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Thursday, June 21, 2012
Inov-8 Bare xf 210 for summer!
Summer is here in full effect and I can't get enough. Working out in the excessive hit may hit the wrong chord for some, but for me, there is not better place to be. Maybe its because it feels like the Games or the Dirty South regional of 2010, but working out with just the sun overhead kicks ass. Its also a good excuse for me to pound Vita Coco coconut water all day long (our local Costco aways has Vita Coco in stock, WIN).
One of the worst problems with the summer heat though is sweaty shoes. Without shoes that breath well, you are stuck with your feet in their own personal sauna. Imagine being in a sauna at a nudist retirement home. Thats what your feet are going through. On the flip-flop side, its not ideal to workout in sandals, frankly I have never seen this and I wouldn't recommend it. Thats asking for broken toes and no one wants to see your yellow nails anyway.
What if you could get that same feeling of wearing sandals while wearing shoes. And what if those shoes were made for Crossfit by a company that has been there since the beginning to support Crossfit. Well the Inov-8 Bare xf 210 are those shoes.
When you first put them on you will notice right away that your feet feel nothing else but the ground There is no cushioning and no nonsense. Just zero drop. If you have never used minimalist shoes before I wouldn't recommend starting in them for runs, but easing into them from some other Inov-8 shoes (F-lite 230, 220, or 195s).
After hours of working out my feet remained cool and I did not have any issues with stank feet.
They work particularly well in most Crossfit workouts and considerably well for rope climbs.
In addition, they are perfect for lounging around, drankin, going to movies, or just living life outside of Crossfit.
All in all, they are versatile and I highly recommend picking up a pair for summer. Get a pic of them poolside while watching the Crossfit Games in July.
Monday, June 11, 2012
This past weekend I had the opportunity to go out of town and train with some high caliber games athletes. I spent a good amount of time on the road (~9 hours) to get there and I'm not one that likes to stop when I need to get somewhere. Driving a stick shift car on the road and not stopping to eat can be a hassle, especially with stop and go traffic.
I'm already a fan of the paleo kits, which include nuts, berries, and jerky, but they can be messy to eat in the car. The paleo stix on the other hand were freaking awesome. You just tear em open and viola. No mess, just meat goodness. The flavor is pretty much a standard beef stick flavor and didn't blow my socks off as something revolutionary but it was good. They are certainly not as chewy as the kits too, so you won't get lockjaw trying to eat them. The empty package didn't have the sticky and juicy mess like the kit, and the wrapper left no stains on the floor mats of my car which was appreciated. I couldn't tell the difference between the grass-fed and regular version so that will be up to you to decide. Word of caution if you hit some beers later on. The potency of those burps will be enhanced. All in all I give them a thumbs up. Less crap than whats in the gas station food isle and better for you.
Thursday, May 31, 2012
Thursday, September 1, 2011
Since I've been gone....
I have eventually found out that it ain't easy to keep up with a blog. You think, "I don't need to write this shit, everyone knows what I'm doing." Then I decided screw it, I'll blog anyway, although I'm pretty sure my fan base of 1 hasn't grown.
Sooooooo, since my last post a lot has happened.
First I got to say that I'm very proud of our team's finish at the 2011 Reebok Crossfit Games. The experience was phenomenal and I wouldn't trade a second of it for the world. Everyone put forth 100% effort and it was awesome to be rewarded for that with a 16th place finish. I'm hoping to continue to train hard and make it back with the team next year. I love the team competition. Even if I was good enough to go as an individual, I'd still do the team. For me the individual stuff is too much pressure, but thats just me. If I screw up someone on the team can step up or if they screw up I can step up. If we both screw up then...its still fun.
Second, Graduate school has made it more difficult to train when I want to, but I think I'm finding a balance which has been great. The added rest has been extremely noticeable on my strength numbers which to me now, seem ridiculous. I don't think I would have thought I would be putting up the numbers I am now, a year ago. I probably would have told you thats impossible. So if you are reading this and you haven't been progressing the way you want and have been Rich Froning your wods, then you may want to back off a little and see what happens. Frankly, I don't know how some of the superstars of Crossfit balance law school, med school, or whatever with high volume training. I just don't have time for shit anymore and have to choose activities carefully. If you are one of those people who hits the gym 10x a day and has a professional career not athletic related, I despise you.
The most important component to my training though is that I have been pushing it for myself, not anyone else. I don't need to prove my strength/crossfit worth by comparing this and that with other people. You need to be thankful for what you have, what you are given to improve, and how you use that improvement. If you only care about beating someone else then you will never be the best that you can be.....G.I. Joe or something.
Aside from training, school has been a welcomed gift. I've been wanting to go back to school for a while and I think I have applied to more programs than anyone I can think of so getting in was like winning the lottery. Now that I'm doing well in the program I can continue to focus on what I need to do to get a good project and less on trying to prove to myself that I belong in the there.
Friday, July 22, 2011
Off to see the wizard!
This is gonna be my last post before the games because I don't want this blog to suck any drop of work capacity that I have left in my body, and staying current with it during this time is about as easy as isolating the Higgs particle with a paper clip, snorkel, and slice of swiss cheese. Just to make it clear I'm saving my body for two things: 1)Destroying the 2011 Reebok Crossfit Games with my Bros and Bras, and 2)Slaying the glutonious orgy of food and non exercise that will ensue afterwards. I've already made a list of food and other stuff that I plan on annhialating so I assure you that I have thought this out thoroughly and calculated it with extreme precision, Crossfit New England eat your heart out. I'm not entirely sure who is all out there, but I'm open to suggestions. I don't even care if you are in a different city, state, country, whatever. If you know of some must have food, post that shit to comments and I will put it on my list. Now, I also understand that as individual human beings, some of us like caramelized sheeps testicles or some scary shit like that....thats not me. If it sounds slightly weird, I'll consider it. If it sounds like you ate that shit to survive just so a lost villager could maybe get you help...I'll pass. This will be both desserts and regular eats (nonpaleo/non zone only)So hit me up.
My list thus far in no particular order:
1) Gooey Louie Buttercake-all of them
2) The Elegant Farmer apple pie in a brown bag- straight from Wisconsin
3) Krispy Kreme- glazed and Bostom Creme
4)Oreo Klondike Sandwich
5)Cheesecake Factory chocolate tuxedo cream cheesecake
6)Deep Fried Pickles-Tylers
7)Whole Foods Harvest Berry Pie
8) The Pit- Sangria, banana bread pudding, honey infused cornbread
9)Mellow Mushroom garlic glazed pretzels
10)An entire pizza from CPK
11)Jeni's salty caramel ice cream
12)Goodberry's Sandstorm
My list thus far in no particular order:
1) Gooey Louie Buttercake-all of them
2) The Elegant Farmer apple pie in a brown bag- straight from Wisconsin
3) Krispy Kreme- glazed and Bostom Creme
4)Oreo Klondike Sandwich
5)Cheesecake Factory chocolate tuxedo cream cheesecake
6)Deep Fried Pickles-Tylers
7)Whole Foods Harvest Berry Pie
8) The Pit- Sangria, banana bread pudding, honey infused cornbread
9)Mellow Mushroom garlic glazed pretzels
10)An entire pizza from CPK
11)Jeni's salty caramel ice cream
12)Goodberry's Sandstorm
Monday, July 11, 2011
When I think of the S.W.A.T. team I think of some serious shit going down. Guns blazin, Seagal kickin, Kano rip your heart out shitstorm with a fu manchu. Now, I'm not one that does illegal activities, unless you can catch me ridin dirty...but seriously being around a S.W.A.T. training facility made me a little nervous. Not sure why, cause it's not like they were gonna break me down for going a little overboard on the tiramasu samples at whole foods last week (get some foos). Anyway, I took one look at the course they had set up for us and I was all smiles. You see, I always wanted to be a navy seal, but never made the commitment. I was always intrigued by the log PT and mini tests they put you through during the de-selection process. Since I never went through BUDS, this was my time to shine.
The first workout we did was a team log carry and tire spin?. 3 of us carried the log, and 2 spun a huge ass tire for a good amount of distance, maybe a mile and a half. I spent most of my time carrying the log, which I really didn't mind at all. Frankly, I thought it was easier. We followed a park trail and were passed by yoggers who proceeded to look at us like we left our minds to bake in our cars. I took a break from the log to roll the tire with Christmas which turned out to be friggin hilarious. As soon as placed my hands on the tire and helped guide it along the trail we hit the top of a hill. I decided, wtf, let's just allow this here tire to navigate itself down the hill, that'll be easier on us........I watched in shear horror as it rolled out of control directly to my unsuspecting teammates. The human bowling game had now commenced. I ran as fast as I could to re-direct the tire, which worked with minimal success. The tire slammed into a close tree and toppled over. I was laughing my ass off in my head. I don't know why it was so funny...wait yeah I do. I quickly switched out onto the log as to not continue my path of destruction. When we finished the workout my shoulders were bruised to shit, but I felt awesome.
The second workout was a hodgepodge of kickass stuff. The S.W.A.T. instructors made it more interesting by creating a story that coincided with what we were physically doing. I don't remember the story verbatim but, my version is just as accurate. So it starts off as such:
There's a serial killer on the loose. This guy is sick, and I mean fucking disgusting. He's known to murder crossfitters left and right and he's got a taste for LuLu's lemons. His victim's bodies are scattered all over the backyard, but they are still alive (6 heavy ass tires). Your job is to stack the bodies one on top of another so the extraction team can pull em out. (This duty was pretty funny because the last tire had to be launched off your body onto the top. I basically still have tire tread imprinted on my thighs from throwing it up there.) Once the bodies are stacked you have to bring the rocket launchers, AK's, C4, and other gear weighing 70# over a 6ft. wall. Throw the gear over the wall, jump over, and continue. Once over the walls you will find a log. Flip that shit. (Not sure where this fit into the story). Next is a disabled cop car. Your job is to push the car to a safe location and bring gasoline jugs over to the cop car and fill it up. The getaway vehicle is now ready to go. Next scale the wall of the compound and setup a sniper post on the roof. When you see the scumbag blow his brains out. The End.
Anyway you get the idea. Probably not the same exact story the S.W.A.T. officer told, but close. It was alot more exciting than most workout stuff I do on a regular basis and so I was pretty thankful to get the opportunity to do that. Which I guess brings up my main point.
I wouldnt have been able to do any of this had it not been for Crossfit. I don't have sponsors or endorsement deals, but the opportunities like the S.W.A.T. team training make life rich. It's experiences on a smaller scale that sometimes mean the most. We all talk about functional fitness but how many of us get broken down in the gym and look like handicap assholes in real life? Get out of the gym and do something different!
Friday, July 1, 2011
Haters need hatin!
One of the highlights of my day is simply checking out all the hatin on Crossfit and Crossfitters. Its really funny shit if you haven't read it. Most of it is clever, but some of it is like Kathy Griffin's plastic surgery, it just doesn't make sense, but is filled with plenty of sarcasm. There's videos, blogs, forums, and even entire websites dedicated to hatin on Crossfit. It's getting more popular and growing faster than your upside down tomato garden. Just google "hate crossfit" and you'll pretty much get most of it. I'm seeing a trend so I anticipate more and more haters to come.
I've found of few of these groups, for instance:
Crossfit Haters group 1 are simply the ex-crossfitters turned lifters. The story starts off similar among this group. They were people who used to be fat or out of shape and by shear chance they found crossfit and got shredded. Sooner or later the testosterone button was getting pushed a little too frequently and finally they decided that met-cons are for weak ass bitches. Their only desire is to be as big, and strong, and scary as possible. They yearn to hit that high-school reunion and Hulk Smash everyone into oblivion. They enjoy only non-paleo 70s BIG meals, hard liquor, porn, non-WADA approved supplements, and yelling. They will continuously knock crossfitters as idiots in the strength department and propose their method as the one that works. I don't need to remind people that Spencer Hendel backsquats 500#+ and squat cleans 335#....yeh he should probably stop being so weak and just focus on strength.
Crossfit Haters group 2 are basically the globo fooz. Back and bi's, chest and tri's etc. You can spot em out from their torn up shirts from football glory days and they periodically will check out their nipples in the mirrors just to make sure they are hard. They don't like crossfit because it's messing with their swagger. Gym, Tan, non-laundry....cuz my smell is musky.
Crossfit Haters group 3 are simply the roided out lifters. They want to rape and pillage, pillage and rape. They wish nothing more than the end of Crossfit, and their sport of....being huge? to come to fruition. They charge ~$150-250 for a phone consult and much more if you decide to take their advice for training/nutrition. They also make fun of crossfitters paying 1k for a cert., but at the same time charge more than Bono for their time.
Crossfit Haters group 4 are actual crossfitters who hate other crossfitters.
I'm trying to figure this group out but am having a hard time understanding these people.
If you know any other haters post to comments.
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Developer's Comments
2.1.2 (12/06/12) - Fixed... (sort of) the weird aspect ratio of videos.
2.1.1 (12/03/12) - Oops spoke to soon, fixed a small error with the detection of the flash player.
2.1.0 (12/03/12) - Fixed script after some changes broke it.
2.0.0 (11/11/12) - Complete rewrite of script to fix changes made at dramafever.com
1.2.3 (10/22/11) - Fixed... stuff; Script now only shows available episodes for shows don't have all episodes available yet; This update is sooooo long overdue I apologize immensely for those who have been waiting.
1.2.2 (08/12/11) - Updated so player controls don't appear.
1.2.1 (08/11/11) - Fixed anything I could find that was broken.
1.2.0 (05/02/11) - Moved script architecture to "PlayOn Premium" API, subtitles for all videos, are now available.
1.1.1 (04/25/11) - Fixed problem caused by change at Google Sites that broke the script and it's auto-update code.
1.1.0 (01/15/11)
-Performed major maintenance and fixed all broken things.
1.0.4 (04/15/10)
-Fixed script after change to site broke it.
-Now gets the longer more indepth show descriptions.
-Very minor update, added ability to see upcoming dramas and see recommendend dramas
1.0.2 (02/06/10)
-Subcategories are now displayed dynamically; meaning that if new categories are added later, the script won't have to be updated to display them.
-Added view types "Year" and "Network" to reflect changes on website.
1.0.1 (01/25/10)
-Fixed duration times so they are actual episode lengths
-Fixed "Lovers in Prague" non-showing episodes
-Added "Episode ... is not available" when an episode is premium and not yet watchable for free.
1.0.0 (01/23/10)
-Uploaded Original Script
Because dramafever.com is a legitamite site with agreements from major Asian broadcasters, access to video content is restricted to the US/Canada only.
Some episodes of more recent shows are not immediately available to free users of the service. They usually become available within a week or two.
nope wrote
works but wish the subtitle issue was solved. ah well.2011-01-18 18:51:59
adonanonymous wrote
Works great. Very awesome.2010-02-23 12:25:50

subset: commoncrawl


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Slowly but surely, three more months passed, and the time had come. Except…..still no baby.
Ila was lying in bed, being a full six months pregnant and full term, but the baby had not made to show himself yet. As uncomfortable as she had been a month ago, that now seemed easy. Her feet had swollen and she could barely walk the short distances she had to make to the chamber pot and the bath tub. Walking was a misnomer; rather she had to waddle now.
No matter how she sat up or lay in bed she simply could not get comfortable. She didn't even want Thorin to hold her while they slept she was so uncomfortable in her skin and no position provided adequate relief. Her back hurt, her feet hurt, she couldn't hold her bladder long at all for her big baby pressing on it, and she was hot all the time. Everything zapped her energy, what little there was, even eating. Getting up was exhausting and she dreaded every time she had to do it. Dwala was required to be close by at all times now and a guard was posted outside her door. He would call for the Oin and then for Thorin when the time came.
If it would ever come. Ila was beside herself with impatience and cabin fever to boot. She had hoped he might come a day or two early but now she hoped he came at all. Thorin too was anxious, but constantly reassured his wife.
"Darling," he would soothe her, "he will come when he is ready and not a moment too soon. Best he is late and healthy than early and sickly."
Ila knew he was right, but that was easy for him to say when he didn't have a giant dwarfling in his belly.
Said dwarfling was rather anxious himself. He had begun to kick more and turn around and roll over inside there, at least it felt like that to Ila. Sometimes she could even see a foot or elbow protruding from her belly, and the first time had been rather shocking. She had hoped that meant he was making his way down to come out finally, but such had not happened yet. Perhaps he was just too comfortable in there, sleeping all day and getting all the food he wanted, to want to come out.
Whatever the case, Ila stared at the ceiling as she was prone to do now when she didn't have visitors. Dwala was tidying the antechamber and humming to herself as she did, and Ila knew she had terrible cabin fever too. She didn't say so but she knew she must and couldn't blame her. And of course she wanted to be with her husband, whom she saw very little of now thanks to her condition.
The only thing that had made said condition bearable in the past month were the visitors that she'd had. Kili had visited a few times a week to 'keep her entertained' as he put it, which he insisted was his most pressing and only real royal duty at the moment. Fili had also surprised her and visited a few times as well, just as she had hoped. And their mother had kept her word about visiting too. Her visits were usually filled with lots of baby talk, telling her what to expect and how to do everything she would have to do with a baby. Ila had not grown up around lots of babies, and dwarves did not reproduce very often anyway being there were so few females of course, so Ila was thankful for all the instruction. Dis gave her advice and Fili and Kili made her laugh. That had been a nice balance through the past few weeks, and she always had her love to look forward to every night, even if they couldn't enjoy each other fully for a while yet.
As Ila stared at the ceiling pondering nothing really, she felt the sudden urge to urinate. She sighed deeply. Not again, she thought. Drinking almost wasn't worth it anymore, given the fact that water went straight through her and the chamber pot may as well be a thousand leagues away.
Sighing again and throwing back the covers, Ila scooted slowly and awkwardly to the side of the bed. Dwala heard her moving from the other room and came to help.
"You are supposed to tell me when you want to get up. What if I hadn't heard you and you'd fallen?" she scolded.
"I declare I will be happy when this baby comes if for nothing else that I won't be treated like a little dwarfling myself anymore." Ila said but without any real irritation.
"I am fine, really." Ila said and waddled her way over to the tiny room at the other end of the great bed chamber. Dwala followed slowly behind anyway and kept an eye on her. While she waited she went to fluff Ila's pillows, though she doubted it would make any difference. Her attention was seized however when she heard Ila whimper loudly.
Dwala dropped a pillow and rushed over to Ila, who had emerged but was slightly bent over and clutching her abdomen.
"Ila, Ila is it time?" Dwala inquired anxiously.
Ila was still bent over, her eyes fixed on nothing and not looking at Dwala. "I….I…." and then her sentence was broken off by a groan of pain.
"Oh, oh, oh, it's time. It's time!" Dwala was repeating since Ila was at a loss for words.
"Can you walk?" Dwala asked but Ila whimpered again. She looked down and noticed fluid, what looked like water dripping onto the floor beneath Ila, and remembered that was a sign Dis had warned of. Dwala then summoned her courage and took Ila by the shoulders from behind and slowly guided her to the bed. Ila was breathing harder now and the journey across the room was painfully slow. When they reached the bed Ila backed onto it and just rested on the side for a moment, still holding her belly and breathing hard, her face strained. Another moment and she calmed a bit, and Dwala took the opportunity to run and tell the guard to fetch Oin.
Ila sat there in that brief moment alone on the side of the bed, absolutely terrified. She had been waiting for this moment for months, had been fretting over when it could finally come for weeks, and now that it was finally here she was scared.
What if I die? What if I can't deliver? What if my baby is not well? Will I be able to bear the pain?
Doubt and fear spun around like a whirlwind in her mind, so intense that she momentarily forgot the pain and discomfort she already felt. She wanted Thorin there, but knew that childbirth was nothing for males to be present for.
Dwala returned and began to rearrange the pillows, trying to make a nice support system for Ila to lie back on.
"Can you move? Can you lay down?" she asked and Ila took a deep breath and nodded that she could. Slowly she pushed herself back into the pillows and onto her back, panting a little and wincing as she lowered her aching and swollen body.
Ila said nothing, only closed her eyes and tried to brace herself for what she knew would be more pain. She thought about trying to control her breathing and tried even harder not to be scared. Dwala brushed the hair out of her face and grabbed a fan to give her a gentle breeze, seeing beads of sweat already forming on her brow.
Dwala silently hoped it would not take ages for Oin to climb the many steps that lay between him and this chamber. He was old and slow, and this was no time for dallying. She didn't know what to do…..perhaps she should run across the landing and get Lady Dis? Or perhaps Ila would not want her there… she just didn't know and now didn't seem the time to ask.
Ila began to whimper in pain again and reached for her belly. She kicked the blankets away, hotter and more uncomfortable than ever now. Worried and scared herself, she looked to her own stomach briefly, wondering if she too were of a certain condition. She didn't look different yet, but she had begun to feel a bit nauseous in the mornings and her body simply felt different, for lack of a better way to explain it. She was now rather afraid of being with child, seeing Ila stretched out in pain.
Dwala looked to the door. Where was that ancient dwarf?!
Downstairs, Thorin sat in council with Fili at his side and his advisors around the table as usual, each briefing the king in turn on items for which they were responsible. Thorin was listening intently as he always did, but not with his usual relaxed posture in which he typically draped himself across this great chair in these meetings. He was more alert, on edge even, as his heir was overdue and could arrive at any second. He was certainly eager for the time to come, first to hold his son but then of course to have his wife back. He actually couldn't say which he wanted more.
As Gloin was reading over the recent acquisitions from the weekly trading trip to Dale, a young guard hastily entered the room.
Thorin's head snapped up and he looked to the dwarf expectantly.
"Pardon my, my lord, but—"the dwarf began as he bowed but Thorin cut him off.
"Enough of that, speak your purpose," he commanded, hoping it was news of his wife.
The young dwarf choked a little after being interrupted by his king and Fili, anxious himself, cut in now.
"Speak!" was all he urged and all eyes were on the poor dwarf.
"My lord the queen is expected to give birth shortly—" he spat out as quickly as he could but Thorin hardly heard it. He practically erupted from his chair, nearly knocking into the table, and the dwarves around him cheered happily. Fili smiled widely and got up too to follow his uncle who was already making for the door.
"Has Oin been called for?" he demanded of the dwarf without looking at him.
"Yes your majesty he is on his way now." He answered.
Fili caught up with his uncle. "Shall I make an announcement?"
"No, not yet. It may be hours yet. Best not to excite everyone." Thorin replied, his face as serious as if he were walking straight onto a battlefield.
"Right. I will find Kili and then go to mother." Fili replied and went off to do just that.
Thorin continued towards their chamber alone now, not noticing a single soul as he did. The time had finally come. His love was about to bring their perfect child into the world. His heir, the heir of Durin, the continuation of the great line. He could not wait to hold him in his arms. And his wife.
His precious wife. He silently pleaded with Mahal to keep her safe. To keep her strong. To help her deliver. Keep her healthy, and keep her with him.
He saw no sign of Oin on his way to his bedchamber and hoped that meant he was already there and tending to his wife. He was taking three steps at a time up the stairs in his heavy boots, and once he reached the landing broke into a jog.
He entered his chamber and slowed as he approached the bedroom, the door open. Dwala peered through the opening and then spoke to Ila, telling her he had come.
Thorin entered the bedroom and was immediately disappointed to see that Oin had not arrived yet. That irritation soon passed though as he beheld his ailing wife.
Ila was moaning and whimpering, sweat on her brow and the covers kicked away from her. Her eyes were closed in pain and she didn't seem to know he was there.
Gently, he sat on the bedside and laid his hand over hers, which he noticed was trembling.
"My love," he said quietly and Ila opened her eyes and turned her head towards him.
Pain etched her face and her eyes were afraid. "Thorin…" was all she said.
"You are alight my love. Just a little bit of courage now." He assured her and then turned to Dwala.
"Fetch my sister, Oin is taking too long." He commanded and Dwala hurried from the room.
"Shhhhh, my love." Thorin tried to comfort Ila. She has closed her eyes again and her breathing was slowing a more normal pace. He held her hand and she didn't grip as tightly. He had never watched a dame in labor, so he honestly didn't know what to expect. This was probably normal but he had no idea. He cursed himself for not making provisional arrangements for Oin closer to their chamber and requiring him to stay there so he could quickly come to aide. Where was he? And where was Dwala with his sister? Thorin made himself remain calm though, knowing he must give his wife nothing to worry over.
Dwala knocked hard and purposefully on the door, hoping Lady Dis was not indisposed. A moment or two later it opened and she was met by her lady in waiting, Layla.
"His majesty asks for his lady sister to come to the queen's side. Her child pains are upon her and Oin is yet to arrive…" Dwala stammered out. Layla nodded and quickly turned to fetch her lady. Dwala stood there in the doorway for a moment, waiting or not knowing what to do, then shook herself and ran back to Ila. Dis knew the way, obviously.
Dwala arrived to see Oin still not there and sighed with annoyance. Ila was panting and moaning in pain again, Thorin looked worried sick at her side. Dwala wished she could tell him not to worry, but honestly she had never seen a baby dwarf being born and couldn't reassure him, or herself.
Dwala went to grab a basin and fill it with water, if for nothing else to cleanse her forehead and chest from sweat. Dis quickly entered the room then and threw her enormous skirts aside to sit on the bed next to Thorin.
He looked to his sister with pleading eyes, asking if all was well and normal. She knowingly placed a hand on his shoulder and reassured him.
"You should go, this is no place for husbands. Or any dwarf other than a healer." She said.
Thorin said nothing but made a face at her that said he didn't want to.
"There dear, you are fine." Dis soothed Ila, brushing her hair from her face.
"Here, you should sit up more, and tuck your legs a bit." Dis instructed gently and Ila slowly tried to do so. Layla had come to stand behind her mistress, waiting for any instruction. Thorin had not moved from where he sat, but knowing he probably did not want to actually see their baby being born. He had seen many difficult and bloody things in his time, but he imagined he'd have no stomach for childbirth. Still, he hated the thought of abandoning his wife in her hour of need.
Though, he wasn't much help. He was afraid to touch her for putting her in more pain, and she barely seemed to register that he was even there.
Finally, they heard footsteps and labored breaths coming near them, signaling that Oin had finally arrived.
He hobbled through the door with a bag in one hand, panting as hard as Ila.
"The stairs harder than they used to be?" Thorin said through gritted teeth, trying not to show his annoyance.
"Aye well I'm not getting any younger now am I? And nor are you." Oin returned.
"Now, now give the queen and I some space." He said gesturing at them all crowded around the bed.
"You should probably go, actually," He said to Thorin and Dis gave him a look to reinforce that statement.
Thorin looked to Ila and bent down to kiss her gently on the forehead. "I will be right outside my love. Be not afraid." He whispered and she squeezed his hand hard, opening her pain stricken eyes to blink pitifully at him.
Oin was right; he did not want to watch his wife in such pain, being powerless to help her. He knew this was the natural way of things but it didn't make him feel any better about what she had to face.
Thorin left the room and went to sit in their antechamber, listening for any progress on the other side of the door. There was no knowing how long this would take, it could take hours or even a couple of days in some extreme cases he'd heard of. He fiddled with his hands, looking at the floor, having no clue what to do with himself.
On the other side of the door, Ila was feeling a little better, the pain easing off as it had before, but she knew it would come again. She could feel the skin of her belly tightening and the baby moving inside her. The pain in her abdomen was worse every time, and she wondered how long before it became absolutely unbearable.
She was so focused on the pain that she hardly cared at what a compromising position she was now in, her gown pushed up to expose her belly that Oin had felt for the position of the baby, then a cloth spread over her knees to provide some privacy. Oin stood at the end of the bed, monitoring any progress, of which there was currently none.
Dis was at one side and Dwala at the other, trying to keep her cool and reminding her to breathe deeply. She wondered if she would know when to push, and Oin said it was not time yet. But when would it be? Would she even know how to push? Ila let a tear slip from the corner of one eye, which Dis wiped away.
"It is alright my dear, you are doing well. You won't even remember this when you see your baby." She encouraged her in a motherly tone, and Ila was very happy to have her there, being the only one in the room that had done this herself.
Dwala was nervous but hid it, staying strong for Ila and assisting Oin as he needed it. Her hand went to her own stomach to see if it felt different, but it didn't. She did however feel a bit sickly, but that could very well be due to current circumstances. She silently prayed this would be a quick delivery, but there was no way to tell yet.
Thorin was so lost in thought and worry that he didn't even hear Fili and Kili walk in.
"Uncle," Fili said and his head shot up. "Anything yet?
"No..." Thorin shook his head and went back to staring at the floor. "But it's not been long."
Fili and Kili sat across from their uncle on the couch that sat in the middle of the room, determined to wait it out with him. They both noted to themselves that they weren't sure they'd ever seen him that worried before, and Thorin had faced my trials and foes.
Kili was worried himself, but made sure to only display the appropriate amount of that. He had a cause to be concerned, that being his aunt and the heir, but no cause to be on the verge of vomiting which is how he really felt. He worried for her safety, as the childbed was a dangerous place for a female of any race. He looked at the floor too, not wanting to give anything away and not knowing what he could possibly say to make anyone feel better. Fili was the only that tried to make conversation, and he failed horribly. As such, the three of them just sat in uncomfortable silence.
As Ila had feared, the pain had gotten worse and worse. She was drenched in sweat and gripping the pillows, even when she wasn't in pain. Her belly continued to feel tight and her back ached and she dug her heels into the bed. It felt like it had been hours but it had only been about thirty minutes.
"You need to get up and walk around a bit between your pains," Oin said and Ila's eyes shot open.
"What?!" she called over her stomach at him.
"The baby is not ready to come out and you are not ready either. It will be a little while yet and it will could make things progress faster."
Ila rolled her eyes at the ceiling. She did not want to try and get up. But with Dis and Dwala's help she did just that, walking slowly in circles around the room, with Dis and Dwala each holding an arm to steady her, until her pains hit again. This was easier than she expected, and made the time pass a little faster, but Ila was growing extremely impatient.
Not that the baby cared, because this went on for another three hours. Ila finally reached the point where she could not walk anymore and she laid down for good. Oin checked and lowered the cloth again. "Almost time to start pushing your highness."
Ila exhaled and groaned as she shifted on her back. Dwala held her hand and fanned her some more. Outside, Kili and Fili were laying over the couch now and Thorin was laid back in the chair. Dwalin had come to sit with his friend as well. No one said much, but twiddled their thumbs or dozed off while they waited. That was until Ila's screams jolted them all alert.
Thorin looked with alarm to the door. He wanted to rush in there but two more screams and she was silent again. No one said anything but looked around nervously. Kili felt sick and Thorin pushed his hair back feeling sick too. Dwalin looked particularly uncomfortable and wondered how he would be able to deal with this when and if Dwala became pregnant.
Oin had declared with Ila's scream that it was finally time to push. With her legs tucked and Dwala and Dis supporting her from behind, Ila tried to do just that. She felt like her lower body was splitting apart. The pains were rapid now and she barely had time to rest between them. Her back was killing her and the pressure in her bottom was intense and painful. She was trying not to scream but she just couldn't help it anymore. Everyone urged her to push and Ila wanted to scream at them to shut-up. Lucky for them she couldn't talk anymore.
The constant chorus of screams had Thorin thoroughly unsettled now, and he had taken to pacing in the hallway but with the door open in case someone called for him. Kili feared that they were listening to Ila dying on the other side of that door and doing nothing to stop it. Fili and Dwalin looked horribly uncomfortable and finally Dwalin got up to join Thorin in the hallway. Kili wanted to break those doors open and help her somehow. He fidgeted in his seat and Fili shifted uncomfortably too before going to stand in the hallway too.
Now alone, Kili clasped his hands and prayed to Mahal to deliver Ila safely from this. He would give anything for her to be alright, take any vow in exchange for her life. He squeezed his eyes shut tight as he heard Ila wail again, pleading for the babe to come into the world already.
Ila suddenly felt a burning sensation and wailed harder than ever.
"The head is coming out," Oin narrated.
"Push Ila, push you're almost there." Dwala urged.
Ila was in excruciating and searing pain. She barely had the energy to scream, but she managed. Dwala and Dis pushed on her back and helped her. As the baby's head passed, Ila screamed so loud she was sure all of Erebor heard it, perhaps even Dale.
"Once more!" Oin cried and Ila fell back into Dis and Dwala's arms, sobbing and gasping for air.
They pushed her back into position and urged her on one last time. Ila summoned just enough energy from where she knew not, then finally pushed her baby free.
Ila immediately collapsed on the bed, chest heaving and breath erratic. Her whole body ached as though she had been trampled by herd of wild horses. She closed her eyes for a split second before she heard it and they snapped back open.
The cries of her baby. Ila looked down to the end of the bed, waiting to see, waiting to hold him. She held her breath as Oin lifted the babe.
Dis and Dwala looked on in breathless anticipation with her, ready to see the royal baby, the heir.
Oin's ancient hands lifted the healthy, squirming, crying, and not small baby dwarf up for them all to see.
"Congratulations your highness, it's a healthy baby girl." He said happily.
Time stopped. There was utter silence, and no one drew a breath. Ila felt like her heart stopped as she beheld the baby. Her baby. Thorin's baby. And not a boy. Not a son. Not an heir.
Dwala looked to Ila, eyes wide and waiting for her reaction. Dis looked to Ila as well. Dwala feared Ila might reject it and weep mournfully for a second, but after what was truly only a split second of hesitation on Ila's part she burst into tears and thrust her arms out for her baby.
Oin placed the babe in Ila's arms and she clutched it to herself as though she feared someone were about to snatch it away from her forever. The little dwarfling had very little blood on her and had shut her eyes as she wailed, not having wanted to leave the comfort of her mother's womb.
Despite wailing her head off, Ila thought the baby was the most beautiful thing she'd ever seen. She only wished she'd open her eyes so she could see them, so she could see her mother. Ila no longer felt any pain and paid no attention to Oin still busying himself with whatever happened after the baby came out. Ila was oblivious to it. The whole world lay in her arms now.
But…..a girl was not what was wanted. Fear seized her heart. Thorin would be disappointed. Erebor would be disappointed. What if Thorin didn't love the baby? What if he couldn't forgive her for not having a son? Ila held her baby even closer now, pressing it to her chest.
Dis smiled warmly and looked in love too, reaching out with a hand to help support the baby.
"She is beautiful Ila, you should be proud," she said. Ila was still crying though, and afraid.
Dwala held Ila from behind, whispering words of praise and encouragement.
The baby was quieting a bit being in its mother's embrace, and finally opened her eyes. Thorin's beautiful blues stared directly up into Ila's hazel ones.
Ila cried even harder, blubbering practically as she fell instantly and madly in love.
Dis moved to get up and retrieve Thorin, but Ila realized what she was doing.
"Please! Not yet…..let me hold her for a moment longer before you tell him that—"
"Shhhhh." Dis calmed her. "I will speak with him. All is well my dear. You have your little one and she is well." Dis assured her, but knew why she felt afraid. She knew her brother however, and knew Ila was overestimating his disappointment.
Ila leaned on Dwala now, cradling her new born and waiting to see how Thorin would take the news.
Kili looked up expectantly as his mother entered the antechamber.
"Where is your uncle?" she asked.
"The hallway. How is Ila?" he demanded standing up.
"She and the baby are fine." Dis assured her youngest.
"Erebor has an heir then?"
"No," Dis said quietly and went from the room. Kili stood dumbfounded as he watched her go.
No heir. So…the line of succession still rested with Fili for the time being then. Not that it mattered, she would surely have a boy next time.
But…..how would uncle take it? He would have to be a little upset, perhaps so much that it would cause a rift between them….
Thorin was pacing all the way down to the other end of the corridor and broke into a run when he saw Dis appear.
Dis raised her hands to stop him and as he reached her concern marked his face with terror ready to follow.
"Calm yourself brother, and listen to me." Dis said as he came to stand in front of her, Dwalin and Fili close as well.
"What of the baby? How is Ila?" he demanded.
Dis took his hands and looked him directly in the eyes.
"Ila is well, as is your daughter." She said calmly.
Thorin took a moment to let that sink in. Ila was well, and so was the baby. His daughter. He had a daughter. Not a son, but a daughter.
Thorin exhaled in relief, and went to move from Dis to go to her, but she stopped him and stood in the doorway.
"Thorin, Ila is terrified. She is afraid because she didn't give you a son, because she didn't produce an heir. You must not let her see you are disappointed." Dis commanded.
Thorin looked at her like she had two heads.
"Disappointed? How could either of you think that? How could you think I would be disappointed?" Thorin asked with sincere disbelief.
Dis didn't know what to say for a moment. This was not the initial reaction she had expected. She didn't think he wouldn't be happy, but that initially he would be saddened by not having a son. Apparently, she had been very wrong.
Thorin didn't wait for her to answer and moved past her swiftly. Fili and Dwalin were left looking at each other in the same disbelief that Dis felt.
Thorin didn't notice Kili as he walked past and threw the bedroom doors open. Ila nearly jumped out of her skin as she looked to her husband.
Dwala braced herself for what she thought was going to be a less than pleasant reunion. Ila did the same, holding her baby and looking down at her, not at Thorin.
Thorin reached the bed, sitting down on the side and drawing Ila into his arms. Her tear filled eyes turned up towards him and he wrapped an arm under her arm to support the baby. The babe looked up at her father, being all calm and quiet now, and Thorin's mouth spread into smile that threatened to break his face.
Turning to Ila with the same smile, he spoke quietly into her ear.
"The world needs princesses too, my love," and kissed her on the temple.
Ila sniffled and let a single tear fall, relaxing into Thorin's arms, all fear and anxiety gone.
"She is perfect," Thorin gushed, "she is perfect my beautiful love." He said as he gently placed a finger in her hand for her to grab.
Dwala's heart warmed to see Ila so happy, but also to see Thorin being the dwarf Ila deserved. She was glad she didn't have to hit him over the head with a vase like she'd planned if he had entered the room being anything less than overjoyed.
Everyone's eyes were on the happy trio, and no one noticed Kili standing in the door. A slight smile played on his lips, relieved beyond words that Ila was well. But his heart was also heavy as he beheld the happy family, wishing beyond reason and infinity that it was him holding Ila, and his baby in their arms.
Ila opened her eyes to see Kili by the door and gave him a weak smile. He returned it and nodded, walking from the room and past his mother, brother, and Dwalin as they came to peak at the royal family too. Dwala went to her husband and he wrapped an arm around her, pulling her close. Kili kept walking, out of the room, down the stairs, and to where he knew not. He just kept walking.
Ila closed her eyes again, ready to fall asleep in Thorin's arms. Their babe was falling asleep too, warm and content in her parent's embrace. As Ila was drifting off, emotionally and physically exhausted, she realized something.
"Her name…..what is her name?" Ila asked Thorin.
Thorin gently stroked her face while looking at their babe. He looked to Ila and smiled.
"Thora?" he asked his wife.
Ila only smiled, too tired to answer any other way, and fell asleep on Thorin's chest. Thorin looked to their daughter and swore that she smiled too.


text: My partner and i stayed here for 3 nights and it was the highlight of our trip to Thailand. Railay is such a beautiful place and is not as busy and touristy as phi phi which is where we stayed the for 3 nights before heading off to Railay. We stayed in the Privacy cottage and the location was very... More


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Along with all the heavy duty iDrive and ConnectedDrive technology, this car also has a very nice line in entertainment. Key components include the high resolution 8.8in LCD panel, 12GB of storage space for music files, support for MP3 and WMA formats, RDS radio tuner, USB connectivity, full iPod support and a single-disc in dash optical drive that not only plays DVDs and CDs but can also rip the latter to hard disk.
Music playback and management is arguably the most important aspect of any in-car entertainment system and here the 330d really delivers. Two USB sockets enable compatibility with MP3 and WMA music files located on a USB memory device. One is located in the glove box, the other in the centre armrest storage bin. Slightly oddly, the first only supports transfer of files to and from the system, while the other is restricted to direct playback of files from the storage device itself and iPod support.
If you choose to transfer files to the internal hard drive, you simply select the folder in question on your USB device and let rip. Any secondary folder hierarchy is maintained and each transfer creates a new entry in the on-board music library. It's a similar scenario for CD ripping, with each album appearing as an item in the library. CD ripping is pretty slow, but on the plus side the system is smart enough to recognise a partially ripped CD and resume the process should you accidentally eject mid-rip.
If you are wondering how the ripping works, CD data is provided by a built-in copy of the Gracenote database, enabling automated track labeling. Along with the static Gracenote database, the system can also poll the online version should you attempt to rip a CD that is newer than the built-in database. It's just a shame that this doesn't happen automatically. It requires that you drill down a few levels in the menu. Likewise, upon insertion of a USB key, the iDrive interface does not throw up any alerts or prompts. In our view, that's a bit of a basic oversight as well as being trivial to fix.
Nevertheless, in terms of overall music management, BMW has done a pretty good job. Not only can you delete ripped CDs and USB uploads, you can also transfer music back onto a USB device. As for playlist support, correctly formatted M3U files are supported, but there's no facility for creating playlists in the car itself. Given how simple that would be to implement, it's a pretty baffling limitation.
Still, viewing the remaining space available for music storage is very simple even if we do rather wish BMW had provided more than 12GB. For sure, that will be enough for many users. But this is the year 2009 and the 330d is a £30,000 motor car. Surely, that's enough to buy 100GB or so and therefore guarantee that 99.999 per cent of users never run out of space?
Of course, if you're a big iPod fan, on board storage will not be an issue. BMW was among the first car manufacturers to jump on the iPod bandwagon and its latest iPod control interface is one of the best. Effectively, all the music and data on your iPod can be played back or presented in the iDrive interface. That includes contacts, calendar entries and notes. The only exception is video files.
Indeed, the iDrive iPod menu offers essentially the same headings and hierarchy as the iPod itself, listing your music library by album, artist, track and so on. Likewise, playlists stored on iPods are also accessible. It's all presented extremely clearly and the iDrive wheel control interface will feel very familiar to any iPod user, which is a big win in our book.
At least, that's the theory. The system refused to pick up the playlists on our first-gen nano test iPod. We queried BMW's Ian Munday regarding this slight glitch and he told us that the early nanos are supported but that the software level of the device can sometimes cause problems. Unfortunately, we had to return the car before we had a chance to update our nano with the latest firmware and retest. However, such is the overall slickness and polish with which the iPod interface works, we're willing to give BMW the benefit of the doubt. Inability to create playlists on the go aside, using your iPod in the 330d is a thoroughly painless experience.
You might expect top notch music playback and iPod support to be a given for the latest revision of iDrive. But what about DVD movies? It's certainly easy to dismiss this as a gimmick - there are no screens for rear seat occupants, for example. But whatever you think about the idea of watching movies in a stationary car (motion video is only displayed when the car is parked) on an 8.8in screen mounted in the dash, there's no doubting BMW has absolutely made the most of the feature.
For starters, the screen boasts excellent interpolation thanks to its very tight pixel pitch. It is a little awkwardly proportioned, admittedly. The ultra-wide aspect means that standard 16:9 video content does not fill the screen horizontally. However, thanks to a zoom function, anamorphic DVD movies with aspect ratios of 2:1 and more can be set to fill the screen very nicely indeed, at which point you begin to appreciate the excellent viewing angles and subtle colour balance.
BMW has included a basic set of colour, brightness and contrast controls, while the large binnacle above the screen does a great job of reducing unwanted glare. Then factor in the superb surround sound imaging from the audio system and you have a remarkably pleasant movie viewing experience. It's so good, you just might find yourself 30 minutes into a test movie when you'd only intended to evaluate it for five minutes, if you catch our drift.
Standard AM and FM RDS radio is your lot, there's no digital radio. But BMW has still done a great job of integrating an antediluvian analogue technology into the iDrive interface. The airways are constantly scanned for RDS enabled radio stations. The results are listed in alphabetical order, allowing you to scroll from station to station with gay abandon. It's hardly a unique system, but it still deserves credit for ease of use.
The sound quality of the BMW Professional Multimedia Package presents something of a conundrum. On the upside, it's extremely precise, the imaging of the sound stage is very well controlled and there's significant dynamic range and clarity.
However, the system is just a little clinical and lacking in warmth. The wow factor that enables the very best in-car stereos to pummel your kidneys while setting your hair on end is also missing. What it probably needs is a bit more bass extension. In the end, it comes down to expectations. For most owners, the audio package will deliver all they will ever need and more. The most discerning users will inevitably be left wanting more. It was ever thus.
A hideously looking monstrosity of a car. BMW's of late look absolutely vile! And don't get me started on the X6....
@Steve - You'll get no argument from me, I think that the 3 Series saloon is an ugly beast. Although at least the rear light clusters on this facelift model aren't as rank as the ones on the original. Of course my trusty coder Rich will argue to the death that the 3 Series saloon is stunning, but at least he's otherwise sane.
Personally I just can't believe how big the 3 Series is now. The 1 Series coupe is about the size that the E30 3 Series coupe was! There's no denying that the technology that BMW is squeezing into its cars is pretty impressive though.
Regarding Information Plus - I'm sure that Fiat had something like this about five or six years ago in the Stilo. Anybody remember? If you went for the satnav option and installed a simcard, you could press some sort of 'Help' button and be put through to a call centre where they could send e.g. a local hotel (or garage more like) to your Nav.
Or am I dreaming again? If not, it's surprising how long it takes for tech to make it's way through cars...
@Mario: Sounds like you're thinking of SmartNav. That's not particularly new or impressive technology and it's not confined to Fiat. It's not quite the same thing either. If you asked the SmartNav lady for the atomic weight of Boron, she'd probably hang up on you whereas a BMW lady would kindly oblige.
The closest I've heard of is the 'concierge' service that used to come with a VW Phaeton, although that was more for restaurant reservations, theatre tickets and the like.
I still think the previous generation 3 series looks far more 'premium' that the current version. The interiors have always been solid but at the same time looked a little old fashioned too, eg. the dials haven't changed much over the years. However, you just can't argue with the way they drive, they are all excellent cars (begrudgingly even the X5 & X6, for what they are), and the latest generation of diesel engines are in a league of their own, returning some amazing Co2 results alongside some serious performance figures.
Anyway, there has to be a point when all this tech gets distracting to the driver, but then seeing as we spend more and more time stuck in traffic jams, having lots of functionality might actually help preserve our sanity!!
"open out the iDrive platform" - now there's a thought. It's one thing to brick your iphone with a dodgy app but messing up your traction control system opens up whole ranges of interesting possibilities...
@ChrisC: The facelift gives it a far more 'premium' look. The new creases in the bonnet lend some aggression and the new rear and light clusters are far more distinctive. The previous rear clusters were ripped straight from a Mitsubishi Charisma, the most inaptly named car in history.
Also, look out for some impressive new diesel engines from Mercedes and Jaguar. Merc's new 4-cyl C250 CDI does 0-60 in 7.1, returns 'over 50 mpg' and pumps out 138g/km CO2.
I drive an 07 335d Coupe, with the earlier generation iDrive. I agree with a lot Jeremy says, I went for upgraded speakers and they make a big difference to the sound, much cheaper than the horrendously expensive Harmon Kardon option. As for the quoted range, you'll find that a bit optimistic. 600-650 miles, certainly, I manage around 500-550 in the 335d at a steady 75-80 on a long run.
Agree with Chris, the new Jaguar XF engine with the twin turbodiesel is getting great reviews, must be worth a look.
A stunning, stunning car. I'd hit it
Thanks for the great info on connected drive. I have just bought a new F10 5 series that includes the same system, and was looking for more in depth tech info that you described.
Overall it is a great system, I am just waiting until the third party hackers start on it so we get things like custom POI (speed cameras) for the NAV, video interface for iPod etc.
Beautiful car man
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Researchers in California, writing in the journal BMJ Online First, gave the results of questioning mothers or carers of 185 babies who died and 312 other parents, reopening the debate about the role of dummies.
Although cot deaths have fallen in recent years, they still end the lives of 300 babies in Britain annually, a rate of about one in 2,000 births. No one cause has been identified although a number of measures are recommended. The Foundation for the Study of Infant Deaths (FSID) says parents should put their baby to sleep on its back and keep it cool with its head uncovered, and should not smoke. Babies should not sleep in their parents' bed but in a cot in the same room, at least for the first six months.
It has been suggested that dummies' bulky external handles might prevent babies suffering accidental lack of oxygen from having their faces buried in blankets or underneath adults' bodies, or that sucking them improves development of babies' airways.
The new study, led by Dr De-Kun Li of the research division at Kaiser Permanente, a not-for-profit health company, said use of a dummy was associated with a "substantial reduction" in risk of death, perhaps as much as 90%.
It might also lessen the impact of other factors linked to cot death such as sleeping face down or on the side, sleeping with a mother who smoked, or sleeping on soft bedding. "These preliminary findings need confirmation ... Funding from our study and others will collectively provide support for the protective effect of dummies, but they are not proof of a causal effect by themselves."
The FSID recommends that if a baby uses a dummy regularly it should not be withdrawn. "That is because two research studies published since 2000 showed that babies who usually use a dummy but then stop it are at increased risk of cot death on the night they don't use it," it said in a statement. The latest study was "very interesting: and appeared convincing. But the statistical analysis is very complicated and needs careful study. FSID is now examining the available data to determine whether parents should be recommended to give their baby a dummy".
One problem was that the new study did not make any reference to the risk identified in previous research if parents forgot one night to give their baby a dummy. There was also no information in the UK as to who used dummies, and the organisation would conduct a study to put that right.
The FSID added: "In the meanwhile our advice is that there is no reason for parents not to use a dummy, but if they do they must use it every time that baby sleeps, and never forget to give the baby the dummy. Other factors to bear in mind are they should not coat the dummy in any sweet solution and they can take the dummy away when the baby is about 12 months old. Also, if they are breastfeeding they might like to wait a month or so before introducing a dummy."


text: Offering many popular games and activities for the children aged 4-12 (kids aged 3 and under are invited when accompanied by parents). Every little discoverer can choose what they are interested in, activities like cookies making, T-shirt painting, kite flying and sandcastle building with many more to choose from. The indoor area has seven different zones of entertainment with its own theme, like story telling, TV viewing, dance hall, block building, arts & crafts, activity and a room for napping.


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Black Crystal Bib Necklace
- 3-5 Hours
- Intermediate
Additional Information
- Black crystal teardrops
- Black crystal marbled rondelles
- Black crystal small rondelles
- Silver flower bead caps
- Silver eyepins
- Silver headpins
- Silver round jump rings
- Silver lobster clasp
- Wire cutters
- Round nose pliers
1. Start by stringing a black teardrop crystal onto an eyepin and forming a simple loop. Repeat to create 100 of these teardrop connectors.
2. String a black small rondelle, a black marbeled rondelle, and another small black rondelle onto an eyepin and form a simple loop. Repeat to make 16 of these triple connectors.
3. Create dangles by stringing a marbeled rondelle bead between two bead caps and onto a headpin. Form a simple loop. Repeat to create 7 dangles.
4. Connect 4 triple connectors together by opening and closing the end loops like a jump ring. To the end connector, add a lobster clasp by using a jump ring.
5. Connect another 4 triple connectors together, connecting a large jump ring on one end (this is what the lobster clasp will clasp on to), and a round jump ring on the other end.
6. With the lobster clasped, to one free end, connect two more triple connectors by using a jump ring. Repeat on the other side.
7. Start adding the teardrop connectors, connecting six of them together in a flower shape with one silver jump ring connecting them all in the center of the flower. Use the picture as a guide to the pattern and layout of these connectors.
8. Once the bib is completed, finalize by adding dangles to the bottom.


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The course is run primarily as a re-sit of the course you followed at school. Gaining a grade 4 (equivalent to the old grade C) or above has proved beneficial to many students enabling them to progress on to more advanced courses here, and gaining the entry requirements for many college and university courses. It is also a requirement for many jobs and careers.
New governmental rules say that students without a grade 4 in GCSE Maths must be working towards getting it. The focus of the teaching on the course will be at Foundation tier in order to aim for a grade 4, with the potential of getting a grade 5 - equivalent to a high C Grade in the old GCSE.
Past students have found the course very rewarding but sometimes difficult. Different students enjoy varying parts of the course. The greatest pleasure comes from gaining in confidence and realising you can do Maths!
Each week you will be taught in a classroom environment which will include time spent on working on exercises or past examination papers and solving real-life problems, in a pleasant and supportive atmosphere. Some lessons will be in a room with IT facilities.
You will take 3 external exams, two calculator and one non-calculator, all at Foundation level. There will also be formal internal assessments at regular intervals, including mock exams, so that you can monitor your own progress.
With a grade 4 Stuart gained entry onto his preferred music course at university and Helen met the entry requirements for the Engineering BTEC level 3 Extended Diploma course she wanted to take after the Level 2 Diploma. With a grade 5, students will find it easier to get to university to study a degree with a higher level of Mathematics content, such as Chemistry. Indeed, the grade 5 will be a minimum requirement for many courses.
Any student with a grade 2 or 3 in GCSE Maths, an A*- E grade in Certificate in Use of Maths, or a pass in Functional Skills Maths Level 1 or 2 can take this course.
All students will be expected to provide their own stationery and geometry set, including a ruler, protractor, compass, pencil and rubber. All students must have a scientific calculator with natural display, such as the Casio FX-83GT+ or Casio FX-85GT+ (both around £10). It is also essential that students buy a Maths Revision Guide, two Revision Workbooks and an exercise book for writing your notes in; these can be purchased at a discounted price of approximately £10 via the College's Online Store. The cost for these Student Course Materials also covers numerous hand-outs, worksheets and ICT resources.
If the costs of equipment, materials and trips may cause you financial hardship, you may wish to read through details of our financial support scheme on our website.
The GCSE is changing
From September 2016 Alton College will be offering the new GCSE in English Language with the grading system 9-1. The first examination will be in June in 2017.
English Language GCSE is a qualification that proves you have achieved a certain standard in speaking, reading and writing the English language. A grade 4 (formerly grade C) or above is required for most advanced or higher level courses in all subjects and many jobs.
The course involves close study of both fiction and non-fiction texts from the 19th, 20th and 21st century. There will also be opportunities for creative and non-fiction writing, as well as group discussions.
Students are expected to take complete responsibility for the production of their own work. This does not mean however that your tutors simply set work and abandon you until the exam date. You will be expected to work closely with your tutor, seeking individual help when necessary. The aim is to help you reach your potential and no effort will be spared to achieve this.
In lessons you will read and discuss media, non-fiction and literary texts. You will develop your skills of comprehension, analysis and comparison. You will produce pieces of creative writing such as descriptions and stories. You will also practise practical writing tasks in various forms such as essays, articles, letters and reports. You will be encouraged to identify any errors in your technique and helped to correct them. There will be many opportunities to develop oral skills in a variety of class situations, including plenty of discussion work in small or larger groups. Overall, the lessons will prepare you for the two examinations in the summer and you will also receive an additional qualification in spoken language. Everything you learn will not only prepare you for getting the qualification but also be transferable – you will be learning life-long skills for use in the real world.
The qualification has two exams which will be in June 2017. In addition, there is a "spoken language" component which is assessed separately and given the markers: pass, merit, distinction. Although this part of the qualification does not count towards your GCSE it does appear on the final certificate and the preparation for it will be a useful life skill.
A high percentage of our students over the last few years have successfully gained their grade 4 or above and gone on to A levels, vocational A levels, Further and Higher Education. It has helped others to gain training places and employment.
Students have enjoyed the confidence gained from improving their English language skills. Please note, this not an ESOL course as it tests different skills. A sound grasp of English would be an advantage.
"Studying English GCSE has been fun and I now have a lot more confidence in my written English and my reading."
"The English GCSE course has been most enjoyable. At first, I was very apprehensive as I had not done English since being at school 21 years ago. The teacher was very supportive and made me feel at ease with the subject."
To qualify for this course, you will normally have achieved a Grade 2 or 3 in English GCSE.
All students will be expected to provide their own texts and stationery (where appropriate). Other course materials will be provided by the teacher.
There are no additional costs.
If the costs of equipment, materials and trips may cause you financial hardship, you may wish to read through details of our financial support scheme on our website.
Edexcel - 1ENO 1 and 2


text:
What it's likeMature
Jack gave a warm smile to the receptionist as he walked into the "Telephone angels" center. Either the girl was getting younger every time or he was getting much older. At forty-one, he was one of the few who always came in. A wisp of ash grey hair fell in his face, he sighed as he brushed it out of his face and tied his shoulder length hair back.
He was getting up in the years, and yet he still had to show any signs of balding. He could feel his left leg was giving him a bit of grief but considering the job he really didn't seem to mind. During the day, he played for the bars. He'd gotten a good reputation, "Sad Jack, the Blues man". He sang songs that most people thought were just made up lyrics. However, anyone who stared into those icey blue eyes of his realize there was something lurking beneath those songs. Pain and sadness, and every night he blew it all away. He gently rubbed a small heart pendant between his finger and thumb as he signed the paper. The receptionist gave a warm grin. Her name was Jenny, right?
"I just downloaded your newest disc off Itunes Jack, I love it!" Jack gave a puzzled grin.
"Itunes? What's that?" Jack hated technology. His agent got him a laptop to answer fanmail and all that nonsense. He didn't really use it much.
"Well it's umm," The young girl gnawed at her lip for a moment before Jack held his hands up.
"Sweety, I really don't think you can dumb it down enough for a coyote like me. I better go man the phones, I heard the new blood already gave up." He referred to the bright eyed college kids coming in thinking they could bring something to these kids. They were wrong and eventually stopped caring. Jack had his share of failures too. However, it never stopped him. He knew these kids had a really bad run at life and for every kid that never called back. There was at least two more asking for Jack. She frowned at the mention of those two.
"Yea, they really weren't very good. Judy overheard em talking and wanted to toss em out already." Jack grinned as he grabbed a steaming cup of coffee and gave her a wave.
"Well then to battle! And all that rabble rousing," He wandered to his designated area. At this point, the regulars like him were given sections to outfit as they want. Jack had his own little studio, he always felt inspired to write when he was here.
The telephone blinked on as Jenny activated Jack in the system. He began to idly plunk at his acoustic while the time began to pass. The first two calls sounded like spoiled little brats, upset that mommy didn't let them do what they wanted. Jack tried his best to explain what their mother felt was in the interest of the kids. Wasn't such a big deal afterall, they hung up after just a few minutes.
Jack sighed as he got to his feet as his back began to ache. Jenny waived to him as his phone went off. She only did that if the call came from a trouble area to begin with. Jack put the guitar down as he adjusted his headset.A girl's voice was on the other end, she sounded faint. Jack knew he needed to keep his voice level.
"Telephone Angels, my name's Jack sweety. What do you want to talk about?" The other voice seemed to hesitate before replying.
"Rhea, I dunno what I want to talk about? I-I found your number at a shelter me and my friend were staying at..." Jack smiled, they had a good success with those shelters. Too many kids find their way there.
"Well Rhea, I've got a really big cup of coffee here. So let's start at the beginning, are you safe?" The line was silent as Rhea sputtered a response.
"Fuck no! I'm in the middle of the damn hood, the shelter's are starting to push us back out. I'm cold and tired and..fuck how do you know what I'm dealing with?"She was already quite agitated, Jack knew he needed to calm her a bit. Show her she has nothing to fear from him.
"Sweety, I don't wanna compare war stories with you. I've got my share of skeletons, and an empty bed where my daughter used to be. So I know what it's like to be when the cards aren't in your favor." The voice calmed down a bit.
"Jack?" she called out quietly.
"I'm hungry.." The voice seemed a bit more eager for him. Jack smiled as he got to his feet.
"That's a good start! A growing girl like you could use a good meal. I know roughly where you are. There's a greasy spoon a few blocks away. Can you be there in ten?" He silently prayed she would accept. None of these kids are ever that easy, at the very least this one can be looked over.
"Ok Jack, don't be late!" The line clicked off. Quickly grabbing a stack of cards with Jack's information and one of his old blues cds. He signed off as Jenny gave him a curious look.
"Got one that needs help, she wants to meet. Poor girl sounds scared out of her wits. Near those damn apartment buildings we got those cases last year." Jenny gave him an encouraging grin.
"Good luck Jack. Be safe ok?"
Jack pulled his leather coat a little tighter as he left the center. He felt hopeful, maybe this girl just really needed someone to sit down and listen. Jack tried to not think down on the parents who failed these kids. He did his best though to realize these kids needed positive encouragement and not his own opinions best left unsaid for now.
Right now there was a girl who could use an ear and a warm meal. Jack made sure he kept his dummy wallet handy. It contained no information, but a wad of bills that could keep a runaway well stocked for at least a week.
Jack pulled up in front of the diner, the food was soulful. The type of meal that could hopefully warm a body for awhile. He tossed the real wallet into the glove compartment and took the dummy containing just money as he locked the door of his vehicle and stepped inside.


text:
And at today's second spring scrimmage, defensive backs coach Scott Lakatos is hoping to take a major step toward finding the right role for each of them.
"The good news is that there's a decent amount of talent here, and all of those guys can help in some way," Lakatos said. "What we've got to do is figure out what they can do and in what situation. That's kind of why you're seeing a lot of mixing and matching guys. Every week we've had a different group in there."
Last week, it was Bacarri Rambo and Jakar Hamilton running with the No. 1 defense, and they're they odds-on favorites to still be handling that role in September. But nothing is set in stone.
"I feel like I've still got to win that job because the depth chart is not set yet, and that job is open," said Rambo, the most experienced of the group in terms of playing time. "I'm working with the first team now, but I'm just trying to take advantage of this great opportunity."
Rambo got more snaps than anyone while backing up Bryan Evans and Reshad Jones at both safety positions last season, and Nick Williams believes Rambo is as close as anyone to locking down a starting gig in 2010.
The competition is intense, however.
Williams, who has switched back and forth from safety to linebacker three times since arriving in Athens has spent some time with the first-team defense, too. As has Shawn Williams, a rising sophomore who got a taste of action on special teams last season but hasn't played many scrimmage downs.
Then there's Hamilton, who displayed an impressive combination of athleticism and physicality while at Georgia Military College the past two seasons. There's Makiri Pugh, a third-year player still trying to find a home. And there's Quintin Banks, the veteran of the group, who has always been a favorite of the coaching staff, but whose career has been mired by injuries.
"You've got different guys bringing in different qualities, so when you put all that together, we're kind of – not bumping heads, but you look at things totally different," Nick Williams said.
That dynamic has made the evaluation process a bit of a challenge for Lakatos. So far, he's rotated the No. 1 units throughout the spring, and the players haven't gotten too much insight as to how the depth chart might look at any particular moment.
"Coach switches it up and different weeks, different guys are going," Nick Williams said. "It's a lot of competition, and he lets everyone know no one has a starting job yet. It's like a job interview, and every day you're interviewing for the job."
Lakatos doesn't mind if his players view the competition that way, but he's not necessarily looking for winners and losers in the competition. It may be a job interview, but he's hiring them all.
The bigger issue, Lakatos said, is finding a combination of players that fit well together, and that's been a big part of the rotations he's employed.
"We're always evaluating guys," Lakatos said. "We talk after every practice to see where guys are, move some people around, try some different combinations."
Rambo said Lakatos is a stickler for technique, and the coach's primary lesson is to ensure that each player is where he's supposed to be at all times.
But that's easier said than done. Even the veterans of the group have minimal experience, and even that experience is undermined a bit because the reps came in the system employed by the previous coaching staff rather than Todd Grantham's new defense.
Getting the safeties excited about learning, however, has not been a problem.
"All those guys have a really good work ethic," Lakatos said. "They're always around asking questions. They want to know what's going in tomorrow, what they need to get better at. They're a good crew."
Just how much his crew has absorbed during the past month will be on display today as the Bulldogs compete in their penultimate scrimmage of the spring. While G-Day still awaits next week, today's practice is likely going to be the only time the players will work with the full breadth of plays the staff has installed this spring before toning things down in front of the cameras and fans next week. That makes today's performance crucial in the battle for playing time.
"This scrimmage, we're trying to figure out who's going to be good at what on game day," Lakatos said. "Who's going to help with communications, who's going to be able to absorb what we've installed and be able to execute it. We're looking at all those things to find who we're going to be able to count on in September."
The significance of that isn't lost on the safeties.
They're a diverse group, but they're all working toward a similar goal, and today marks a crossroads in that journey.
"Every day (Lakatos) is coaching us to get better, and he said whoever starts is up to us," Nick Williams said. "That's basically what it is. He's here to coach, and we're here to prove to him we're ready to play. It's your own destiny."


text:
Strand-specific transcriptome profiling with directly labeled RNA on genomic tiling microarrays
With lower manufacturing cost, high spot density, and flexible probe design, genomic tiling microarrays are ideal for comprehensive transcriptome studies. Typically, transcriptome profiling using microarrays involves reverse transcription, which converts RNA to cDNA. The cDNA is then labeled and hybridized to the probes on the arrays, thus the RNA signals are detected indirectly. Reverse transcription is known to generate artifactual cDNA, in particular the synthesis of second-strand cDNA, leading to false discovery of antisense RNA. To address this issue, we have developed an effective method using RNA that is directly labeled, thus by-passing the cDNA generation. This paper describes this method and its application to the mapping of transcriptome profiles.
RNA extracted from laboratory cultures of
An efficient method was developed for mapping transcriptome profiles specific to both coding strands of a bacterial genome. This method chemically labels and uses extracted RNA directly in microarray hybridization. The generated transcriptome profile was free of cDNA artifactual signals. In addition, this method requires fewer processing steps and is potentially more sensitive in detecting small amount of RNA compared to conventional end-labeling methods due to the incorporation of more fluorescent molecules per RNA fragment.
The oligonucleotide microarray has become a powerful and popular tool for comprehensive transcriptome mapping. Closely tiled probes are suitable for high resolution transcript detection and have been extensively used for discovering novel RNAs including coding, non-coding, antisense, and structural RNAs. However, the conventional methods for transcriptome mapping involve sample processing steps such as reverse transcription, amplification, and ligations which introduce biases and thus may not reveal the true transcriptome profile
In most microarray assays for studying gene expression, RNA is converted to cDNA by reverse transcription and the signal intensities detected from labeled cDNA hybridized to the probes on the microarray are interpreted as the level of transcription. However, reverse transcription has been known for its tendency of generating artifactual cDNA through various mechanisms
To circumvent problems introduced by the reverse transcription, several methods of using RNA directly in microarray hybridization have been developed. For example, Hu
In this paper we describe a simple, efficient, and sensitive method that chemically labels and uses RNA in microarray hybridization experiments, which can be used for both prokaryotic and eukaryotic transcriptome profiling. The chemical labeling covalently alkylates the RNA on guanine, adenine, and cytosine residues
Tiling microarray design
The genomic tiling microarray probe set consisting of 385,000 oligonucleotide sequences was dynamically designed from both forward and reverse-complement strands of the target genome
Bacterial culture preparation
Total RNA extraction
Lysis of bacterial cells was performed according to the protocol provided with the MasterPure RNA Purification Kit (Epicentre, Madison, WI, USA). The lysate was treated with Proteinase K at 65°C for 15 min and placed on ice. To increase the recovery rate of small RNA, 0.1 volume of 5 M NaCl was added to the lysate. Acid-phenol: chloroform (5:1) (v:v) extraction was then done using Phase-Lock Gel Heavy tubes (Eppendorf, Hauppauge, NY, USA). Total RNA in the upper aqueous phase was purified with a solid-phase extraction filter supplied in the mirVana miRNA Isolation Kit (Applied Biosystems/Ambion, Austin, TX, USA) according to the recommended protocol. The filter-trapped RNA was washed and eluted with the provided wash solutions and elution buffer. To completely remove genomic DNA, the RNA extract was treated twice with Turbo DNase (Applied Biosystems/Ambion) at 37°C for 30 min, and purified again with the mirVana miRNA Isolation Kit.
RNA direct labeling and microarray hybridization
RNA-DNA hybridization was performed on microarray slides covered with a HybriWell chamber (Grace Bio-Labs, Bend, OR, USA). Prehybridization was carried out for 90 min at 42°C in a 400 μl solution containing 343 μl Long Oligo hybridization buffer [94 mM Tris/HCl pH 7.0, 9.4 mM EDTA, 29.15% formamide, 5.83X SSC, 0.12% SDS], 0.5 mg/ml BSA, and 0.1 mg/ml salmon sperm DNA (Applied Biosystems/Ambion). The salmon sperm DNA was denatured at 95°C for 5 min before added to the prehybridization and hybridization solutions. After the prehybridization the HybriWell chamber was removed and the slide washed with nuclease-free water. The slide was then spin-dried at low speed for 2 min, and a new HybriWell chamber was sealed onto the slide. For each microarray 3 μg of labeled RNA was denatured at 65°C for 5 min in a final volume of 300 μl hybridization solution containing 257 μl Long Oligo hybridization buffer, 3 μl Alignment Oligo (NimbleGen), and 0.7 mg/ml salmon sperm DNA. Denatured RNA was then loaded in the chamber and hybridization was carried out at 42°C for 16 hours with 10 rpm rotation in an oven (Labnet, Edison, NJ, USA). The slide was washed according to NimbleGen's protocol (NimbleGen Arrays User's Guide Gene Expression Analysis).
cDNA labeling and microarray hybridization
Synthesis of cDNA and biotin end-labeling were performed according to NimbleGen's protocol (Prokaryotic Biotin-Label Procedure). First-strand cDNA was synthesized from 10 μg total RNA. The RNA together with 3 μg random hexamer primers (Invitrogen, Carlsbad, CA, USA) in a 12-μl volume was denatured at 70°C for 10 min. The solution was then cooled to 25°C and mixed to a final concentration of 0.5 mM dNTP, 20 mM DTT, 1X first strand buffer and 0.75 U/μl RNaseOUT (Invitrogen) followed by heating to 42°C. A total of 1200 U SuperScript II reverse transcriptase (Invitrogen) was added to a final volume of 54 μl and the mixture incubated overnight at 42°C. The cDNA product was treated with 0.02 U/μl RNase H (Invitrogen) and 0.01 μg/μl RNase A (Epicentre) in a 100-μl volume to eliminate RNA contaminants. The cDNA was then purified with standard procedures for phenol: chloroform extraction using Phase-Lock Gel Light tubes (Eppendorf) followed by ethanol precipitation. Purified cDNA was fragmented to 50-200 nucleotides in size with DNase I (Applied Biosystems/Ambion), and labeled at the 3'end in a 100-μl volume containing 0.5 U/μl Terminal Deoxynucleotidyl Transferase (Promega, Madison, WI, USA) and 0.025 mM Biotin-N6-ddATP (Enzo Life Sciences Inc., Farmingdale, NY, USA). The labeling reaction was carried out for 2 hours at 37°C. The biotin-labeled cDNA was then concentrated using Microcon YM-10 filters (Millipore, Billerica, MA, USA). Microarray hybridization was carried out with the customer service provided by NimbleGen.
Genomic DNA extraction and hybridization
Genomic DNA for DNA reference microarray hybridization was extracted with the MasterPure DNA Purification Kit (Epicentre) and RNA was removed using the RNase A supplied in the kit. Genomic DNA was fragmented with DNase I (Applied Biosystems/Ambion) to 100-200 nucleotides in size and then labeled either with biotin 3'end-labeling (i.e., cDNA procedure) or with the
Data acquisition and normalization
After washing and drying, the microarray slides were immediately scanned in a GenePix 4000B Scanner (Axon Instruments, Union City, CA, USA) using the provided GenePix Pro 6.1 software. NimbleScan v2.5 software was then used for spot features extraction from the scanned images. Each of the microarray hybridization results hybridized with RNA, cDNA, or genomic DNA, consisted of at least two biological repeats. Two types of normalizations were performed - normalization by DNA reference array and between-array normalization. For DNA reference array normalization, intensities of cDNA and RNA signals were normalized with signals from DNA reference arrays that were hybridized with fragmented genomic DNA labeled in the corresponding way, i.e., biotin end-labeling and chemical labeling for cDNA and RNA, respectively. Both normalizations were done using the Bioconductor R package "tilingArray"
Data availability and deposition
Original and normalized microarray data used in this paper were deposited in the NCBI Gene Expression Omnibus (GEO) database
Strand-specific RT-PCR using tagged primers
In conventional reverse transcription PCR (RT-PCR), false positive PCR artifacts have been reported in absence of primer used in the reverse transcription step, due to self-priming of RNA or non-specific small DNA/RNA oligonucleotide contaminants that can also be used as primers in RT reactions
Additional file 1
The additional file includes a table listing the primer sequences used in this work and four figures showing additional results including the efficiency of RNA labeling, the effects of blocking reagent on the background signals, and the effects of actinomycin D added in the reverse transcription reaction.
Click here for file
Comparison of transcriptome profiles revealed by labeled RNA and cDNA
RNA isolated from
Figure 1
Comparison of RNA- and cDNA-based hybridization signals
Comparison of RNA- and cDNA-based hybridization signals. A: Histograms of log2 probe signal intensities obtained from the RNA-based (left panel) and cDNA-based method (right panel). Probe intensities from coding and non-coding regions highlighted red and blue, respectively. B: Scatter plots of signal intensities between those from the coding (x axis) and corresponding antisense (y axis) regions of the genome. RNA-based signals were plotted in the left panel and cDNA-based in the right.
The transcriptome profile was compiled by plotting the normalized hybridization signal intensities on the genomic coordinate based on the probe positions. Forward and reverse-complement probe signals were plotted separately and reflect the genome-wide level of RNA transcribed in the cells. Figure
Figure 2
Comparison of RNA- and cDNA-based transcriptome profiles
Comparison of RNA- and cDNA-based transcriptome profiles. Normalized log2 probe signal intensities (y axis) from a 100-kbp region of
Determination of sensitivity and optimization of method conditions
In a time-series experiment we observed that the incubation time was linearly proportional to the labeling density of the RNA molecules during the first four hours (Figure S1, Additional file
In our hands, optimal signal-to-noise ratio of the probe intensity distribution was achieved in a hybridization solution with 25% formamide at 42°C. RNase-free BSA was added to the pre-hybridization solution and salmon sperm DNA was included in both pre-hybridization and hybridization solutions to block non-specific binding of RNA molecules to the probes and surface of the microarray slide. We achieved optimal effect when adding 0.7 mg/ml blocking reagent (i.e., salmon sperm DNA) to the hybridization (Figure S2, Additional file
Validation of RNA signals by strand-specific RT-PCR
To verify the transcription signals detected by either the RNA- or cDNA-based method, especially in the antisense regions where cDNA artifactual signals were often reported, strand-specific RT-PCRs using tagged RT-primers were performed. The antisense strand of five highly expressed ORFs was targeted: PG0279, PG0933, PG1069, PG0559, and PG0775 (marked A1-5, respectively). Strong antisense signals were detected in these regions from the cDNA-based profile while only near background levels of signals were found in the corresponding areas of the RNA-based profile (i.e., Figure
Figure 3
Strand-specific RT-PCR targeting
Strand-specific RT-PCR targeting
We have described a comprehensive experimental procedure with conditions optimized specifically for studying strand-specific transcriptome profiles using genomic tiling microarrays. Our results, including the comparison of RNA- and cDNA-based transcriptome profiles and the detection of transcripts by RT-PCR, confirmed that the use of fluorescently labeled RNA generated a profile without artifactual antisense signals, revealing a better transcriptome profile than the use of cDNA. However, RNA is labile and susceptible to degradation, thus extra cautions are needed to prevent the degradation of RNA and reduction of signals. In addition, strong secondary structures of RNA molecules may reduce hybridization efficiency
The fluorescent labeling density directly affects the signal intensity, and thus the overall sensitivity for detecting RNA, especially RNAs transcribed at low levels. The sensitivity of the labeling technique we used (i.e., one fluorescent label per 20 nucleotides) is potentially five times greater than that of single dye end labeling
The most important advantage of using labeled RNA directly in microarray hybridization for detecting transcription signals is the elimination of cDNA artifacts. The generation of artifactual cDNAs in reverse transcription is due to several possible mechanisms including self-priming or non-specific oligonucleotides priming to the newly generated first-strand cDNA
In addition to the experimental procedure, post-scanning data processing is also important in maximizing the quality of the transcriptome profile
The final step for compiling a transcriptome profile is the determination of transcription boundaries between expression and non-expression signals. We have developed a dynamic algorithm specifically for this purpose and have used it to annotate the transcriptome profiles obtained in this study. The detailed description of this algorithm has been published elsewhere
A comprehensive procedure for mapping transcriptome profiles specific to both strands of a genome was developed. Chemically labeled RNA was used directly in the microarray hybridization. Hence, experimental artifacts induced by cDNA synthesis were eliminated and the generated transcriptome profile was free of cDNA artifactual signals. In addition, this method requires fewer processing steps and is potentially more sensitive in detecting low level RNA expression compared to conventional end-labeling methods due to the incorporation of more fluorescent molecules per RNA fragment. The complete RNA-based
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
WY and HH performed the experiments in this study. All authors contributed to the study design, data processing, and the writing of the manuscript. All authors read and approved the final manuscript.
We thank Dr. Mary-Ellen Davey at The Forsyth Institute for improving the writing of this paper. HH was supported by a Ph.D. stipend and mobility grant from the Faculty of Dentistry, University of Oslo, Oslo, Norway. This project was funded by the National Institute for Dental and Craniofacial Research (grant No. R21 DE018803-01A1).
Direct RNA sequencing
Properties of the reverse transcription reaction in mRNA quantification
Antisense transcripts of V(D)J rearrangements; artifacts caused by false priming?
Sense or antisense? False priming reverse transcription controls are required for determining sequence orientation by reverse transcription-PCR
Reverse transcriptase template switching and false alternative transcripts
An RT-PCR artifact in the characterization of bacterial operons
Potential pitfalls in the accuracy of analysis of natural sense-antisense RNA pairs by reverse transcription-PCR
Fidelity of two retroviral reverse transcriptases during DNA-dependent DNA synthesis in vitro
Antisense artifacts in transcriptome microarray experiments are resolved by actinomycin D
An antibody-based microarray assay for small RNA detection
Dynamic transcriptome of Schizosaccharomyces pombe shown by RNA-DNA hybrid mapping
Gold nanoparticle probe-based gene expression analysis with unamplified total human RNA
Direct labeling of RNA with multiple biotins allows sensitive expression profiling of acute leukemia class predictor genes
Direct and sensitive miRNA profiling from low-input total RNA
Nonradioactive 3'-end-labeling of RNA molecules of different lengths by terminal deoxynucleotidyltransferase
Prevalence of transcription promoters within archaeal operons and coding sequences
Directly labeled mRNA produces highly precise and unbiased differential gene expression data
Array comparative genomic hybridization with cyanin cis-platinum-labeled DNAs
Comparative genomics of wild type yeast strains unveils important genome diversity
MicroRNA Labeling Methods Directly Influence the Accuracy of Expression Profiling Detection
Dynamic probe selection for studying microbial transcriptome with high-density genomic tiling microarrays
Interactions of Porphyromonas gingivalis with epithelial cells
Transcript mapping with high-density oligonucleotide tiling arrays
Strand-specific, real-time RT-PCR assays for quantification of genomic and positive-sense RNAs of the fish rhabdovirus, Infectious hematopoietic necrosis virus
Secondary structure in the target as a confounding factor in synthetic oligomer microarray design
Expression profiling using microarrays fabricated by an ink-jet oligonucleotide synthesizer
Effects of formamide on the thermal stability of DNA duplexes on biochips
Troubleshooting microarray hybridizations
Impact of surface chemistry and blocking strategies on DNA microarrays
Basis of actinomycin action. I. DNA binding and inhibition of RNA-polymerase synthetic reactions by actinomycin
Issues in the analysis of oligonucleotide tiling microarrays for transcript mapping
Prioritized selection of oligodeoxyribonucleotide probes for efficient hybridization to RNA transcripts
A hidden Markov support vector machine framework incorporating profile geometry learning for identifying microbial RNA in tiling array data