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The setting of this story is not long after Naruto and Sasuke's fight on the hospital roof; for the sake of story, Sasuke has not run away from the Konoha Village to join Orochimaru.
Transition Anchor
Part One
The man adjusted his headband. There were a lot of people in this world, most he had never met, many he likely would never meet. Few gave him the notion to be intimidated. But this woman? Yeah. She could be absolutely frightful.
"I don't know." He said. Because he didn't know. He stuck his hands in his pockets; posture, expression, voice tone -- they spoke indifference. "He's alive. It was mostly exponential luck, but we can't complain."
She was kneading her forehead, skewing the diadem-shape located in her central brow. He considered taking a step back; she kept a cool head, but in particular moods was rather volatile. "He can't keep surviving on luck, Kakashi. None of us can."
He smiled, which was only notable through the single eye visible on his face, "Isn't luck what it all boils down to, Hokage-sama? Every good gambler knows that."
When she didn't take up the challenge to answer, he took it as signal to exited the room. Best not to tempt fate anyway...
The tingling, prickly sensation of a waking body. Fuzzy mind and... aching head... Damn. Ow. He was lackadaisical and careless, but even he knew the procedure for regaining consciousness in an unknown situation. First... and... foremost: Check for injuries.
The left side of his face was swollen (hn, nothing permanent there, no worries. Moving on)... possibly a few crushed knuckles (that was more concerning; there was implication then that he'd been fighting, then?)... Crap; his left ankle might be broken. Might. No need to panic yet. Um. He'd twisted it... running. Tree jumping, actually. No. No, no, it was roof-jumping.
His eyes slipped open to the rather familiar ceiling of the Leaf Hospital. Which meant he was safe. Or... had been saved by someone else. Crap. Beatings he could deal with. Death... well, he wasn't terribly afraid of much. But being indebted to someone -- that, he didn't tolerate very well. Yes, yes, he was remembering a bit more now, and there was a powerful voice in his mind telling him to relax and stay still so his body could heal. As he'd been doing quite a bit of lately.
... this was, what, the third time this month?
"Dammit!" He hurled his pillow across the room, where, judging from the sound, it overturned a tray of utensils. He lay there, quaking, glaring at the light above him. Silence. Crap. "Dammit!" Now other pains were taking shape from the abrupt movement. Swollen joints, rather stiff. Perhaps he'd been slammed into a few things before losing consciousness? "Dammit!" Ah, that familiar burning of flesh shredded raw. So likely he'd also had the fun of skidding across a few unforgiving surfaces (rock? tree bark? no, no, he thought he was remembering cobblestone. Or was that another time?)
He thrashed for a while in impotent rage, twisting, a fish caught up in a net of sheets, accomplishing very little aside from stripping the bed, kicking the blankets off into a huddled mass on the floor. It would have been nicer to find someone sitting there. It would have been relieving for someone to grab him by the shoulders and force him down to the bed, make him be still so he wouldn't injure himself.
No one was there, of course; there were things to be done, missions to fulfill, trades to trade, orders to be given and obeyed. For a short time, even the Genin apprentice groups had been disbanded to free up individuals for missions. Forces were spread thin. There wasn't time to visit a single lonely kid in a hospital. ... his friends probably didn't even know he was here, right? (... of the few people he was going to tentatively label friends, anyway...) It was possible... Sakura was in the next town over; the hotel district of the Hidden Leaf village. He thought she'd said that. And surely his sensei's Iruka and Kakashi were out and about on some important errand... Jiraiya wouldn't have come either way, being repelled by public places altogether (selfish bastard.)
... and Sasuke was likely still mad at him, prone to grudges as he was. For whatever reason. (They had run into each other a few times since the fight on the hospital roof, but the other had showed no feigned attempts at hospitality. The treatment was reciprocated.)
Damn. Though he loved attention, he wasn't yet willing to give up the life of one accustomed to solitude, and thus wouldn't admit so out loud, but he was severely beginning to miss the company of Team Seven. The interaction had been somewhat therapeutic. He'd never been forced to get along with others before; prior to graduation, the academy usually gave the option of teams, but he never bothered participating, since no one wanted him in their group and in all honesty he didn't exactly want to join them. But... he'd become accustomed to his fellow teammates.
Up to the point that Iruka sensei gave him his own headband, he'd accepted blindly that he was destined to be apart. He'd burned every figurative bridge available (among other more literal things -- heh, he'd gone through a short pyrotechnic stage, but the punishments had been too severe to be gratifying.) So the company of those he dared believe to be genuinely concerned about him would have been very comforting right now.
He wanted to be on a stupid mission. Not the more recent ones he'd had to do solo. He could handle people trying to kill him or kidnap him to torture on a later date or whatever it was they wanted with him so long as he was with people he could trust and talk to.
... damn. He wanted to storm across the town and slam Sasuke right in the face. Bastard, trying to ruin their comradery (since he wasn't dense enough to call it friendship exactly.)
Irritated and aching, he finally crossed his arms and sent his disapproving scowl into the world, wishing misfortune upon all.
Uh... this most recent attack... ah. Yes. Stupid renegade ninja. Akatsuki or whatever the hell they were called. It seems since that last unfortunate run-in with the older Uchiha brother (which he wouldn't be forgetting any time soon) he'd been dealing with underlings instead; Missing-nin, ferocious and determined. The Nine... he hadn't heard much from since.
So he wasn't top priority (yet? anymore? relief? indignation? bastards.) For now, he was fodder for hired Missing-nin of the Chuunin and possibly Jounin variety. And boy, was it starting to ache.
His pride was suffering as deftly and painfully the knowledge was sinking in that, clever and tricky as he was, it wouldn't have taken much for them to have killed him. His survival had been attributed to their ambition to get a hold of him still warm and alive. Or rather, still providing a warm, alive nest to house the Kyubi. It was getting unpleasantly obvious that they didn't particularly care if he was wholly in tact, however. If a broken ankle was the worst of his troubles, he would be grateful (last time they'd crushed a few ribs and both collar bones. )
Not prone to remain still (ever a body in motion) he tentatively tried sitting up. Ow. Hurt his stomach muscles but... yeah. Okay, it was tolerable. Try dangling his feet off the bed -- ooh, he felt that. Something had popped in his knee (he wasn't clear-minded enough to recall where each of these injuries came from. Terror and desperation tended to cloud the mind.) Slowly. Slooowly. He slid off the bed and onto his good foot. His shoulders and back strained under the weight of his body, and he was finding his neck rather sore.
Well! He was nothing if not a veteran to physical hardship!
Okay! Great, he could walk. Maybe the ankle wasn't broken after all -- either way, it was well wrapped. So he was mobile. He needed to go exploring. Screw sitting here and waiting for someone to come and tell him what happened (they never told him everything anyway, neglectful bastards.)
Ow, shit. Limping...
Out the door, down he hall, down a flight of stairs, another hall, possibly a little lost, thoughts jumbled...
There! Voices. Someone who could fill him in, yes?
He slowed down before pushing the door open, trying to recognize what voices were within (as, though he'd begun to trust a few people, he was still somewhat on pins and needles with the general Hidden Leaf populous and not looking forward to get a few solid cuffs and a lecture from an embittered villager in his current state.)
Oh! There was Iruka sensei's voice! That man above all others he would follow like a sheep and trust entirely with his life and safety... but still he held back. There was tention in his former instructor's tone; tension tempered by a pinched tone of respect. As dear as the man was, he was only a Chuunin, a young one, and thus had a good few superiors to defer to.
He held his breath and pressed his ear to the door; there were many people in the room and all of their voices were overlapping, and somewhat hushed. Complete statements were hard to make out.
"-n't know for certain it was the Akatsuki-"
"-the issue isn't who it is, it's what do they want with-"
"-whether he's a boy or not doesn't matter. If they do get ahold of him and harness the power of the Kyubi-"
"-it's a valid point; perhaps it was sentimental folly that he's been allowed to live-"
"-should all lower your tone; this is a private-"
"-not really interested in who is at fault; I've said from the begining we were making a mistake in not ending the spirit's existance the instant it was in a body that could actually be ended-"
"-'s true, though he's still young and petty now he's obviously still interested in destroying our happiness and peace-"
"-those horrible, destructive pranks of his-"
"-'re all being absurd -"
"-painted obscenities on the wall of my bathhouse-"
"-switched the road signs-"
"-have my full confidence that he's entirely harmless-"
"-maybe at first, but he's becoming more dangerous. In these desperate times can we afford an unstable element to the machinations of our society-"
"-look what he did to the Hyuuga boy! A genius amoung even the advanced bloodlines-"
"-could only be a short time before he starts-"
"-we've had this discussion millions of times before, you can't-"
"-this is different! Think for a moment, even if he is truely innocent-"
"-do you think they will do to him if they catch him? They'll try every means possible to remove that seal-"
"-one small body can't take a series of anti-seals, plus they'll likely try adding a few of their own for safety-"
"-worse than torture; his mind and body would be ripped to shreds-"
"-couldn't it be more humane to simply-"
A hand clamped down on his shoulder, another quickly covering his mouth firmly to prevent the startled yelp forthcoming. His hand was already going to his thigh to retrieve a knife, which weren't there of course, since his weaponry had been removed along with his outter layer of clothes as standard procedure for patients in the hospital. Getting dragged down the hall, heels scraping on the hard ground. Damn it all to hell!
He'd resolved himself to bite when he was released on his own; he spun, curled little fist arcing along with him--
Of course he did no damange. He never was very good at reacting to surprises.
"You scared the hell out of me, Kakashi sensei!" He pulled his hand free from his instructor's, where it was being held stationary.
The single eye conveyed uselessly little, "It's unbecoming to eavesdrop, Naruto."
He wondered if there was someway to escape the man. He knew which pouch Kakashi kept his smoke bombs; if he could just snag one... Yeah right, and somehow escape on his mangled foot, already aching, from a Jounin level ninja. So he spat, "I think I should be allowed to hear when they're talking about me. Do you know what they were even saying? They want to kill-"
The angular man only rubbed his chin disinterested, gaze slipping off to examine a poster encouraging hygene, and said bluntly, "They've been having debates like that for the past twelve years. It's not even an official meeting, so stop worrying." He offered a irritating smile, "Besides, even if they did manage to rally a meeting, it would be... a very large debate. It would mean involving every competant Leaf Ninja's opinion, and you don't just have Iruka and me in your defense. They won't dare act on anything without the Hokage's consent."
There wasn't much one could respond to that with. He wasn't terribly happy, but in all honesty not entirely surprised to know there were also conspirators against him much closer to home. Thank goodness the Third and now Fifth Hokage spoke in his favor...
"Come on, then, if you're well enough to be up let's get your things. I'll walk you home."
He jerked his shoulder out of the man's reach, "I can get home by myself."
Expectant stillness.
"... Oh." It took valiant effort to keep his shoulders from slumping. It wasn't a matter of company. It was a matter of insuring he actually reached home, for the safety of the village. Kakashi... you... can kiss my ass.
The damn Jounin didn't bother to respond; useless mass of flesh and bone. He wondered sometimes about this man; wondered about his motives -- attached as he'd personally gotten, there was an eternal air of formality that left him distanced. It wasn't blatant disreguarde, just... a reminder, through ever word and action, that he was nothing more than a student. On a good day, maybe a comrad. On a bad day: little more than a responsibility. But in all cases, not a friend.
Well. He could just go to hell, anyway.
Fists balled in pockets, he marched appropriately down the hall, doing his best to ignore the rather silent echo of his footsteps behind him. He would have muttered but didn't want to open an invitation for his escort to try talking to him. Crap.
Through the trees, at rapid speed; higher, above the branches, sailing through the air, now. Up, up, until it's all a long field of broccoli below. Faster, still; breaking all known records of traveling the Earth, and then still within the life-giving reach of the atmosphere. Land and water rush by, for miles and miles.
Finally, far beyond the lush reach of the Hidden Leaf shinobi, over a chain of mountains that form a natural barrier against potential rain, is spread a vast, wide desert of sun-bleached sand. Perhaps at one time it had been a sea, leaving behind a glittering of salt that burns the eyes of an ill-preparted traveler. The terrain changes little as it whips by, the endless rise and fall of unforgiving dunes until...
There. A large, abrasive stony wall of cliffs. Scale the outside and stop before you plummet to your death -- look down at a very sudden, devastating drop. It's a naturally made wall, and nestled deep down, as far as the eye can see, partially protected by the rocky shade for most of the day, is the colony of Those Hidden in the Sand.
A more gradual descent down the inner slope; where long series of stairs have been carved -- archers could stand along the rugged wall and easily pick off approaching attackers.
Lazily drift by over the clay roof tops of the dwelling, look there: a well, with a thick lid to pull over it when the sun hits its scorching zenith. The village, warped in vision by the heat rising from the streets, looks abandoned. Most have retreated indoors for this excruciating portion of the day. A dog, it's flanks hot, lays on its side in the meagre shade of a house.
Travel these forsaken streets, hearing snatches of lazy voices drifting through windows thrown vainly open, hoping to coax a breeze inside.
At the center of the village now; a tall structure, pillars supporting its four corners. Here sits the new Kazikage, leader of the Sand, and he is trembling.
"We... have confirmed in... every way possible..."
Cold, red eyes staring into his.
"... that your father... is..."
"Dead?" Though he lacked eyebrows, the ridges where they would have occupied rose. The black bags around his eyes, product of unnumbered sleepless nights, did not make him look vulnerable or exhasted.
"I'm... afraid so."
There wasn't as much reaction as was expected. "Oh."
The Kazikage squirmed in his robe, pulling his overhanging hat a fraction lower. Hiding behind it.
It was duly noted.
"What else are you afraid of?"
A hesitation seized the older man, so powerful that his mouth was stuck open, like a dying man failing to suck in his last gasping breath, "Defeat."
He was studied.
"Currently, we have surrendered to the Konoha Village for... I suppose I can call it 'accidentally' attacking them. We were manipulated by Orochimaru but... it has currently left the Leaf in a vulnerable position. Soon, we will attack on our own command. Will you join us, and help us win?"
But thoughts had gone elsewhere. To the Leaf shinobi. To the warriors there. To the blood he'd extracted there. To the giant toad he'd fought there.
To the boy who defeated him there.
He shrugged, "I don't care. I don't want to fight the Leaf anymore. Whatever."
He departs, a great shadow of golden sand overlapping his head to protect him from the scalding sun. He hissed up at it, and it did not respond.
In his absence, the Kazikage muses. He is old; not much younger than the recently deceased Third Hokage. His memory is long, his mind is sharp, and his ambition is restless. The uncle to the Sand's own recently killed Kazikage, he stares out at the heat stricken village and taps fingertips together. More than any other shinobi, the Sand and Leaf had the most dealings together. They had the most trade, the most shared resources and the most competition and, though it had been difficult to keep up with the Leaf's inherent power (they were one of the only villages that continuously embraced and encouraged the growth of Enhanced Bloodlines), Those Hidden in the Leaves were also the village most notorious for bad luck.
They were a closely knitted village of single parent families; orphans buried their siblings, a father is known to outlive three or four of his children.
Look back for just a moment, over the long distance traveled to reach this secluded enclosure, back through the trees and look through a window with a view of the mountain bearing the four faces of the great Hokages. Sit inside the academy teacher Iruka's office and note a sparse series of photographs, their edges burned, suffering water damage, bent and folded. This is a man who not only lost his parents but also his childhood home, which had long ago collapsed under the great typhoon of the Kyubi's tails.
This man is not so significant. For two houses down is another man who lost a mother and two brothers to the same creature, only he also has lost one arm.
Across the ally is a father who had two lungs punctured to save a daughter from a caving roof, caused by the same tragedy. Few were left untouched by the destruction of the creature and the first few years after its disappearance were devoted to dragging the almost destroyed civilization out of poverty and despair, out of times the few Jounin left were all that stood between the hovering, vulture-like shadows of smaller, ambitious shinobi settlements looking to pick off competition. With their old generation nearly lost, pressure was placed upon the surviving children to excel, to train, to fight and become powerful.
Sweltering heat. A dry, arid breeze, chapped lips like peeling pain.
The Kazikage knows the situation of the Leaf shinobi. He had been a spectator in the third round of the Chuunin exams, and he had seen the power of their barely graduated Genin. These children were barely rookie class, and most did not even have a full year yet to get accustomed to being called 'ninja'. They were twelve, thirteen years old.
And they had reached the final rounds.
That... was not comforting to the Sand. A village nearly crushed into oblivion a decade ago was now producing a race of super children.
Thank the gods for the absolute genius that created Gaara.
He was the previous Hokage's jewel, and his bane, his greatest weapon and his hated creation; not his son, not even his blood. Sealing . The absolute power and absolute efficiency of the boy was soul shattering. ... If only he were actually controllable.
While retreating to an indoor area, the man was considering the creation of the monster boy -- it was also been to compete. The rumors that the Fourth Hokage had managed to seal a demon into an infant couldn't be left unchallenged after all... And if the Leaf could do it, so could the Sand. Had to keep up. First and formost, always, always keep up.
Well. Once they had smashed their rivals, they could take the next few years dissecting the children to figure out just how they had gotten so powerful...
To boil it down, what he disliked most about having a social life was letting people get away with things. A stranger he could ignore. A contemporary, he could deck full in the mouth. A superior he could sass.
But a person he'd spent days in the presence of? They... got away with things. The time spent with someone, for him at least, merited some sick obligation to tolerate their antics, even when you didn't want to. Even when you wanted to crawl into the knot of a tree and meet every curious face with a kunai to the eyeball. Just to piss him off, Kakashi insisted on putting a hand on his shoulder as he walked. What, to be... supportive? Like hell. It was the touching that nettled him. Ill-accustomed as he was to the unfamiliar sensation of physical contact, and already somewhat humiliated about once again finding himself hospitalized...
It was times like these that he could really stand to be hidden away in his stuffy little apartment.
The lighting to the receptionist's desk was dim and useless. He carried his jacket slung over an arm, wrapped inside was his headband. Over a shoulder he kept his belt and weapons pouch; it wasn't particularly heavy, but at the moment it was a little cumbersome -- it was full of metal weaponry. For just a moment, he wondered what he would have done were his sensei to offer helping him carry his things. Die of shock, first. But he probably wouldn't let him do it anyway. So he dismissed it.
He was in a ripe bad mood. He would deal with it.
"Naruto!" Startled, he refocused on the world, taking a moment to comprehend that a powder pink shape was moving towards him. And talking, "What happened? I heard you were attacked-"
"Nothing I couldn't handle." He spread his face wide with a semi-artificial grin. If he wasn't smiling, no one else would either. Except Kakashi, but he didn't count. Crazy bastard.
So someone had come to visit him. Odd how life turns out; two years ago, a pre-pubescent version of himself would very skeptically ask what she wanted. Last year, in the first budding throw of hormones, he would likely have back flipped into Kakashi's arms and started singing. What... was he feeling now? Touched. Glad to see a face of someone he had confirmed more than once at least worried about him sometimes.
But in all honesty... what was Sakura to him these days? Outside of Iruka-sensei, he could call her the person he trusted most. Without question he would (and had before) willingly enter a fight to the death to protect her. She talked to him; confided maybe even. She listened to him if he was trying to be serious (which was rare, thank god. There were too many miserable people in this world and he was determined to not be one of them.) She got on his case. These past weeks when they were required to accomplish missions on their own, she would nag and remind him to write his report summary. Did he still want to kiss her? Well, yeah, but... how to word it? It was in the same way he wanted to hug Iruka sensei. And throw Sasuke through a wall.
He'd never had a sibling, never had any sort of relation by blood, or close ties to refer to from his childhood. She was Sakura. And were he more comfortable with the word, and more concrete in understanding its definition, he would say she was like a sister. A part of the small, patch-work quilt family he was putting together for himself.
"What are you doing with your things?" She was moving closer and, despite his extra level of tolerance allotted to her, he was feeling penned in with her in front of him and Kakashi still unmoving behind him. Being around a lot of people was scary sometimes. "You're still covered in bandages -- you can't have been here more than a few hours. Get back into bed right you, you moron!"
"Sakura, if Naruto wants to go home, he can go home," their sensei assured her.
The girl didn't seem mollified and called to the doctor behind the desk, fat and riffling through his papers like a pig rooting in a trough, "Hey! Tell him he has to return to his room until he's recovered!"
It took only a second, a gleam of comprehension. Just a matter of looking at the doctor, and the doctor looking back. A twitch of an eyebrow. A silent understanding. A recognition. A comprehension. An inert curiosity: had this man lost someone to the Kyubi? Was he a doctor, treating victims of the monstrous fox's attack? Ponder, ponder...
"If he wants to leave, he can leave." Was all the doctor said, and returned to his grunting, snorting, ruffling of paper.
Kakashi pushed him along from behind, "See? Nothing to worry about."
"Thanks for trying to visit me!" He called over his shoulder as he was ushered out, "Hey, stop by later and tell me about the hotel district-!" The door was shut behind them.
Now out in the open, to the friendly smells of a close-living community, they walked in silence. And observed. It was night, and the stars and moon cast more than enough light to see by, and the larger streets had lamp posts to guide the way.
Most of the town had been repaired from the battle with Orochimaru and his allies. The bodies had all been buried within the first week, once they had been identified and claimed by grieving families. It was times like these he felt both regret and relief that he would never have to identify the body of a brother, an aunt, a grandfather. Loss was part of what being a shinobi was all about, and this was not the first, the last nor the greatest loss of life the Hidden Leaf had suffered. (Um... though perhaps the battle with the Kyubi had the largest body count to date.)
The majority of property damage had been patched up with strips of sheet metal and the broken windows at least had boards over them (supplies were slower in coming these days; already the neighboring towns were skeptical in getting involved with shinobi on the potential edge of war.) There were still deep pocks in the streets, where maybe exploding notes had detonated, or perhaps a skilled Jounin had done some impressive feat with a good portion of Chakra.
The towering walls of the buildings ran in asymmetrical directions; streets made little impact to a people accustomed to leaping from one root top to another. Not wanting to expand the boundaries of the great wall surrounding the village, the natives were quite content building upward, in whatever direction they could, and not often in a tidy way. Alleys ran for miles sometimes, only to stop at a splintery wooden wall; some slender walkways narrowed out until only the residents of slighter stature could glide through sideways (an amenity he was still able to enjoy.) There were many stripes of shadow on the ground as different levels of elevated paths, sometimes just planks of wood, allowed villagers to travel across the street without having to set foot on the dirt.
Unlike the neighboring commercial districts, it was more silent here. Ninja moved with stealth by instinct, and habitually watched their surroundings -- more so than ever now, in these darker days. Children were kept indoors at night. Some even during the day. With the academy temporarily closed to free up instructors for missions, parents were now pressed to home school. To keep even the youngest prepared. Just in case. Always just in case.
In the distance he saw the top-heavy building where he lived; on a blustery day, it looked like it was on the eternal edge of blowing over; his back patio was precariously suspended by randomly placed pipes and planks of wood. He would know; he'd built it himself a few years back, after begging permission from the landlord.
Oh, crap, the landlord. His rent was due this week. When he was younger (far back as he could recall, actually) he paid for living through labor; between studying for the academy, practicing his jutsu and running laps around the village, he would sweep and scrub and feed animals while the other tenants were away. He tended the small flower boxes jutting out of windows, as though the building itself had acne. He was even pretty decent at mending, and had made a few extra bucks by darning socks for the busy women when in a pinch.
That was the plus side about being a full-fledged Genin -- it was a real job, with real wages. He was even finding himself with enough extra money to buy frivolous things like pictures to hang on his walls and a carpet to decorate the wooden floor. (Though having money was also teaching him the value of not flashing it around; more than once Jiraiya had weaseled his cash away from him. Bastard.)
Finally, he was home and, holding his weapon pouch in his teeth, he searched his pockets for the key.
"Well...," his escort said, eyes wandering as though anxious to leave. "Looks like you got here safely."
He let his door swing open, showing a long narrow flight of stairs leading up to his studio-style flat. When he felt the other turn to leave, he found himself saying, "Ah... Kakashi-sensei?"
The man paused. "Hm?"
He was speaking to the stair case, not sure what expression was on his face, "If for some reason... Tsunade-Hokage did agree with those guys about... not letting the Kyubi fall into the hands of the enemy...," he was speaking quieter now; the taboo on the topic still pressing in on him despite his growing accustomed to it, "Would you, ah, y'know... If she ordered, um, that... would you, if you were ordered to, y'know, kill-"
A hand ruffled his hair, and he didn't have to look to know the Jounin was smiling, "Let's just hope it never comes down to that."
Sometime later he found himself in his apartment, going through and making sure all the windows were locked, setting up a few noise-traps to the latches to alert him if anyone was trying to break in over the night.
He didn't even bother feeling paranoid; a kid living alone had to take care of himself.
Changing into pajamas, he pauses to finger the pendant around his neck, wondering. Always wondering.
He stuffed a few extra pillows beneath the blanket of his bed to look like a human shape was beneath.
And then he went to his closet, where he already had a extra clothes and towels piled up inside to make it softer and, after nestling into them to get as comfortable as possible, he made sure his leg pouch was secured to his thigh and full of shiruken and kunai. And then he closed his closet door, and fell promptly asleep.
There was something utterly astounding and eternally unpredictable about the boy's thought process.
It was... the only explanation for it. The only plausible reason a Jounin and two Chuunin could fail to capture a rather pathetic, ill-trained, terrified and rather young Genin brat. He was mindlessly unobservant, which should have made him easy to subdue; somehow, it didn't work like that. His skeptical temperament had leveled out his obliviousness -- though he failed to notice a group of three men watching him from the table to his right, close enough they could have touched his elbow, grabbed his cheek, he had the mental conditioning to instantly spring off and away at the cue of a single blatantly intentional step in his direction.
And he wasn't just fast. He was little. Less wind resistance. Could fit through tighter alleys. To have him suddenly take off across the rooftops of the familiar environment of his home, it was akin to standing in a field and watching a single horsefly hover like a spot of reflected light in front of you, so close you had to go cross eyed to even fully register it, but nearly impossible to successfully swat. And the winding chases he could lead! His demon lent him endurance, and his fear lent him desperation, and his years of trickery lent him experience in running from repercussion.
Not too proud to drop to all fours in closed-in areas. Not beneath long series of transformations; first replicating shadows, and then each one taking on the form of a different person. That was something no one had warned them about; Bunshin no Jutsu they could handle. It was elementary; an intangible replication can't do much. But Kage Bunshin... each clone, each with a portion of mind to itself -- they did not have to be within sight of their creator. They were true, living creatures. And they could perform jutsu of their own, with enough Chakra.
More than once a clone was mistakenly pursued; transforming first into a woman, and then a man, and then a rabbit, and then a stone. And, out of breath, irate, frustrated, when finally it looked like the boy was tiring... poof.
It had taken... hours... of haggard pursuit. Heading for the fish shop, going in, -- damn, it was only another shadow replication! The pat-pat-pat as sandaled feet scurried over the rusted metal roofing; out the window, after him, from roof to roof, ricochet off a lamp post, bursting through a stranger's window, darting through their house, a foreign living room, a bathroom -- where? The window too small to fit a full-grown man's shoulders through. The sounds of pursuit as others take up the chase. They catch up, they send him skidding; he should have been down for hours, but instead he's using the momentum to run up the side of a fence, vaulting over a railing into a strip of second-story porches, bursting a flair bomb, temporarily blinding them.
Wanting to kill him, to wring his skinny little neck, to jerk his arm out of its socket, to slam a fist through his stomach to squeeze his warm little guts -- where?! Three men can't keep track of a -- one shouts, "There!" The flash of an orange coat, dammit, it wasn't him; withdrawing to the shadows, scanning the people as they pass, looking for a short blonde head...
Brain-jarring impact from behind; this was the boy's only feature of consolation: if he was running, he would only do it for so long. He was mad enough to think he could fight back.
It ensued, lacking the breath to call the others; the gleaming ringed hilt of a kunai, buried in chest; the organic crunch of tree bark where the boy's little body hits. Not supposed to kill him but having long since abandoned the kid gloves -- just get a good hold on one arm, just one firm grip and then it would be no struggle, swing him into a neighboring post -- like beating out a dusty rug!
If that could just be accomplished, if that could just be accomplished--
... that had been a few days ago, now. Somehow, the quarry had evaded escape long enough to reach the densely populated heart of the city, where they dared not go.
So now, the three of them remained huddled in the treetops a few miles outside the village limits. A few strong breezes offered tantalizing hints of what the natives were cooking in their nice gas stoves, the smell of well prepared meals, of seasoned meats, of anything, anything better than ninja rations. Odorless, tasteless, textureless; so bland and useless not even insects found it interesting, marauding predators that caroused their fireless camp at night disregarded it. Wild animals, squirrels, raccoons, they would steal shiny shiruken, they would steal loose buttons but nothing, nothing outside the logic of men in hiding would consider putting a ration in their mouth.
They couldn't even kill a fish. They had to avoid killing insects. Just in case. There could be no sign they were here; they slept on moist earth without fire, they could not bathe upstream, but the town used the passing river as a means to dispose of waste, so they couldn't bathe downstream either. As a result, they had the warm, heavy organic smell of unwashed hair and the sour, rather acrid odor of sweaty body.
The village, still recovering from a previous assault a few months earlier, was on edge. They couldn't risk being discovered, but they also couldn't risk returning without results. Damn the Hidden Sand and Sound! Damn Orochimaru for his inopportune time to begin tormenting the Leaf Shinobi -- like beating a hive of bees with a stick! He could decimate them if he wanted, any other time but now! Their security was strung so tight, their sentry so active, their watchmen so alert it would be difficult to kidnap any child from within the village gates.
"I... hate..."
The other two looked to him.
"... everything."
They looked back down at their laps again.
The wet sound of chewing and swallowing, squishy bricks of unspecific brown and gray junk. Full of nutrients and vitamins to keep their system up.
"Well." A different one speaking, "I'm going to be grateful."
More silence.
"Consider our options.We got the less-dangerous demon."
That, they could nod to. The three of them had been haunting the skirts of the village for quite a while, miserable and craving creature comforts but alive and, aside from Tomo, who was still healing from the brat's well-placed kunai, they were healthy. No, the Leaf's demon was by far the best charge they could have gotten. It was common knowledge that the Missing-nin after the Sand's demon weren't nearly so fortunate.
And the amount of money being offered for these rising demon brats... whatever the Akatsuki were planning to do with them had to be important.
"... I still hate... everything."
Pause. "
"... yeah." They nodded with him.
To be continued.


text: >new jag that boasted "the only > >vehicle in its class with permenant all wheel drive." Yeah, its class is leaky, overpriced de-contented British sh*tboxes which depreciate faster than Audis, and won't run anywhere near as long. Problem is just exactly how do you define the "class" of a car? It sounds like sports statistics - "highest number of touchdowns by a Republican Amish third baseman for games played between 6 and 10 PM on odd-numbered Thursdays" - possibly true, but utterly meaningless. It's also permanent all wheel drive because they haven't figured out how to switch it on and off yet. (Don't mind me this AM guys, I'm just mightily PO'ed about something else today . . . ) Best Regards, (And yes, even to our friends across the pond) Mike Arman

subset: commoncrawl


text: I'm sure many of you have the same issue, over the years we have allowed many a customer to just call the center to have a truck dispatched for a pick up. I can't even begin to imagine how much money has been lost in drivers wages and fuel, not to mention lost revenue from service charges. Fri in our PCM our sup told us that this practice is no longer allowed. Customers have to call the 800# to schedule a pick up. I have always been an advocate of this strategy and someone has finally seen the light. Now the problem is getting the customers past their, 'it is my right' mentality to call the center. I was done with my rte yesterday at 4:20. As I was pulling into the center I received an ODS saying a customer out on the edge of town had a pick up and would I stop by and pick it up. NOPE - I'm done and in the center.


text:
http://www.tastetv.com/theme2/wp-content/uploads/2010/05/4627693026_456953a40c.jpg 488 500 TasteTV http://www.tastetv.com/theme2/wp-content/uploads/2014/06/enfold-logo-test2.jpg TasteTV2010-05-25 22:21:002010-05-25 22:21:00Judges Invitations issued for 2011 TASTY Awards
Judges Invitations issued for 2011 TASTY Awards
Preparations for the 2011 TASTY Awards are underway, and one of the first orders of business is at hand: the presentation of invitations to the industry gurus and tastemakers who will be this year's judges.
Who will be a judge? How many? Where are they from, what will they say?
The answers are coming soon.
See the full televised show from 2010 on Hulu.com/tasty-awards


text:
This backpack, offering a generous volume, has lots of pockets, a large and secure flap and adjustable shoulder straps.
S@FARI-SUR-SEINE embraces a Lodge theme that teams comfort with luxury. Crafted from cotton-effect nylon canvas, it has a raw and authentic feel. To further accentuate its smart casual style, the models have leather patches on their shoulder straps and leather trimmings. The smooth, pure style of the leather and the aged copper finish on the rivets play up its adventurous spirit. Everything you need to brave the office jungle!
Reference : L1695689806


text:
Welcome to WebmasterWorld Guest from 126.96.36.199
Forum Moderators: open
I've seen numerous messages on the forum where people are talking about 'PR3', 'PR4' etc.
I assume this relates to some kind of rating as to how well ranked the site is on one or more search engines, but I could be wrong (not for the first time!).
Could someone please explain what these numbers mean and how I can tell which number(s) I should use to rate a site in the future. It's all a bit confusing to a new guy :)
and [toolbar.google.com ] ;)


text:
So the boys have a new toy. Something I would shudder to buy, but one that the hubby is super excited about. A bug vacuum. Yeap. Two boys. Two bug vacuums. With a sucking action, and two mini canisters to house the poor insects. (Excuse the mess, them insect-busters are too busy to pick up… Read more »
Posts Categorized: Tales from mommydom
Zoie at 6 months
Oh my. I cannot believe i let the day pass without a blog post, so this is making it up to my little princess. Who turned 6 months as at 30 October 2013. How my little baby has grown. She is sweet and sassy all at the same time (as should all girls, doncha think?)… Read more »
From a FTWM to a SAHM
A couple of months back, I hosted a blog linky entitled "FTWM: Motherhood Madness" where together with 29 other full time working blogger mummies, we shared our ups and downs of juggling both work and family. And now, as I've more or less eased into life on the opposite end of the spectrum – being… Read more »
Meet my kids – quirky, cheeky and erm… hungry
The loves of my life. It's been awesome, and awesome tiring at the same time, juggling my new role as a stay at home mom. No longer are my days spent in the office, but picking up Lego bricks and endless laundry cycles. I miss so much about working life, it really takes another post… Read more »
Zoie – at two months
Dear Zoie (or Zozo as your brothers call you), Happy second month, babe! Has it been two months already? I guess all that sleep deprivation has caused Mummy to lose track of the days, hours and minutes. If we were still in Singapore, this would mean the end of my maternity leave and the start… Read more »


text:
Ms. Bonnie Boothe and Mr. and Mrs. Pete Boothe, all of Danielsville, announce the engagement of their daughter, Tonya Boothe, to Jason Ramsey, son of Kathy Duncan of Hull.
Miss Boothe is the granddaughter of Mrs. Sue Gaddis of Danielsville and the late Mr. John Gaddis, and Mrs. Susie Boothe of Danielsville and the late Mr. Leroy Boothe.
Mr. Ramsey is the grandson Mr. and Mrs. Ede Harike of Athens and the late Mr. Hoyt Haynes.
The future bride is a 1998 graduate of Madison County High School. She is currently employed in the operations department of the National Bank of Georgia.
The future groom is a 1996 graduate of Madison County High School. He is currently employed as a driver for University Moving and Storage.
The wedding is planned for 5:30 p.m. June 29, 2002, in Gatlinburg, Tenn.
Athens Banner-Herald, Sunday, June 2, 2002
Published in the Athens Banner-Herald on Sunday, June 2, 2002.
© 2018. All Rights Reserved. | Contact Us


text:
So me and my gf go to different colleges. Last night she went out with her roommates nothing out of the norm. We were talking earlier today and she sent me this message saying that she
"had a rugby player who seemed very interested in becoming the bae last night. He was like following me around and asking me if I was okay and making sure I was comfortable hahaha like he offered me a sweatshirt at one point and Kristin and Megan were like "ookaaayy time to go" haha I was trying to be polite but when he was like "I'm sorry, but you have really bright lipstick" I almost told him to stay away from my lips haha
When she was telling me this I was glad but also a little angry at the same time. I was glad that she was telling me this because she felt conferrable and that I wouldn't freak out. I don't really know why I felt angry, I kinda guess because I wasn't with her at the time. Is this a normal feeling to have?
Is this a normal feeling to have?
So me and my gf go to different colleges. Last night she went out with her roommates nothing out of the norm. We were talking earlier today and she sent me this message saying that she
Have an opinion?
What Girls Said 2
I think most men have a protective instinct concerning their women, so it's natural to feel a little 'overwhelmed' to know someone was trying to enter your territory when you weren't around. But you shouldn't let things like those stay in your mind because as you said, the fact that she is comfortable enough to tell you and joke about it to boot is a good thing. You want to keep that momentum going so she doesn't feel like she can't talk to you or you would freak out and get mad at her, when she is clearly only for you. Good luck!00|00|0
Ya it really is I mean its uncomfortable to hear a story about someone who was kinda hitting on your girlfriend while you weren't there... just be glad she loves you enough to tell you and that she's clearly not interested! :p00|00|0
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text:
BLUE Earth's Essentials™ Harvest Table™ in Gravy
Bring the best the earth has to offer to your canine companion's meals. Made with real farm-raised chicken and nutrient-packed quinoa, these delectable tubs offer your best friend a true farm-to-table dining experience with simple, honest and delicious ingredients.
And like all BLUE foods, these delicious recipes are made with the finest natural ingredients and enhanced with the vitamins and minerals to help your dog thrive.Available in 8-oz. tubs.
Guaranteed Analysis
|Crude Protein
|8.5% min
|Crude Fat
|3.5% min
|Crude Fiber
|1.5% max
|82.0% max
Nutrition Statement
BLUE Earth's Essentials Harvest Table in Gravy is formulated to meet the nutritional levels established by the AAFCO Dog Food Nutrient Profiles for maintenance.
Feeding Guidelines
Feed 1 to 1¾ containers per 10 lbs. of body weight per day.
Refrigerate unused portion.
Fresh Water
Be sure to have a clean bowl of cool, fresh water available at all times.
Note: Individual dog's requirements may vary due to age, breed, environment, and activity level. Adjust food as required to maintain optimal body condition, and ask your veterinarian if you are unsure.
Calorie Content:
919 Kcals/kg, 210 Kcals/tub


text:
HR Solution is a recruitment and consultancy company, specialized in the IT sector. We are a part of an international group of companies with more than 13 years of experience, recognized as a reliable partner by both clients and candidates.
On behalf of one of our clients, exciting cloud based Software Company, we are looking for a talented Web developer to join their team in Belgrade.
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Confidentiality of all applications is assured! Only short-listed candidates will be contacted! The candidates do not pay any taxes to the agency – the services are FREE OF CHARGE!


text:
Hoping to find somewhere in the CNP or on the outskirts where I can do some winter hind stalking initially.
Looking for somewhere I can visit perhaps 6 times per year, to supplement existing woodland Roe stalking.
Not so interested in trophies, just want good quality meat to utilise at home.
Will pay a reasonable price.
Sent from my LG-D855 using Tapatalk


text:
Elaine Weinstein is the wife of Warren Weinstein. Weinstein was an American contractor in Pakistan working for J.E. Austin Associates, a firm that oversees new business projects in developing economies. In 2011, Warren Weinstein was kidnapped by al-Qaeda members and held hostage for seven months before he had the chance to speak with his wife by phone. She sent the kidnappers $243,000 but Warren was not released. He was handed over to another terrorist group in Pakistan.
On January 14, 2015, Weinstein was accidentally killed by a US drone strike along with al -Qaeda leader, Ahmed Farouq. Elaine Weinstein, who remains angry at US government efforts to secure her husband's release, is releasing never before seen or heard chat messages and phone calls she had with the terrorists while negotiating for the release of her husband. She talks with 60 Minutes which airs Sundays at 7pm on CBS.


text:
Benefits of the Rainbow® Head Stabiliser:
- It secures to the board with screws for reliable stability.
- Adjustable side panels accommodate a wide range of patients and tilt in at a slight angle for a snug fit.
- The thick cushions on the sides allow for a bit of pressure, while maintaining patient comfort.


text: This AMR papercraft is designed by Juke. The Rabbit Doll (or Rabbit Toy) is a raggy stuffed rabbit toy made out of cloth, and wears a loose waistcoat, which makes it slightly resemble the White Rabbit. During the years, it has needed a few repairs and is missing a eye. It is Alice Liddell's dearest possession, and even as an adult she still wishes to keep it. It is one of, if not the only object from the Liddell household that survived the fire. Although it appears in both American McGee's Alice and Alice: Madness Returns, the doll's current whereabouts is unknown. You can download this alice paper toy from here: Alice: Madness Returns - Rabbit Doll Free Paper Toy Download


text:
The mammalian SIN3 complex consists of histone deacetylases (HDAC1, HDAC2), several known proteins (SAP30, N‐CoR) and as yet unidentified proteins. Here we show that the mouse tetradecanoyl phorbol acetate induced sequence 7 (TIS7) protein is a novel transcriptional co‐repressor that can associate with the SIN3 complex. We have identified tis7 as a gene that is up‐regulated upon loss of polarity in a mouse mammary gland epithelial cell line expressing an estrogen‐inducible c‐JunER fusion protein. In unpolarized cells, TIS7 protein levels increase and TIS7 translocates into the nucleus. Overexpression of tis7 causes loss of polarity and represses a set of genes, as revealed by cDNA microarray analysis. We have shown that TIS7 protein interacts with several proteins of the SIN3 complex (mSin3B, HDAC1, N‐CoR and SAP30) by yeast two‐hybrid screening and co‐immunoprecipitations. TIS7 co‐immunoprecipitated HDAC complex is enzymatically active and represses a GAL4‐dependent reporter transcription. The transcriptional repression of endogenous genes by tis7 overexpression is HDAC dependent. Thus, we propose TIS7 as a transcriptional co‐repressor affecting the expression of specific genes in a HDAC activity‐dependent manner during cell fate decisions, e.g. scattering.
The complex regulation of gene expression at the transcriptional level is controlled by the combinatorial action of sequence‐specific DNA‐binding proteins, transcriptional co‐regulators and components of the basal transcriptional machinery (Chi et al., 1995; Goodrich et al., 1996; Mannervik et al., 1999). Acetylation of internal lysine residues of core histone N‐terminal domains by histone acetyltransferases (HATs) and deacetylation by histone deacetylases (HDACs) are associated with transcriptional regulation (Hebbes et al., 1988; Braunstein et al., 1993). Several HATs and HDACs have been shown to also acetylate and deacetylate non‐histone protein substrates (e.g. transcription factors), implicating internal lysine acetylation as a possible mechanism of rapid and reversible regulation (Kouzarides, 2000; Humphrey et al., 2001). The two major class I‐containing histone deacetylation complexes are NuRD (nucleosomal remodelling and histone deacetylation) and SIN3 (reviewed in Knoepfler and Eisenman, 1999; Ahringer, 2000). The SIN3 complex shares four core proteins with NuRD (HDAC1, HDAC2, RbAp46 and RbAp48) and additionally contains Sin3, SAP18 and SAP30 (Ayer, 1999). RbAp46 and RbAp48 proteins interact with histones and histone deacetylases HDAC1 and HDAC2. The SIN3 complex does not bind to DNA directly, but through sequence‐specific DNA‐binding proteins: passive repressors, e.g. nuclear hormone receptors (Nagy et al., 1997). Sin3 proteins, mSin3A and mSin3B, act as adaptor molecules linking repressive transcription factors to accessory proteins. This facilitates targeting of the HDAC complexes (Hassig et al., 1997; Laherty et al., 1997; Knoepfler and Eisenman, 1999; Kyrylenko et al., 2000). However, SIN3‐mediated repression is not exclusively HDAC dependent, suggesting that the SIN3 complex either contains an intrinsic repressor activity or it associates with other co‐repressors (Laherty et al., 1997; Wong and Privalsky, 1998).
To study transcriptional regulation during cell fate decisions, we have used the previously established c‐JunER mammary epithelial system (Fialka et al., 1996). Activation of the estrogen‐responsive c‐JunER fusion protein results in reduced transepithelial resistance, disruption of intercellular junctions, their barrier function and in the formation of irregular multilayers. The reversible loss of epithelial polarity resembles a scattering phenotype with redistributed apical and basolateral proteins to the entire plasma membrane. Activation of c‐JunER mimics the c‐Jun activation in vivo and transcriptionally regulates a variety of AP‐1 target genes (Fialka et al., 1996).
In this study we identified, by RT–PCR‐based differential display (DD RT–PCR), the TPA induced sequence 7 (tis7) gene (Varnum et al., 1989a) to be up‐regulated upon the loss of epithelial polarity in c‐JunER cells. TIS7, also known as PC4 (Tirone and Shooter, 1989) or IFRD1 (interferon related developmental regulator 1) (Buanne et al., 1998), was described previously as an immediate early gene (Tirone and Shooter, 1989; Varnum et al., 1989b; Guardavaccaro et al., 1994, 1995) that is possibly involved in the differentiation of various cell types (Tirone and Shooter, 1989; Guardavaccaro et al., 1994, 1995). However, the molecular function of TIS7 has not yet been elucidated.
Here we present evidence that TIS7, up‐regulated after c‐Jun activation, translocates into the nucleus and acts as a transcriptional co‐repressor. We identified by cDNA microarray analysis several genes specifically repressed by overexpressed TIS7. TIS7 can associate with the mammalian SIN3 complex and HDACs, and thereby affect the expression of specific genes.
TIS7 is up‐regulated and translocates into the nucleus of c‐JunER cells losing epithelial polarity
We were interested in the identification of genes whose expression changed as a result of activation of c‐JunER. In the present study we performed DD RT–PCR with RNA samples isolated from untreated and 48 h β‐estradiol (E2)‐induced c‐JunER cells. One of the several differentially expressed genes was tis7. A full‐length TIS7 cDNA was subsequently cloned, which was identical to the one described in Varnum et al. (1989a). This nucleotide sequence encodes a protein of 449 amino acids that migrates on SDS–PAGE at ∼53 kDa. After 48 h of E2 treatment, the tis7 gene was up‐regulated 5‐ to 10‐fold (Figure 1A). Upon removal of E2, as the cells regain their epithelial polarity, the tis7 mRNA expression returned back to basal levels. Since estrogen hormones are potent mitogens for mammary epithelial cells, EpH4 parental cells of the c‐JunER cell line were treated with an identical concentration of E2 as a control. The mRNA levels of the tis7 gene were not affected (Figure 1A, right panel).
We generated a rabbit polyclonal antiserum against a peptide comprising 18 N‐terminal amino acid residues of TIS7. The TIS7 antiserum recognized the GST–TIS7 protein with the predicted size of 80 kDa on western blots (Figure 1B, middle panel). It did not cross‐react with GST protein loaded in comparable amounts, as documented by the α‐GST antibody blotting (Figure 1B, left panel). Next, we tested the affinity‐purified α‐TIS7 antibody in immunoprecipitations of lysates from HeLa cells transfected with the His6Xpr‐TIS7 expression construct. The α‐TIS7 antibody specifically immunoprecipitated the 63 kDa His6Xpr‐TIS7 band recognized by the α‐Xpress monoclonal antibody. The specificity of the immunoprecipitation was confirmed by its peptide inhibition (Figure 1B, right panel).
In accordance with the DD RT–PCR results, TIS7 protein levels in total cell lysates were also up‐regulated upon c‐JunER activation (up to 3‐fold) (Figure 1C, graph). TIS7 protein levels were induced 4 h following the onset of the E2 treatment and remained up‐regulated for >48 h (Figure 1C). Effects of c‐JunER activation on the subcellular distribution of TIS7 were analysed by confocal laser scanning immunofluorescence microscopy. In fully polarized cells [4 days in culture; epithelial polarity documented by the ring‐like staining of the tight junction protein ZO‐1 and high transepithelial electric resistance (TER) values], the majority of TIS7 localized in the vicinity of the lateral plasma membrane (Figure 1D, −E2). Upon c‐JunER activation, cells lost epithelial polarity (as documented by the disrupted ZO‐1 staining and low TER values), the lateral staining of TIS7 was reduced and the TIS7 protein accumulated in the nuclei (Figure 1D, +E2), similar to sparsely seeded undifferentiated cultures (data not shown).
In summary, the loss of polarity caused by the activation of the c‐JunER fusion protein in mouse mammary epithelial cells was accompanied by an increase in both tis7 mRNA and protein levels, and by translocation of the TIS7 protein into the nucleus.
TIS7 overexpression changes the polarity of epithelial cells
Next we examined whether overexpression of TIS7 protein itself would affect the polarity of the mammary epithelial cells. We constructed a recombinant TIS7 chicken adenovirus (CELO) and expressed TIS7 by infection of fully polarized monolayers of c‐JunER cells. Virus expressing eGFP was used as a control. Immunofluorescence analysis revealed that the majority of TIS7‐overexpressing cells lost the polarized epithelial phenotype, as assessed by the distribution of junctional proteins β‐catenin and ZO‐1, respectively (Figure 2A and B). These cells were higher, rounded up and protruded from the monolayer (Figure 2A and B, x–z section; note the difference in the thickness of the x–z section, reflecting the number of cell layers). The cumulative effect of prolonged TIS7 overexpression on the epithelial cell polarity also reflected the fall in the transepithelial resistance, as shown in the graph (Figure 2C). Although eGFP was overexpressed in similar amounts and also accumulated in the nuclei of infected cells, it did not affect epithelial cell polarity. We conclude that overexpression of TIS7 may have interfered with the polarized epithelial phenotype since contact with substratum and adhesion to neighboring cells were impaired. The nuclear localization of TIS7 in unpolarized cells suggested a role in the regulation of gene expression.
Analysis of TIS7 effects on gene transcription using cDNA microarray analysis
In order to find out whether nuclear TIS7 had the capacity to affect transcription, we performed a cDNA microarray analysis. The experimental set‐up of this analysis is explained in detail in the Supplementary data (available at The EMBO Journal Online). The expression data were ranked using a web‐based software tool developed in our laboratory to query a relational database (MicroarrayViewer; see Supplementary data). Ninety‐three percent of genes were specifically down‐regulated by TIS7 overexpression, while only two genes were up‐regulated. The expression differences corresponded to an average expression ratio of 2.2. Transcriptional effects of TIS7 were independently, quantitatively, reconfirmed for several of the candidate genes using Real‐Time PCR (Light Cycler™) (Table I). Western blot analysis also showed that protein levels of two candidate genes (CRABP II and OPN) were down‐regulated in cells overexpressing TIS7 (Supplementary data).
In conclusion, the cDNA microarray experiment revealed that TIS7 overexpression preferentially caused down‐regulation of a specific set of genes and this finding, together with TIS7 nuclear localization, suggest a potential role for TIS7 as a transcriptional co‐repressor.
TIS7 interacts with the SIN3 HDAC complex
In order to identify the mechanism by which TIS7 affects transcription, we decided to identify proteins interacting with the mouse TIS7 protein. In a yeast two‐hybrid screen, we used a mouse embryo cDNA library and the full‐length cDNA (amino acids 1–449), as well as N‐terminal (amino acids 1–227), C‐terminal (amino acids 262–449) and central domains (amino acids 240–280) of TIS7 as baits. We identified mSin3B, a member of the SIN3 HDAC complex (four positive clones), as a protein potentially interacting with the N‐terminal domain of the TIS7 protein.
Having established the interaction in yeast two‐hybrid screens, we tested whether the TIS7 interaction with mSin3B could also be demonstrated in vivo. First, HeLa cells were transfected with a FLAG‐tagged mSin3B long form (FLAG‐mSin3B LF) (Naruse et al., 1999) and immunoprecipitated using α‐FLAG antibodies. Endogenous TIS7 protein co‐immunoprecipitated with FLAG‐mSin3B (Figure 3A).
Next, we mapped the mSin3B interaction domains on the TIS7 protein. FLAG‐tagged mSin3B LF was transfected into HeLa cells either alone or together with a series of Myc9‐tagged TIS7 deletion constructs and immunoprecipitations were performed using α‐Myc antibodies. Two TIS7 constructs encompassing the N‐terminal amino acids 1–321 and 1–367 co‐immunoprecipitated mSin3B. However, a partially overlapping construct expressing the protein encoded by amino acid residues 140–449, lacking the N‐terminal domain of the TIS7 protein, was not capable of binding mSin3B (Figure 3B). The results of this domain mapping experiment confirmed our initial finding that TIS7 interacted in the yeast two‐hybrid screen with the mSin3B protein through its N‐terminal domain. Thus, we concluded that the region necessary and sufficient for the interaction with mSin3B is the TIS7 sequence between the amino acids 1 and 140.
Some of the specific domains of mSin3B important for its interaction with interacting partners are already well characterized, e.g. interaction of the paired amphipathic helix (PAH)2 domain with the transcription factor MAD (Ayer et al., 1996; Harper et al., 1996; Laherty et al., 1998), or of PAH1 and PAH3 domains with N‐CoR (Alland et al., 1997; Heinzel et al., 1997). Therefore, we examined the in vivo interactions between the series of mSin3B deletion constructs and full‐length TIS7. Among the series of mSin3B deletion constructs examined (Figure 3C), mSin3B LF as well as constructs containing the PAH2 domain and the intervening region between the PAH1 and PAH2 domains represented significant binding to TIS7. The PAH1 domain alone also bound TIS7, though less efficiently. In contrast, constructs containing the C‐terminal half of mSin3B only, encompassing PAH3 through PAH4 domains (mSin3B P3‐4), showed no interaction with TIS7 (Figure 3C).
It was shown previously that mSin3 proteins (A and B), as part of the SIN3 complex, recruit HDACs to specific transcription factors (e.g. Mad or NRSF) and thereby repress transcription (Ayer et al., 1995, 1996; Korhonen et al., 1998; Naruse et al., 1999). To test whether TIS7 is associated with HDAC1 in vivo, TIS7 immunoprecipitates were assayed for HDAC1 by western blot analysis. TIS7 antibody specifically co‐immunoprecipitated HDAC1 and formation of this precipitate was blocked with the cognate immunogen. Endogenous HDAC1 was co‐immunoprecipitated in a dose‐dependent manner with increasing amounts of the α‐TIS7 antibody (Figure 4A). In the reciprocal experiment, endogenous TIS7 was co‐immunoprecipitated with the monoclonal antibody against the endogenous HDAC1 (Figure 4B). To test whether TIS7 has the capacity to associate with other HDACs besides HDAC1, we performed in vitro pull‐down assays using GST–TIS7 fusion protein. Radioactively labelled HDAC1, HDAC2 and HDAC3 specifically interacted with the GST–TIS7 protein (Figure 4C). Similarly, GST–TIS7 interacted with N‐CoR nuclear receptor co‐repressor, a component of HDAC complexes. These interactions were specific since, first, products of in vitro coupled transcription/translation reaction did not interact with GST control protein alone and, secondly, GST–TIS7 did not interact with an unrelated nuclear protein, human cohesin subunit SCC1, synthesized in an identical way (Figure 4C).
In the next experiment, we tested whether TIS7 may associate with some of the currently known subunits of the SIN3 complex in mouse epithelial c‐JunER cells. We analysed c‐JunER nuclear extract fractions after separation by continuous sucrose gradients. In E2‐untreated cells, part of the TIS7 pool co‐sedimented with mSin3B, HDAC1 and SAP30 during sucrose gradient centrifugation (Figure 5A, fraction 9). Another part of the TIS7 pool in these normally polarized cells appeared in lighter fractions from the gradient centrifugation representing monomers. However, following 4 days treatment of c‐JunER cells with E2, there was a significant increase in the amount of TIS7 that co‐sedimented with mSin3B and HDAC1 during sucrose gradient centrifugation (fraction 10). At the same time, TIS7 sedimenting in the lightest fractions of the sucrose gradient (fractions 2 and 3) disappeared. We have confirmed that TIS7 associates specifically with the SIN3 complex in an independent sucrose gradient separation of a nuclear extract by blotting with antibodies against MTA2, a component of a different HDAC complex. MTA2 fractionated in different fractions than TIS7 (data not shown).
Performing the immunoprecipitation with the affinity‐purified α‐TIS7 antibody, we found that besides mSin3B, other endogenous protein components of the SIN3 complex, e.g. HDAC1 and SAP30, also co‐immunoprecipitated specifically with the endogenous TIS7 (Figure 5B). These interactions were detectable both in polarized (−E2) as well as unpolarized (+E2) c‐JunER cells. Although there was also a minor increase in the amount of immunoprecipitated TIS7 in the +E2 sample, the amounts of co‐immunoprecipitated SIN3 complex proteins did not increase. These immunoprecipitation results confirmed our initial two‐hybrid screen findings, namely the interaction of TIS7 with the SIN3 complex subunits in vivo.
The TIS7 HDAC complex is enzymatically active
Our data demonstrated that TIS7 can interact with mSin3B and HDAC1, as well as other members of the HDAC complex. To establish that TIS7 associates with a functional HDAC, TIS7 immunoprecipitates from HeLa and c‐JunER cells were assayed for HDAC activity. Endogenous TIS7 immunocomplexes contained substantial HDAC activity when compared with control precipitates using an antibody directed against an unrelated epitope (Myc) (Figure 6A). The specificity of the immunoprecipitation was confirmed by the absence of HDAC activity from the immunoprecipitate where the antibody was blocked by the cognate immunogen peptide. Importantly, HDAC activity also co‐immunoprecipitated with TIS7 in the c‐JunER cell line. Since immunocomplexes with HDAC activity could also contain enzyme isoforms other than HDAC1, we decided to compare TIS7 immunoprecipitates that were derived from wild‐type and HDAC1−/− mouse embryonic stem cells. The total HDAC activity in the HDAC1−/− cells was only partially, but not completely, decreased because of the up‐regulated activity of the remaining HDAC isoforms (Lagger et al., 2002). The HDAC activity co‐immunoprecipitated with TIS7 in the HDAC1−/− sample was significantly decreased (Figure 6B). On the other hand, the HDAC activity was not totally abolished, showing a possible interaction between TIS7 and other HDAC proteins, as suggested by our GST pull‐down assays (Figure 4B). Subsequent western blotting confirmed that TIS7 interacted with HDAC2, which is more abundant in HDAC1−/− cells (Figure 6B). These results demonstrate that TIS7 associates with a functional HDAC complex in vivo.
TIS7 represses transcriptional activity in an HDAC‐dependent manner
It was shown previously that targeting HDACs to the vicinity of a promoter resulted in transcriptional repression (Yang et al., 1996; Alland et al., 1997; Hassig et al., 1997; Zhang et al., 1998). Therefore, we tested whether directing TIS7 to a particular promoter would inhibit transcription through recruitment of HDAC activity. HeLa cells were transfected with a construct containing a minimal thymidine kinase–luciferase template regulated by five consensus Gal4‐binding sites in the promoter‐proximal region (Alkema et al., 1997). Co‐transfection of the Gal4‐dependent reporter with a mammalian expression vector encoding the Gal4 DNA‐binding domain fused to full‐length TIS7 sequence resulted in up to 75% repression of the transcriptional activity. Furthermore, the mechanism of this transcriptional repression was HDAC dependent since repression was partially relieved by HDAC inhibitor [trichostatin A (TSA)] treatment of TIS7 co‐transfected cells (Figure 6C). Several Gal4‐fused TIS7 mutants (expressing the same parts of the TIS7 protein as used for the interaction study in Figure 4A) were generated and their repression activities were analysed in order to determine minimal regions required for the repression activity of TIS7. Both the N‐terminal TIS7 constructs 1–321 and 1–367 repressed transcription (Figure 6C), although less strongly than the C‐terminal construct 140–449 or wild‐type full‐length TIS7. Interestingly, transcriptional repression by the N‐terminal constructs of TIS7 was almost completely de‐repressed by the TSA treatment. In contrast, transcriptional repression by the C‐terminal construct (140–449) was not significantly affected by even high TSA concentrations. These results suggest that histone deacetylation is involved in the transcriptional silencing via the N‐terminal domain of TIS7. On the other hand, the C‐terminus of TIS7 exhibits repressive activity independent of histone deacetylation (Figure 6C).
Finally, we asked whether the transcriptional regulation of genes regulated by TIS7 overexpression, identified by the cDNA microarray analysis, was HDAC dependent. c‐JunER cells were infected with CELO‐eGFP and CELO‐TIS7, respectively. TSA treatment substantially reverted the TIS7‐mediated repression of one tested gene: crabp II (Figure 7). Importantly, TSA treatment did not have an effect on the crabp II RNA levels in the control, eGFP‐overexpressing cells or on the expression of the house‐keeping gene gapdh. These data suggest that TIS7 specifically down‐regulated the expression of crabp II via a HDAC activity‐dependent mechanism.
According to studies in several different biological systems, TIS7 has been postulated to be involved in the regulation of various cell differentiation processes (Tirone and Shooter, 1989; Guardavaccaro et al., 1995; Rubin et al., 1998). Our data provide a molecular mechanism by which TIS7 can regulate gene expression and might be involved in differentiation processes. Here we show that, under specific conditions, TIS7 associates with the SIN3 repressor complex possessing HDAC activity. This interaction suggests a possibility that TIS7 may repress transcription of specific genes via a HDAC activity‐dependent mechanism. The Supplementary data for this study provide a list of genes specifically regulated following the overexpression of TIS7, accompanied by online links leading to their characteristics (http://www.imp.univie.ac.at/lh/chip_10_5_01/index.html).
TIS7 shuttles between cytoplasm and nucleus
It was previously shown that TIS7 translocates between the cytoplasm and nucleus (Kujubu et al., 1987; Tirone and Shooter, 1989; Guardavaccaro et al., 1994). Similarly, in sparsely seeded and unpolarized c‐JunER cells, TIS7 localized preferentially in the nucleus. As the density of the cultures increased, TIS7 redistributed to the cytoplasm and in fully polarized and contact‐inhibited cells localized mainly in the vicinity of the plasma membrane (data not shown). Upon the loss of epithelial polarity, the TIS7 protein translocated from the plasma membrane into the nucleus. Our data suggest that the nuclear localization of TIS7 is essential for transcriptional regulation.
TIS7 is a transcriptional co‐repressor
We have demonstrated that the ability of TIS7 to attenuate transcriptional activity is associated with HDAC activity. Our co‐immunoprecipitation data suggested that TIS7 can act as a co‐repressor that can associate with the SIN3 complex. According to sequence analysis, TIS7 does not contain a DNA‐binding domain and, therefore, we do not expect TIS7 to directly bind to DNA, thereby tethering the HDAC activity‐containing complex to a particular promoter on its own. However, via its interactions with other components of a multi‐molecular complex, TIS7 may interact with DNA‐binding proteins and thereby co‐repress expression of genes. Since our GST pull‐down assays were performed using rabbit reticulocyte lysates without purification of the in vitro transcribed/translated, labelled proteins, one can not rule out the possibility that interactions between GST–TIS7 and HDACs are indirect, through additional proteins in the complex. Our data show that TIS7 has the capacity to interact with several members of the HDAC class I. The experiment showing that TIS7 co‐immunoprecipitated HDAC activity in cells originating from HDAC1−/− ES cells opened the possibility that TIS7 interacts with several different HDAC activities, which was confirmed by the co‐immunoprecipitation of HDAC2.
By mapping experiments, we identified two distinct domains at the N‐ and C‐terminal ends of the TIS7 molecule exhibiting repressive effects in the Gal4–TK‐luciferase reporter assay. Importantly, the only TIS7 domain necessary for the direct interaction with mSin3B resided at its N‐terminus. We also showed that the transcriptional repression caused by the overexpression of the N‐terminal TIS7 constructs could be relieved by TSA treatment. However, the repression of the C‐terminal construct was not TSA sensitive, suggesting that this part of the TIS7 molecule participates in the HDAC‐independent transcriptional repression mechanism. This result is analogous to the effects of the neural restrictive silencer factor (NRSF) where the N‐terminal repression domain, but not the second, C‐terminal domain, is also TSA sensitive (Naruse et al., 1999). We show here that TIS7 interacts with the intervening region between PAH domains 1 and 2 of mSin3B. This binding site was also similar to a domain encompassing the region of PAH1 and PAH2 domains of NRSF, which was shown to have the strongest interaction with the mSin3B SF (Naruse et al., 1999).
Role of TIS7 in HDAC activities in vivo
In summary, the studies presented here demonstrate that TIS7 represents a novel transcriptional co‐repressor interacting with the functional HDAC complex in vivo and in vitro. We found expression of a TIS7‐regulated gene to be de‐repressed by TSA treatment. However, because of a very general HDAC inhibitory effect of TSA, one can not test whether HDAC inhibition would revert loss of epithelial polarity caused by TIS7 overexpression. We propose a model for TIS7 function in which an increase in the amount of nuclear TIS7 protein (e.g. as a result of transfection or loss of epithelial polarity) is capable of transcriptional repression of particular genes in a HDAC‐dependent manner. Our future interests remain in confirming the role of TIS7 as a specific transcriptional co‐repressor in vivo at the level of the whole organism.
Materials and methods
Cell culture
EpH4 and c‐JunER cells were cultured on permeable filter culture inserts, as described in Fialka et al. (1996). HeLa cells were grown in DMEM supplemented with 5% FCS (Gibco BRL), 10 mM HEPES pH 7.3, 1% penicillin/streptomycin (Gibco BRL) and 1% l‐glutamine.
Full‐length mTIS7 cDNA was amplified by PCR and cloned into pcDNA3.1/HisC, pcDNA3.1(−)/MycHis (Invitrogen), pcDNA3.1(−) with a C‐terminal Myc9 tag and in a pGEX2 GST vector (Pharmacia). For the Gal4 transcriptional activity assays, mTIS7 fragments were PCR amplified and cloned into the pPuroGal4‐DBD vector (Alkema et al., 1997). The pLGCLuc‐E351 luciferase reporter construct was described previously in Braselmann et al. (1992); pCIneoHDAC1‐Myc plasmids in Bartl et al. (1997) and Doetzlhofer et al. (1999); and mSin3B constructs in Naruse et al. (1999). The hSCC1 product of in vitro coupled transcription/translation reaction was previously characterized in Waizenegger et al. (2000).
Northern blot analysis
Northern blots were performed as described previously (Vietor et al., 2001). Hybridizations were normalized to gapdh expression on the same blot. Radioactivity of specific bands was measured and evaluated by the ImageQuant software (Molecular Dynamics).
Differential display
Total RNA samples isolated from the controls and E2‐treated c‐JunER cells were analysed with the Differential Display Kit (Display Systems) according to the manufacturer's recommendations. Analyses were repeated five times and sequences analysed by the Blast search program at the NCBI home page. Differential regulation of identified genes was confirmed by northern blot analysis.
Yeast two‐hybrid analysis
The two‐hybrid screen was performed as described in the Clontech MATCHMAKER LexA manual, using full‐length, N‐terminal (amino acid 1–227), C‐terminal (amino acid 262–449) and central domain (amino acid 240–280) constructs of TIS7 obtained by PCR. The LexA DBD‐containing pBTM116‐TIS7 vector was co‐transformed with a mouse embryo cDNA library fused to the activation domain of Gal4 of the pACT2 vector into a yeast strain L40. Positive clones were re‐analysed in a β‐galactosidase (β‐gal) overlay assay and prey clones were directly PCR amplified and sequenced.
Real‐Time PCR
The primer sequences and protocol of these analyses are available in the Supplementary data.
Construction of recombinant CELO adenovirus vector
A recombinant CELO virus expressing the TIS7 gene was prepared as a derivative of CELO AIM46, previously described in Michou et al. (1999). Full‐length TIS7 cDNA, tagged at the 3′ end with Myc epitope and His6 tag sequences, was cloned into the plasmid pPM7 flanked upstream by the CMV IE promoter/enhancer and downstream by a rabbit β‐globin intron/polyadenylation sequence. The CMV/TIS7/β‐globin cassette was excised and recombined into pAIM46, thus replacing the luciferase cDNA. Construction of a recombinant CELO‐eGFP virus (CELO AIM53) was described previously (Michou et al., 1999).
The rabbit antiserum to TIS7 was raised against a peptide comprising 18 N‐terminal amino acid residues (TIS7; DDBJ/EMBL/GenBank accession No. X17400; Swissprot P19182; AA 30–47; GGPHRTVQPFSDEHA SIC) and affinity purified. Blocking of the antibody with the peptide used for immunization was performed by 60 min incubation at room temperature with 1 μg of peptide/ml of antibody solution. Rabbit polyclonal affinity‐purified α‐Myc antibodies were from Gramsch Laboratories (Schwabhausen, Germany); mSin3B (E‐20), HDAC1 (C‐19) from Santa Cruz and HDAC2 from Upstate Biotechnology; α‐GST from Chemicon; α‐Flag antibodies (M1) (Eastman Kodak) from Sigma and Alexa 488 goat α‐mouse IgG‐conjugated secondary antibodies from Molecular Probes. Mouse monoclonal antibodies against Xpress epitope were from Invitrogen, ZO‐1 and actin from Chemicon, β‐catenin from Transduction Laboratories and GFP from Roche Molecular Biochemicals. Mouse monoclonal α‐HDAC1 antibodies were prepared in the laboratory of C.Seiser. Rabbit polyclonal antibodies against mouse osteopontin were a generous gift from Toshimitsu Uede, Hokkaido University, Sapporo; rabbit anti‐cellular retinoic acid binding protein II antibodies were from P.Chambon, CNRS Strasbourg. H68.4 mouse monoclonal antibodies against α‐transferrin receptor were kindly provided by I.S.Trowbridge.
Immunoprecipitation and immunoblotting
Cells were lysed on ice in buffer containing 50 mM Tris pH 7.5, 150 mM NaCl, 1% Triton X‐100, 10% glycerol, 0.5 mM EDTA, 0.5 mM EGTA, 50 mM NaF, 5 mM NaPi, 0.2 mM Na3VO4 and a cocktail of protease inhibitors. Lysates were sonicated and centrifuged in order to separate insoluble proteins. Equal amounts of soluble proteins were immunoprecipitated with 5 μg of the respective antibodies followed by 20 μl of (1:1) protein A− or protein G–agarose slurry (Ultralink; Pierce). Following four washes with the lysis buffer, immunoprecipitated proteins were eluted with the sample buffer and boiling. Western blots were performed as described previously (Vietor et al., 2001).
Cell extracts and fractionation
For sedimentation analysis, HeLa cells were harvested in exponential phase. The nuclear extract was prepared according to Dignam et al. (1983), snap‐frozen in liquid nitrogen and stored at −80°C. The final buffer composition was 20 mM HEPES, 100 mM KCl, 0.2 mM EDTA, 0.5 mM DTT, 0.2 mM PMSF and 20% glycerol. Nuclear extract of HeLa cells containing 0.5 mg of protein was centrifuged for 18 h at 30 000 r.p.m. and 4°C through a 10–35% continuous sucrose gradient (in a buffer containing 150 mM NaCl, 20 mM Tris pH 7.4, 0.2% Triton X‐100 and 0.5 mM DTT) using a SW40 rotor (Beckman), as described previously (Waizenegger et al., 2000).
Preparation of samples and laser confocal scanning immunofluorescence analysis were performed as described previously (Vietor et al., 2001).
Transient transfections and luciferase assay
Transient transfections were performed using the Lipofectamine Plus reagent (Life Technologies). In luciferase assays, pCMV‐β‐Gal plasmid was used to normalize for transfection efficiency. The total amount of transfected DNA was equalized by addition of empty vector DNA.
HDAC activity assay
HDAC activity assays were carried out as described previously (Lechner et al., 1996; Bartl et al., 1997; Doetzlhofer et al., 1999). To measure HDAC enzymatic activity, equal amounts (4% of whole‐cell lysate) of immunoprecipitate obtained from 375 μl of whole‐cell lysate were used.
GST pull‐down assay
In vitro expression of radiolabelled proteins was performed in a TNT‐coupled reticulocyte lysate system (Promega) in the presence of [35S]methionine. GST–TIS7 recombinant protein was expressed in Escherichia coli BL21. GST pull‐down assays with glutathione–Sepharose 4B beads (Pharmacia) coated with GST fusion proteins (2 μg) were performed as described previously (Karlseder et al., 1996; Doetzlhofer et al., 1999). Bound proteins were eluted with loading buffer, resolved by SDS–PAGE and visualized by autoradiography. In order to confirm equal loading, gels were stained with Coomassie Blue.
cDNA microarray experiment
Technical details and the exact protocol concerning this experiment are described in Supplementary data.
Supplementary data
Supplementary data are available at The EMBO Journal Online.
Supplementary Information
Supplementary data [emboj7594678-sup-0001.pdf]
We would like to thank M.Busslinger for helpful comments and discussion on the manuscript. We would like to thank C.K.Glass (UCSD) for providing us with the N‐CoR expression plasmid. We would also like to thank C.Pasquali, R.Kurzbauer, M.Bister, C.Gieffers and K.Paiha for excellent technical assistance. This work was supported by Boehringer Ingelheim and by the Austrian Science Foundation grant (FWF, P13577‐GEN).
- Copyright © 2002 European Molecular Biology Organization


text:
GraphicRiver Fashion Certificate 10140478
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text:
Croatian failways: just when you thought it's as bad as it can get...
So, new train schedule is in effect from today. It's changed yearly so this shit is here to stay for another 12 months, yay! This is a small sample of that I discovered today, just on the route I'm taking. I expect there's a lot more, but I didn't have the time to study it.
First, an image so you can refer back to it:
A is where I work
A, B and C are major stations where train lines usually end. There are some exceptions, but in general, these are the only stations in the graph where there's actual storage for the trains and such so the line can end there.
D is where I live.
E is a small-ish station that will be relevant later
So, first of all, I messed up this morning, unaware that the new schedule is in effect. The train I usually take got moved from 06:11 to 06:08. I arrived at pretty much 06:08 sharp and missed my train.
So, there used to be a train from C to A that used to arrive to D at 06:27. It would then just continue up to A from there and it was the train I used to take in case that I was late or something. Now? The train only travels from C to D, arriving at 06:33, then turns around and goes back the way it came. Then, everyone going to A have to get off and wait for a train from B to A that leaves at 06:42. Which is great in this weather, too!
Now, the kicker about this? The train from B to A has the same number of cars that it used to have before! Meaning most of the people making this new connection can't find a seat. As a bonus, today one of the cars was without power and heating! I'm not sure what the actual temperature was this morning, but it was damned near freezing point in any case.
Now you're saying, oh Onyx, it's just one inconvenient connection, you're blowing it out of proportion. Hold on, I just got to work, how do I get back home?
Well, there used to be two trains: I usually took one at 16:50 from A to B, and another one at 17:27 from A to C. Since they both went through D where I live, it was all fine and dandy. Now? Both GONE.
But fear ye not! There is now a train from E to B, which makes no god damned sense, at 17:57. But, I have to get to E first. There's multiple trains at earlier times that can do that, but lets take a look at the two closest ones to that time: one leaves A at 17:05 and arrives at E at 17:28, meaning you have to wait for half an hour extra. The next one leaves A at 17:42 and arrives at E at... 17:58, making you miss the connection by a minute! YAY!
And I have it easy. Lines to C were crappy before, I imagine it's even worse now. So those people are fucked.
So yeah, probably the best thing I can do it pay extra for an express train at 16:40. Meanwhile, my friend that works until 17:00 instead of 16:00 as I do is pretty much fucked. In fact his first IRC message this morning was linking this video with the comment "this is how they made the new schedule":
South Park-Cable Company full scene – 01:58
— 666asianpride
Ok, it's cars then? Those terms never made sense to me in English, I mix them up all the damned time.
But technology is at about that level, yes.
Deadfast last edited by
Alternatively a "carriage" in UK English. Probably doesn't help with your confusion ;).
Cars or carriages.
There, fixed, I think, unless I missed any.
I just thought Croatian trains might have been a bit old fashioned.
Some of the trains still have bathrooms you shouldn't use at stations because pushing the pedal next to the bowl just opens a flap that allows the stuff to fall out to the ground.
Old fashioned enough?
Old fashioned enough?
Yes but I'm not convinced some of the trains here aren't still like that.
keigezellig last edited by
It sounds very much like the Dutch railways...
Nocha last edited by
Pretty sure that still happens in the UK... In fact, it took me a moment to work out this thread isn't about the UK Railways
blakeyrat last edited by
Cars or carriages.
If they're all-steel, you can also call them Pullmans in the US.
Pretty sure that still happens in the UK... In fact, it took me a moment to work out this thread isn't about the UK Railways
I thought that too - having checked, it seems it is indeed still the case.
ixvedeusi last edited by
Filed under: Somehow I think the image for "cart" was closer to reality, English, how does it work?
Yeah, they were kinda similar, no?
ixvedeusi last edited by
Parse error: Line 1:
Parse error: Line 1:
my jorb here is done.
Shoreline last edited by
Nowhere near as bad as this, but while I was going through a particularly stressful period of contracting, we also had a few years of snow.
In the UK, if it snows, the trains stop. It will snow for maybe 1-2 days and be fully melted in a week. You may as well call in sick on the first day. This is probably what I should have done, rather than walk (no buses) to the train station and wait on an increasingly crowded train for 4 hours to be taken short of my destination and have to find an alternative route.
Either that or I'm getting confused with one of the bi-monthly copper signalling wire thefts.
I don't live out that far anymore so it's fine I guess, but then it doesn't snow lately either.
Zecc last edited by
a particularly stressful period of contracting
How many weeks pregnant?
According to the train I'm on now, the official term is coaches.
Don't think I've ever heard anyone actually call them coaches outside of the train company though.
blakeyrat last edited by
I like croutons.
Update: under the pressure of customers and the media, one of the trains has been reinstated. It means I'll probably stay at work until 5PM instead of 4 (it's a nicer place to wait, anyway), but at least it's something...
I'm honestly surprised that the pressure worked. They are a monopoly after all. But hey, I won't complain... as much.
Shoreline last edited by
@Shoreline said:
a particularly stressful period of contracting
How many weeks pregnant?
I'm using that.


text:
CHICAGO — Wrigley Field became Wrigley Healed on Thursday, a shrine to all Cubs fans, a place to find peace and happiness that had eluded this franchise for 108 years.
Bathed in the warm sunshine of victory after the Cubs won their first World Series since 1908, the fans once again swarmed to Wrigley the day after the magnificent 8-7, 10-inning win in Game 7 over the Indians at Progressive Field.
The streets overflowed with fans wanting to share this once in a lifetime moment with each other.
For those Cubs fans who have passed on, their spirits were present as well.
Brandon Bender, 25, walked around the plaza carrying a picture that showed him sitting with his grandmother, Rita Mary Nugent, who passed away in January at the age of 92. He was giving her a Harry Caray-like play-by-play of the gorgeous day.
"They came close last year,'' Bender said of the Cubs. "I was hoping they would do it for her, but they did it this year for her and now she is up there with grandpa and Ernie Banks, Ron Santo and all the other loved ones that are smiling right now.
"I'm just letting her know what a special and beautiful day this is, the day we've all been waiting for. They can go 108 years without winning and my life would still be complete.''
As Bender talked he walked past a tribute message written on the cream colored wall of Wrigley in Cubs blue chalk.
It read: "Dad, your Cubs have won — R.I.P. Now! Robert F. Salinas.''
Yes, the writing really is on the wall about what these Cubs mean to their fans. It all started after Game 4 of the World Series when fans started writing tributes in chalk on the Wrigley brick walls along Sheffield Avenue. Wrigley has become a message board of love.
Katie Compton, a special education teaching assistant, wrote a dozen names on the wall. She lent her chalk to Lashawn Bennett, a fan since 1976 and throat cancer survivor.
There are heartwarming messages like this: "For all the Cubs fans that didn't get a next year.''
Linda Simeone, 60, was here with her son, daughter-in-law and two grandkids, generations of Cubs fans. Linda's mom, Louise, died seven years ago at the age of 82.
"This is overwhelming,'' Linda said. "We had to come here today.''
They all watched the final win together with Linda's mom's Cubs brick on the table: "And the brick says,'' Linda explained with a smile, "I couldn't wait any longer.''
Incredible joy was everywhere.
Aaron Post was so happy he teared up after thinking about the final game and looking at the marquee that read: Wrigley Field, Home of Chicago Cubs World Series Champions.
Post and his husband, Tom Welch, had just taken a picture in front of the marquee. "This is much more meaningful than winning a World Series,'' Post said, thinking about generations of fans.
Post and Welch were married two years ago and they live in Wrigleyville.
"A couple friends of ours got married [Wednesday],'' Welch said, "knowing that if the Cubs win, they will never forget this date.''
A good walk up Clark Street brings you to beautiful Graceland Cemetery. Way in the back by Lake Avenue is Ernie Banks' grave. He died in January 2015. It's a lovely spot, and Cubs fans visited Thursday, leaving a baseball with this inscription: It finally happened! Cubs win! There were several "W'' flags and a copy of the Chicago Tribune with a picture of the victory celebration and the headline: What A Wonderful World.
"He's only a few blocks away from his home field,'' said gravedigger Ron Danish. "Cubs fans young and old come.''
One such fan, John Kaney, 58, made his way to pay his respects.
"As a kid, you used to hear: 'Let's Play Two,' '' Kaney said. "I always wanted to be No. 14. I saw this team in spring training, and I just had a magical feeling about them. I'm a runner, and three years ago I ran the Race to Wrigley that benefits Cubs charities and Ernie sent us off, saying, 'Let's Run Two.' ''
Finally, the Cubs have won the race.
Their fans, living and dead, are at peace.
— With AP


text:
Credits and Contact Information
YaPiG was initially developed by NaTaSaB, that is, me. My electronic mail is email@example.com and my personal home page is http://natasab.merlos.biz (Spanish Only)
I would like to thank:
- Manu for his help testing this program in first versions
- Matt Thompson, Thomas Orieux and Miikka Koskinen for their help sending bugs and patches.
- All people who sent Feature Requests and Bugs
- All the people who translated YaPiG into their language (view translations)
- Sourceforge.net for their resources, hosting etc...
At this momment, YaPiG is a personal Project, but as it is GNU, I hope any other people would like to join this project. Everybody is welcome :D.
YaPiG Home page is:
YaPiG Project Page is: http://sourceforge.net/projects/yapig/