The Lust Lizard of Melancholy Cove Chapter 9~10

Nine When Mikey â€Å"the Collector† Plotznik wheeled into town and saw that the Texaco station had blown up, leaving a charred circle two hundred yards wide around it, he knew that it was going to be a great day. It was a shame about the burger stand going up too, and he'd miss their spicy fries, but hey, you don't often get to see the toasting of a major landmark like the Texaco. The fire was all out now, but several firemen were still sifting through the wreck-age. The Collector waved to them as he wheeled by. They waved back, somewhat reticently, for the Collector's reputation preceded him and made them nervous. Today would be the day, Mikey thought. The Texaco was an omen, the star in the sky over his lifelong dream. Today he'd catch Molly Michon naked, and when he did (and brought back the proof), his reputation would grow to mythic proportions. He patted the disposable camera he carried in the front pouch of his hooded sweatshirt. Oh yes, he'd have evidence to back up his story. They would believe him – and bow to him. At this point in his life, the Collector was more interested in explosions than in naked women. He was only ten, and it would be a couple of years before his interests moved to girls. Freud never identified a stage of devel-opment known as â€Å"pyrotechnic fascination,† but that was only because there wasn't an abundant supply of disposable lighters in nineteenth-cen-tury Vienna. Ten-year-old boys blow shit up. It's what they do. But today a strange new feeling had come over Mikey, a feeling he couldn't put a word to, but if he could, the word would have been â€Å"horny.† As he Rollerbladed through town, tossing the Los Angeles Times into the shrubs and gutters of businesses along Cypress Street, he felt a tightness in his shorts that until now he had associated with having to take a raging pee in the morning. Today it signified a need to see the Crazy Lady in a state of undress. Paperboys are the carriers of preadolescent myth. On every paper route, there is a haunted house, a kid-eating dog, an old woman who tips with twenties, and a woman who answers the door in the nude. Mikey had never actually seen any of these things, but that never stopped him from spinning wild stories for his buddies at school. Today he would get proof, he could feel it in his loins. He skated down the driveway into the Fly Rod Trailer Court, chucked a paper into the rose bushes in front of Mr. Nunez's trailer, then made a beeline for the Crazy Lady's house. He could see a blue glow coming through her windows, a TV. She was home and awake. Yes! He pulled up a couple of doors down and noticed that a new trailer had moved in next to the Crazy Lady. A new customer? Why not give it a try? The Crazy Lady didn't receive the paper, so his pretense for knocking on her door was to get her to subscribe. He could practice on these new people. As he skated up to the front door of the new trailer, lights came on in the two front windows. Yes! Someone was home. Strange curtains – they looked like cat's eyes. Through a part in the curtains, Molly watched the kid come down the road into the trailer park. She liked kids, but she didn't like this kid. At least once a week he knocked on her door and tried to get her to subscribe to the paper, and once a week she told him to go away and never come back. Sometimes he would bring one of his little buddies along. She could hear them skulking around her trailer, trying to peek in the windows. â€Å"Swear to God, she's got a dead guy in there that she does it with. I've seen him. And she ate a kid once.† The kid was heading for the monster trailer. In the background, a videotape was playing on her TV – Mechanized Death: Warrior Babe VII – and THE SCENE was coming up. Molly looked away from the window and watched THE SCENE for the thousandth time. Kendra is standing in the back of a jeep, manning a rack of net guns as the jeep pursues the Evil Warlord across the desert. The driver turns, as he is supposed to, throwing up a fishtail of dust, but the front wheel of the jeep hits a rock and the jeep rolls. Kendra is thrown fifty feet in the air and lands in a heap. The steel bra she is wearing cuts deep into her chest and blood sprays out across the dust. The bastards! Every time she watches THE SCENE she can't believe the bastards left it in. The accident was real, the blood was Molly's, and when she returned to the set ten days later, a security guard escorted her to the producer's trailer. â€Å"I can pay you extra's wages as a mutant,† the producer said, â€Å"but let's face it, babe, you didn't get your billing because of your acting ability. You think I'm gonna hold up filming for ten days when the whole schedule is only three weeks long? We got a new Kendra. Wrote the accident and the facial reconstruction into the script. She's a cyborg now. Now you can get in line with the mutants to pick up your bag of rags, or you can get the fuck off the set. My audience wants perfect bodies, and you were getting up there anyway. With that scar you don't sell anymore.† Molly had just turned twenty-seven years old. She pulled herself from THE SCENE and looked out the window again. The kid was there, right there in front of the monster trailer. She should warn him or something. She pounded on the window and the kid looked up, not startled, but with a dreamy expression on his face. Molly gestured for him to move away. The window she was looking out of didn't open. (Trailers built in those days were designed so people would burn up in case of a fire. The manufacturers thought it would keep the lawsuits down.) The kid just stood there, his fist poised before the door as if he were frozen in the middle of knocking. As Molly watched, the door began to open. Not on the hinges, but vertically, like a garage door. Molly pounded furiously on the window with the hilt of her sword. The kid smiled. A huge red tongue snaked out of the door, wrapped around the kid, and slurped him in, Rollerblades, paper satchel, and all. Molly screamed. The door slammed shut. Molly watched, stunned, not knowing what to do. A few seconds later the mouth opened and expectorated a soccer-ball-sized wad of newspaper. Theo The hours of Theo's day had moved like slugs crawling on razor wire. By four in the afternoon, he felt as if he'd been awake for a week and the cups of French roast he'd been drinking had turned to foaming acid in his stomach. Mercifully, there hadn't been a single call for a bar fight or do-mestic dispute, so he had spent the entire day at the scene of the fuel truck explosion, talking to firemen, representatives from Texaco Oil, and an arson investigator sent up from the San Junipero Fire Department. Much to his surprise, going all day without a hit from his Sneaky Pete pot pipe had not sent him into fits of anxiety as it usually did. He was a little paranoid, but he wasn't sure that that wasn't just an informed response to the world anyway. At a quarter past four, the arson investigator crossed the charred parking lot to where Theo was leaning on the hood of his Volvo. The investigator was in his late twenties, clean-cut, and carried himself like an athlete, even in the orange toxic waste suit. He carried a plastic space helmet under his arm like a tumorous football. â€Å"Constable Crowe, I think that's about all I can do today. It'll be dark soon, and as long as we keep the area closed off, I'm sure everything will still be here in the morning.† â€Å"What's your call so far? â€Å"Well, we generally look for evidence of accelerants, gas, kerosene, paint thinner – and I'd say there were definitely some flammable liquids involved here.† He smiled a weary smile. â€Å"So you don't know what happened?† â€Å"Offhand, I'd say a fuel truck blew up, but without further investigation I'd hate to make a commitment at this time.† Again the smile. Theo smiled back. â€Å"So no cause?† â€Å"The driver probably didn't seal the hose correctly and a cloud of fumes got set off. There wasn't much wind last night, so the fumes would have just clung to the ground and built up. Anything could have set it off: the driver could have been smoking, the pilot lights at the hamburger place, a spark in the truck exhaust. Right now I'd say it was totally accidental. It was a company-owned store, and it was turning a profit, so there really isn't a financial motive for arson. Texaco will definitely be building your town a new burger stand and probably paying off some nuisance settlements from people claiming trauma, duress, and irritation.† â€Å"I have the information on the driver,† Theo said. â€Å"I'll check to see if he was a smoker.† â€Å"I asked him. He's keeping quiet† came a voice from a few yards away. Theo and the arson investigator looked up to see Vance McNally coming toward them holding up a Ziploc bag full of white and gray powder. â€Å"I've got him right here,† the EMT said. â€Å"You want to interrogate him?† â€Å"Very funny, Vance,† Theo said. â€Å"They're going to have to do the autopsy with a flour sifter,† Vance said. The investigator took the Ziploc from Vance and examined it. â€Å"You find any remains of a cigarette lighter? Anything like that?† â€Å"Not my job,† Vance said. â€Å"The fire was so hot it turned the seat springs to liquid. Even incinerated the bones, except for those little bits of calcium in there. Honestly, this might not all be our boy. We might be giving his wife a bag full of burnt-up truck parts to put in an urn on the mantel.† The investigator shrugged and handed the bag back to Vance. Then to Theo he said, â€Å"I'm going home. I'll come back tomorrow and look around some more. As soon as I give the okay, the oil company will send in a crew to drain the ground tanks.† â€Å"Thanks,† Theo said. The investigator left in a county car. Vance McNally turned the Ziploc bag of truck driver in the air. â€Å"Theo, this ever happens to me, I want you to get all my friends together, have a big party, and snort me, okay?† â€Å"You have friends, Vance?† â€Å"Okay, it was just an idea,† Vance said. He turned and carried his bag to the waiting ambulance. Theo sipped his coffee and noticed something moving in the charred brush beyond the Texaco. It looked as if someone was holding up a TV antenna and getting altogether too close to the yellow tape he had run around the perimeter. Jeez, was he going to have to stay here all night guarding the scene? He pried himself off the Volvo and headed for the offender. â€Å"Hey there!† Theo called. Gabe Fenton, the biologist, emerged from the brush, indeed holding up some kind of antenna, followed by his Labrador retriever, Skinner. The dog ran to meet Theo and greeted him with two muddy paw prints on the chest. Theo rubbed Skinner's ears to hold him at bay, the classic slobbering Labrador control move. â€Å"Gabe, what in the hell are you doing down here?† The biologist was covered with burrs and foxtails, his face striped with soot from the charred brush. He looked exhausted, yet there was a note of excitement bordering on ecstasy in his voice. â€Å"You won't believe this, Theo. My rats moved en masse this morning.† Theo tried, but couldn't match Gabe's enthusiasm. â€Å"That's swell, Gabe. Texaco blew up last night.† Gabe Fenton looked around at the surrounding area as if seeing the destruction for the first time. â€Å"What time?† â€Å"About four in the morning.† â€Å"Hmmm, maybe they sensed it.† â€Å"They?† â€Å"The rats. Around 2 A.M. they all started moving west. I can't figure out what caused it. Here, look at the screen.† Gabe had a laptop computer strapped into a harness around his waist. He turned it so Theo could see the screen. â€Å"Each of these dots represents an animal I have implanted with a tracking chip. Here's their location at 1 A.M.† He clicked a key and the screen drew a topographical map of the area. Green dots were scattered pretty much evenly along the creek bed and the business district of Pine Cove. Gabe hit another key. â€Å"Now here they are at two.† All but a few of the dots had moved into the ranchland east of Pine Cove. â€Å"Uh-huh,† Theo said. Gabe was a nice guy. Spent too much time with vermin, but he was a nice guy. Gabe needs to talk to humans occasionally, Theo thought. â€Å"Well, don't you see? They all moved at once, except for these ten over here that moved to the shore.† â€Å"Uh-huh,† Theo said. â€Å"Gabe, the Texaco blew up. A guy was killed. I was talking to firemen in space suits all day. Every paper in the county has called me. The battery is almost out on my cell phone. I haven't eaten since yesterday and I only slept an hour last night. Help me find the significance in rat migration, okay?† Gabe looked crestfallen. â€Å"Well, I don't know the significance yet. I'm tracking the ten that didn't move east, hoping the anomalies will give a clue to the behavior of the larger group. Strange thing is, four of the ten disappeared off my screen a little after two. Even if they were killed, the chips should still transmit. I need to find them.† â€Å"And I wish you the best of luck, but this area may still be dangerous. You can't be here, buddy.† â€Å"Maybe there were fumes,† Gabe said. â€Å"But that doesn't explain why they all moved in the same direction. Some even came through this area from the shore.† Theo couldn't bear to express to Gabe how little he cared. â€Å"You had any dinner, Gabe?† â€Å"No, I've been doing this since last night.† â€Å"Pizza, Gabe. We need pizza and beer. I'll buy.† â€Å"But I need to†¦Ã¢â‚¬  â€Å"You're a single guy, Gabe. You need pizza every eighteen hours or you can't function properly. And I have a question to ask you about footprints, but I want you to watch me drink a few beers before I ask so I can claim diminished capacity. Come, Gabe, let me take you to the land of pizza and beer.† Theo gestured to his Volvo. â€Å"You can stick the antenna out the sunroof.† â€Å"I guess I could take a break.† Theo opened the passenger door and Skinner leapt into the car, leaving sooty paw prints on the seat. â€Å"Your dog needs pizza. It's the humane thing to do.† â€Å"Okay,† Gabe said. â€Å"I want to show you something over by the creek bed.† â€Å"What.† â€Å"A footprint. Or what's left of one.† Ten minutes later they sat over frosty mugs of beer at Pizza in the Pines, Pine Cove's only pizza parlor. They'd taken a window table so Gabe could keep an eye on Skinner, who was bouncing up and down outside, giving them an ever-changing view of the street, then the street with dog face (ears akimbo), then the street, then the street with dog face again. Other than to order a beer, Gabe Fenton hadn't said a word since they'd gone to the creek bed. â€Å"Will he just keep doing that?† Theo asked. â€Å"Until we take him a slice of pizza, yes.† â€Å"Amazing.† Gabe shrugged. â€Å"He's a dog.† â€Å"Always the biologist.† â€Å"One needs to keep the mind limber.† â€Å"Well, what do you think?† â€Å"I think that you obliterated most of what you thought was a footprint.† â€Å"Gabe, it was a footprint. A talon or something.† â€Å"There are a thousand explanations for a depression in the mud like that, Theo, but one of them is not an animal track.† â€Å"Why not?† â€Å"Well, for one, there hasn't been anything that large on this continent for about sixty million years, and for another, animals tend to leave more than one track, unless it's a creature especially adapted for hopping.† Gabe grinned. The flying dog head pogoed by the windowsill. â€Å"There were a lot of people and vehicles around there, the other tracks might have been wiped out.† â€Å"Theo, don't let your imagination run away with you. You've had a long day and†¦Ã¢â‚¬  â€Å"And I'm a pothead.† â€Å"I wasn't going to say that.† â€Å"I know, I'm saying it. Tell me about your rats. What will you do when you find them?† â€Å"Well, first I'm going to keep searching for the stimulus of their behavior, then I'll catch a few of the group that migrated and compare their brain chemistry to those that headed toward the shore.† â€Å"Does that hurt them?† â€Å"You have to blend up their brains and run the liquid in a centrifuge.† â€Å"I guess so then.† The waitress brought their pizza and Gabe was severing cables of cheese from his first slice when Theo's cell phone rang. The constable listened for a second, then stood and dug into his pocket for money. â€Å"I've got to go, Gabe.† â€Å"What's up?† â€Å"The Plotznik kid is missing. No one's seen him since he left on his paper route this morning.† â€Å"Probably hiding. That kid is evil. He rigged up something with his remote control car that affected the chips in my rats once. I spent three weeks trying to figure out why they were running figure eights in the parking lot outside the grocery story before I found him lurking in the weeds with the controller.† â€Å"I know,† Theo said. â€Å"Mikey told me that if he wired ten of your rats together, he could pick up the Discovery Channel. I still have to find him. He has parents.† â€Å"Skinner is a pretty good tracker. Want to take him?† â€Å"Thanks, but I doubt that the kid had a pizza in his pocket.† Theo folded his phone, snagged a slice of pizza for the road, and headed out the door. Ten Val Riordan leaned against her office door, trying to catch her breath and maintain her temper. Nothing in her clinical experience compared to the sessions she held on the day after the Texaco exploded. She had seen twenty patients in ten hours, and every one of them had wanted to talk about sex. And not abstract sex either, not issues or attitudes about sex, just squishy, thumping sex itself. It was unnerving. She'd anticipated a spike in libido among her patients (it was a common symptom of withdrawal from antidepressants), but the books said not more than five to fifteen percent would have a reaction – about the same number that experienced a loss of libido upon taking the drugs. But today she'd hit one hundred percent. It was as if she were running a kennel for hopeless horndogs rather than a psychiatric practice. After the last patient, she'd come out of her office to find her new receptionist, Chloe, furiously masturbating, her feet hooked into the edge of the desk, her steno chair squeaking like a tortured squirrel. Val had excused herself, turned on her heel, walked back into her office, and shut the door. Chloe, twenty-one, had maroon hair, an entire wardrobe rendered in black, and a sapphire nose ring. Val had begun treating the girl in her teens for bulimia, then hired her when the volume of appointments skyrocketed after the placebo went into effect. Chloe worked in exchange for therapy; Val had thought it would be a good financial move. Frankly, she'd liked her better when she just threw up a lot. Val was still trying to figure out exactly what to do when there was a soft knock on the door. â€Å"Yes?† â€Å"Sorry,† Chloe said through the door. â€Å"Uh, Chloe, that is not appropriate office behavior.† â€Å"Well, your last appointment had left. I thought that you would be working on your notes or something for a while. I'm really sorry.† â€Å"That's it? My last appointment leaves, so let the wild rumpus begin?† â€Å"Am I fired?† Val thought for a second. There were twenty more patients to see tomorrow and twenty the day after that. If the weirdness didn't kill her, the workload would. She couldn't afford to lose Chloe now. â€Å"No, you're not fired. But please, no more of that in the office.† â€Å"Do you have time to talk? I know my next session isn't until next week, but I really need to talk to you.† â€Å"Wouldn't you prefer to go home and, uh, think about things?† â€Å"You mean finish? No, I'm finished for now. That's what I want to talk to you about. That wasn't the first time today.† Val gulped. It was highly unprofessional to talk to a patient through a door. She steeled herself and opened it. â€Å"Come in.† She returned to her desk without looking at the girl. Chloe took a seat across from her. â€Å"So this wasn't the first time today?† Val was the psychotherapist now, not the boss. If she'd been the boss, she would have come over the desk and strangled the little slut. â€Å"No, I can't seem to get enough. I, well, it started about two in the morning, and I went straight though until time to get ready for work. Then once or twice while each patient was in session.† Val's jaw dropped. Sixteen hours of intermittent masturbation? The other patients she had seen had cited two in the morning as when their sexual adventures had started too. She said, â€Å"And how do you feel about that?† â€Å"I feel okay. My wrist hurts a little. Do you think I could have carpal tunnel?† â€Å"Chloe, if you think that you're going to file a workmen's compensation claim for this†¦Ã¢â‚¬  â€Å"No no no, I just want to stop.† â€Å"Did something happen to set this off? Something at two in the morning? A dream perhaps?† Her other patients had described various sexual dreams. Winston Krauss, the pharmacist with the sexual obsession for marine mammals, confessed to dreaming of having sex with a blue whale, riding it through the depths like Ahab with a hard-on. Upon awakening, he'd abused his inflatable Flipper until it would no longer hold air. Chloe shifted uncomfortably in her chair. Her long maroon hair hid her face. â€Å"I dreamed I was having sex with a tank truck, and it blew up.† â€Å"A tank truck?† â€Å"I came.† â€Å"Sexual dreams are completely normal, Chloe.† Right, a tank truck? That's normal. â€Å"Tell me, was there fire in your dream?† Pyromaniacs de-rived sexual pleasure from setting and watching fires. That's how they caught them, look in the crowd for a grinning guy with a woody and gas stains on his shoes. â€Å"No, no fire. I woke up at the explosion. Val, what's wrong with me? All I want to do is, you know, do it.† â€Å"And you feel that you might do something impulsive?† Chloe put on her cynical Goth-girl face. â€Å"If you mean something like buffing the muffin while I'm at work, yes, Dr. Riordan, I'm a little worried. Can't you adjust my medication or something?† There it was. In the past, that would have been the answer. Increase the Prozac to eighty milligrams, about four times the dose for the average de-pressed patient, and let the side effect of reduced libido do the work. Val had used the method to treat a nymphomaniac when she was an intern and it had worked marvelously. But what now? Duct tape oven mitts to her receptionist's hands? Although her typing probably wouldn't suffer much, it might make the patients nervous. Val said. â€Å"Chloe, masturbation is a natural thing. Everyone does it. But obviously there are appropriate times and places. Perhaps you should just cut back. Allow yourself to masturbate as a reward for controlling your urges.† Chloe's face went slack. â€Å"Cut down? I'm worried about driving home safely. I have a stick shift. I need both hands to drive, but I don't think I'm going to have them. Do you have a patch you can prescribe, like they do for smoking?† â€Å"A patch?† Val suppressed a laugh. She imagined a twitching, moaning line of people around the block at the pharmacy, there to pick up their prescriptions for the orgasm patch. It would make heroin look like Gummi Bears. â€Å"No, there's no patch, Chloe. You're just going to have to try to control yourself. I have a feeling that this is a side effect of your medication. It should pass in a day or two. I want to hear more about this dream of yours. We'll talk tomorrow, okay?† Chloe stood, obviously not satisfied with the help her therapist was offering, which was none. â€Å"I'll try.† She left the office, closing the door behind her. Val let her head fall to the desk. Jesus, Joseph, and Mary, why didn't I go into pathology? she thought. It would be so peaceful sitting around, boiling up beakers of urine and culturing bugs. No wackos. No stress. Okay, occasionally you'd be exposed to some deadly anthrax spores, but at least other people's sex lives stay in the bedroom and the tabloids where they belong. Her appointment with Martin and Lisbeth Luder rose in her head. They were in their seventies, had been in counseling because they hadn't had a decent conversation since 1958, and today they had come in and dumped a half hour of explicit sexual narrative on her, an account of perversions they'd indulged in the night before, starting at around 2 A.M. The visual conjured in Val's mind – all that parched, wrinkled flesh in furious fric-tion – culminated in flames, as if some giant cosmic Boy Scout had decided to rub two old people together to make a fire. The worst of it, the absolute worst of it, is that she'd found herself getting turned on while listening. She'd had to change her panties between appointments four times today. She considered pouring herself a hefty tumbler of brandy and settling down in front of the television, but that wasn't going to do it. Batteries; she needed four C-cell batteries and she needed them now. Then it was time to dig through her lingerie drawers and find a long-forgotten friend – and hope that it still worked. Molly Long past dark and Molly was still staring though the gap in the curtains at the trailer that ate the kid. The problem with being nuts, she thought, is that you don't always feel as if you're nuts. Sometimes, in fact, you feel perfectly sane, and there just happens to be a trailer-shaped dragon crouching in the lot next door. Not that she was ready to go out and pro-claim that fact to anyone, because no matter how sane you feel, some stuff just sounds too crazy. So she watched, still wearing her Warrior Babe outfit, hoping someone else would come along and notice. Around eight, someone did. She saw Theophilus Crowe going from door to door in the park. He came into view two trailers down at the Morales home, spoke briefly with Mr. Morales at the door, then headed for the dragon trailer. Molly was torn. She liked Theo. Yes, he'd taken her to County once or twice, but he'd always been kind to her – warned her about the guy in the day room who cheated at Parcheesi by eating the marbles. And he never spoke to her like she was a crazy woman. Theo was a fan. As Theo was raising his black Mag lite to tap on the dragon trailer's door, Molly saw the two windows on the end slowly open, revealing the cat'seye pupils. Theo obviously didn't see them. He was looking at his shoes. She threw up the aluminum sash and shouted, â€Å"They're not home!† The constable turned toward Molly. â€Å"Just a second,† she said. She bolted out the door of her trailer and stopped by the street where Theo could see her. â€Å"They aren't home. Come here a second,† she repeated. Theo tucked his Mag lite into his belt. â€Å"Molly, how are you?† â€Å"Fine, fine, fine. I need to talk to you, okay? Over here, okay?† She didn't want to tell him why. what if the eyes weren't there? What if it was just a trailer? She'd be on her way to County in a heartbeat. â€Å"They're not home then?† Theo said, pointing over his shoulder to the dragon trailer. He was staring at her now, at the same time trying not to stare. He had a goofy grin on his face, the same sort Molly had seen on the kid right before he got slurped. â€Å"Nope, gone all day.† â€Å"What's with the sword? Oh shit! She forgot she'd grabbed the sword on the way out. â€Å"I was just making some stir-fry. Chopping up some veggies.† â€Å"That ought to do it.† â€Å"Broccoli stems,† she said, as if that explained everything. He was looking at the leather bikini, and she watched his eyes stop on the scar above her breast, then look away. She covered the scar with her hand. â€Å"One of my old Kendra costumes. Everything else is in the dryer.† â€Å"Sure. Hey, you don't get the Times, do you?† â€Å"Nope. Why?† â€Å"The kid that delivers it, Mikey Plotznik, left for his route this morning and no one has seen him since. Looks like the last paper he delivered was a few doors down. You didn't happen to see him, did you?† â€Å"About ten, blond kid, Rollerblades? Kinda evil?† â€Å"That's him.† â€Å"Nope, haven't seen him.† She watched the eyes of the dragon trailer close behind Theo and took a deep breath. â€Å"You seem a little tense, Molly. You okay?† â€Å"Fine, fine, just wanted to get back to my stir-fry. You hungry?† â€Å"Did Val Riordan get hold of you?† â€Å"Yep, she called. I'm not nuts.† â€Å"Of course not. I'd like you to keep an eye out for this kid, Molly. One of his buddies fessed up that Mikey had a little bit of an obsession with you.† â€Å"Me? No kidding?† â€Å"He might be creeping around your trailer.† â€Å"Really?† â€Å"If you see him, give me a call, would you? His folks are worried about him.† â€Å"I'll do that.† â€Å"Thanks. And ask your neighbors when they get home, would you?† â€Å"You betcha.† Molly realized he was stalling. Just staring at her with that goofy grin on his face. â€Å"They just moved in. I don't know them very well, but I'll ask.† â€Å"Thanks.† He said, still just standing there, like a twelve-year-old ready to make an assault on the wall-flowers at his first dance. â€Å"I'd better go, Theo. I have broccoli in the dryer.† No, she had wanted to say she had to get back to dinner, or to her laundry, not both. â€Å"Okay. See ya.† She ran into her trailer, slammed the door, and leaned against it. Through the window she could see the dragon trailer open an eye and close it quickly. She could have sworn it was winking at her. Theo A niggling voice in Theo's head told him that finding the Crazy Lady attractive – extremely attractive – was an indicator that he was less than sane himself. On the other hand, he didn't feel that bad about it. He didn't feel bad about anything, not since he'd walked into the trailer park anyway. He had to deal with an explosion, a lost kid, the recent increase in general nuttiness in town – a virtual shit storm of responsibility – but he didn't feel all that bad. And in that moment outside of Molly's trailer, reflecting and waiting for the tide of lust to ebb, he realized that he hadn't smoked any pot all day. Strange. Normally this long without nursing from his Sneaky Pete and his skin would be crawling. He was heading back to his Volvo to resume the search for the lost boy when his cell phone rang. Sheriff John Burton didn't say hello. â€Å"Get to a land line,† Burton said. â€Å"I'm in the middle of trying to find a lost kid,† Theo replied. â€Å"A land line now, Crowe. My private line. You have five minutes.† Theo drove to a pay phone outside the Head of the Slug Saloon and checked his watch. When fifteen minutes had passed, he dialed Burton's number. â€Å"I said five minutes.† â€Å"Yes, you did.† Theo smiled to himself in spite of Burton's tone, which was on the verge of screaming. â€Å"No one goes on the ranch, Crowe. The lost kid is not on the ranch, do you hear me?† â€Å"It's standard procedure to search all the ranchland. Emergency services has the area gridded out. We have to cover the whole grid. I was going to call in some deputies to help us. The volunteer fire guys are exhausted from the explosion this morning.† â€Å"No. None of my guys. Don't call the Highway Patrol or the CCC either. And no aircraft. If the grid on the ranch has to be checked off, then check it off. No one goes on that land, is that clear?† â€Å"And what if the kid actually is on the ranch. You're talking about a thousand acres of pasture and forest that won't be searched.† â€Å"Oh bullshit, the kid is probably in a tree house somewhere with a stack of Playboys. He's only been missing for what, twelve hours?† â€Å"What if he's not?† There was silence on the line for a moment. Theo waited, watching three new couples leave the Head of the Slug in less than a minute. New couples: in Pine Cove everyone knew who everyone else was dating, and these were people who didn't go together. Not that unusual a phenomenon perhaps on a Friday night at 2 A.M., but this was Wednesday, and it was barely eight o'clock. Maybe he wasn't the only one feeling a wave of horniness. The couples were groping each other as if trying to get all the foreplay out of the way before they reached the car. Burton came back on the line. â€Å"I'll see that the ranchland is searched and call you if they find the kid. But I want to be the first to know if you find him.† â€Å"That it?† â€Å"Find that little fucker, Crowe.† Burton hung up. Theo got into his Volvo and drove to his cabin at the edge of the ranch. There were at least twenty citizen volunteers searching for Mikey Plotznik. The effort could spare him long enough to catch a shower and change his smoke-saturated clothes. As he parked the Volvo, an expensive, tricked-out red pickup truck pulled into the ranch entrance and rolled slowly by. As they passed, a Hispanic man sitting in the bed laughed and saluted Theo with the barrel of an AK-47 assault rifle. Theo looked away and walked to the dark cabin, wishing that there was someone there waiting for him.

Preferred language style Essay

Your questions should focus on how the counselor develops with clients. For example you may ask questions such as If a client asked you how you work or about your orientation to helping, what would you say? Or simply` How do you build relationships with clients, and what suggestions do you have for a new counselor in training? Or ` How do you deal with people who don’t want counseling or aren’t open to the process. And are mandated to be there by state. Please read the page from the text about building relationships it will help provided ideas Thanks Lethax In my session, I would be interviewing a psychologist who would be performing psychotherapy or ‘talk therapy’ for his patients or clients. The main purpose of the interview is to get an idea of the services the psychologists would be providing and the manner in which they would be managing the patients. These are the set of questions to be asked: – What are your professional qualifications and do you have a professional license to practice psychotherapy? How many years have you been practicing? What is your area of specialization and interest? In your practice, do you ensure that you stick to a professional code of ethics? What is the nature of your clients? What are the problems they come with? Do you provide treatment on the advice of another professional? Do you have to maintain confidentiality of information of your clients? Do you share this information with anyone else? If you have to secure this data on a system, how do you do it? Do you maintain relationships with the client or the family/parents? Do you read books or other literature in order to gain more information regarding better development of relationships with the clients? Whilst conversing with the clients do you make assumptions or judgments? How do you gain the trust of your clients? What are the means by which you would be able to develop a rapport with the patients (formal and informal interactions)? What are some of the challenges that you face whilst establishing a relationship and how do you cope with them? How do you motivate your clients (monetary or non-monetary means)? How do you come to a treatment plan for a particular patient? Do you follow any guidelines that you would be using whilst treating patients? Do you publish the information so that patients visiting you can know before hand? What are your likes and dislikes regarding the profession? Do you experience similar patients or patients having different types of problems? References: Kaye, K. (1998), Interviewing a Psychologist, Retrieved on October 17, 2007, from

Health Care Roles Essay Example | Topics and Well Written Essays - 3750 words

Health Care Roles - Essay Example It can also help sick individuals to find support from being a part of groups comprised of other patients who are also in similar situations and thereby influence or reinforce their attitudes to the illness and aid them in developing coping mechanisms.(NCI, 1989) Communication may be verbal, which is the spoken communication, or non-verbal which contributes meaning to the verbal expressions or may occur independently of verbal communication – such as the body motions and gestures, facial expressions, smiles, nods, which also function as a medium of communication. While communication itself is the process of transmission of messages and the interpreting of their meaning, during therapeutic communication, a care giver seeks to communicate with empathy and elicit such responses from the patient which will be beneficial to his or her well being. Therapeutic communication is geared towards promoting the well being of the patient through communication and interpretation of both verbal and non verbal cues. It is especially useful in the field of nursing and in the care of patients who may face chronic health conditions, because it is an aid in facilitating positive clinical outcomes. The given scenario concerns the patient Vivian, who has been suffering form abdominal pains for some time but has not been able to avail of health services due to a lack of health insurance. However, as soon as she gets a job and secures health insurance, she schedules an appointment with the doctor. The dà ©cor of the waiting room is stark stark, with only two notices up on the wall – one stating â€Å"Turn off cell phones† and the other â€Å"have co-pay ready.† She signs in at the front desk but before she can even begin to state her problems and mention the pain she is in, the assistant tells her to take a seat and calls another patient who

Art Criticism Essay Example | Topics and Well Written Essays - 1000 words

Art Criticism - Essay Example The recent cleaning of this painting is what ignited the interest of those who made the Constable exhibition possible. (3) The White Horse is an oil painting of a landscape of a cottage and its grounds, with a white horse as its focal point. The horse is harnessed and standing in the corner of a fence which is a prison him. Wanting to be free, in its stillness, the horse illustrates a contrast against the movement shown by the rest of the painting. Pulling the viewer into the scene are the paintings rounded forms, especially, in its trees and foliage. The energy depicted in the painting makes it not only have a harmonious feel but excitement as well. "Toward the end of his life, in the 1830s, Constable's art became more emotionally charged. He increasingly regarded the sky as 'the chief organ of sentiment' in landscape painting, and very likely looked to his cloud studies more for their expressiveness, than for their empirical or scientific content."(4) Constable often used paper rather than canvas to paint upon because it dried faster and allowed him to more freely express his emotions. It is the energy of his paintings that pulls me into them, while drawing forth the excitement of my emotions. Van Gogh's painting, The Olive Orchard, a 28 by 36 oil painting, is much smaller in size than the size of Constable's paintings. However, the sky in The Olive Garden has the same energy as that depicted in the skies paintings by Constable. Constable, like Van Gogh, was a lover of pastoral settings, and there is something about the combination of their subjects; animals and nature, that presents the romantic sides of their natures. It is through the pastoral theme in their paintings that I'm drawn away from the excitement of them, to a calmer side of myself. Prior to being exhibited in the Royal Academy of Art, The White Horse was part of the estate of Peter A.B. Widener. Later, it became a part of the Widener Collection on loan to the academy. (5) Other works in the Constable exhibition include The Hay Wain (1820-1821), View on the Stour near Dedham (1822), The Leaping Horse (1825), and Hadleigh Castle (1829). There are six paintings by Van Gogh in the gallery's permanent collection. Among them is The Olive Orchard, which is a 28 by 36 oil painting. Its size is much smaller than that of Constable's paintings. Yet, in many ways, their paintings are alike. The subjects in The Olive Orchard, two young women on a ladder, picking olives from an olive tree, like the subjects in Constable's pastoral scenes are similar. They act as contrasts against the energy of the skies both painters depict in their works. The Olive Orchard painting was completed in 1889, and is now a part of the gallery's permanent collection. The gallery also has in its permanent collec

Build a Learning Managment System Essay Example | Topics and Well Written Essays - 4000 words

Build a Learning Managment System - Essay Example decentralisation, prices have remained relatively low and simple making it very cheap for many other networks, such as big companies, telecoms firms and educational establishments to connect to the Internet (Economist, 2003). Education is one area that has largely embraced the location-free advantage of the internet with innovations such as e-learning. Educational establishments have always strived to reach out to all sections of society by offering distance-learning modules to students who could not always travel to the teaching location, for example, adults with children. This was not optimal as the success for both the establishment and the student was based on access to a central location as the traditional teaching methods were centralised. The student came to the classroom, and not the other way round. E-learning on the other hand, brings the classroom to the student. The internet has been instrumental in the development of e-learning, web blackboards and has totally changed the nature of distance learning. The internet has resulted in an increase in sales of personal computers and record internet subscriptions. As a result, students of almost every age are far ahead of their teachers in computer literacy. Students are also aware that they can do everything faster, and gain access to a wealth of information (Zuckerman). Students can now take virtual trips and collaborate with other students around the world and access the best libraries (Zuckerman) such as the Athens database and the British Library for current information on various topics. The benefits are also available to tutors as they can compare techniques with colleagues around the country and create innovative teaching modules. When it comes to the internet, the possibilities seem limitless. Schools, universities and colleges are facing increasing pressure to enter the digital world. However this has cost implications, especially when the pace of IT development is growing

Sula in her Society Essay Example | Topics and Well Written Essays - 1250 words

Sula in her Society - Essay Example When describing Sula’s hometown, the author offers the environment as one in which the local community is facing unemployment and the pain from not having work for an extended period of years. Morrison describes the community who watches Sula as being secretively pain-riddled where humor is utilized in an attempt to not have to focus on their poverty. Morrison describes a â€Å"shucking, knee-slapping, wet-eyed laughter that could even describe and explain how they came to be where they were†¦a nigger joke† (Morrison, 4). This describes a community which is well-aware of their social status, however they feel virtually powerless to break free from this lifestyle and utilize humor to gain a sense of control and to cope. Sula is powerfully-different from this lifestyle belief and manages, over her lifetime, to develop quality friendships and find the positive self-identity she is searching for. In Sula’s household, the women use door-slamming and other extreme behaviors such as throwing objects as a means to gain a perception of control over a very chaotic household environment and poverty living standards. Toni Morrison illustrates these wild behaviors in Sula’s household to describe how her upbringing taught Sula that relief from stress and unhappiness in the form of these extreme behaviors were positive methods to gain power over the boundaries caused by their gender, racial and financial statuses. Toni Morrison likely described the environment where Sula was raised as a means to highlight the sociological problems of African-American women during the novel’s time-period. Her hometown maintains many more-affluent white citizens which also impacts the view of the self which less-privileged people experienced from 1920-1960. Morrison is clearly using Sula, and her somewhat unorthodox belief in living life to its fullest, as a representation that an individual does not

Definition of Human Rights Essay Example | Topics and Well Written Essays - 1000 words

Definition of Human Rights - Essay Example It is clear from the discussion that the development of human rights is supported by many institutions but the major source of conception is the Universal Declaration of Human Rights. The treaties and laws instituted under this statute are later adopted by several international organizations. The human rights philosophy has answers to the queries regarding the creation, concept, nature, universal approach, justification and legality of each and every human right. Human rights are considered to be universally applicable to people from all societies, to provide legal and independent status to them. However, this concept it somehow produces continuous doubts in the mind of critics and, thus, a philosophical difference occurs at many stages. A wide range of literature, interpreting and illustrating the philosophy of human rights, has been published. Thus, if people analyze and understand the concept in its right perspective, such doubts can be eliminated. In 1948, the Universal Declarati on of Human Rights has proposed a list of more than twenty-four specific rights that every member nation has to accept to guarantee protection to its citizens (Morsink 12). The division of the formulated human rights also has been done in six different categories. The first category is right to security, which gives protection to citizens from criminal activities like annihilation, bloodshed, violence, victimization, and rape. The second category includes the rights that guarantee complete safety from atrocities of the legal system like punishment without trial, unfair or secret trials and harsh punishments. The third category includes liberty rights that provide protection and liberty of belief, speech, right to assemble and participate in political or social movements.  

Organic Chemistry Testing Markovnikov's Rule Lab Report

Organic Chemistry Testing Markovnikov's Rule - Lab Report Example This is practically true considering a case in which an alkene is meant to react with water in the addition reaction with alcohol that involves the carbocations formation as the ultimate result. Arguably, the group of hydroxyl (OH) combines with the atom of carbon with the highest amount of carbon-carbon bonds, with the hydrogen bonding to the carbon possessing on high carbon-hydrogen bonds. Certainly, it is apparent that for Markovnikov Rule, the chemical basis is the establishment of a carbocation often considered most stable during the process of addition. As widely cited, the hydrogen ion addition to a unit atom of carbon within the alkene establishes a charge that is positive on another carbon forming an intermediate carbocation (Lehman 7). With increasing substitution of the carbocation, the bonds to carbon or substituent to electron donating increases, as well making it an intermediate, which is somewhat stable given the hyper conjugation and induction. What this means is that the component of the reaction of addition is a product that is made from an intermediate considered most stable. In this case, one key product of HX addition to an alkene contains an atom of hydrogen in a position of minimal substitution whereas X is in a position that can easily be substituted. The less stable, less substituted carbocation will eventually be established to a certain degree and will end up establishing a product that is minor with the X attachment on the opposite side. With a view to test the practicality of Markovnikov’s Rule, this experimental was set up with a focus on Markovnikov’s Rule A solution containing 0.80mmol of iodine was prepared in 1.0mL of THF in a vial dram. 4.00mmol of 1-hexene was charged into a 10ml round-bottom flask, a 4.0 mL of THF was added, and a stir bar used to stir and dissolve the mixture. 1.6mmol of

Freud on Civilization and Religion Essay Example | Topics and Well Written Essays - 750 words

Freud on Civilization and Religion - Essay Example Naturally, civilization and religion have held the interest of many philosophers and sociologists alike. The well-known Austrian psycho-analyst and neurologist Sigmund Freud (1856 – 1939) was one of them, who held the view that mankind would be better off without belief in God - "It would be an undoubted advantage if we were to leave God out altogether and admit the purely human origins of all the precepts and regulations of civilization." (PBS 2). This essay shall briefly discuss the basis on which Freud places his argument to denounce civilization and religion, in the book â€Å"The Future of an Illusion† (1928). Freud scorns at any attempt to differentiate between culture and civilization; they provide on with two aspects of the same thing – the mastering of natural forces by collectively acquired knowledge and power to satisfy man’s individual needs and, in order to facilitate and distribute/regulate â€Å"attainable riches† (Freud 8-9). And cul ture or civilization selectively permits certain actions and condemns some others which lead to â€Å"privations† – in other words they are the frustrations caused by unfulfilled instincts, like incest, cannibalism etc (Freud 17). These â€Å"external compulsions† become â€Å"internalized† in the process of forma â€Å"super-ego† (Freud 18) which then judges within a wide range of such acquired information as to what is permissible and what isn’t permissible. This then becomes the greatest tool in the hands of culture or civilization.... Religion, Civilization and Freud According to Freud, man has devised the concept of God - a father-figure whom he dreads and always reveres, seeks to propitiate, and looks to for constant protection as the consequence of his own human limitations and weaknesses. Religion, therefore is born out of culture, primarily due to "the necessity for defending itself against the crushing supremacy of nature;" secondly, there was a pressing need to correct the perceived "imperfections of culture" (Freud 37). His perception of religion and its origins thus explained, he gradually builds his arguments against religion and civilization, stating that despite having had thousands of years to show what it can achieve, it has hardly done enough for mankind (Freud 60); and unlike scientific theories and laws, the dogmas or religion discourage testing and probe for proof, on grounds that they deserve to be believed, and they have been already tested and handed down since times of yore, and they were unquestionable (Freud 45). He deno unces these arguments stating for example, while the scientific theory that the earth is globe can be proved and its accuracy ascertained, religious dogmas were full of "contradictions, revisions, and interpolations; where they speak of actual authentic proofs they are themselves of doubtful authenticity" (Freud 46). Freud questions that if religious truths solely depended "on an inner experience" for proof, "what is one to make of the many people who do not have that rare experience" (Freud 49) He concludes of religions that they are "illusions, they do not admit of proof, and no one can be compelled to consider them as true or to believe in them." (Freud 55). Freud states

Community Empowerment and Motivation to Healthcare in an Acute Care Se Essay - 13

Community Empowerment and Motivation to Healthcare in an Acute Care Setting - Essay Example Motivation, however, involves triggering interest for application of developed potentials. Healthcare based empowerment and motivation within communities, therefore, involves facilitating self-reliance in improving health standards through prevention of diseases and infections without the direct involvement of care personnel. This aids self-care that, according to World Health Organization, is people’s potential, at either personal or communal level, to improve health standards, â€Å"prevent disease, and maintain health, and cope with illness and disability† without input from care personnel (Barlow, n.d., p. 1). Such empowerment and motivation are essential in the scope of acute care that majorly involves injuries and injury-related complications such as central nervous system and cerebrovascular accident complications, and â€Å"respiratory diseases,† â€Å"head injury,† â€Å"hemorrhage,† (Asha, 2013, p. 1). Community empowerment and motivation is an essential need that requires attention in acute care because of its preventive measure and its importance in providing emergency care in absence of care personnel. Its involved awareness and development of strengths for care initiatives ensure communities’ ability prevent the need for acute care and ability to meet rising health care needs. Consequently, the need ensures a healthy society regardless of the scarcity of health care facilities and personnel. It also means that community empowerment and motivation will relieve pressure on care facilities and promote efficiency in acute care services (See, 2007). My developed knowledge from the Doctor of Nursing Practice has diversified potentials to contribute to a nursing professional organization. My most significant contribution could be in the field of nursing research in which I could investigate aspects of self-care in an acute care setting. My research could explore people’s behavior and actions, and their possible effects on health.

Conflicting Perspectives Essay Example for Free

Conflicting Perspectives Essay Composers are able to evoke in the audience certain reactions to characters or events in their texts by presenting conflicting perspectives on different issues through the manipulation of the language forms and features of their medium, often communicating their own ideas about issues in question, which results in the creation of meaning within their texts. (?). David Guterson in his 1995 novel Snow Falling on Cedars (Snow) and Henry Bean in his 2001 film The Believer (Believer) demonstrate conscious choices made regarding structure and techniques in the construction of their texts in order to represent conflicting perspectives exploring ideas on racial prejudice and hatred and cultural contrasts and thus engage the audience. Composers can examine racial/religious prejudice brought on by war by using form specific techniques to present conflicting perspectives on the same event, designed to incite certain audience responses. Guterson, in Snow, purposely presents conflicting perspectives between Arthur Chambers and Hatsue and other members of the white community on San Piedro, particularly Etta Heine, in order to draw sympathy for the treatment of the Japanese after Pearl Harbour is bombed. Arthur is empathetic towards them, saying in his local paper the San Piedro Review, â€Å"†¦ those of Japanese descent on this island are not responsible for the tragedy at Pearl Harbour. Make no mistake about it.† The high modality language and short, direct sentences used by Guterson highlights Arthur’s deeply-held opinion of the innocence of the Japanese on the island. In support of Arthur’s argument, Hatsue, through the narrative’s non-linear structure, recalls her pain and confusion at the treatment of her people, saying, â€Å"It just isn’t fair – it’s not fair. How could they do this to us, just like that?† The emotive appeal in addition to Arthur’s article triggers audience support of the Japanese community. Guterson, however, also presents the contrasting racial hatred of the white islanders towards the Japanese. Etta Heine justifies the deportation of the Japanese with blunt, monosyllabic sentences – â€Å"They’re Japs†¦ We’re in a war with them. We can’t have spies around.† The use of the derogative term â€Å"Japs† and the distinct differentiation between â€Å"them†, the Japanese, and â€Å"we†, the white people, illustrates her bigoted hatred of the Japanese. Through the conflicting perspectives of Etta against Arthur and Hatsue, Guterson sways the audience to feel for the ill treatment of the Japanese, and shows them his own opinion on the negative effect of racism in wartime on the perceptions and conduct towards certain groups. Conflicting perspectives are established by Bean in Believer between Daniel, a neo-Nazi who is paradoxically a Jew himself, and a number of Holocaust survivors pertaining to the strength of their actions during WWII which aims to convey a pro-Jewish sentiment to audiences. At a sensitivity training session, Danny is enraged at a Jewish man’s lack of action while watching his son being murdered by a Nazi during the Holocaust. Rapidly cutting over-the-shoulder shots between Danny and the Jews indicate their opposing views. A close-up of Danny when he is asked by the Jews what he would have done in the situation shows his contempt and incredulous disbelief of the Jews’ weakness as he replies â€Å"Not what he did. Just stand there and watch?† Bean immediately employs a close-up reaction shot of the female Jew who rebuts with, â€Å"How do you know? You’ve never been tested like he has. Here in his rich, safe, stupid country it is so easy to imagine oneself a hero.† The personal address through 2nd person and the accumulation of adjectives to build a negative image of America strongly opposes Danny’s prejudiced conviction that Jews are pathetic, and also appeals to audiences the idea that religious prejudice towards Jews is unjustified. As Guterson does in Snow, conflicting perspectives are represented by Bean in order to sway his audience to respond negatively to unfounded sentiments of prejudice. Conflicting perspectives between characters can be used by composers to control the way in which an audience perceives them by exploring the cultural clashes that exist in the text as a reflection of societal (or social?) behaviour. In Snow, Guterson presents conflicting perspectives between Kabuo and the jury during his murder trial. In the opening chapter, a vivid description of Kabuo’s posture and expression is given from the jury’s perspective; he is shown as â€Å"proudly upright†¦ rigid†¦ detached.† This initial portrait portrait of Kabuo makes him suspicious not only to the jury but also to the audience, as Hatsue tells Kabuo using a simile that he â€Å"looks like one of Tojo’s soldiers.† However, Guterson, through the novel’s non-linear structure, refutes this perspective by explaining Kabuo’s behaviour to the audience via a flashback. Through his father’s teachings that â€Å"the greater the composure, the more revealed one was†, the audience learns the reason behind Kabuo’s unemotional stance. Third person omniscient allows the audience to sympathise with Kabuo’s emotive explanation that â€Å"he sat upright in the hope that his desperate composure might reflect the shape of his soul.† Guterson, through conflicting perspectives, influences his audience to understand Kabuo and the impact of contrasting cultural values on the perception of an individual. In Believer, Bean likewise shows contrasting opinions between Danny, who cannot fully repress his secret Jewish identity, and his anti-Semitic ‘skinhead’ friends to create audience sympathy for Danny’s inner struggles with the opposing aspects of his identity. When Danny and his friends break into a synagogue, Daniel shows a surprising respect for his religion which clashes with those of the other neo-Nazis. This directly conflicts with Danny’s character established at the film’s opening, when he violently beats up a Jew for no apparent reason. Wearing a brown shirt symbolising the Nazi SA (brown-shirts), Danny’s dark costuming contrasts with the light coloured one of his Jewish victim, highlighting the evil in his nature. Bean, however, challenges the audience’s view of Danny in order to allow them to understand his conflicting identities. In one frame, Danny is in the foreground walking down an aisle, which is juxtaposed with the othe r Nazis vandalising the synagogue. Their loud, raucous whooping contrasts to that of Danny’s respectful silence, highlighting their different treatments of the Jewish culture. When one of the Nazis tears up a Torah, a sacred Jewish text, after much opposition from Danny, a reaction shot of him shows sadness and pain accompanied by melancholy music, underlining Danny’s unspoken deference for Judaism. Bean’s portrayal of conflicting perspectives on Jewish culture incites the audience to respond more sympathetically towards Danny, and to understand that his veneer is a product of cultural differences in his society. The composers in Snow and Believer have effectively utilised techniques within their medium to represent conflicting perspectives about racial or religious prejudice and cultural differences in order to provoke certain audience responses to the characters, events or situations in their story. This includes reactions of sympathy for a certain perspective or disbelief and even dislike of opposing perspectives. In this way, the composers connect to the audience and generate meaning within their texts. In Snow, Hatsue is confined by the traditions of her culture, as shown when her mother Fujiko says to her â€Å"don’t allow living among the hakujin to become living intertwined with them. Your soul will decay†¦ rot and go sour.† The change in language to refer to the Americans as hakujin and the emotive metaphor of Hatsue’s breakdown of purity highlights Fujiko’s dislike of American culture. This

Rapamycin And Cisplatin In Breast Cancers

Rapamycin And Cisplatin In Breast Cancers Recent gene expression profiling has identified five breast cancer subtypes, of which the basal-like breast cancers are the most aggressive and possess serious clinical challenges as there are currently no targeted therapies available. Although there is increasing evidence that these tumors confer specific sensitivity to cisplatin, its success is often compromised due to its dose-limiting nephrotoxicity and development of drug resistance. To overcome this limitation, our focus is to maximize the benefits associated with cisplatin therapy through drug combination strategies. Using a well-validated kinase inhibitors library, we showed that inhibition of mTOR, TGFà ¯Ã‚ Ã‚ ¢RI, NFà ¯Ã‚ Ã‚ «B, PI3K/AKT and MAPK pathway sensitized the basal-like MDA-MB-468 cells to cisplatin treatment. Further evaluation demonstrated that combination of mTOR inhibitor, rapamycin, and cisplatin generated significant drug synergism specifically in basal-like cells (MDA-MB-468, MDA-MB-231 and HCC1937). Th ese synergistic effects were not observed in the luminal-like T47D and MCF-7 cells. We further showed that the synergistic effects of rapamycin and cisplatin is mediated through p73. Treatment of rapamycin induced p73 upregulation and synergized cisplatin activity through activation of the p73 pathway. Depletion of endogenous p73 in basal-like cells abolished these synergistic effects suggesting that p73 is required for the rapamycin and cisplatin synergism. In conclusion, combination of mTOR inhibitors and cisplatin may be a useful therapeutic strategy in basal-like breast cancers. INTRODUCTION Recent identification of novel breast cancer subtypes with distinct biological features promises a more specific, effective and less toxic therapies to the patients. Through gene profiling, breast cancer can be categorized into five different subtypes with distinct clinical outcome. The five major subtypes of breast cancer are luminal A, luminal B, human epidermal growth factor receptor-2 (HER2) overexpressing, normal-like and basal-like breast cancer [1, 2]. Of particular important is the basal-like breast cancer which accounts for 15-20% of breast cancers overall and confers a remarkably poor prognosis compared to other subtypes. Majority of basal-like breast cancers exhibit a triple-negative phenotype, characterized by the lack of expression of estrogen receptor (ER), progesterone receptor (PR) or HER2 amplification, and often have high frequency of p53 mutation [3, 4]. Due to the lack of expression of these receptors (ER, PR and HER2), patients with basal-like breast cancers usua lly do not response to hormonal therapy, Herceptin or chemotherapy [5, 6]. As a consequence, the mortality rate of basal-like breast cancer is relatively high in comparison with the non-basal subtype [1]. Numerous clinical studies are currently ongoing to identify novel therapy for treatment of basal-like breast cancers. These include the use of specific targeted therapeutic agents (e.g. Cetuximab, Dasatinib, Bevacizumab, Abraxane and Erlotinib) or conventional chemotherapeutics agents (e.g. cisplatin, doxorubicin, and paclitaxel), either as single agent or in combination, as first line therapy for basal-like breast cancers [7-9]. Cisplatin, a chemotherapeutic agent not commonly used for breast cancer, come to light in the management of basal-like breast cancer on account of evidence that breast cancer cells with basal-like phenotype confer a selective sensitivity towards cisplatin as compared to other chemotherapeutic agents. A variety of evidence suggests that basal-like breast cancers may share defects in BRCA1-associated pathways, of which DNA repair mechanism has been compromised [10]. Indeed, recent clinical studies have demonstrated the clear advantage of cisplatin in treatment of basal-like breast cancer compared to other chemotherapeutic agents [11, 12]. Nevertheless, dose-limiting toxicity including nephrotoxicity, neurotoxicity and ototoxicity have withold the wide-spread use of cisplatin in treating breast cancers in the clinic. To address this problem, we developed a high-throughput screening assay to rapidly identify new therapeutic agents that could synergize the antitumor effects of cisplatin in basal-like breast cancers. Through the use of a small chemical library that targets some of the most relevant oncogenic pathways in basal-like breast cancer, we show that inhibition of mTOR by rapamycin incurred a specific synergistic effect with cisplatin in basal-like breast cancer cells. This synergistic effect is mediated in part through the induction and activation of p73 in the presence of rapamycin and cisplatin, respectively. Together, our findings demonstrate evidence of a synergistic relation between rapamycin and cisplatin in both inhibition of cell growth and induction of apoptosis. This suggests that rapamycin and cisplatin may be a rational combination of a targeted therapy for the refractory basal-like breast cancers. Materials and Methods Cell lines and cell culture The human breast carcinoma cell lines MCF-7, T47D, MDA-MB-231, MDA-MB-468 and HCC1937 were obtained from the American Type Culture Collection (Manassas, VA) and maintained in RPMI 1640 medium containing 10% fetal bovine serum (FBS), 100 IU/ml penicillin and 100 ÃŽÂ ¼g/ml streptomycin (Sigma-Aldrich, St. Louis, MO, USA) at 37 °C with 5% CO2. MTT cell proliferation assay Dose-response curves and IC50 values were determined using the methyl thiazolyl tetrazolium (MTT) cell viability assay as described previously [4, 13]. Cells were seeded into 96-well plates for 24 hours at a density of 5 ÃÆ'- 103 cells/well. Serial drug dilutions were prepared in medium immediately before each assay, and viable cell masses following 3 days of drug exposure were determined by cell-mediated MTT reduction. Cell growth as well as drug activity was determined by measuring absorbance at 550 nm using an Anthos systems plate reader. Construction of IC50 mean graph The IC50 mean graph was constructed as defined by the Developmental Therapeutics Program of the National Cancer Institute (http://dtp.nci.nih.gov). The mean graph consists of positive (more sensitive) and negative (less sensitive) delta values, generated from a set of IC50 values by using a three-step calculation. The IC50 values for each of cell line against the tested compound were converted to log(IC50) values. For each tested compound, the log(IC50) values are averaged. Finally, the individual IC50 value is then subtracted from the average to generate the delta value. Positive delta values project to the right of the vertical line and represent cellular sensitivities to the test agent that exceed the mean. Negative values project to the left and represent cell line sensitivities to the test agent that are less than the average value. Library screening The Inhibitor Select„ ¢ chemical library which consists of 160 well-characterized, cell-permeable inhibitors was purchased from EMD Chemicals, USA. MDA-MB-468 cells at the logarithmic phase of growth were seeded into 96-well plate at a density of 5 ÃÆ'- 103 cells/well. Each compound was added to a final concentration of 10  µM in the absence or presence of 1  µM cisplatin. Plates were incubated for 72h at 37 °C. Cell proliferation was examined using MTT assay as described previously. Combination treatments that induce growth inhibition higher than those of the same doses used alone (p Drug interaction analysis Drug combination analysis was performed by using the method as described by Chou and Talalay [14]. Briefly, cells were seeded at 5 ÃÆ'- 103 cells/well in 96-well plates and treated with various concentrations of cisplatin and compound alone or in combination for 72h. Cell proliferation was measured in each well by MTT assay. Multiple drug dose-effect calculations and the combination index plots were generated using Calcusyn software (Biosoft, Cambridge, UK). Combination index, CI 1 indicate synergism, additive effect and antagonism, respectively. Apoptosis assays Quantitation of apoptosis by annexin V/PI staining was performed as described previously [3, 4]. Briefly, both floating and attached cells were collected 72h after drug treatments. Apoptotic cell death was determined using the BD ApoAlert annexin V-FITC Apoptosis Kit (BD Biosciences, USA) according to the manufacturers instructions, and cells were analyzed on a FACSCalibur flow cytometer using CellQuest Pro software (version 5.1.1; BD Biosciences, USA). Quantitative PCR (qPCR) analysis Total RNA from cells was extracted using Qiagen RNA isolation kit (Qiagen, Valencia, CA, USA) according to the manufacturers protocol. First-strand cDNA was synthesized from total RNA using random hexamer primers and the SuperScript II system for RT-PCR (Invitrogen, Carlsbad, USA). Gene expression levels were measured by qPCR using the iQ SYBR Green Supermix reagent and an Biorad iQ5 real-time PCR detector system (Bio-Rad, Richmond, CA, USA). Data analysis was performed using Opticon Monitor Analysis Software V1.08. The expression of each gene was normalized to ÃŽÂ ²2M as a reference. The relative copy numbers were calculated from an 8-point standard curve generated from a 10-fold serial dilution of full-length cDNA constructs as described previously [3, 4]. Specific forward and reverse primer sequences are as follows : TAp73fwd, 5-GCACCACGTTTGAGCACCTCT-3; TAp73rev, 5- GCAGATTGAACTGGGCCATGA-3; ÃŽÂ ²2Mfwd, 5-AGCTGTGCTCGCGCTACTCTC-3; ÃŽÂ ²2Mrev, 5-CACACGGCAGGCATACTCATC-3; PUMA fwd PUMArev NOXAfwd NOXArev. The conditions for all QRT-PCR reactions were as follows: 3 minutes at 94 °C followed by 40 seconds at 94 °C, 40 seconds at 60 °C, and 25 seconds at 72 °C for 40 cycles. All PCR products were confirmed by the presence of a single peak upon melting curve analysis and by gel electrophoresis. No-template (water) reaction mixtures and no-RT mixtures were performed on all samples as negative controls. All experiments were performed in duplicate. Protein isolation and Western blot analysis Protein lysates from cells were extracted in ice-cold lysis buffer (0.75% NP-40, 1 mM DTT, and protease inhibitors in PBS). Total protein (25 ÃŽÂ ¼g) was subjected to SDS-PAGE followed by immunoblotting with the following antibodies: p73 (diluted 1:1,000, Ab-2; CalBiochem); pS6K (diluted 1:1,000; Cell Signaling Technology); S6K (diluted 1:1,000, Ab9645; Abcam); and ÃŽÂ ²-tubulin (diluted 1:2,500, D-10; Santa Cruz Biotechnology). Lentiviral production and infection The shRNA lentiviral constructs were created by transferring the U6 promoter-shRNA cassette into a lentiviral backbone, and high-titre lentiviral stocks were generated by co-transfection with packaging vectors into 293T cells as described previously [3, 4, 13]. The shRNA target sequences for TAp73 was 5-GGATTCCAGCATGGACGTCTT-3. The TAp73 targeted sequence is found within p73 exon 3. Therefore, this shRNA does not target ΆNp73 [4]. RESULTS Selective sensitivity of basal-like breast cancer toward cisplatin To gain an overview of the selectivity of chemotherapeutic agents for basal-like breast cancer cells, we compared their antiproliferative properties in a panel of basal-like and luminal-like breast cancer cell lines which has been validated previously through gene profiling [15]. All cells were treated with increasing concentrations of cisplatin, paclitaxel or doxorubicin for 72 hours and growth measured using the MTT assay. Figure 1A and B summarizes the results from these breast cancer cell lines in which basal-like breast cancer cells demonstrated selective sensitivity to cisplatin. This selectivity was absence in cells treated with paclitaxel or doxorubicin suggesting that basal-like breast cancer cells confer selective sensitivity towards cisplatin (Figure 1A, B and Supplement Table 1). Small chemical library screening identify rapamycin as synergistic agents for cisplatin Although cisplatin is currently one of the most used agents in the treatment of cancer, the use of cisplatin is hampered by its side effects, especially neurotoxicity, nephrotoxicity and rug resistance [16]. Hence, the present study was aimed to identify chemosensitizers that could synergize the effects of cisplatin for treatment of basal-like breast cancers. To identify small molecules that enhance sensitivity of basal-like breast cancer cells to cisplatin, a cell-based high-throughput screen was performed using MDA-MB-468 cell line and a small chemical library consisting of 160 well validated specific inhibitors. The screens were done in 96-well plates to which compounds were added at 10  µM, followed by cisplatin at 1  µM. Cell viability was measured 72 hours later by MTT assay. Each plate included controls of untreated cells, cells treated with compounds or cisplatin only, and cells treated with a combination of both agents. Combinations of the treatments that induced growth inhibition higher than those of the same doses used alone (p The molecules identified in this screen includes rapamycin, [3-(Pyridin-2-yl)-4-(4-quinonyl)]-1H-pyrazole (LY364947), 4-(3-Chloroanilino)-6,7-dimethoxyquinazoline (AG1478), (E)3-[(4-Methylphenyl)sulfonyl]-2-propenenitrile (BAY11-7082), 2-(4-Morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002) and 4-(4-Fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)1H-imidazole (SB203580). Structures of these compounds and their growth inhibitory effects were shown in Figure 2. The molecular target of these compounds was listed in Table 1. The 6 compounds identified are specific inhibitors of the mTOR, TGF-à ¯Ã‚ Ã‚ ¢, EGFR, NFà ¯Ã‚ Ã‚ «B PI3K/AKT and MAPK pathways which have been previously reported to be upregulated specifically in basal-like breast cancers [7, 17-20]. However, when tested for synergism with cisplatin at fixed concentration ratio, only rapamycin showed synergism when combined with cisplatin in MDA-MB-468 cells using the isobologram method that simulate the median-dose combination index (CI) [14]. The mean combination index at ED50, ED75, and ED90 of rapamycin (10:1 cisplatin:rapamycin) when combined with cisplatin was 0.52 à ¯Ã¢â‚¬Å¡Ã‚ ± 0.06, where combination index Specific synergistic effects of rapamycin and cisplatin in basal-like breast cancer cells Next, we sought to investigate if combination of cisplatin with rapamycin exhibit specific synergism in basal-like breast cancers by comparing their effects in a panel of breast cancer cell lines. The antiproliferative effect of treatment was evaluated using MTT assays and drug interaction was assessed by the isobologram method as described in the previous section. The results demonstrated that combination of rapamysin and cisplatin exhibited selective synergistic effects only in the basal-like MDA-MB-468, MDA-MB-231 and HCC1937 cells, but not in the luminal-like T47D or MCF-7 cells (Figure 3A and Table 2). To ensure that the lack of synergistic effects of rapamycin and cisplatin observed in luminal-like cells is not due the general lack of sensitivity of luminal-like cells to cisplatin per se, we compared the apoptotic effects of cisplatin and rapamycin alone or in combination in MDA-MB-231 and T47D cells using an equal potent dose of cisplatin (IC50 dose). Both cell lines exhibited similar amount of apoptosis following treatment with their respective IC50 dose of cisplatin. Interestingly, when both cell lines were treated with combination of cisplatin and rapamycin, synergistic effects were evidenced only in MDA-MB-231 cells but not in T47D cells. The potentiation effects of rapamycin observed in MDA-MB-231 cells were further enhanced by sequential treatment with rapamycin for 6 hours followed by cisplatin (data not shown). These results demonstrated that rapamycin is capable of sensitizing basal-like breast cancer cells to cisplatin, suggesting that the synergistic effects of rapamyc in and cisplatin may be mediated through a common pathway. Rapamycin sensitizes basal-like cells to cisplatin through p73 up regulation Previous study has shown that inhibition of mTOR by rapamycin up regulate p73 in breast cancer cells [21]. We and others have also demonstrated that p73 is overexpressed in a subset of triple-negative breast tumors and that p73 is required for cisplatin sensitivity in basal-like breast cancer cells [4]. On the basis of these findings, we ask if the synergistic effects of rapamycin and cisplatin combination could be explained by the activation of the p73 pathway. To test this notion, MDA-MB-231 cells were treated with 10  µM of cisplatin and 100 nM rapamycin alone or in combination for 48 hours. mTOR inhibition was confirmed using phospho-S6K as a marker of mTOR activity. Quantitation of the mRNA and protein expression of the TAp73 was performed using qPCR and immunoblotting, respectively. The results showed that treatment of rapamycin or cisplatin alone did not change the expression of TAp73 mRNA but induced significant up regulation of TAp73 protein expression in MDA-MB-231 cells (Figure 4A and B). Next, we evaluated the expression of the two potent pro-apoptotic BH3 only proteins, PUMA and NOXA, which has been identified as specific p73 target genes, using qPCR [22, 23]. As expected, treatment of cells with cisplatin for 48 hours induced mRNA expression of PUMA and NOXA in both MDA-MB-231 and MDA-MB-468 cells. Significant induction of PUMA and NOXA were also observed in MDA-MB-231 cells but not in MDA-MB-468 cells following treatment with rapamycin alone. When cells were treated with combination of cisplatin and rapamycin, the expression of PUMA and NOXA was further enhanced, corroborated with the massive induction of apoptosis as shown in Figure 3B. Together, these results suggest that rapamycin synergize cisplatin activity in basal-like cells through induction of p73 pathway. p73 is required for the synergistic effects of rapamycin and cisplatin in basal-like breast cancer cells To further evaluate whether p73 is the effector of the synergistic effects of rapamycin and cisplatin in basal-like breast cancer cells, we generated a series of isogenic cell lines that have been depleted for TAp73 by stably expressing a shRNA species that target specifically human TAp73. Unlike MDA-MB-231 cells which express one predominant TAp73 isoform (TAp73à ¯Ã‚ Ã‚ ¢) isoform, MDA-MB-468 cells express high levels of two p73 isoforms, TAp73à ¯Ã‚ Ã‚ ¡ and TAp73à ¯Ã‚ Ã‚ ¢ [21]. Figure 5A showed efficient knock-down of TAp73 isoforms in MDA-MB-231 and MDA-MB-468 cells. As expected, treatment of cisplatin alone induced significant amount of apoptosis in MDA-MB-468 vector control cells. This apoptotic effects were further enhanced in the presence of rapamycin, consistent with our previous observations (Figure 3B). In stark contrast, depletion of TAp73 not only reduced the amount of apoptosis following treatment of cisplatin alone but also completely abrogated the synergistic effects of rapamycin (Figure). This result is further supported by the isobologram analysis which showed a lack of synergism of rapamycin-cisplatin treatment in the TAp73 depleted cells. Together, these results suggest that TAp73 is required for the synergistic effects of rapamycin and cisplatin in basal-like breast cancers. DISCUSSION By gene profiling, breast cancers can be classified into 5 molecularly distinct subtypes: luminal A, luminal B, HER2+, basal-like and normal breast cancers. The basal-like subtype, which represents 15-20% of breast cancers, has been subjected to extensive investigation in recent years due to its association with poor patient survival [1, 2, 20, 24]. Unlike many breast cancers, patients diagnosed with basal-like breast cancers are not eligible for molecular targeted therapy that target ER (e.g. tamoxifen, aromatase inhibitors) or HER2 (e.g. Herceptin) as they do not express the estrogen receptor (ER) or progesterone receptor (PR), nor do they have amplified HER2 [1, 24]. The treatment option therefore is relied on aggressive conventional chemotherapies which have limited efficacy, many side effects and often high rate of relapse. Hence, development of an effective therapeutic strategy remains an important goal in the management of basal-like breast cancer. Several lines of evidence has suggested a link between basal-like breast cancers and BRCA1 deficiency [7, 25, 26]. In most cases, the clinical features and outcomes for women with sporadic basal-like breast cancers are broadly similar to those with BRCA1-related cancers including high tendency of developing high grade, high mitotic index tumors, shorter time of relapse, similar pattern of metastatic spread and cytogenetic changes associated with frequent loss of X-chromosome inactivation [7, 27-32]. The majority of BRCA1-associated cancers are also triple-negative (ER, PR and HER2 negative), express basal cytokeratins and other markers commonly seen in basal-like breast cancers (e.g. p53, P-cadherin and EGFR) [7]. Gene expression profiling also demonstrated that BRCA1-associated cancers segregate strongly with basal-like breast cancers [2, 29, 33, 34]. Although BRCA1 somatic gene mutations are uncommon in sporadic basal-like cancers, these tumors have been shown to have a dysfunction al BRCA1 pathway due to BRCA1 gene promoter methylation and/or BRCA1 pathway transcriptional inactivation [7, 25, 26]. The fundamental biological similarities between hereditary BRCA1-related breast cancers and basal-like cancers suggest that strategies targeting the dysfunctional BRCA1 pathway may be effective in basal-like breast cancers. There is increasing evidence that the DNA repair defects characteristic of BRCA1 related cancers, especially defective homologous recombination, confer sensitivity to certain systemic agents, such as platinum-based chemotherapy and poly(ADP-ribose) polymerase (PARP) inhibitors [34-38]. Indeed, recent clinical studies revealed that sporadic basal-like cancers responded to platinum-based chemotherapy and were associated with a high rate of complete pathologic response [9, 11, 38]. Consistent with the clinical data, our in vitro study also reveals that basal-like breast cancer cells confer specific sensitivity to cisplatin as compared to other chemotherapeutic agents (e.g. doxorubicin or paclitaxel) (Figure 1), further support research into the utility of platinum-ba sed agents in basal-like breast cancers. Given the high specificity and response rate of basal-like breast cancers toward platinum-based therapy, our focus is to maximize the benefits associated with this therapy through drug combination strategies. Using a small chemical library consisted of 160 well-validated and specific inhibitors that target the human kinome, we have identified 6 compounds that significantly potentiate the antiproliferative effects of cisplatin in basal-like breast cancer cells. These compounds include rapamycin, LY364947, AG1478, BAY11-7082, LY294002 and SB203580 which targets the mTOR, TGFà ¯Ã‚ Ã‚ ¢RI, NFà ¯Ã‚ Ã‚ «B, PI3K/AKT and MAPK pathway respectively. Of note, these pathways have been reported previously to be over activated in basal-like breast cancers [7, 17, 18, 20]. To further investigate the mode of interaction between these compounds and cisplatin, we performed a drug combination study using the isobologram approach as described previously (Ref). Out of the 6 compounds identified, rapamycin showed the strongest synergistic effects with cisplatin while others (LY364947, AG 1478, BAY11-7082, LY294002 and SB203580) showed mainly additive effects. This result is consistent with other studies which show that inhibition of mTOR by RNAi or small molecules (e.g. rapamycin, CCI-779, RAD001) enhances cisplatin chemosensitivity in ovarian [39-41], endometrial [42], head and neck [43, 44], lung [45], skin [46, 47] and liver [48] cancers. We next compared the synergistic effects of rapamycin in combination with cisplatin in a panel of luminal-like and basal-like breast cancer cell lines that has been previously validated by gene profiling [15]. Intriguingly, the synergistic effects were observed only in MDA-MB-468, MDA-MB-231 and HCC1937 basal-like cells, but not in MCF-7 or T47D luminal-like cells. Several models have been proposed to explain the synergistic effects of rapamycin and cisplatin in cancer cells. Beuvnk et al., 2005 showed that RAD001 (Everolimus), a rapamycin derivative, dramatically enhances cisplatin-induced apoptosis in wild-type p53 but not mutant p53 tumor cells by inhibiting p53-induced p21 expression [49]. Wangpaichitr et al., 2008 demonstrated that inhibition of mTOR by CCI-779 decreased levels of the anti-apoptotic proteins, BCL2/BCLxL, and increasing apoptosis in lung cancer cells that is resistance to cisplatin [50]. Although these models provide important evidence for mTOR inhibition and cisplatin synergism in cancer cells, it fails to explain the specific synergism we observed in basal-like breast cancer cells, as the basal-like cells that we tested are p53 mutated and do not express high level of BCL2/BCLxL (data not shown). This led us to postulate that a common signal transduction pathway inhibited by rapamycin may be an important component that sy nergizes cisplatin sensitivity in basal-like cells. Since p73 has been reported to mediate cisplatin sensitivity in a subset of triple-negative breast cancer cells [4] and that inhibition of mTOR by rapamycin or RNAi lead to upregulation of p73 [21], we postulated that activation of the p73 pathways might be important for the synergistic effects of rapamycin. To test the role of p73 in rapamycin and cisplatin synergism, we first evaluated the expression of p73 mRNA and protein levels following treatment with cisplatin or rapamycin alone or in combination in MDA-MB-231 cells. Consistent with previous studies, treatment of cells with cisplatin or rapamycin alone induces p73 protein expression followed by transcriptional activation of the 2 potent pro-apoptotic p73 target genes, PUMA and NOXA. When MDA-MB-231 cells were co-treated with rapamycin and cisplatin, the elevation of p73 and its pro-apoptotic target genes were synergistically enhanced. The observed changes in p73 protein in MDA-MB-231 cells, however, were not due to parallel changes in p73 RNA levels, suggesting that inhibition of mTOR might lead to inactivation of a yet unknown p73 specific protein degradation pathway. To validate that the rapamycin and cisplatin synergism is mediated by p73, we generated isogenic MDA-MB-231 and MDA-MB-468 cells that were depleted for p73 using a lentiviral-shRNA that target specifically the transactivating isoform of p73 (TAp73). Indeed, depletion of TAp73 in MDA-MB-231 and MDA-MB-468 cells completely abrogated the synergistic effects of rapamycin suggesting that the synergism between rapamycin and cisplatin required p73 function. Although the combination of cisplatin and rapamycin has not been previously investigated in clinical study, it is worth noting that a phase II neo-adjuvant clinical trial of cisplatin and RAD001 (Everolimus), in patients with triple-negative breast cancers has recently open for recruitment (ClinicalTrials.gov Identifier: NCT00930930), and will be able to address the potential of cisplatin and mTOR inhibitors combination therapy directly. It would be equally intriguing to determine the role of p73 related pathway as potential biomarkers that might predict response to treatment given the pivotal role of p73 in the synergistic effects of mTOR inhibition and cisplatin sensitivity. In conclusion, combination of mTOR inhibitors and cisplatin may be a useful therapeutic strategy in basal-like breast cancers.

China and the Central African Republic

China and the Central African Republic I-Introduction Central African Republic is a landlocked country bordered in the south by the Democratic Republic of the Congo and the Republic of the Congo, in the north by Chad, Sudan in the east and Cameroon in the west. It gained its independence in August 13th 1960 and Bangui is the capital. The two official languages are French and Sango. Francois is Yangonvouda-Bozize current the President took the power during a coup in March 2003 [The Economist US (2005)]. Since its independence, the Central African Republic has been destabilized and further impoverished due to a cycle of political-military uprisings and violence [HAC, (2011); UNICEF, (2008); The Economist, (2008)]. Constitutional order was restored in 2005[UNICEF, (2008)]; however, the overall situation until now in the country remains volatile, marked by tensions among and within political parties, a precarious socio-economic situation, deteriorating humanitarian conditions and insecurity in the north of the country due to activities of rebel and criminal groups [UNICEF, (2008)]. The United Nations implemented a Peace-building Office in the Central African Republic (BINUCA), in an attempt to promote peace and stability in the country and reduce cross-border insecurity [UNICEF, (2008)]; This was carried out to encourage national dialogue and reconciliation and furthermore to help the Central African Republic enact military reforms, implement human rights policies, train civilian police, and disarm, demobilize and reintegrate former fighters. Besides that, the CAR is surrounded by countries like Chad, Sudan and DRC, which have been destabilized for a long time by civil wars. These lead many refugees to seek for shelter in CAR [The Economist, (2005)].All those characteristics create a volatile political situation and an uncertain climate for foreign investment. The CAR is blessed with many natural resources such as diamonds, timber, uranium, oil, iron, gold and hydropower [Africa.com, (2008)].However, it is still rated among the poorest countries in the world. Its economy is disadvantaged due to its landlocked position, which isolates it from foreign suppliers and markets and contributes to high import prices. It has the potential to become a major agricultural product exporter. It also has hydroelectric potential that could be developed for export to neighboring countries that have power shortages. The relations between China and the CAR have evolved saw tooth since the early 60s. On several occasions, the CAR broke and re-established its ties with the People Republic of China. Since the arrival in power of President Bozize, it seems that the CAR and China are enjoying steady and smooth relations. Yet the country is still facing to challenges regarding to its socio-economic situation. Nothing is so much said nor written about China and the Central African Republic. What is really going on between the two countries? China has invested in it, especially in terms of infrastructure. Does the government gain from it? What about the population? Is CAR really benefited from that relation? In what extent China can help the CAR to come out from the cycle of poverty? II-Central African Republic (C.A.R): 2.1Facts and figures According to the United Nations Human Development Index, the CAR now ranks 172 out of 177 least developed countries in the world [UNDP,(2006)]. Two-thirds of the population live under the line of poverty, less than half are literate, and the average life expectancy hovers around 40 years, resulting in a very young population with almost fifty percent under the age of 18, and only four percent aged 60 and over[UNDP, (2006)]. The CAR is really affected by AIDS that is why the life expectancy is constantly falling and dropped below 40 according to the UNDP Report on Human Development 2006. Per capita income is USD 456. POPULATION 4,422,000 SIZE 622,984 km ² CAPITAL CITY Bangui LANGUAGES French and Sango (officials languages);TRIBAL LANGUAGES LARGEST CITIES Bangui and Bimbo RELIGION 35% Tribal beliefs, 25% Protestant, 25% Catholic, 15% Muslim MONETARY UNIT Franc NATURAL RESOURCES Diamonds, uranium, timber, gold, hydropower AGRICULTURE Cotton, coffee, tobacco, cassava, yams, millet, corn, banana; timber EXPORT COMMODITIES diamonds, timber, cotton, coffee, tobacco IMPORT COMMODITIES food, textiles, petroleum products, machinery, electrical equipment, motor vehicles, chemicals, pharmaceuticals INDUSTRY Gold and diamond mining, logging, brewing, textiles, footwear, assembly of bicycles and motorcycles 2.2 Political and economic situations The CAR is blessed with natural resources to use and develop in order to come out of its cycle of poverty. However, three decades of mismanagement, from the 60s when it gained its independence until 1993 which seen the organization democratic elections for the first time, most of the time under military rule and one decade of chronic unrest and political fights (1993- 2003) led the country to bankruptcy [International Crisis Group, (2007)]. Francois Bozize gained the power in March 2003 by a coup and overthrown Ange-Felix Patasse [International Crisis Group, (2007)]. Due to the chronic instability since 10 years this makes very difficult for CARs governments to follow-up a long term economic development strategy that is why the country is currently facing a lack of modern transportation network which represents the chokepoint for the development of its economy. For instance, the length to reach Bangui the capital city is about 12 to 14 days of river transportation [CAR, (2009)]. Why is CARs economy always under threat of strangulation? Geographically, the CAR is trapped in an unstable triangle and surrounded by the Democratic Republic of the Congo, Sudan and Chad that which are destroyed by many years of civil wars [IMF, (2010); The Economist, (2008); UNHCR, (2000)]. Furthermore, shipment arrivals are often delayed or canceled because of rebels attacks [HAC, (2011)] that are causing so much trouble in the region. The nation of CAR is principally agrarian [International Crisis Group, (2007)]. In 2003, agriculture accounted for 60.8% of the countrys GDP [UNCTAD, (2005)]; crops primarily including cotton, food crops such as cassava, yams, bananas, maize; coffee and tobacco. In 2004, the agricultural labour force accounted for 69% [UNCTAD, (2005)]. The population of CAR is engaged in subsistence farming with 56% attributed to the agricultural sector in 2006. In 2002, timber accounted for about 30% of export earnings. The CAR is blessed with a huge unexploited natural resources in the form of uranium, gold, diamonds and others minerals [Investment Climate Report, (1995)]. In 2010, its main exports were diamonds, cotton, timber, coffee and tobacco. Diamonds are the main natural resources being currently developed [Investment Climate Report, (1995)]: in 2002, the CAR was able to export 50% of its diamonds earnings [Crisis Group Africa, (2010)]. In 2003 and 2006, industry contributed for respectively 24.9% and 15% for the latter of the countrys GDP; breweries, diamond mining and sawmill represent the bulk of the sector. Due to the oversized government bureaucracy and its landlocked position, services accounted only for 29% of GDP in 2006. The same year, its main importer of goods was France and represented for 17.6% and they mainly exports goods toward Belgium (40.4%) [Investment Climate Report, (1995); UNCTD. (2005)]. In 2000, the percentage of population with access to electricity accounted for 5% only [UNCTAD, (2005)]. The CAR really lack of electricity supply. The main and only hydroelectric plants are based in Boali. Mostly, fuel supplies are transported in via the Ubangui River or trucked through Cameroon, resulting in frequent shortages of gasoline, jet fuel and diesel. The countrys transportation and communication network is limited; it does not possess a railroad, however it has only 650 kilometers of paved road. It does not possess any domestic air service, except charters, so very limited internationally. Furthermore, commercial traffic on the Ubangui River is very rough even impossible from December to May or June, due to ongoing conflicts in the region that sometimes prevented shipments from moving between Kinshasa (capital of DRC) and Bangui. The telephone system functions, albeit imperfectly. The CAR possesses only one television station that has been built since the independence, an d four radio stationsare currently operating in the country. Numerous newspapers and pamphlets are published on a regular basis, and at least one company has begun providing Internet service. In terms of economic, the C.A.R. has made slow progress; its budget and external trade are deficiency due to many constraints like the poor infrastructure that it is facing, economic mismanagement and scarce private investment, as well as a limited tax base and adverse external conditions have led the country to a desperate situation. Its debt burden is considerable, as well as its per capita gross national product (GNP). The World Bank and the International Monetary Fund (IMF) have implemented some structural and interest-free credits as well to support investments in the agriculture, livestock, and transportation sectors, unfortunately these have had limited impact. The World Bank and IMF are now encouraging the government to concentrate exclusively on implementing much-needed economic reforms to jumpstart the economy and defining its fundamental priorities with the aim of alleviating poverty. As a result, many of the state-owned business entities have been privatized and limited e fforts have been made to standardize and simplify labor and investment codes and to address problems of corruption. The C.A.R. Government has adopted the Central African Economic and Monetary Community (CEMAC) Charter of Investment, and is in the process of adopting a new labor code. Furthermore the CAR received debt relief in June 2009 by the World Bank and the IMF [IMF, (2010)].It reached the final completion-point stage of the Heavily Indebted Poor Countries Initiative [IMF, (2010)]. Since its independence, the CAR has been unstable and endured several coups; David Dacko ended the Bokassa era in a coup in 1979 [International Crisis Group, (2007)]. David Dacko was overthrown by a coup led by the General Andre Kolingba two years later. For the first time, he allowed multi-party presidential elections in 1993 and was defeated in the first round by his successor Ange-Felix Patasse. However, 1997 the discontent of unpaid soldiers resulted in a successive wave of strikes and uprisings from the latter [International Crisis Group, (2007)]. . The same year the French troop pulled out and Paris financed a group of French-speaking African countries to create a peacekeeping force. This leads to the creation of the UN Mission to the Central African Republic, or Minurca [International Crisis Group, (2008)]. In 1999 Ange-Felix Patasse won the elections a second time however there were allegations of electoral fraud. He went into exile in Togo after being overthrown in a coup in 2003 by General Bozize the current president of the CAR. The legacy of years of unrest has generated an illegal weapons proliferation across the CAR. Rebels armed groups are active in the volatile north. The unrest has displaced tens of thousands of Central Africans; many of them have crossed the border into Chad. Under Bozize era, some progress towards ending the conflict was made in 2008, when peace talks led to an agreement committing two of the main rebel groups to disarm [IMF,(2010)]. The process culminated with the creation of a national unity government incorporating two rebel leaders in early 2009. 2.3 A Modest recovery Since 2004 the Central African Republic has been able to achieve a modest economic growth [IMF, (2010)]; firstly, it gained full debt relief from the IMF and the World Bank in June 2009. Secondly it concluded a current Poverty Reduction and Growth Facility programme in June 2010. This successfully led to the achievement of a three-year programme with the IMF especially after the success of the Emergency Post-Conflict Assistance Programs. Furthermore, the CARs government implemented a macroeconomic management policies and necessary structural reforms that led to a modest recovery from the economic shock in 2009. The government worked hard to further strengthen the credibility of public financial management, enhance public infrastructure, and intensify foreign donors and the private sector investment climate [US department, (2010)] as well. Since President Bozize leads the CAR some encouraging result has been noticed by the Internationals institutions like IMF or World Bank. The CARs government understood that it was its priority to restore security and peace in the country. The government established what it called the â€Å"inclusive political dialogue† that gathered the political opposition and the government together to seek for security and peace for the country. That initiative was successful in the extent that it was part of the demobilization, the disarmament and reintegration program which led to the smooth conduct of the elections in 2010 [IMF, (2010)]. In terms of building infrastructure and climate investment the CARs government has a lot to do in order to unlock the countrys economic situation. The government has no other choice than on the one hand relying on foreign partners and on the other hand must engage coherent and filmy policies over the long term to come out of its cycle of poverty [IMF, (2010)]. III- Chinese engagement with the CAR 3.1- The political relations AT many occasions the CAR cut off and resumed its diplomatic relations with the PRC and Taiwan. The diplomatic ties between China and the Central African Republic experienced ups and downs: In 1962, the CAR recognized Taiwan as part of Chinas territory and established for the first time diplomatic ties with the island. Two years later, it broke off its diplomatic link with Taiwan and established diplomatic relations with China. The CAR resumed diplomatic ties Taiwan third times. Leaders of both side exchanged high levelcontacts. These underline the importance a state attaches to its relations with another. 3.2 Trade and Economic Relations In November 2000, China and Central Africa Republic signed an agreement on trade, economic and technical cooperation [Chinese Foreign Ministry, (2006)]. In August 2004, both countries signed a bilateral cooperation agreement in fields such as energy, infrastructure and agriculture[Holslag, (2006)]. Today, China is an important commercial partner for the CAR.In 2002, trade volume between both nations accounted for US$ 1.944 million with US$ 0.687 stemming from Chinas export and US$ 1.257 million from the Chinas import. The CAR mainly exports timber and cotton to China and imports principally footwear, textiles and electrical and mechanical products from China [Chinese Foreign Ministry, (2006)]. China has been an active investor in the Central African Republic, especially in construction, and has been involved in a number of joint cooperative ventures with the government, covering health, agriculture, stock-breeding, communications and trade .It undertook numerous projects in CAR and built for instance the Boali agricultural technological station, a training center for straw-bamboo weaving handcrafts, the new stadium, a clinic in the presidential house, the Bangui hospital and agricultural project, the extension work of the Pingpo Radio Transmitting Station, etc[Chinese Foreign Ministry, (2006)]. During his visit in China in 2009, President Francois Bozize called on China to invest in its country 3.3 Exchanges in other fields After signing a joint communiquà © in August 20th, China and the CAR signed in November 1976 an agreement on sending Chinese medical teams to Central Africa Republic. In June 1980 both countries signed an agreement on cultural cooperation and renewed it in June 1998 look for the number of students in China. Since 1997, China has begun to accept Central African students for study and on the job training. Today both parties acknowledge the necessity for the CAR to enhance its self-development ability as well as its local Human Resources training IV- China- CAR: Why the slow progress? One wonders what is happening in Central African Republic as the country is blessed with natural resources but still ranking as one the poorest in the world. International news portrays it as a â€Å"phantom state† [African report, (2007)]. Since the resuming of bilateral cooperation with China in 1998, the relation is experiencing smoothly and slowly evolution. In 2009, during a high level visit to China President Francois Bozize spoke to his Chinese counterpart Hu Jintao and expressly called for Chinese investment in its country [BBC, (2009); AFP, (2009); China consulate, (2011)]. China is now an important economic partner for the CAR [BBC, (2009)]. However in an article published in 2009, the BBC wrote that the â€Å"CAR is a much less valuable economic partner to China†. Considering that statement one wonders whether China really considers the CAR as a less valuable partner. How the Chinese governments perceive the CAR? How does the CAR perceive the Chinese involvement in the country? The CARs government welcomes the Chinese aid in spite the political instability in the country, especially in the north [African Report, (2007)].However, China doesnt deter to invest in it by building the new 20.000 seats-stadium, schools, hospitals, etc. Regarding to that assistance, can it be stated that the CAR is â€Å"a much less valuable† for China? The CARs situation is peculiar and really needs from the scholars, journalist, central African leaders, the civil society, all the stakeholders as well, further attention.