Drive for Diversity program,'' Jay Frye, the MB2 chief executive and general manager, said in a telephone interview yesterday. ''Everything that we did last year was all brand-new equipment and basically, when the season ended last year, we had a meeting with the Hendersons. They wanted us to graduate him to the next level. We didn't feel he was ready for that. At that point, we said, 'If you can find another deal, find another deal, and we'll do the same thing.' ''
Joe Henderson Jr. is not the only one critical of the Drive for Diversity program. Tommy Gray, a mechanic who has worked with the diversity drivers Allison Duncan, Chris Bristol and Davis, said some race teams were not using the funds properly.
''Some of the people have taken advantage of the diversity program,'' Gray, a teacher in Concord, N.C., said during a telephone interview Saturday. ''Let's just say they never did spend the money wisely. I don't think some of the people that had diversity rides the last few years had all the resources used for them.
''I think the program needs some looking at.''
But Nascar is happy with the performance of the program so far.
''I think that from the onset, our goal in working with Access Marketing on this program was to create opportunity for young African-American, Hispanic and female drivers,'' said Marcus Jadotte, the head of Nascar's public affairs department, which oversees the diversity efforts. ''I think we're doing that. And so, by that measure, I would absolutely consider the program a success.''
Sam Belnavis, one of the few African-American executives in the sport who has run his own race team and currently oversees diversity efforts at Roush Racing, said the Nascar diversity program had succeeded in developing drivers at the lower levels by improving their skills. But asked if the failure to promote drivers was a red flag, Belnavis said the program needed more owners to participate if it was to succeed, and more sponsors to recognize there is an African-American fan base.
''Nascar's not unlike any other sport,'' said Belnavis, who compared the integration of stock-car racing to Jackie Robinson's breaking the color barrier in major league baseball. ''We have to go through our growing pains as the sport becomes adapted.''
Despite complaints, a number of drivers in the diversity program have won races. Jesus Hernandez, who replaced Joe Henderson III in the MB2 diversity program, had two victories in Late Model competition at Hickory using what Frye said was the same equipment. Hernandez will move up to some Craftsman Truck races next year, Frye said.
Davis notched six victories in the Limited Late Model series at Hickory this year. Davis, though, was one of several drivers subjected to racial epithets as they tried to move up in the sport.
Joe Henderson Jr. said his son had an incident in 2005 at Hickory with another driver who uttered a racial epithet. And 13-year-old Darrell Wallace, an African-American driver who is being developed by Belnavis, was subjected to a racial epithet by a competitor in October after a race at Lanier National Speedway in Braselton, Ga.
The Davis incident followed an on-track confrontation between Davis and another driver.
''It was 100-plus fans charged down to the fence or even where they were sitting just yelling racial slurs,'' said Tracy Dean, a local reporter for a Web site who was covering the race at Hickory. ''I thought we'd come a long way. Apparently, we have a long way to go. He was just about to come to tears.''
A handful of fans were ejected from the racetrack.
''It's not tolerated at Nascar events and it won't be tolerated at Nascar events,'' Jadotte said.
Photo: Marc Davis, right, with Woody Howard, has found success this year, but he has been subjected to racial epithets as he has tried to move up. (Photo by John Russell/Associated Press)


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Streetwise: High speed rail
Don Spalding, Retired postal worker: I feel high speed rail is important. It's either pay for it now or pay for it later. However, by the time rail gets built all us retired folks will be dead. You have to look at the California population projections for 2025 and determine whether the infrastructure we have now will support it, and I say it won't. Rail compared to highway or air is much more environmentally beneficial.
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Richie Beyer, County Engineer Lee Colley, Maintenance Superintendent
Highway Department 155 County Shop RoadWetumpka, Alabama 36092(334) 567-1162 Phone(334) 567-1100 email@example.com Hours: Monday - Thursday 6:00am - 4:30pm
The Subdivision & Land Development Regulations, Subdivision FAQ, and Access Management Policies are in Adobe Acrobat Reader (tm) format. If you don't have Reader, you may download it free from Adobe.
The Elmore County Highway Department manages the County�s Road and Bridge network. Management of this network is done through the unit system, a system by which the County Commission employs a County Engineer who supervises and directs the daily operations of the County�s Highway Department. The County Engineer is responsible for the direction of all maintenance, construction, engineering and administration activities pertaining to the County�s road and bridge system.
The Highway Department personnel perform routine maintenance such as mowing grass, cutting brush, blading dirt roads, patching potholes on paved roads, cleaning ditches, and maintaining traffic control signs. The Highway Department operates its maintenance functions with the county�s allocation of gasoline tax resources. These revenues are earmarked under state law, limited in use, and must be used on the county�s road and bridge right-of-ways. The county utilizes this revenue stream to maintain its 750+ miles of paved road, 250+ miles of unpaved roads and the 127 bridge structures located in the county.
Construction projects play a large role in the Elmore County Highway Department. Engineering personnel plan, design and inspect construction activities on all federal, state and county funded projects. These include resurfacing and rehabilitation of our paved road network and replacement and rehabilitation of our county bridges.
The County Highway Department, through its land development division, is also responsible for regulating the subdivision and development of land in the county�s unincorporated areas. All division of land is subject to the county�s subdivision regulations. These regulations are made available to the public at the Elmore County Highway Department and are designed to provide citizens/taxpayers with well-built, safe, and sustainable developments.
Elmore County, also through its County Highway Department, utilizes an Access Management Policy to help aid in congestion reduction and to monitor and attempt to provide safe access to county roads. All entrances or drives (both residential and commercial) are subject to this policy and shall be approved prior to installation on the County right-of-way. Unapproved entrances are subject to removal and/or closure by the department in an effort to provide safe and efficient roads.
Any citizen who has a problem request for maintenance on the county road system is encouraged to contact the Elmore County Highway Department at (334) 567-1162. The County Highway Department operating hours are Monday through Thursday, 6:00 a.m. to 4:30 p.m. After hours, non-emergency requests can be left on the department�s answering service. Emergency requests can be made by calling the Elmore County Sheriffs Dispatch at (334) 567-5441. The dispatch office forwards all requests or notifications to the Highway Department�s on call personnel to address the problem.
County Engineer, Walter Richard "Richie" Beyer, IVCounty Engineer, Richie Beyer, holds a B.S. in Civil Engineering from Florida State University, Tallahassee, Florida and is a Registered P.E. in Alabama, Florida and Georgia. He is a certified Bridge Inspector and Herbicide Applicator. Mr. Beyer was awarded the Association of County Engineers of Alabama (ACEA) County Engineer of the Year Award for 2012. He is a former President, Vice President, Secretary/Treasurer, Sixth Division Board Representative and EIT Representative of the ACEA. Currently, Mr. Beyer is the President of the National Association of County Engineers (NACE) Board of Directors. Previously, he served as NACE President-Elect, Secretary/Treasurer and Alabama State Director.
In addition to serving as speaker for numerous ACCA education events, Mr. Beyer has served on numerous committees for the ACEA addressing Subdivision Regulations, Floodplain Management, Joint Equipment Bidding, Highway Safety, Education and Training, ATRIP Liaison, Pre-event Disaster & Monitoring Contract and Logging Advisory Committee.
Mr. Beyer has been employed as Elmore County Engineer since April 2003. Previously he served five and half years as Chambers County Assistant Engineer and two years as Barbour County Assistant Engineer. He served two summer internships in the Coffee County Engineering Department.
Mr. Beyer is married to the former Kelli Gant and together they have three children, Gracie, Aubrey and Colby.


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Ben Affleck trusted his instincts before signing on to play the Caped Crusader in "Batman v Superman: Dawn of Justice," and he's hoping fans do too.
"Everyone's entitled to their opinion. I wouldn't have taken the part if I didn't trust my instincts in terms of the filmmaking. I think Chris Terrio wrote a terrific script. Zack's a great visual director. And there's an interesting take. I wouldn't have done it if I didn't think I could do it," he told Entertainment Weekly.
The Oscar winner doesn't appear to mind some fans' fervor surrounding the role.
PHOTOS: 'Batman V. Superman: Dawn Of Justice': Pics From Scene In Detroit
"It's great that people do care that much. They want to see the movie that much. And it is incumbent on you to honor the story. There are the Greek myths and these are the American myths. The American myths are these superheroes. People care about 'em a lot. And it's incumbent on you to do a good job and make it as excellent as you possibly can. At the end of the day, the movie's all that matters," he said.
The 41-year-old also revealed to the mag that he worked hard to get into superhero shape after being in not-so-super hero shape for another role.
"It's the responsibility of an actor to get their physical self as close to the role as possible. And Batman's obviously got a set of expectations that are tough. So I spent a lot of time working out. And it's a far cry from Gone Girl where my character is described as 'puffy and hungover,'" he said. "I want you to know I worked equally hard at both!"
"Batman v Superman: Dawn of Justice" will debut in 2016.
-- Jesse Spero


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Mellanox® Technologies, Ltd. (NASDAQ: MLNX), a leading supplier of high-performance, end-to-end interconnect solutions for data center servers and storage systems, today announced it ranked number 251 on Deloitte's Technology Fast 500™, a ranking of the 500 fastest growing technology, media, telecommunications, life sciences and clean technology companies in North America. Award winners were selected based on percentage fiscal year revenue growth from 2008 to 2012. Mellanox grew 365 percent during this period. Mellanox has been a Technology Fast 500™ award winner in each of the past 7 years. "We are proud to be named on the 2013 Deloitte's Technology Fast 500, and are pleased that our efforts continue to gain recognition within the financial community," said Eyal Waldman, president and CEO of Mellanox Technologies. "Our growth represents the increased worldwide adoption of our industry-leading, end-to-end FDR 56Gb/s InfiniBand 10/40 Gigabit Ethernet interconnect solutions in the high-performance computing, Web 2.0, cloud, storage, database and data center markets." "The 2013 Deloitte Technology Fast 500 companies are exemplary cases of those spurring growth in a tough market through innovation," said Eric Openshaw, vice chairman, Deloitte LLP and U.S. technology, media and telecommunications leader. "This year's list is a who's who of companies behind the most exciting and innovative products and services in the technology space. We congratulate the Fast 500 companies and look forward to what they do next." Supporting Resources:
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Brenda Frese, Alyssa Thomas earn ACC Player, Coach of the Year awards
The Maryland Terrapins women's basketball team will begin their run toward an ACC title on Friday, but on Thursday, they prefaced the postseason by earning two major awards from the conference.
Head coach Brenda Frese was named the ACC's 2013 Coach of the Year on Thursday, her first such honor in her 11 years at the helm of the Terrapins. At the same time, junior forward Alyssa Thomas was named the league's Player of the Year.
Maryland enters the 2013 postseason boasting a 23-6 overall record with a 14-4 mark in the ACC. That's good enough for the #2 seed in the conference tournament, which begins Thursday in Greensboro.
Frese, who led the Terrapins to their first and only national championship in 2006, previously won the AP's National Coach of the Year award as the head coach of the Minnesota Golden Gophers.
"This is an individual award, but it's really about every person in our program," Frese said. "I couldn't be prouder of how we've fought to get better each and every day, no matter the circumstance."
Thomas, who also won this award last year, leads the ACC in rebounding, grabbing an average of 10.4 boards per game in 2012-13. She's also third in scoring with 17.7 points per game and second in assists with just over 5 per contest. She's leading the league in each of those categories during conference play and has recorded 17 double-doubles.
"To be voted this honor again is really special because there a lot of really good players in our league," Thomas said. "I just work hard every day to make my team better and be the best I can."
Maryland opens up ACC Tournament play at 6 p.m. Friday. They'll play the winner of Thursday's Georgia Tech/Wake Forest first-round game.


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Loeffelbein, Gustav A.; florist; born, Germany, Jan. 3, 1845; son of August and Augusta Kewinski Loeffelbein; educated in the Government schools of Germany; married, Germany, March 6, 1870, Fransika Liebensiki; issue, six children, but one, a son; living; landscape gardener, Townsend & Fleming, Buffalo, 1857; started with Carl Smith, with him till 1860, twelve years with Linthope Tenny; came to Cleveland, and went to work for Martinetz; then for Theo. Shear; started business in Glenville, then in E. 49th St.; present location since 1886; designer and decorator; member Presbyterian Church; member Florist, Lasalle, Union.
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onions wrote:So that we're all on the same page: if they are indeed limiting the number of greetings sent per ip address, then there is no way that a sender is going to know this. There is no error message, no notification whatsoever. If you are reading this and are still sending multiple messages, please stop until further notice. I really don't want anyone to waste their time if unnecessarily.
deco_droid wrote:i think lego's intent was to donate the toys based on a large group of fans participating -- not based on a few afol's uber-spamming tactics.
follow the rules and be happy with the results.
onions wrote:probably a good idea to turn off any bots you guys have running until we can confirm that messages are once again being sent out. like i said, they may be figuring out other measures to prevent bot spamming.
buriedbybricks wrote:Better than the 100 a day it was crawling at when we started.
lostone wrote:Just wondering what the figure is at now
lostone wrote:and is there anything else we can do to help?
onions wrote:i'll be making a post on the front page later today with an update.
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It was the strangest dream. I had been thinking about that "what five historic people would you invite to dinner?" question as I drifted to sleep. Next thing I knew, this happened:
Ernest Hemingway: Can I get a martini? Very dry.
Me: What?
Hemingway: A martini, dry.
Joan of Arc: And I'll have a red wine.
Me: Why are you telling me this?
Vincent Van Gogh: Because we're here for dinner. You invited us.
Me: Oh, of course.
Abraham Lincoln: I'd like a nice cold beer. I don't get many of those where I am now.
Me: Really? I figured you of all people would be up. . . you know.
Lincoln: Oh, I am, I am. But Mrs. Lincoln doesn't approve. Says it's a sin. Even Jesus has tried to get her to lighten up, but she's having none of that, and tells me so several times a day. I sometimes visit Jefferson Davis just to get some peace and quiet.
Lincoln: Just down the street from me.
Hemingway: I have to say, I'm a bit surprised to hear that.
Lincoln: You and me both, brother!
Me: So this is a unique opportunity. I wanted to meet you all and to hear your stories, so I thought—
Joan: Not now, waiter. I am hungry. Some beef bourguignon, s'il vous plait.
Hemingway: Beef bourguignon? That's a little fancy for a soldier, isn't it?
Me: I'm not actually the waiter, I'm—
Joan: Don't be a chauvinist, you bearded old fool.
Hemingway: Ha ha! I like this one. I think I found the fifth ex-Mrs. Hemingway.
Joan: Don't flatter yourself. I could never be with such an ungodly man.
Van Gogh: Maybe she goes for the sensitive artist types, Heer Hemingway.
Joan: Maybe she doesn't appreciate being spoken about in the third person.
Van Gogh: Maybe she should learn that the last 600 years haven't done her any favors!
Joan: Go choke on a sunflower, you one-eared ginger!
Van Gogh: I know what I want for dinner. Steak, burned and crispy.
Joan: Ass!
Lincoln: I thought you were a vegetarian.
Van Gogh: What?
Lincoln: I SAID, I THOUGHT YOU WERE A VEGETARIAN.
Me: If we could return to the topic at hand. I'd like to discuss a few things before we—
Lincoln: Barbecued ribs! That's what I want. Man, nothing says summertime cookout like ribs and a few cold ones with friends.
Hemingway: The wife not let you have those either?
Lincoln: No. She keeps yammering about my cholesterol. I keep telling her, 'Mary, we're already dead,' but she doesn't listen.
Van Gogh: What?
Lincoln: What do you want for dinner? Were you serious about the steak?
Van Gogh: No, I was just—argh, now I don't know. Maybe pizza? Come back to me.
Hemingway: Steak sounds like a grand idea. And can we get another round of drinks here, barkeep?
Me: I'm not actually the barkeep either. I'm the—
Joan: Yeah, yeah, we didn't ask for your life story.
Lincoln: Hey, Hem, you got any cigars?
Hemingway: Sure thing, Abe. Vinnie, Joanie, you want one?
Van Gogh: Sure, why not?
Joan: Well, I usually try to stay away from fire, but I think I can make an exception.
Me: You're going to have cigars before dinner? I thought they came after.
Lincoln: Doesn't look like we're in any danger of that happening anytime soon.
Me: Actually, the dinner has already been planned and prepared, and should be here shortly.
Hemingway: Then why don't you make like a tree, and go get it.
Me: I keep telling you. I. Am not. The waiter. I'm your host, and the entire purpose tonight was to gather five notable people and have some engaging conversations about the world today.
Van Gogh: Five? I only see four of us. You aren't the fifth, are you?
Me: Oh, no. I am merely a humble—
Hemingway: Thought so. I mean, I'd never heard of you.
Me: Our final guest has yet to arrive.
Amelia Earhart: Sorry I'm late everyone. I seem to have gotten lost. Waiter, could I have a gin and tonic, please? I'm parched. There's a good lad.
Me: I give up. You people can just feed yourselves. And I hope you all choke on it!
Van Gogh: Alright, pizza time! Somebody get Alexander Graham Bell in here, and let's order some pies! Waiter, where's our drinks?
There's a lot of laughter and talking as I leave, and I wake up in my own bed.
With a voicemail from my credit card company asking whether I meant to order 12 large pizzas, two cases of beer, and a side of barbecued ribs.
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Friday, September 05, 2014
Five People You Meet At Dinner
Posted by Erik Deckers at 08:00
Labels: Abraham Lincoln, Amelia Earhart, dinner party, Ernest Hemingway, Joan of Arc, Vincent Van Gogh
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We have previously reported that 5 days of a high-fat diet followed by 1 day of high-carbohydrate intake (Fat-adapt) increased rates of fat oxidation and decreased rates of muscle glycogenolysis during submaximal cycling compared with consumption of an isoenergetic high-carbohydrate diet (HCHO) for 6 days (Burke et al. J Appl Physiol 89: 2413–2421, 2000; Stellingwerff et al. Am J Physiol Endocrinol Metab 290: E380–E388, 2006). To determine potential mechanisms underlying shifts in substrate selection, eight trained subjects performed Fat-adapt and HCHO. On day 7, subjects performed 1-h cycling at 70% peak O2 uptake. Muscle biopsies were taken immediately before and after exercise. Resting muscle glycogen content was similar between treatments, but muscle triglyceride levels were higher after Fat-adapt (P < 0.05). Resting AMPK-α1 and -α2 activity was higher after Fat-adapt (P = 0.02 and P = 0.05, respectively), while the phosphorylation of AMPK's downstream target, acetyl-CoA carboxylase (pACC at Ser221), tended to be elevated after Fat-adapt (P = 0.09). Both the respiratory exchange ratio (P < 0.01) and muscle glycogen utilization (P < 0.05) were lower during exercise after Fat-adapt. Exercise increased AMPK-α1 activity after HCHO (P = 0.03) but not Fat-adapt. Exercise was associated with an increase in pACC at Ser221 for both dietary treatments (P < 0.05), with postexercise pACC Ser221 higher after Fat-adapt (P = 0.02). In conclusion, compared with HCHO, Fat-adapt increased resting muscle triglyceride stores and resting AMPK-α1 and -α2 activity. Fat-adapt also resulted in higher rates of whole body fat oxidation, reduced muscle glycogenolysis, and attenuated the exercise-induced rise in AMPK-α1 and AMPK-α2 activity compared with HCHO. Our results demonstrate that AMPK-α1 and AMPK-α2 activity and fuel selection in skeletal muscle in response to exercise can be manipulated by diet and/or the interactive effects of diet and exercise training.
- acetyl-coenzyme A carboxylase
- fat oxidation
- muscle glycogen
- skeletal muscle metabolism
short-term (<1 wk) manipulation of dietary macronutrient intake is associated with marked changes in skeletal muscle gene expression (1, 5, 24), substrate stores (36), metabolic flux, and fuel oxidation (10, 22, 23). Exercise training also results in striking modifications in muscle gene expression (14), energy reserves, and the relative contribution of fuels to the energetic demands of muscle (9). Accordingly, the extent to which acutely altering substrate availability might modify the training impulse has been a key research area among exercise physiologists and sport nutritionists for several decades (for review, see Ref. 19). Indeed, evidence is accumulating that nutrient manipulation can serve as a potent modulator of many of the acute responses to both endurance (15) and resistance exercise (7, 11).
During recent years, our laboratory has undertaken a series of independent but related studies into the effects of a practical "dietary periodization" strategy in well-trained endurance athletes on selected aspects of metabolism and exercise performance (3–6, 32, 33). We have shown that short-term (5-day) exposure to a high-fat diet, while undertaking high-volume, intense training, followed by 1 day of rest and "carbohydrate restoration," results in greater rates of whole body fat oxidation and decreased rates of muscle glycogenolysis during submaximal [70% peak O2 uptake (V̇o2 peak)] cycling than when subjects undertake the same training and consume an isoenergetic high-carbohydrate (HCHO) diet (3, 32). The decreased rate of muscle glycogenolysis during exercise following fat adaptation with carbohydrate (CHO) restoration can, in part, be explained by decreased pyruvate oxidation via pyruvate dehydrogenase flux (32). However, it is not clear what precise metabolic signal(s) accounted for the marked shifts in substrate utilization during exercise. In this regard, it has been suggested that AMP-activated protein kinase (AMPK) may provide a direct link between intracellular signaling events and subsequent substrate selection in exercising muscle (16, 34). In support of this hypothesis, diet-exercise manipulation that results in low muscle glycogen content is associated with increased resting AMPK activity (31, 35) and elevated phosphorylation of AMPK's downstream target, β-acetyl-CoA carboxylase (ACC-β) Ser221, compared with high glycogen stores (35). Furthermore, the degree of AMPK activation during submaximal exercise was also shown to be dependent on the fuel status of the contracting musculature, with AMPK activity elevated to a greater extent in muscle with low compared with high glycogen levels (35).
While the findings of Wojtaszewski et al. (35) imply that glycogen availability per se may modulate AMPK activity, it cannot be ruled out that the diet-exercise regimens used to differentiate glycogen concentration in that study also resulted in significant changes in muscle lipid availability [i.e., increased muscle triacylglycerol (TG)]. Accordingly, the observed changes in AMPK activation may have been more a consequence of lipid-induced rather than CHO-induced effects on this enzyme. In this regard, we have recently shown that endurance training in high-fat-fed rats was associated with increased AMPK-α1 activity, whereas AMPK-α2 activity was elevated by high-fat feeding alone and was not increased further by exercise training (25). Accordingly, the aim of the present study was to investigate the effect of a diet-exercise protocol known to be associated with elevated rates of fat oxidation, independent of muscle glycogen availability on AMPK signaling at rest and during subsequent submaximal exercise. We hypothesized that fat adaptation followed by CHO restoration would increase resting muscle triglyceride concentrations and that changes in lipid availability per se would modify AMPK signaling at rest and during exercise.
Subjects and Preliminary Testing
Eight endurance-trained male cyclists or triathletes [body mass (BM) 70.2 ± 1.8 kg; age 31.6 ± 2.3 yr; V̇o2 peak 61.5 ± 1.5 ml·kg−1·min−1, peak sustained power output (PPO) 339.85 ± 5.8 W; values are mean ± SE] were recruited to participate in this study, which was approved by the Human Research Ethics Committee of RMIT University. Subjects were fully informed about the possible risks of all procedures before providing their written consent. One week before experimental testing, each subject undertook an incremental cycling test to exhaustion on an electronically braked cycle ergometer (Lode, Groningen, The Netherlands). The initial testing protocol has been described in detail previously (20). During the maximal test and the subsequently described experimental trials, subjects breathed through a Hans-Rudolph two-way non-rebreathing valve and mouthpiece attached to a gas analysis system (Parvomedics) interfaced to a computer, which calculated the instantaneous rates of O2 consumption (V̇o2), CO2 production (V̇co2), minute ventilation (stpd), and the respiratory exchange ratio (RER) every 15 s from conventional equations (20). Before each maximal test and all experimental trials, the analyzers were calibrated with commercially available gases of known O2 and CO2 content. V̇o2 peak was defined as the highest V̇o2 a subject attained during any 60 s of the test, whereas PPO was calculated from the last completed work rate plus the fraction of time spent in the final noncompleted work rate multiplied by 25 W. This value was used to determine the power output corresponding to 70% of each subject's V̇o2 peak (63% of PPO) to be used in the subsequently described experimental trials. The maximal test session and all experimental trials were performed under standard laboratory condition (18–22°C, 40–50% relative humidity), and subjects were fan cooled during all these sessions.
Overview of Study Design
The experimental protocol is shown in Fig. 1 and has been described in detail previously (3). In brief, each subject undertook two 7-day trials in a randomized, crossover design, separated by a 2-wk washout period. The interventions consisted of prescribed supervised training while undertaking either a 5-day high-fat diet followed by 1 day of HCHO restoration (Fat-adapt) or a 6-day HCHO diet. During Fat-adapt, the subjects were prescribed a high-fat (4.6 g·kg−1·day−1 fat, 68% of energy), low-CHO (2.5 g·kg−1·day−1 CHO, 17% of energy) diet supplying 0.25 MJ/kg BM. HCHO was an isoenergetic diet providing 10.3 g·kg−1·day−1 and 70% of energy from CHO and 1.0 g·kg−1·day−1 and 18% of energy from fat. Protein content was maintained at 2.3 g·kg−1·day−1 during both trials, and diets were constructed to maximize, or at least match, absorbable energy. Fiber intake was kept to a daily mean intake of <50 g and matched to within 5–10 g each day between dietary treatments. All meals and snacks were supplied to subjects, with diets being individualized for food preferences as well as BM. Subjects received their food in prepared packages and were required to keep a food checklist to note their compliance to the dietary instructions and their intake of any additional food or drinks. Every 2 days, they met with a dietician to receive new food parcels and check their adherence to the previous days' diet. The HCHO diet prescribed in this study is similar in composition to that recommended by sports nutritionists for endurance-trained athletes (2). However, the high-fat diet is far removed from the nutritional energy sources typically consumed by our subjects. While an ad libitum diet group may provide insight into diet-training interactions, the addition of such a group was not considered within the current experimental design.
On the morning of days 1 and 5 of each trial, subjects reported to the laboratory after a 12- to 14-h overnight fast and undertook a 20-min steady-state (SS) ride at 70% of V̇o2 peak. Pulmonary gas exchange data were collected for the last 5–6 min to calculate the rates of whole body fat and CHO oxidation. This ride was immediately followed by a high-intensity interval training session consisting of eight repetitions of 5-min work bouts at 85% of V̇o2 peak with 1 min of recovery at 100 W. The metabolic demands associated with this interval training session have been described in detail previously (33). The intent of this ride was to lower the muscle glycogen concentrations on the first day and initiate a rapid differentiation between dietary treatments on the basis of their ability to restore depleted muscle glycogen stores. On day 6 of both the Fat-adapt and HCHO trials, subjects were required to rest and ingest a HCHO diet (10.3 g/kg BM).
Experimental Trial
On the morning of day 7, subjects returned to the laboratory after a 12- to 14-h overnight fast to undertake a 1-h SS ride at 70% of V̇o2 peak. On arrival in the laboratory, a single leg was prepared for muscle biopsies, and two incisions were made 2–3 cm apart along the vastus lateralis muscle. A resting muscle sample was then taken using the percutaneous biopsy technique with suction applied. After resting for 10 min, subjects began the 60-min SS ride, and a second biopsy was obtained within 10 s on completion of the ride while subjects remained seated on the ergometer. All muscle biopsies were rapidly frozen in liquid N2 within seconds of the designated time point for collection and stored at −80°C until subsequent analysis.
Analytic Procedures
Rates of fat and CHO oxidation.
Whole body rates of CHO and fat oxidation (g/min) were calculated from the respiratory data collected during the last 5–6 min of the 20-min SS ride on days 1 and 5 and during the last 5 min of every 20 min of the SS ride at 70% V̇o2 peak. The calculations were made from V̇co2 and V̇o2 measurements, assuming a nonprotein RER value, according to the following equations (27). Total fat and CHO oxidation during the 60-min SS ride were estimated by calculating the area under the oxidation (g/min) vs. time curves for each subject. Rates of CHO oxidation (μmol·kg−1·min−1) were determined by converting the rate of CHO oxidation (g/min) to its molar equivalent, assuming 6 mol of O2 are consumed and 6 mol of CO2 produced for each mole (180 g) oxidized. Rates of fatty acid (FA) oxidation (μmol·kg−1·min−1) were determined by converting the rate of triglycerol oxidation (g/min) to its molar equivalent, assuming the average molecular weight of human triglyceride to be 855.3 g/mol and multiplying the molar rate of triglyceride oxidation by 3, because each molecule contains three molecules of FAs.
Muscle glycogen and TG concentration.
Approximately 40–50 mg of muscle were freeze-dried and powdered, with all visible blood and connective tissue removed. The freeze-dried muscle sample was then divided into two aliquots. The first aliquot (∼3 mg) was extracted with 250 μl of 2 M hydrochloric acid, incubated at 100°C for 2 h, and then neutralized with 750 μl of 0.67 M sodium hydroxide. Glycogen concentration was determined via enzymatic analyses, as described previously (7).
The second aliquot (∼7 mg) was used for the determination of muscle TG content, as previously described (13). In brief, lipid was extracted by Folch extraction (12), the TG was then saponified in an ethanol/potassium hydroxide solution at 60°C, and glycerol content was determined fluorometrically and expressed as micromoles per gram dry weight.
AMPK Signaling
Approximately 60 mg of wet muscle were homogenized in buffer A (50 mM Tris·HCl, pH 7.5, 1 mM EDTA, 1 mM EGTA, 1 mM dithiothreitol, 50 mM NaF, 5 mM sodium pyrophosphate, 10% glycerol, 1% Triton X-100, 10 μg/ml trypsin inhibitor, 2 μg/ml aprotinin, 1 mM benzamidine, and 1 mM phenylmethylsulfonyl fluoride), and the homogenate was then centrifuged at 20,000 g for 25 min. The supernatant was aliquoted, and the total protein concentration was determined by bicinchoninic acid method (Pierce Chemical, Rockford, IL). The aliquots were then stored at −80°C until further analysis.
Approximately 2.5 mg of protein from the supernatants were incubated with AMPK-α1 or AMPK-α2 antibody bound protein A-Sepharose beads (6 MB; Amersham Biosciences, Uppsala, Sweden) for 2 h at 4°C. The polyclonal antipeptide antibodies to AMPK-α1 and AMPK-α2 were raised to nonconserved regions of the AMPK-α1 (rats 231–251) and AMPK-α2 isoforms (rats 351–366). The immunocomplexes were washed with PBS and suspended in 50 mM Tris·HCl (pH 7.5) buffer for the AMPK activity assay. The AMPK activities in the immune complexes were measured in the presence of 200 μM AMP. Activities were calculated as picomoles of phosphate transferred to the SAMS peptide per minute per milligram of protein subjected to immunoprecipitation (pmol·min−1·mg−1). The post-AMPK immunoprecipitation supernatants were then incubated in streptavidin-Sepharose high-performance beads (Amersham Biosciences) for 1 h at 4°C for affinity purification of ACC.
Western Blotting
The affinity-purified ACC fraction was electrophoresed on 7.5% SDS-PAGE and detected by immunoblotting with anti-phospho-Ser221-ACC polyclonal antibody. The blots were then stripped (50 mM Tris·HCl, 2% SDS, 115 mM β-mercaptoethanol) at 55°C for 20 min, blocked, and incubated with horseradish peroxidase conjugated streptavidin (Amersham Pharmacia Biotech UK, Little Chalfont, UK) to determine the total ACC content. Aliquots of the immunoprecipitated AMPK-α1 and AMPK-α2 proteins were electrophoresed on 10% SDS-PAGE and detected by immunoblotting with antibodies specific for the AMPK-α1 and AMPK-α2 isoforms. The post-AMPK immunoprecipitation supernatants containing 60 μg of total protein were electrophoresed on SDS-PAGE {acrylamide concentration either 15% [glucose transporter (GLUT)-4] or 10% [Fat/CD36, peroxisome proliferator-activated receptor-γ coactivator (PGC)-1α], respectively} and detected by immunoblotting with antibodies specific for GLUT-4 (Biogenesis, UK 4670–1704), Fat/CD36 (AbCam, Cambridge, UK), or PGC-1 (Millipore, AB3242). The immunoreactive proteins were detected with enhanced chemiluminescence (Amersham Biosciences) and quantified by densitometry. α-Tubulin was used as a loading control to ensure that the protein content was the same in all wells.
Statistical Analysis
Data from the two experimental trials were compared by using a two-factor (diet and time) ANOVA with repeated measures. Separate analyses were undertaken to compare data from 20-min rides on days 1, 5, and 7, and data were collected at different time points during the experimental ride. Holm-Sidak post hoc tests were undertaken when ANOVA revealed a significant interaction. Muscle glycogen and muscle TG concentration between trials were compared by using Student's t-tests. Relationship between muscle fuel stores (muscle glycogen and muscle TG) and AMPK activities were studied using Pearson product-moment correlation coefficient. All values are expressed as means and SE, with the critical level of significance established at P < 0.05.
Substrate Oxidation During Exercise
Figure 2A shows the rates of whole body CHO oxidation during 20 min of cycling at 70% maximum V̇o2 (V̇o2 max) (20-min SS) on days 1, 5, and 7. Rates of CHO oxidation during the 20-min SS were similar during both trials on day 1. There was a significant effect of diet on rates of CHO oxidation, such that HCHO was higher than Fat-adapt at all time points after baseline (P < 0.05). During HCHO trial, 1 day of rest on day 6 increased the rates of whole body CHO oxidation during 20-min SS significantly compared with baseline (P < 0.05). During Fat-adapt, 5 days of high-fat diet and exercise training decreased the rates of whole body CHO oxidation, such that it was significantly lower than all of the other time points.
Figure 2B shows the rates of whole body fat oxidation during 20 min of cycling at 70% V̇o2 max (20-min SS) on days 1, 5, and 7, along with the average RER data during the 1-h SS ride (60-min SS) on day 7 (Fig. 2C). Rates of fat oxidation during the 20-min SS were similar during both trials (P < 0.05) at baseline (day 1). The effect of diet on rates of fat oxidation was significant at all time points after baseline, with Fat-adapt being higher than HCHO (P < 0.05). One day of HCHO diet and rest decreased rates of fat oxidation, such that, on day 7, values were not different from day 1 (30.20 ± 3.42 vs. 25.79 ± 3.72 μmol·kg−1·min−1, respectively; P < 0.05). During HCHO, rates of fat oxidation were similar on days 1 and 5 (23.03 ± 3.61 vs. 18.85 ± 2.61 μmol·kg−1·min−1; P < 0.05) but decreased on day 7 compared with day 1 (23.03 ± 3.61 vs. 14.44 ± 1.19 μmol·kg−1·min−1; P < 0.05). The average RER over the 1-h SS ride on day 7 was also lower during Fat-adapt (0.87 ± 0.004 vs. 0.93 ± 0.004; P < 0.01; Fig. 2B), such that fat oxidation accounted for 40.3 ± 3.6% of total fuel oxidation in this trial compared with 21.9 ± 1.5% in HCHO (P < 0.05).
Glycogen and Muscle TG
Table 1 displays muscle glycogen concentrations before and after the two experimental trials. As intended, there were no differences in preexercise muscle glycogen levels between dietary treatments (547 ± 40 and 694 ± 55 mmol/kg dry wt for Fat-adapt and HCHO, respectively). Glycogen utilization during 60-min SS was significantly lower after Fat-adapt (179 ± 35 vs. 264 ± 45 mmol/kg dry wt, P = 0.04).
Figure 3 displays muscle TG concentrations before and after 60-min SS cycling at 70% V̇o2 max. Muscle TG concentrations were higher on day 7 after Fat-adapt compared with the HCHO trial (69 ± 10.2 vs. 39 ± 3.4 μmol/g dry wt; P = 0.006). Postexercise muscle TG levels were similar between Fat-adapt and HCHO trial (Fig. 3). There was a small but nonsignificant increase in muscle TG concentration with HCHO after exercise.
AMPK-α1 and -α2 Activity
Figure 4 displays resting (preexercise) and postexercise AMPK signaling [AMPK-α1 and -α2 activity and the phosphorylation of ACC (pACC) at Ser221] after the two dietary treatments and resting total protein content of AMPK-α1, AMPK-α2, and ACC. Resting AMPK-α1 and -α2 activities (Fig. 4, A and B) were higher after Fat-adapt compared with HCHO (P = 0.02 and P = 0.05, respectively). There were no differences in the protein content of either AMPK-α1 or -α2 in response to either treatment (Fig. 4C). Exercise increased AMPK-α1 activity (Fig. 4A) after HCHO (P = 0.02), but did not have any further effects on either AMPK-α1 or -α2 activity after Fat-adapt. AMPK-α1 and -α2 activity postexercise were not different between dietary treatments.
There was a significant relationship (r = 0.53; P < 0.05) between resting muscle TG concentration and AMPK-α2 activity. When this relationship was expressed as the difference in resting muscle TG for each subject between the two dietary treatments vs. the difference in AMPK-α2 activity between treatment, the correlation improved to r = 0.82 (P = 0.04). Resting muscle glycogen concentration was significantly associated with AMPK-α1 (r = −0.53; P < 0.05), but not AMPK-α2 activity. There was a trend for a relationship between muscle TG utilization and changes in AMPK-α2 activity (r = 0.52; P = 0.06), but no relationship between muscle TG utilization and AMPK-α1 activity.
pACC Ser221
The phosphorylation of the AMPK downstream target ACC at Ser221 (relative to total protein content) on day 7 tended to be higher at rest after Fat-adapt compared with HCHO (P = 0.09). Exercise increased the pACC Ser221 during both HCHO (P = 0.042) and Fat-adapt (P = 0.008), with the postexercise pACC Ser221 being higher during the Fat-adapt compared with the HCHO trial (P = 0.02; Fig. 4D).
Protein Expression
Total protein abundance for FAT/CD36, GLUT-4, and PGC-1 was not different between trials (data not shown).
We have used a human model of short-term (5 days) adaptation to a high-fat diet followed by 1 day of CHO restoration (i.e., a HCHO diet) in well-trained athletes undertaking high-volume, intense training to study the effects of diet-exercise interactions on muscle signaling and metabolism. The novel findings from the present study were that, despite the brevity of the intervention period, 1) the Fat-adapt protocol resulted in elevated resting muscle triglyceride concentrations that were independent of glycogen availability; 2) changes in muscle triglyceride levels were associated with increased basal AMPK activity without concomitant changes in protein levels; and 3) Fat-adapt attenuated the exercise-induced rise in AMPK-α1 and AMPK-α2 activity compared with an isoenergetic HCHO diet.
In accordance with our laboratory's previous investigations (3, 4, 6, 32), we again report that rates of fat oxidation during 60 min of submaximal cycling (70% of V̇o2 peak) were elevated approximately twofold after fat adaptation followed by CHO restoration compared with when subjects performed identical training and consumed an isoenergetic HCHO diet throughout the 6-day intervention period (Fig. 2). Furthermore, muscle glycogen utilization was significantly lower after Fat-adapt (Table 1). In our laboratory's earlier studies (3, 4, 6, 32), it was not clear what metabolic signal(s) accounted for the shift in muscle substrate use during exercise. Here we show that one of the residual effects of the short-term, high-fat diet was to increase resting muscle TG stores (Fig. 3). The resting TG levels observed in our well-trained athletes after just 5 days of consuming a high-fat diet are in close agreement with those reported by Helge et al. (21) for previously untrained subjects who completed 7 wk of endurance training while consuming a high-fat diet (21). They are also similar to the values of Zderic et al. (36) for endurance-trained cyclists after 2 days of a high-fat diet and Zehnder et al. (37) in muscle from trained athletes after just 1.5 days of lipid supplementation (37). Taken collectively, these findings suggest that muscle TG stores are prone to rapid fluctuations in response to changes in nutrient composition and physical activity levels.
In a previous study from our group (5), we showed that 5 days of a high-fat diet coupled with a rigorous training program resulted in a small (17%) but significant increase in the protein abundance of FA translocase (Fat/CD36). These data (5) indicated that an increase in dietary lipid availability was, in part, responsible for the rapid upregulation of Fat/CD36 protein expression, suggesting a role for this FA transporter in facilitating greater skeletal muscle FA uptake. In that investigation, we only assessed the effects of 5 days of a high-fat diet without CHO restoration on Fat/CD36. In the present study, we measured Fat/CD36 protein content after fat adaptation followed by 1 day of rest and a HCHO diet. Under these dietary conditions, we were unable to detect any differences in the abundance of this FA transporter between the fat adaptation protocol and the HCHO diet intervention. To the best of our knowledge, no previous studies have measured GLUT-4 or PGC-1 protein expression in response to the short-term fat adaptation and CHO restoration protocol utilized in the present study. However, Steinberg et al. (31) reported higher skeletal muscle GLUT-4 mRNA expression immediately after an acute bout of endurance exercise commenced with low (vs. normal) glycogen content. With regard to PGC-1, Sparks et al. (30) reported that PGC-1α mRNA was reduced by 20% in muscle from healthy young males following 3 days of a high-fat diet, whereas Mortensen et al. (26) found no differences in PGC-1α mRNA expression after a chronic training intervention in previously untrained subjects in which 50% of the training sessions were performed under conditions of low starting muscle glycogen concentration.
A second important finding from the present study was that the high-fat diet was accompanied by increased AMPK-α1 and -α2 activity (Fig. 4, A and B). Wojtaszewski et al. (35) have also demonstrated that resting AMPK activity is sensitive to fuel status (35). These workers reported that AMPK activities for both α1- and α2-isoforms were higher in the face of low- (∼160 mmol/kg dry wt) compared with high-resting (∼900 mmol/kg dry wt) muscle glycogen content. While their results (35) suggest a relationship between muscle glycogen content and AMPK activity, it should be noted that glycogen does not inhibit AMPK activity in vitro (28). Indeed, it is unlikely that altered muscle glycogen availability is the only factor regulating muscle AMPK activity. Further support for this hypothesis is that the diet-exercise intervention used to differentiate resting muscle glycogen stores in the study of Wojtaszewski et al. (35) is also likely to have resulted in elevated muscle TG levels, particularly as arterial plasma free FA concentrations were significantly higher in the low- vs. high-glycogen condition. Although we did not measure plasma FA levels in the present investigation, our laboratory has previously shown no differences in resting concentrations between the two diet interventions (3, 4, 6).
Our laboratory has recently reported an increase in AMPK-α2 but not -α1 activity in rodent skeletal muscle in response to chronic high-fat feeding (25). Of note in that study was the observation that AMPK-α1 activity was only increased in high-fat-fed rodents when they concurrently undertook an intense endurance training program with a high-fat diet (25). At the time, we suggested that there may be distinct roles for the AMPK-α subunit isoforms in skeletal muscle, with AMPK-α1 activity being linked to exercise training-induced adaptations and muscle glycogen storage, and AMPK-α2 activity being responsive to increased lipid availability (25). In the present study, we provide further data in support of this premise. We observed a significant relationship between resting AMPK-α1 activity (but not -α2) and muscle glycogen content (r = −0.51, P = 0.05). In contrast, AMPK-α2 (but not -α1 activity) was positively associated with resting muscle triglyceride content (r = 0.53, P = 0.05). Perhaps the most robust evidence to corroborate the isoform-specific modification of resting AMPK activity and muscle lipid status comes from the paired data from each subject's resting biopsy samples: compared with a HCHO diet, fat adaptation significantly increased muscle triglyceride concentration (Fig. 3), with such changes being strongly correlated to difference in basal AMPK-α2 activity (r = 0.82, P = 0.04). Raney and Turcotte (29) have also reported a positive association between FA uptake and oxidation and AMPK-α2 activity in the perfused rat hindlimb model. However, it should be noted that correlational data cannot determine causality, and it is possible that factors other than muscle lipid status may play a role in modifying AMPK-α2 activity. In this regard, while resting muscle glycogen concentrations were not statistically significant between the two dietary groups, values in the HCHO trial were, on average, 27% higher than after the high-fat diet. It is well accepted that preexercise muscle glycogen availability influences subsequent rates of muscle glycogenolysis (17, 18). Indeed, it is likely that a 25–30% difference in resting glycogen content is of physiological significance (17).
While it is tempting to attribute a causal role for altered muscle substrate availability (i.e., glycogen and/or triglyceride stores) on subsequent changes in AMPK signaling, it should be acknowledged that differences in AMPK responses in the present study could, in part, be due to our well-trained subjects performing prolonged, strenuous training on a high-fat diet (i.e., the interactive effects of diet and exercise). In this regard, Hansen et al. (15) have previously reported that, compared with daily training with normal glycogen reserves, commencing 50% training sessions with low muscle glycogen (and presumably higher triglyceride) levels resulted in a more pronounced increase in citrate synthase activity. The potential mechanism(s) for this augmented training response after "low glycogen" training is hard to define, but it is possible that commencing exercise with reduced glycogen (i.e., high-fat) availability may promote training adaptations through perturbation in circulating systemic factors (i.e., increased catecholamines), altered muscle substrate availability, or a combination of both. Although we did not measure catecholamine levels during training in the present study, Hansen et al. (15) have reported that the catecholamine response to exercise performed with low muscle glycogen levels is higher than when exercise is undertaken with normal glycogen stores, demonstrating a higher stress response. Further work will be needed to determine the precise roles of local (muscle) vs. systemic factors in modifying AMPK responses to diet/exercise manipulations.
As noted previously, one of the residual effects of 5 days of a high-fat diet in combination with intense training is that rates of whole body fat oxidation during submaximal exercise remain elevated above those observed after a HCHO diet, even when CHO stores are restored (3, 6, 32). Accordingly, one would predict that the pattern of activation of the downstream target of AMPK, ACC, would reflect such perturbations. This is indeed the case. pACC Ser221 tended to be higher at rest after fat adaptation compared with the HCHO diet (P = 0.09, Fig. 4D), and while there was an exercise-induced increase in pACC Ser221 for both treatments, the phosphorylation state of ACC Ser221 was significantly higher at the end of exercise after fat adaptation (P = 0.02; Fig. 4D). Wojtaszewski et al. (35) have previously reported that resting ACC-β Ser221 phosphorylation was higher in subjects with low compared with high resting muscle glycogen stores. In that study (35), concentrations of creatine phosphate and adenine nucleotides in resting and exercised muscles were not altered by the glycogen manipulation, leading these workers to suggest that, in basal conditions at least, fuel-dependent mechanisms independent of energy status may regulate AMPK signaling. Due to insufficient muscle sample in the present investigation, we could not measure high-energy phosphate contents and muscle metabolites, but, using the identical diet-exercise regimen and subjects of comparable training status, our laboratory has previously reported similar resting values for muscle adenine nucleotides (32). However, in contrast to the findings of Wojtaszewski et al. (35), we found that, after 20 min of cycling at the same intensity as employed in the present study, fat adaptation was associated with a lower accumulation of free ADP and AMP after 20 min of submaximal exercise compared with the HCHO intervention (32).
In conclusion, we have used a human model of "dietary periodization" (i.e., fat adaptation followed by CHO restoration) to study the interactive effects of alterations in fuel availability (i.e., muscle lipid stores) and exercise training on AMPK signaling and metabolism. We found that, compared with an isoenergetic CHO diet for 6 days, 5 days of a high-fat diet followed by 1 day of rest and a HCHO diet resulted in higher resting muscle triglyceride levels. The altered muscle fuel status at rest was associated with increased AMPK-α1 and -α2 activity. During exercise, Fat-adapt increased rates of whole body fat oxidation and "spared" muscle glycogen. Accordingly, the increase in ACC Ser221 phosphorylation was greater after fat adaptation compared with the HCHO diet. The changes in AMPK activation observed in the present study could be due to altered fuel status (increased muscle lipid availability), a greater adaptive response when training with low muscle glycogen availability or, most likely, the interactive effects of both of these factors.
This study was supported by research grants from GlaxoSmithKline (UK) to J. A. Hawley and the National Sports Council of Malaysia (to W. K. Yeo).
The authors thank Emmanuel Churchley for technical assistance in the study, and Clare Wood for supervision and implementation of all dietary control.
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
- Copyright © 2008 the American Physiological Society


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