Thứ Tư, 1 tháng 2, 2017

Waching daily Feb 1 2017

Cooking Celebration Cake Princess | Games for Girls - Beautiful and Delicious Cake

For more infomation >> Cooking Celebration Cake Princess | Games for Girls - Beautiful and Delicious Cake - Duration: 5:32.

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Handcuffed for handing out the Constitution - Duration: 1:23.

WE JUST ANNOUNCED YESTERDAY,

AT KELLOGG COMMUNITY COLLEGE? STUDENTS WERE ARRESTED,

HANDCUFFED, HAULED OFF TO JAIL AND BOOKED FOR HANDING OUT

CONSTITUTIONS. >> AS WELL THEY SHOULD-NO, THAT'S WRONG, THAT'S

BAD, ISN'T IT? THAT'S- >> (LAUGHS) YEAH. >> FOR HANDING

OUT THE CONSTITUTION. 'CAUSE IT'S SUCH A HATEFUL DOCUMENT. >>

ABSOLUTELY. SO IT'S- >> THAT'S JUST STAG-OKAY, THAT KIND OF,

LIKE, MAKES YOU GO "BOING!" ANOTHER CASE WHERE IT WOULD BE,

LIKE, "YOU'VE GOTTA BE KIDDING," CAN YOU THINK OF ANOTHER? >> OH,

ABSOLUTELY. LIKE EXAMPLES TO WHERE YOU HAVE CHRISTIAN

BUSINESS OWNERS ARE BEING TOLD THEY HAVE TO PARTICIPATE IN

SAME-SEX WEDDINGS. >> RIGHT. >> AND IT'S, LIKE, THEY CANNOT, YOU

KNOW, REFUSE A CLIENT. IT'S NOT THAT THEY HAVE ANY ANIMUS

TOWARDS ANYONE IN THE HOMOSEXUAL LIFESTYLE, AND SOME OF THESE

INDIVIDUALS EVEN HAVE HOMOSEXUAL EMPLOYEES. THEY HAVE NO ANIMUS,

BUT THEY CAN'T PARTICIPATE IN THAT ACTIVITY BECAUSE THEY FEEL

IT'S A DENIAL OF THEIR FAITH. >> YEAH. BUT, SEE, THAT, THOUGH,

THAT LITTLE...RULE RIGHT THERE APPLIES ACROSS THE BOARD, WHICH

IS WHY THE CONGRESS PEOPLE WHO DIDN'T WANT TO ATTEND THE

INAUGURATION HAD TO GO. >> EXACTLY. (LAUGHS) >> YEAH.

ALRIGHT. KEEP UP THE GREAT WORK.

For more infomation >> Handcuffed for handing out the Constitution - Duration: 1:23.

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Prime Minister's Questions: 1 February 2017 - Duration: 39:19.

For more infomation >> Prime Minister's Questions: 1 February 2017 - Duration: 39:19.

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Topeka votes to hire out of state firm to look for new city manager - Duration: 0:31.

COUNCIL VOTED

TONIGHT

TO HIRE AN

OUT-OF-STATE FIRM

TO

SEARCH FOR ITS

NEXT

CITY MANAGER.

THE GOVERNING

BODY VOTED 7 TO 2

TO

NEGOTIATE A

CONTRACT

WITH EXECUTIVE

RECRUITING FIRM

"STRATEGIC

GOVERNING

RESOURCES" OUT

OF

TEXAS.

COUNCIL MEMBERS

"KAREN HILLER" AND

"ELAINE SCHWARTZ"

CAST

NO VOTES.

DURING

DISCUSSIONS..

HILLER

SAID SHE SHE

WANTED

THE CITY TO CHECK

REFERENCES

CHECKED

BEFORE MOVING

FORWARD.

SCHWARTZ SAYS

SHE

WANTS THE SEARCH

TO BE

CONDUCTED BY

SOMEONE

MORE FAMILIAR

WITH THE

NEEDS OF

TOPEKA.###

For more infomation >> Topeka votes to hire out of state firm to look for new city manager - Duration: 0:31.

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HOT NEWS! 2017 Subaru BRZ Performance Package - Duration: 4:09.

there's something to be said for using

the right tool for the job

it's not always possible but you know

that when you will have the vice grip

deep down inside it just feels wrong

their satisfaction joint even dignity

and using the proper instruments to

execute a task accurately on the first

tribe our attempt will be updated 2017

super brz on fuji speedway last year

were stymied by summer college

enrollment at the base of Mount Fuji

reducing visibility to just a couple of

car line but it's an extremely can't

circuit with a nearly mile-long front

straight so even on a clear day it's not

the ideal venue for the BRZ which is why

we accepted Subarus invitation to take

another go at it this time at Turkey to

aquatic and off-the-beaten-path track

near granada spain if you go trying to

pawn ella the performance packages the

highest spec available on the chassis to

date in the US and that includes toyota

and science presents the setup is

exclusive to the ER is a line and has no

toyota counterparts with the price of

1195 dollars on top and only available

lon the 20 8465 dollars limited trim

level with a manual it gets you a host

of upgrades to its unsprung component

all of which would cost several times

more is procured piecemeal in the

aftermarket sorry there are no power

upgrade save for the bump of five

horsepower and five pound feet of torque

on manual transmission 2017 modules the

most noticeable of the enhancements are

sharp gunmetal finish 17 by seven got

five inch wheel inspired by the same

Doris watanabe designed the door in

Japanese turning racers of the nineteen

seventies

the extra half inches with accommodate

larger grandma break four-piston

calipers upfront biting down on rotors

that have grown by 0.95 inches in

diameter and thickness to 12.8 by 1.18

and to pot calipers pinching 12 dot

forward by zero dot 79 inch rotors up

from 11.4 buying 0.71 how that these are

the same great dimensions that you'll

find on the car subaru still considers

the flag bearer of its enthusiasts

lineup the rally ready WRX STI

performance pack the are these are

suspended by fax EF tampers and curry

weight penalty of just 20 pounds over

the limited the series dot yellow seen

here takes all the goodies of the

performance package and adds exclusive

yellow paint this through is quick to

point out is entirely different from the

yellow that appeared on the 2015 lioness

RS release series 1.0 will concede that

it's less boy racer but only slightly

other visual cues include black versions

of the performance a callaway all black

side mirror housings and smokes imagine

For more infomation >> HOT NEWS! 2017 Subaru BRZ Performance Package - Duration: 4:09.

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Things That Make Me Happy - Winter 2017 - Duration: 6:06.

Lately, I've been feeling like I need to

add a little bit of positivity to my

life. It's easy to let your outlook get a

little bit darker whatever things aren't

really going your way so I thought that

I would bring back up this idea for a

series that I had a long time ago. We'll see if I

stick with it this time. Because I

think now more than ever, it's important

to focus on the things that make me

happy.

Hot cocoa!

I like tea. I've never really been a

coffee drinker but whenever I'm making a

choice regarding which hot drink i'm

going to have in the winter, it's hot

chocolate. My step-brother got me this

amazing hot chocolate that I can make

over a stove. It's these little blocks of spicy

chocolate that you melt in milk or in my

case, coconut milk and it is so good. So

especially when I'm having a bad day,

there are a lot of things that a cup of

hot cocoa will fix. [stressed out laugh]

Lately, I've been finding a lot of joy

in making art for people. I consider

myself an artist in a kind of broad

definition of the term. And I've enjoyed

making art but I've never really

considered myself to be much of a

traditional artist as far as drawing and

painting goes so I've been practicing a

lot lately and this past holiday season,

I made a bunch of custom portraits for

families that are important to me and

I'm actually really happy with the way

that they came out and more than

anything else, the reaction that I've

been getting to them has completely made

my stressful holiday season worth it.

There was something about watching them

look at this piece of paper that had a drawing

I had done of them. Before my dad even got

the bubble wrap off of his portrait, he

was crying. Giving gifts is always a

wonderful experience like that but I

feel like given how much time I put

into it and how personal it was, it

really just made me feel so good inside

to see how much they loved that so I'm

gonna try and do it more often if I have

the time because it really made me feel

good and I really needed that this

season.

Even though I'm all grown up, even

though I hate driving in it, I really love

big fluffy snowflakes. Just when there's

a blizzard and they're, the snowflakes are like

this big and they kind of float through

the air instead of falling down, they're

just so beautiful and I just want to take pictures of

them but the photos never quite capture

just how light and airy and fluffy and

wonderful they are and they make me very

happy.

I know it's a silly thing, but you have to

focus on the silly things sometimes.

Another thing that has made me very

happy

is being able to sleep through the night

for the first time since last summer.

When you have a baby, sleep just usually

doesn't happen and for a very very long

time

our son was not sleeping and lately we've been

working extra hard with him on

sleeping and he's done it a few times

this week

And it has been so nice! I've gotten to hang out at night

instead of going immediately to bed. I've had

the opportunity to wake up in the

morning and not feel like I've been hit

by a truck

it's been beautiful and wonderful. It's very

good. doesn't happen all the time but when it

does, I am very happy about it.

Pokemon! Pokemon has been making me very

happy lately. Yes, I'm still playing

Pokemon Go. I don't get to go out for as

many walks because it's really cold

outside but I bought myself Pokemon Moon for

Christmas and I've been enjoying it

mostly because I actually have time to

play it once my baby goes to bed.

It's been really nice to just do

something for me that's not stressful, that

I've actually enjoyed since I was a

little kid and it's just

yeah! It makes me happy. That's been a thing

lately that has made my life just a little

bit better.

I definitely go through stages of my

life where I'm really obsessed with

Polaroid or Instax photos sometimes they just

come back into my life and I realize, man

they're cute! What is so cute about

these little instant photos with the little

white plastic frames?

I don't know! There's something about them. My

nephew brought an Instax camera out to

dinner with us and took a bunch of

pictures of everyone in the family and

gave them to us to keep and we have them up on

our fridge and I just, I love them! They're adorable.

The newly illustrated versions of the

Harry Potter books are beautiful.

We have the first two since that's all that's been

released so far and I have not read them

yet because I want to experience them for the

first time with my son when he's old

enough to understand what I'm saying

to him but I flipped through them and they

are spectacular. I just, we didn't have them

when I was younger obviously because

they're new and I am really excited to

get to enjoy this new version of the

stories I love so much with my own child.

I hope he likes them. I might be a little bit sad

if he hates Harry Potter.

another thing that makes me happy is

supporting handmade artists when I'm

financially able to. In addition to

being a handmade artist myself, I really

really love buying other people's handmade work.

It makes me super happy and knowing how

important it is for me to make a sale,

I really think it's important to support

the handmade artists that I love

Lately money's been a little hard to come

by but this past holiday season, whatever

anybody asked me what I wanted for

Christmas, I just basically sent them the

link to these artists that I love and some

people bought me some work that I've

been admiring for years and years and so

event though I didn't have the means to

support the artists myself, I was able to

direct somebody who did to check their

stuff out and now I have these wonderful

little handmade products and these

beautiful pieces of art and I get to

enjoy them and I love it! and to wrap up

this episode, the fact that I have a place

where I can share the things that I

create with people who will listen and

who will tell me nice things if they enjoyed

it is really wonderful and amazing!

I make these videos because I really

enjoy making these videos. I like having

something to look back on to document

different parts of my life but the fact

that all of you are here makes me

incredibly incredibly happy and I feel

really fortunate to have the opportunity

to show you the things that I'm making.

Thank you so much for being here and

supporting me. If any of these things

made you smile too, definitely like this

video and I'll see you soon!

For more infomation >> Things That Make Me Happy - Winter 2017 - Duration: 6:06.

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Games for Kids Talking Tom Gold Run Vs Despicable Me Vs Talking Hank Kids Video Android Youtube Kids - Duration: 9:04.

Games for Kids Talking Tom Gold Run Vs Despicable Me Vs Talking Hank Kids Video Android Youtube Kids

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For more infomation >> Games for Kids Talking Tom Gold Run Vs Despicable Me Vs Talking Hank Kids Video Android Youtube Kids - Duration: 9:04.

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When will the IPO floodgates open for tech's unicorns? - Duration: 1:06.

The floodgates are probably open today.

Our team talks about a difference between a demand-side problem and a supply-side problem. Floodgates

suggest that the demand has not been there, I'd actually argue the demand

has certainly been there, but because of the preponderance of capital available in the

private markets for the last several years, companies have not needed to come public.

We've seen the private markets shift quite a bit from '15 to '16 and we've seen

activity pick up in terms of companies preparing to go public, so I would certainly anticipate

that '17 will be the year in which we address the supply side.

We've seen some really encouraging signs here in the latter part of '16 in terms

of investor interest in technology companies, in growth companies, that's very evident

to us today, it's really just a matter of companies coming to the fore and wanting to

be public companies.

For more infomation >> When will the IPO floodgates open for tech's unicorns? - Duration: 1:06.

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Wrangler Barricade Trailforce HD Front Bumper w/ LED Lights & Winch (2007-2017 JK) Review & Install - Duration: 8:26.

I'm Ryan from extremeterrain.com, and this is my review and installation of the Barricade

Trailforce HD Front Bumper with LED Lights and Winch Combination, fitting all 2007 and

up JKs.

Today, we're gonna talk through the installation of this kit, which is going to be a fairly

simple one out of three wrenches.

Even though there are lights on this bumper and you have the winch, you're really only

making a couple of connections at your battery.

So again, one out of three wrenches for this install, but do give yourself around two hours

to get it done, maybe even a little bit more.

We'll talk more about the install in just a second.

We're also gonna talk through the construction and a few of the features of this kit.

This combination includes, of course, both your bumper and your winch.

You know that these two pieces are going to work together, you're gonna save a little

bit of money by buying them at the same time, and you're going to end up with a package

that is both very useful and will also drastically transform the look of the front of your Jeep.

Now, this kit is available with a couple of different bumpers, and this one, in particular,

has the LED lights built into it.

These lights are a little bit more of a marker light than they are an auxiliary light that's

actually gonna throw a lot of light in front of you on the road or on the trail.

But if you're looking for that, if you like that look of an integrated, built-in flush

mount light, and you want a winch that's gonna be very functional as well, all while saving

a little bit of money, buying them together in this combo pack is a really good option.

So obviously, there are two separate pieces here, and first we'll talk about the bumper.

This is the Barricade Trailforce HD Front Bumper with integrated LED lights.

This is going to be 5/32nds-inch steel covered in a textured black powder coat finish.

It does, of course, have the winch mount right up on top and your fair lead mount as well.

It has two D-ring mounts that are welded and bolt directly into your frame horns, giving

you a really solid connection for either a snatch strap recovery or using a snatch block

with your winch in order to double the pulling power of the winch.

And this does also come with the 3/4-inch D-rings as well, so it's not a piece that

you have to purchase separately, which is really nice, something you don't have to think

about.

This, of course, has a 2-inch hoop up on top of the bumper, which is going to offer a little

bit of protection to your winch from branches and brush.

And it also two light mount tabs up on the top, allowing you to mount any traditional

single post light.

Whether you're looking to mount some additional auxiliary lights or even a light bar up here,

you do have a couple of different options.

Now, this is a mid-width bumper from Barricade, so it is going to be slightly shorter than

that factory front bumper.

But even if you have a factory fender flare on your Jeep, I still think that this is going

to look pretty good.

Sometimes, when you go with a stubby bumper, you really want an aggressive fender flare

that jets in to give you a nice, clean look from the front.

With this, you can get away with running a bunch of different styles of fender flare

because it is a mid-width bumper, so you are going to get some protection and you're gonna

get those additional features that we just talked about.

Of course, this is the kit that comes with the winch as well, and this is Barricade's

9,500-Pound Winch with Synthetic Line.

Now, the general rule of thumb for the pulling capacity of a winch is that you want it to

be roughly double the loaded trail weight of your vehicle.

So if you have a really heavy, armored-up 4-door JK that you plan on burying up to the

axles in mud, maybe you wanna consider the 12,000-pound winch.

However, if you have a little bit of a lighter 4-door or certainly a light 2-door rig, 9,500

pounds is going to be more than enough for you.

Now, this is gonna be a 5.5 horsepower motor.

This does have a three-stage planetary gear set, so it is going to give you a good bit

of pulling power.

Now, in general, I really like a synthetic line.

It's going to be more expensive up front, you're going to have to replace it and maintain

it more often, and it is going to be more expensive every time you purchase a new one.

However, the benefit to a synthetic line is that it's lighter-weight, it's not going to

kink, it's easier to drag up those muddy hills, and it doesn't store energy like a steel cable

will.

This is going to stretch just like steel will, but when steel is under load and it breaks,

it snaps back.

It can hit somebody, causing an injury, or just some damage to your Jeep.

With the synthetic line, again, it doesn't store that energy, so if it were to break,

it falls directly to the ground fairly harmlessly.

You can mount your solenoid box right up on top here, or you could, of course, mount it

in back if you chose to.

Now, there are winches on the market that have some additional features, that if you're

gonna be doing some really long, hard pulls every single weekend, I might recommend spending

a little bit more on one of those other winches.

However, if you're going to be using this occasionally, this is going to be a really

nice winch that's gonna save you some money over those other options that are available.

This install is going to a one-out-of-three-wrench install that will probably take you right

around two hours, and the first step in getting this installed is going to be removing your

factory front bumper.

You'll do that by removing the splash shield on the bottom and the frame cover on the top,

removing the fog light sockets and the fog light wiring harness, and finally unbolting

that bumper and removing it from the Jeep.

Now, at this point, I would recommend installing the wiring harness for the lights on your

new bumper.

These are not going to plug directly into where your factory fog lights do, but this

does come with a really nice harness.

That includes the switch, the relay, the fuse and will plug directly into these lights,

so all you have to do is pick up power at the battery.

So you're gonna have to pull the switch wire through the firewall, get the switch mounted

inside the cab.

You're gonna run the two power wires, positive and negative, up to the battery where you'll

pick up power there.

And then you'll run the actual connection ends down to the front of the frame where

the bumper is going to be.

Once you get that into place, I would recommend bolting your winch up.

First, you're going to bolt the solenoid box on top of the winch if that's where you choose

to mount it, attach your wires from the box to the winch itself, so all you're going to

have are those two long leads that go to the battery, and finally bolt your winch onto

the bumper.

For that, you're gonna get your fair lead in place.

So once you have the winch bolted up to the bumper, you're gonna have a big, heavy package.

Why not install the bumper first and then the winch?

Well, it can be a little bit difficult to get to those winch mounting bolts if you do

it that way.

So again, I recommend the winch gets mounted to the bumper and the bumper and winch as

a package get mounted onto the Jeep.

Once you do have everything bolted up, you simply lift the bumper and winch onto the

frame rails and use the included hardware to get it bolted down tight.

Because you've already run all of the wiring for your lights, you simply plug those lights

in.

You'll want to stretch the two positive and negative wires from your winch through your

grille, up along the fender, and attach those to the battery as well as attaching the two

leads that you ran for your wiring harness to the battery as well.

And that's why I'm still giving this a one out of three wrenches.

Even though you do have the lights and the winch to wire up, it's really just four wires,

two that get attached to the positive terminal of your battery, two that get attached to

the negative terminal of your battery.

So even if you're not someone who's comfortable with automotive wiring, that's something that

really anybody can do.

As far as the tools necessary to get this installed, really just your traditional hand

tools.

No wiring tools or anything like that are necessary.

This is gonna be a pretty straightforward and a pretty easy install for anyone to do

in their driveway.

Overall, I think that this bumper and winch combination is fairly priced.

Of course, you're going to save some money over purchasing the winch and the bumper separately,

and if you're looking for the features that are offered in this kit, I do think this is

gonna be a really nice option for those of you who are going to be occasionally using

your winch.

This is gonna save you some money over those top-tier winches that are much more expensive

that has additional features, and of course, you're going to have the bumper for its features

and its protection.

Overall, these are gonna work really well together, they're gonna drastically change

the look of your Jeep, and I think that they're gonna be a pretty nice option for the price.

So if you're looking for a front bumper and winch combination for your Jeep to offer some

more function and drastically change the look of the front of your Jeep, I think this is

going to be a fairly-priced option.

So that's my review of the Barricade Trailforce HD Front Bumper with LED Lights and Winch

Combination, fitting all 2007 and up JKs, that you can find right here at extremeterrain.com.

For more infomation >> Wrangler Barricade Trailforce HD Front Bumper w/ LED Lights & Winch (2007-2017 JK) Review & Install - Duration: 8:26.

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Hilarious Cruise Ship Pier Runners - ROASTED! - Duration: 4:37.

I love that he's like trying desperately to get this man up, but when he does, what is

his plan?

He's going...

I don't know...

Does he think he can disguise him?

I think it's "Weekend at Bernie'."

I think his plan is he's going to "Weekend at Bernie's" it, just kind of shove his arms

underneath and like "hey!"

Let me grab my phone... hold on, it's ringing... my phone's ringing.

Jimmy that's not a phone.

Mom!

Mom!

Jimmy, that's your shoe.

Mom!

Jimmy, that's your shoe.

Jimmy.

Jimmy.

Hang your shoe up.

Jimmy.

Now we've got some... they gave him a pillow to rest on, and just go to sleep.

If they could just let him sleep stand up...

Thank you, you're the best.

Your my buddy.

I love you Guinness shorts, hey Guiness shorts, you're my best friend.

Come on come on I gotta tell you something.

Can I tell you something? Hold on. Nah, forget it. Bye.

I feel like this is making the behavior acceptable.

Like hey, we're going to get you this tiny boat - we'll figure it out - we'll get you

this tiny oil rig... like, what is that?

If you had missed a boat, and that was the boat that came up to help you out, I'd be

like, no, I'm good.

No, this is the boat that Mark Wahlberg never came back on.

What was that movie?

The Perfect Storm?

Yes!

That is The Perfect Storm Boat!

Number 1, I was nuts enough to even get on that boat, then when they said to me what

we were about to do, like, not only are you going to get on this boat but hey, see that

guy that just opened this strange door on the cruise ship... this magical doorway appears...

we're going to figure out, we're going to throw you in that.

I'd be like, throw me in that?

And they're like, no, don't worry, we're going to give you this rickety strange ladder.

Oh gosh, she's flashing everyone. Wait, why would that be thing?

Also, we don't have... her husband must be like... uh, we don't have enough time to get

on this boat, we definitely don't have enough time for you to flash people.

She's probably one of those people that like she's really conservative when she's sober

but then you get a couple cocktails in Natalie and forget it, her bottoms come off.

That's just her instinct.

And her husband's so used to it, he's like pfff... that's my wife!

That's how she is! That's who I married!

They gave us extremely strict instructions and we said "I'm not listening! Phooey on you!"

They gave us instructions but they also let us get off on an island and 17 rum

cocktails.

Mama needs a minute.

I have a caftan, several caftans... that I need to purchase for discount prices at this

straw market.

Do you know how inexpensive?

First of all, my dollar goes twice as far, and they're knocking off, they're slashing

prices... left and right... in mid-conversation.

We're going to get to the edge of the dock and they're going to help from here.

I'm going to get to the edge of the dock and I'm going to let someone know who's standing

there "We're the group... they left... um, my dinner seating is at 5:30.

I don't wish to be late."

[WHISTLES & CAT CALLS FROM SHIP] "Here come the ***holes!"

"Stop!"

Stop!

Stop!

Stop!!

Stop!!!

Daniel, you're not going to yell curse words from our boat window, alright, Daniel?

This could go on the YouTubes and everyone's going to think suddenly that you're this guy?

For more infomation >> Hilarious Cruise Ship Pier Runners - ROASTED! - Duration: 4:37.

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treatment for diabetes in urdu – hindi sugar ka ilaj - Duration: 3:37.

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For more infomation >> treatment for diabetes in urdu – hindi sugar ka ilaj - Duration: 3:37.

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Kids First FY17 X01 Pre-Application Webinar - Duration: 47:33.

hello everyone and welcome to the kids

first webinar for the x1 pre-application

discussion

my name is wretched boy and I'll be

doing introductory part of the webinar

um I'd like you to go over a few

instructions so that you understand how

we're going to operate the webinar on

the program is being hosted by the

Gabriella Miller kids first pediatric

research program and several of the

people involved in the program here at

NIH will be talking with you today right

now all the participants are music upon

entry for the webinar and we ask that

you please remain seated until the

question or discussion period at the end

of the presentation and that's how you

can unmute yourself by selecting start

six on your telephone or clicking on the

light symbol under the audio connection

on the webinar you can ask technical

questions at any time using the chat

service through the webinar to the host

if you are having technical difficulties

please feel free to use this means of

communication please save any questions

related to the program or funding

opportunity announcement for the

question period at the end and it's my

chance you have additional questions

after the webinar is over please feel

free to email them six his first Cody's

at iah not go and just a reminder we are

recording the webinar and the recording

will be posted to the kids first website

in case you want to review any aspect of

it or you have colleagues that were

unavailable this time to listen to live

so this is the pre-application webinar

for the discovery of the genetic basis

of childhood cancers and structural

birth defects the Gabriella Miller his

first TD after research programs

excellent one initiative and before we

get into the details of the funding

opportunity announcement but I want to

provide a little bit of background

information and introduce to you the

people that comprise the church

leadership group here at NIH we have a

number of programs involved isn't common

fund projects as you know it was

initiated in 2014 2015 with the overall

goal of establishing its first a

resource for the pediatric research

community we find we're very excited by

this opportunity both for Ali a chance

for the PDF research community programs

focusing on structural birth defects in

pediatric cancers and this is a comment

on program leadership responsibilities

are being shared across for NIH

Institute's Wallace college but the

people listed on this slide are the

folks that are involved in coordinating

the program as i mentioned i'm erection

boy i'm a program officer and I chg

Jonathan called mentions from NHLBI and

he and I are the points of contact for

special birthday sex questions of

themselves solve is an NHGRI and along

with James cool

nichd there the point of contact for

people in questions

James the project officer for the first

statement punters Nelson S&J goodra all

our program officers of the National

Cancer Institute and there are the point

of contact for ideas

PS answer questions

and Martin lurkey is the is involved

with our data resource project which

essentially funds this year will be the

process by which coordinating our

efforts from the common funds we have

very serious who is our program officer

and my instructor who is the operations

and budget person we also have a person

who handles policy planning evaluation

indication for us it was Josie honest we

recently lost them to another position

here at NIH more the process of getting

someone new and you all are probably

familiar with Valerie cotton who is our

administrative points1 coordinated

application and webinar today I just

want to take a moment to point out that

there's more than usual interest in this

program from both Congress and the other

50 communities partly because the way

the program change to be and today we

have stepped ishe of the Common Fund on

the line with us shoot the team leader

for the policy planning evaluation and

communication boxes and she's filling in

in the role of communications person

today during the webinar but you have

any questions on this aspect of the

program should be able to answer them

during our discussion period so the

people highlighted involved on this

slide myself and John Colin and Valerie

hot and will be the people talking with

you today during the course of the

webinar can XYZ I just want to go

through a bit of background as to how we

got where we are today so the program

was initiated and respond to the 2014

Gabriella Miller its first research act

which was signed into law and a third

2014 this law actually did three things

it ends attacks their contribution to

the presidential nominating conventions

and look at the small graphic at the

bottom of the slide visit the snap of

the other portion of your IRS 1040 and

the area highlighted in red was actually

a little checkbox that existed on

1040 back and your prior to 2014 and

2014 which allows taxpayers to

contribute tree dollars towards the

presidential nominating conventions fun

turned out a lot of money with cities

and the funds wasn't being best but the

times long past what it did was it

authorize the transfer of a hundred

twenty-six million dollars which were

sitting in the fun into a new pediatric

research initiative but it also

authorized congress to appropriate 12.6

million dollars a year for each of 10

years directly to the nih common fund

for a new pediatric research program and

the first appropriation was accomplished

in fiscal year 2015 think from our

perspective in bunny the program the

most important word on the slide is it

authorized the appropriation on an

annual basis so very important to

remember that this program there is a

mandate for 10 years of funding were

dependent upon congressional

appropriation annual basis to get the

funds for the program which artists back

to my earlier statement that there's a

lot of interest on the part of advocacy

group to see the captain's on an annual

basis

ok so if we look more closely at his

first working group as i mentioned as a

transit is effort supported by nih

common fund the Institute's that Sheriff

his first working group are the Child

Health Institute hard ones laws cancer

institute and the genome Institute it's

very important that you understand there

are a number of other is that you

working on this program and they're

listed here on the slide so just briefly

reiterated fidelis to choose the drug

Institute alcohol diabetes and kidney

Environmental Health Sciences the eye

and tissues are prized the

musculoskeletal diseases infectious

disease and the office research

instruction program

and we also have an ex-officio member

from the CDC's birth defects division

sitting on the working group so we've

got a lot of people involved in this

program and want you to be aware of this

because if you're interested in applying

to the exelon program it may be that you

your mission align more with one of

these other institutes and you should

feel free to contact program officers

you know what these Institute's that you

have questions about how relevant what

you want to do is to their mission for a

complete list of our working remembers

you can go to the kids first Common Fund

West fight and look at working group

members of the link there now I'm going

to turn the webinar over to John call

then who's going to review the program

goals

alright thank you so the overall goal of

his first program is to develop a data

resource which we are calling the

Gabriella Miller kids first p educated

resource where pediatric research

community can access well-curated

clinical genetic sequence data that will

allow them to identify genetic pathways

that underlies childhood cancer and

structural birth defects a greater

understanding of the genetic

underpinnings of these pediatric

condition is critical for developing and

improving preventive measures for

diagnostics is the character

intervention there are three main

edition is supported his first program

they include cohort identification DNA

sequencing the cancerous data resource

and data analyses

we'll talk about the first one goes late

today although i'll i'll briefly

mentioned what that the other two will

be doing swell cover identification the

DNA sequencing is accomplished during

the excellent program which will be the

topic of discussion to a rule is to

identify cohorts that children with

either childhood cancer or structural

birth defects more bone and their

families for whole-genome sequencing

concerns before the whole genome

sequence take of these cohorts to

elucidate the genetic contribution to

childhood cancers and a genetic etiology

of special birth defects the pictures

pediatric data resource will be awarded

an established later this year

the goal of this reasonable to develop a

web-based portal that will aid

researchers in identifying cohorts that

are available and have a sequence data

and that will also facilitate out cross

cohort analysis and it finally street

analyses conditions which is further

down the rows will be set up after the

resources of the naughty and will

further support analysis of kids

regenerated as well as knowledge it is

generated data to uncover new insight

into the biology of childhood cancer she

shows us now to talk about the extra

watches need a third cycle of our extra

1i it's currently called discovery of

the genetic basis to challenge cancers

and instructional 30 secs the first one

was released in 2015 and

jun 2015 x12 childhood cancer cohorts

five structural birth defects covert

sequence in 2016 three childhood

Cantrell chords and five structural

represent cool ones you want and then

the current exit one is par-70 dash 63

we hope you plan to continue sequencing

for several more years provided the

funds are appropriated as right

discussed and that cohorts are available

funding announcement has slightly

evolved over the three years of the

program in particular lesson emphasis

being placed on the minimum number of

samples also less than s is being placed

on trio design as long as there is a

valid analysis plan that accompanies the

overspeed proposed just a word about the

actual one back isn't the extra one is

not a typical award mechanism or typical

grant mechanism there is no award that's

given to the recipients and in fact

there is no notice of award recipients

are selected for the opportunity to have

their co-worker sequence again no funds

are provided more analysis but there is

an accompanied ro3 which is PR 16

dentistry 48 which originates from nichd

but has the number of partners that

supports analysis of data that is

generated by the excellent project also

these awards are not listed the is

reporter but abstracts from our prior

awardees are listed all the kids first

website the

web addresses on your street no

alternative about Valerie so i'm not

really tired and at laura is my a

program analyst 4-h program and a large

part of my job is to be able to speak

with the intent of fat as well as the

sln investigators will be selected

through PR 70 degrees i'm going to go

over some details that will hopefully

help you with your application to this

half fish and some of the information

about to cover is already addressed an

RFA Hughes if you are considering

applying for this at all i highly

encourage you to look at her if you

think you can find on our Facebook first

combines website at this link here we

can offer constantly updating faq will

update them after today of the questions

that we get the discussion that we have

and we see them on an ongoing basis as

we get everything for you see us

becoming centers that are providing the

UC printing services so that i described

in the so one away

they were selected for cooperative

agreements and we will be working with

such as for three years starting in

fiscal year 16 those cohorts have

already been selected and the samples

are being shipped and they're answering

the secret life around now I'm gonna be

working with them

physical you're 18 the first answer is

because been outside Institute provide

technology but I thought we'd be in

partnership with speak to children's

research hospital but by my exact the

other sensor is the grossest issue of

harvard and MIT like myspace Behavior

all photos of the sensors have the

capacity to sequence both cancer process

and birth defects projects

however our plan is to assign all kinds

of projects to the sequencing center

husband else's shoes to take advantage

of the analytical expertise for

identifying somatic variants that was

offered by the center and but depending

on the application as we can

exposes better proposed by with a

coworker left behind a process may not

work out exactly this way but this is

our general framework to see screen

centers are tests with 40 the sequencing

as well the variant calling I've already

ate things of DNA and RNA so that means

that will generate the variant calling

vials as well as bands and in some cases

that cute smile because cohorts will

receive holiness sequence paying an

average of thirty x coverage of cancer

called board will also receive 3x

coverage for whole-genome sequencing for

journalists sample but one change in

this my way that we are really not

introducing the submission of nucleic

acids from tumor samples I to go to the

husband elbows Thank You Spencer was it

was generate hold my own secret thing as

well as all that something RNA

sequencing because this is an

information thanks for their analytical

pipeline you may propose other methods

or technological approaches pretty one

thing in your applications for example

this past I stole someone's for proposed

higher coverage than 30s in the end

after working with us and our sequencing

centers they determined that 30s was the

most reasonable they would have free

sample size they wanted higher coverage

and they decided that the higher

simplifying the more valuable so on this

is an example of how you can propose

other things but the final product

design will be decided among discussions

with excellent investigators sequencing

center shaft and a nice program staff

another possibility that you might be

interested in proposing is a lynx long

new sequencing this is actually a

technological approach that was a pro

proposed by mostly sequencing centers

but since this is a new technology we've

actually asked them to work together to

run a small pilot study and this is so

that we can evaluate and better

understand the added value of utility of

this approach for these types of comfort

for birth defects and pediatric cancer

those they're doing this using 10x

platform so based on the results of a

pilot studies this is something that may

become more available for future

opportunities including this one so this

interests you at all

I encourage you to describe this in your

application and you invited application

for why you think it might be useful

your code for I just want to take a

moment and talk about where data lives

and how it's accessible so all this

hurts that you will have to be

registered in DB gap if you're not

familiar with unique a pin and I a

system for facilitating control access

of genetic data to other researchers so

you would be the time to research

research area generating the data but a

goal is to make this accessible to other

researchers as well and the end pts was

sort of a keeper out that process so I

projects well beyond the certain TV gap

sumx and i will also be submitted a gap

of sequence Dana will be submitted to

ncbi sequence read archive actually the

sequencing center will be responsible

for them but i just wanted to make sure

you guys need to wear daily living

intertitles of cancer process they also

have to be resurrecting PDF but all data

my assessment make that and sequence

data will be submitted to NCIS genomic

data Commons this is a database that was

launched within the last year it's still

a little bit new show NCIS that working

with us and our investigators and see

sequencing centers to make sure that we

get the necessary information from each

project properly submitted so now i'm

going to address a few of our

expectations are requirements in terms

of what his first program goals are so

from the beginning we've been very

focused on whole genome sequencing to

discover Connecticut

is contributing to these pts condition

but we had the opportunity for full deck

so it's just transfer zone season of

humor as well based on the proposal and

the value of this argued by uzma thank

you

so given these are just because they

develop them be performed our efforts

expectation is that the DNA and in the

case of tumors are named as well

exceptions that you plan to submit are

sufficient quality and concentration of

these technologies will talk a little

bit later about how we ask you that

information is the application another

goal of the program is justin silicate

cross-court analyses base and these be

done based on common development one

very large task of our upcoming his

first date of resource will be to

harmonize phenotype across all of our

process so to this end we really need

some basic element but the better the

more details happy that you have the

better

um for birth defects projects we don't

have established set of standards for

phenotypes data but since the candidate

will be going into NCIC economic data

comments they have already clearly

established the elements that they

expect for my assessment and clinical

and you know Vegas so you can find this

swing here and also in our FAQ to look

at that you use data dictionary another

goal of ours is programmed out and so we

meet to run the diversity of both PSR

cancer and special birth defects data

says that will initially be part of our

kids first data resource this means that

if we get a lot of applications of me

hopes that we knew and we are faced with

the choice of cute very similar

applications but one of the proposals of

sequencing of the condition that we

haven't covered before we are more

likely to select that cord

this doesn't mean we can serve you broke

lying if you have a cohort of similar to

what we've got the board that we want to

be a parent

office and we in previous life if you

link to where you can find the list of

quickly statements before and we

encourage you to look at that huge goal

ours is data sharing so to make the

animated by our program as acceptable to

be this community as possible so in line

with NIH's no mysterious errand policy

cohort participants must have your

physics consent to allow the sharing of

individual level sequence and the

associated seen effective at through

high approved repository the classic

example EE gap another priority of ours

is to make sure this data to go to as

many as are constantly service possible

so we are most interested in doing that

is impressive to share as broadly as

possible though something that submitted

for general issues something that is

constructive for health medical or

biological research is preferred over

busy specific data use restrictions and

it's just because we want to promote

collaboration long investigators as much

as possible and then just because a

little bit more complicated with data

sets are restricted success researchers

again it doesn't mean we discourage you

from applying that you have is the

specific restrictions of but we do ask

that you previously that your

application and some of you like us is

recruiting you court or even be

confessing for every anyway as me and I

have guidelines two guys using who have

made with that allows for grass sharing

sample size and family structure as John

mentioned in previous years we really

focus on his trio base designs of what

we realized that there are other family

structures and designs out there that

can lead to discovery but this needs to

be very clearly communicated in your

application of obviously the larger the

sample size the better

um in some cases we have birth defects

of in Hawaii we talk about the birth

defects minimum 100 trios

but if you less than that I'm we would

like to see you so I clear scientific

justification for how you will be able

to identify the Americans if your sample

size not that large very strongly

encourage you to consider collaborating

with other investigators cool samples

together for larger sample size and of

course in the case that you don't have

trios use this week you on show us an

argument for how us that you have enough

program and toss that you will see Chris

enough effective and/or unaffected

family members to be in any discovery

and this type and very closely with

analysis Lance the goal asking for

analysis plans in these applications if

you have investigators seven days that

the proposed project has an adequate

research design for genetic discovery

and that usps one investigators are in

addition to responding analyses so one

of modification of this iteration of the

F away is that we do communicate that we

recognize the analytical power of each

dataset will increase once incorporated

with all the other datasets that are

part of his first no class it is

important that you demonstrate the data

will be usable on its own

if you feel that your team does not have

the analytical and little call our

statistical expertise to perform these

now is we highly encourage you to

partner with another group who may be

able to work with you on an analysis

plan so now i'm going to move on to

adjust a part of b funding announcement

called other attachments entire years

we've always discussed the institutional

education of indiscretion but from this

year we greatly expanded this part of

the application of to ask for sample

information description of the penis

penis

any description of the family structures

and even after this here that we want to

enable you to submit supplemental

additional material all the explaining

and response to make me a sport here

without really cutting into your limited

user strategy so we have also created a

optional downloadable form that is

available on earth a fuse and that is a

template where you can give us the

information that was asking or this is

awesome

you may choose to respond this

information that we need and in another

format whatever way is best for you by

the Board says as a place where both

applications and reviewers can look at

work with the uniform document

information can be organized and I'm

going in the next slide right over

easily use one by one so the first

attachment would be the official

certification or criminal certifications

or sample consent form institutional

certifications are documents that sure

is that these innovative new BFF pieces

and i use genomic data sharing policy of

many team cannot usually get a full

official official certification from

their ir e until after the board or the

state after you left yet so one program

so you need to remind the original

certification or the simple consent form

instead we use after the presentation

please use the current and I Swiss can

be found at this link that's where you

can find both institutional education

and the cell certain cases that's what

no matter what you submit it needs to be

clear need to cover all the sites that

contribute simple to your project nice

to specify whether being a competitive

presented the energy gap or other and i

use repository and organizations need to

be very clear bass playing

this snapshot below is a msg from the

institutional certification but you can

see how automatically you will have to

organize and label that spreads arms but

if you're not submitting a full official

certification with your application you

need to describe them somewhere else the

application and i just want to mention

that no matter what you submit with your

application if your cousin is selected

you will have to provide the school and

Cisco certification using this template

shortly after we notify you know your

project has been selected and this is

the first step of that he got

registrations I talked about we're not

going to get into the details here just

know that i SAT here to help you with

this but this would be the first step on

the other questions we asked for our

sample information so we really need to

understand the total number of samples

that you are proposing for sequencing

very clearly again you can modify this

table you can submit this the

information in the s4 whatever way works

for you but this is one way you can

organize these details though this table

of the rows are organized by sample type

or or DNA or RNA source this hot part of

this table would be for both birth

defects cohorts and cancer cohorts and

the bottom part is for tumor sample for

testing project we just need you to

describe the number of samples from heat

exhaustion message that was used the

concentration some never got quality and

tell us which metra could use a method

of quantification and then we need to

know how how many samples will be ready

to ship by August 2017 and if you slide

I'll talk more about the timeline of

this program but we anticipate that we

will be notifying crew being selected

under this so17 team iterations in the

life so we are hoping that we will be

getting bigger than our samples of

pipeline by august but we realized that

types there are some lingering

extractions pencil penis off and

sometimes there are some shipping little

script coordinate so we asked for how

many samples are ready now and have

examples already six months from now

just to get a sense of what is ready to

go again you can modify this slightly if

you know that you'll have everything

ready by November of 2017 that you can

change the January 218 number 170 and

tell us when all your will be submitted

by we also ask you to describe what you

no text information and demographic

information you collected from these

participants have minimum we expect a

good diagnosis maybe maybe you have some

other Asian formation sex race/ethnicity

and then just tell us what else you

collected describe you

primary uh clinical description and I

purchase a kind heart tumor spice up and

tell us what other phenotype information

whether it's clinical or otherwise that

you've collected over also interested in

knowing what family medical history you

have this is a very general table again

we just need to who you give us a sense

of what being a kata you have available

again for cancer code words this is a

little bit more clearly laid out of the

standards provided funds you see we

highly encourage you to look at those

and again for those cohorts that are not

feel designs we ask that you describe

what the family structure bar you might

have a program parents siblings of that

you might have multiple family might

have something with his family's we

don't ask me to submit enough

information to make it clear out any

samples for family you're planning on

sequence and whether they're a bit and

then he doesn't like to see simplest to

submit a pedigree information but again

tell us who you're proposing for

sequencing if you don't have a degree

information we ask that you find another

way

makes you really make it clear what

you're proposing is a handful of people

that makes us to work from with one

group they have a trios although they

also have situations where a program and

another person three family member was

infected with some defective family

members so because needed to be clear

how many of these family-type if you

have many different family types of and

how many samples from unaffected and

family number of your proposal when a

couple other things that didn't quite

fit into other slice

we just want to emphasize that another

change to lose this iteration of this

program is that we are cruising the

submission of application up guess who

has already sequence a case or programs

through other funding but have fun

sequencing play NASA's problem of

parents four parents and siblings for

tumor if you do this you ask that you

pretty quickly just ride the sequencing

technology that uses into the program

and I was but we are certainly open

because Google we also an SS opportunity

is open soon is real research program

I'm and that includes collaborators of

that is real research program is because

it doesn't come with formal fund is that

opportunity for pain so for the cycles

in 2017 cycle of you made for me how

about a lot of things that we prefer a

fact but really no matter what we heard

any investigators that have a co-worker

that meets the minimum requirements to

consider flying as you so we're working

with our current because others until

2018 on but we also have plans to

continue sequence being in later years

so we encourage you to start collecting

samples of your requirements now but

stay and just with us about this again

what will continue continue to

sequencing as long as funds are

appropriated and so forth are now also

be you

stay aware of what cohorts are out there

this is the timeline for this cycle it

is on an earlier time lines in our

previous two years but we wanted to make

sure but he was clear on this

this funny as that was published in

December today's category 5 30 this is

the weapon are the letters of intent is

to separate them as you know these are

not required but they do help us prepare

for their review of their helpful

mark 7 is the application deadline we

anticipate that some review will take

place sometime in April but we're not

going to be between unions alive we will

work with our working group to make a

selection there are no funds associated

with this program it doesn't go to an

institution helpful to work with our

working group and then we get a

recommendation approved by the Institute

directors at Coachella or a group to

make our dividend and then we plan to

know either as of yet sighs line and we

hope to get these comforts multiplying

by august so now I'm logins the floor up

for questions

I just want to reiterate that you can

continue to reach out to us this is the

analogous memberships for which we are

monitoring you also have the hot

information for program officer Barry

who are leaders of this program in the

study and ask myself my email addresses

on this life so we'll be publishing the

TV

PS of this side effects of all

publishing the webinar recording of the

webinar and just a reminder will take

the questions now we like you have some

vocally and so you'll have to unmute

yourself if you can call them through

telephone you can select start six on

your phone to unmute yourself and if you

call them through your computer you'll

have to go to the audio connection and

select the likes of all who won't need

yourself

sounds like something yummy to

themselves and her friends

so we're ready to receive your questions

hey thanks for the presentation my name

is Craig 1 i'm calling from the

university of new mexico i had a

question regarding existing coverage

have been already uploaded to DB gap but

with a net level genotyping data what a

doesn't make it simpler to have that

already done ahead of time if we want to

get a whole genome sequencing for this

data set go ahead

not sure I fully understand the

questions you don't get a full so I've

already done uh sniff genotyping and had

already uploaded cohort data to DB gap

however we would like to get whole exome

or whole genome sequencing for this

existing cohort on that and this is data

or this is DNA out of the DNA repository

so regarding the certificates and all

the paperwork and registering for the

you know DB gap and and and the kind of

administrative requests that were

presented earlier in the presentation at

zacks exact stuff gets grandfathered in

or do you have to go ahead and you know

we do a lot of the paperwork you

probably have to work with the folks who

helped you

registers as the original trial but

probably you can piggyback this data

onto that trial we've done that it

other other scenarios that are somewhat

similar where a cohort has already

undergone friend is X equal to and then

underway whole genome was a part of this

so so that data was sort of attached to

the original code word data and

obviously that will likely simplify the

registration process that you've already

done terminated and downloaded to take

it as well thank you very much

other questions

tight

Alex writing from Pittsburgh and the

question regarding the phenotypes does

it have to be one particular phenotype

or 10 and actually the collection

includes several phenotypes

right so yet the the short answer is yes

it can

are you talking about birth defects of

war camp every four right back on it it

certainly can be a part of the idea here

was that from a genetic standpoint you

take to find a fair bit of overlap

between genetics of different types of

birth defects obviously i think an

application would be stronger from

analysis standpoint if you could make an

argument about how these different

phenotypes might be analyzed together

but but certainly that possible

and what about prenatal sample and that

would be DNA obtained from what source

ubi it will be a million pieces and when

you have ultrasound data to support the

fact that the fetus has a deep at birth

because yeah the point is that it would

be the phenotyping would be down like

prenatal ultrasound that he would be a

very unique phenotyping data sets

it's not precluded from being such as we

haven't had anything I guess we had

really consider the possibility of

something prenatal oh yeah I they're

happy scientific justification for why

we would devote resources to a prenatal

cohort as opposed to pediatric cohort it

i can imagine reviewers might wondering

about the sort of validity / accuracy of

prenatal diagnosis of which obviously

could potentially be bolstered by

postnatal confirmation of the diagnosis

but you know I think you have to make a

strong argument for why be done this way

as opposed to our exit waiting

thank you

Daniella Silva Washington I was just

wondering you know you're really quiet

my I wonder if you're not coming here my

friend boner you find another way to

make your foot ladder her outright that

better

ok so i was i I was wondering if there's

an emo concentration for DNA the new

position yet you go through a huge we

have some information about what are you

can be under the best if you want more

details feel free to contact me Valerie

or distress that loss and we talked

about that

so there is some information on our

issues thank you and my other question

was about the results so when do you

expect we would get the results back so

and and are they going to be VCF files

or are you going to send us the damn

styles

yeah so like a few people generate BCS

and band and those will be somehow

transferred back to the investigator and

the investigator will have six months to

work with their own data before it gets

released to the public through DB gap so

yes we have ways to get the vaccine

investigators a timeline for data

generation

well depend on a number of factors i

have 15 days I mind but for example of

colors that are going into the pipeline

now are expected to finish the pipeline

between four and seven minutes four and

seven months

ok and so it is expected that we have

capacities to receive vampires we do I

just want to make sure if i have to

state that on the application that

emergency or not we weren't sure you can

handle a couple words in there so have

you spoken sure so we want to know if we

have to put in the application that we

have space to receive them files for

example here we weren't sure if we're

going to receive those fires are going

to access them promote yes you you will

need to talk about your capacity of your

institution to store those vampire

okay great thanks

yeah so that's a good point of one thing

as well as but once you are electric

service program and assigned to a

requesting better

there's a lot of room to collaborate

with the sequencing center on analyses

again your application you're going to

have to tell us what what your resources

you have but just know that I'm you know

this is a partnership and collaboration

and they're really great resources and

state-of-the-art sequencing centers that

were working with you know from

everything to analysis is mostly best

but so I'm you know we're really excited

about working with us and help that

makes you and and the road so you can

keep that in mind but don't forget to

have analysis plan

well it doesn't not like we have any

other questions

I'm so I think we're going to go ahead

and wrap this up but please continue to

ask questions of us three rules all of

our lives and resources and email

addresses again this webinar will be

posted on our website and as the slides

will be posted for the links and

information as well so i'm going to be

myself now and the housetop the webinar

thank you thank you

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NCBI Minute: New Version of E-utilities Supports Accession.version Identifiers - Duration: 28:00.

All right everyone.

Welcome to the webinar today.

We will be talking about a new functionality within the E-utilities API that supports accession.version

identifiers.

We have materials for this webinar, the PowerPoint and a few bells and whistles that will be

available for you at the FTP site and you can get to that through the coursesandwebinars

link on the screen here and there is a direct link also, the go.usa.gov URL.

With all of that said, I will start the webinar.

What we're talking about today is actually the fourth webinar in a series of webinars

about changes that have been going on with the way that NCBI deals with sequence identifiers.

Today we are happy to announce that the E-utilities, our main API for the Entrez system, is now

able to fully support accession.version identifiers, both as input and output.

What I want to do is give you a quick overview of how that works and point you to some examples.

As I said we have been doing webinars about this since the past summer starting back in

July.

there are three webinars we put out there about various topics relevant to the changes

in identifiers for sequence records here at NCBI.

Feel free to go back and take a look at those if you haven't had a chance to view them.

Very short and sweet, the change that is happening is that gradually NCBI is phasing out an identifier

called G.I. number.

These are integer identifiers that have been used for many years for sequences but for

a number of reasons we are phasing those out and transitioning to a situation where we

will be using accession.version identifiers as the primary identifier for sequence records.

Since so many of our tools, including the E-utilities, have been dependent upon GI numbers

in the past, it was critical for us to be able to offer these APIs in a form that would

not be dependent upon GIs but would be dependent upon a different identifier set, accession.version.

The main thing that we introduced as a parameter called idtype that will control this.

It works a little bit differently in the various e-utilities.

I will go through esummary, efetch, esearch, elink . And then, the final prize package

at the end is how you get sequences that don't have GIs and what is that about and how you

can think about that problem going forward.

Let me review quickly again a little bit more detail about what's changing so everyone is

on the same page.

The GI numbers are being phased out.

What this means is they are not being deleted.

It's important for people to understand we are not deleting GI numbers but we are not

going to assign GI numbers to a growing number of newly deposited sequences.

If you have a sequence that has a G.I. number it will continue to have that G.I. number

and will always be retrievable by that number for the foreseeable future.

What's happening now is, as more and more new studies come in, those will not be assigned

GI numbers so if you stick with just G.I. space you won't have access to more and more

of the new data coming in.

A few things that have already been happening are that the GI numbers are no longer visible

on the flat file formats on the web or in FASTA definition lines, including in the BLAST

databases.

So you won't see the GI numbers in those presentations and again just to be clear those numbers are

still in the raw data files, for example, the ASN.1 or the XML.

They will not be shown in the more public web friendly presentations.

After March 15 of this year none of the releases of GenBank or RefSeq will contain GI numbers

in their presentations in the flat file or in the FASTA that you get off the ftp site.

That's another thing to be aware of.

That's what's happening.

Let's talk about E-utilities and what effect it has on them.

For those of you not as familiar I wanted to go over what the utilities are and what

they do and their function and how they depend upon the unique identifier.

The E-utilities work in all of the Entrez databases, from PubMed to the sequence databases,

medical databases, whatever, the e-utilities function across a wide variety of identifier

spaces.

What they do is a few functions.

ESearch can take a text query and return back to a set of these unique identifiers, that

correspond to the result set.

Those unique identifiers, uids, can be passed to additional E-utilities to retrieve those

records.

For example, I can pass a UID to ESummary to get the record summary, or docsum, which

is a short abbreviated metadata about a record that you commonly see as search results on

the web.

Alternatively I can take a UID and pass it to EFetch which would download the full data

record in a variety of formats.

PubMed you can get abstract, citation, Medline, etc., ASN.1, XML.

For sequences you can get FASTA, flat file, XML, ASN.1, etc.

Efetch is a great utility for getting the data down once you have the identifiers in

hand.

ELink is another powerful utility that lets you take identifiers from a given database

and find linked records in another database.

For example I want to find the PubMed literature associated with a sequence.

I can link from the sequence records to PubMed.

I can link from nucleotide to protein, etc., etc.

Elink also talks in this UID space.

One set of UID's in and another out.

Finally, epost is essentially an upload utility that takes UIDs and puts them onto a temporary

storage facility that we call the Entrez history.

And this is a server that stores temporarily your search results.

Then I can take the results from that history and pass those to fetch or to summary or to

link for further processing.

I think it's pretty clear as you look at the diagram that all of these utilities either

output UIDs or accept UIDs as inputs.

It's critical to the function of this API that the UID space be well determined.

For sequence records, talking about nucleotide, protein, EST and GSS databases, and Popset,

the UID has always been the G.I. number, the integer identifier.

Not anymore.

You can continue doing that but now all of these utilities will do the same thing for

accession.version.

It's a very powerful upgrade in the functionality of these APIs, it extends their scope significantly

and we'll talk more about that towards the end.

The set of UIDs is now a set of accession.version identifiers.

You have the option, it's important to understand that you can continue to use GI numbers but

you have the option to use accession.version.

I listed there the primary databases that this affects.

Let's step through them and talk about what would happen.

Luckily ESummary and EFetch get an upgrade that is silent.

You use the ID parameter as input for the UID's.

So in the first URL showing there, ESummary, and the database is nuccore, there are two

UIDs there.

That's a traditional ESummary call.

For EFetch it would be the same kind of call, same UIDs, but I might put rettype equals

fasta to specify I want the FASTA format.

What you can do now is instead of using those integer identifiers, the gi's, you can put

accession.veresion.

If you put in accession without a version you get the most recent version of that accession.

That just works.

You don't have to do anything else to your call.

You don't have to add an ID type parameter or anything else.

It just works.

This is great and you can just start using this

For those of you who've been using these APIs, you may realize that EFetch has been able

to do this for many years.

It has been able to accept accession.version for several years along with GIs, it continues

to do that.

In this regard EFetch has not changed.

ESummary has changed.

It has never been able to accept accessions as input but it can now.

That's what's happening with ESummary and EFetch.

Pretty simple.

You don't have to do anything, you can just use accessions now.

ESearch is more interesting because what ESearch can do now is output data as accession.version.

What ESearch does is takes a text query, like foxp2[title] AND human[organism].

I want the human sequences for foxp2.

Before, I would assemble this call.

What that would give me is a list of GI numbers back.

What it does now is you have the option, if you add the ID type parameter, and give it

the value acc, so &idtype=acc is appended to that same search call, you will get something

different.

This is what you get different.

The default without using ID type is a list of the top 20 GI numbers.

Unique identifiers that match your query.

Those should be nucleotide GIs.

If you use ID type equals acc, you get back the accession.versions.

This is cool.

You get the accession.version as if the GIs don't exist.

You get them back directly and can use them for further processing.

Elink is going to behave in much the same way.

It takes sets of identifiers as input and gives you sets of identifiers as output, in

the same or a different database.

A very simple example is here's a nucleotide G.I. number and I want to find the protein

that it encodes so I say dbfrom=nuccore, db=proetin, and there is my ID.

I would expect to get back the protein G.I. that is associated with this.

If I use ID type equals acc, append that to the URL, now it allows me to do two things.

I can have an accession number as input.

So I don't have to use a G.I. number and I will get back accessions in the output.

Here's a comparison of the XML that I would get back from those two calls.

If the ID type is not set, the default is essentially GI numbers.

Here's my nuccore dbfrom, here is the id in nuccore, the link is to protein, here is the

ID number for protein, 34577063.

That's the G.I. number of the protein linked to the incoming nuccore record.

But if I use ID type equals acc.

What you see on the right side, is that the NM_ record, 001126.3, is now linked to NP_001117.2.

so I get the accessions back immediately.

ID type equals acc is required for ELink and ESearch if you want to get the accessions

in the output.

Before we do a little bit of playing with this I want to talk about some expanded scope

that the e-utilities now have access to because of this change.

It's important to understand this so you don't run afoul of the way the data are restructured

now and be aware that these changes will continue happen as things progress.

we have sequences that don't have GIs.

That's an issue for a number of reasons because a sequence without a G.I. is not part of the

Entrez system.

It's not part of Entrez Nucleotide, nuccore, or Entrez Protein or any other Entrez database.

That has several consequences.

ESearch cannot find them.

They are not indexed in Entrez, so esearch has no access to them.

They do not have document summaries, they are not in Entrez so ESummary cannot retrieve

them.

They have no Entrez links so ELink cannot perform links between gi-less objects.

Also they cannot be placed on the Entrez history.

For example, EPost cannot handle or process them.

For those of you who are e-utility aficionados, if you use ESearch with usehistory=y, it won't

work for a gi-less sequence because they can't be put on the history, they are not part of

Entrez.

But EFetch can now download them.

EFetch has become an odd man out in that it now has a special function, that it has access

to the whole world of GI-less sequences through their accession.version.

All NCBI sequences, whether they have a gi number or not, have a accession.version identifier

and EFetch now can access all of them.

This is brand-new functionality that we have never had before, that API access to all of

these GI-less sequences that still have a accession.version.

So what are we talking about?

A lot of WGS and TSA.

So whole genome shotgun datasets and transcriptome shotgun assembly datasets.

Many of these data sets do not have GIs for their contigs.

They may have a G.I. number for their master record but they will not in many cases have

GI numbers for all the congtigs.

Ultimately most new sequences, eventually almost all, will be like this.

They will not have GI numbers but they will have accession.version.

It's important to understand that this is the future and where things are moving so

that accession.version becomes the main identifier that you can depend on.

A cardinal example currently is the Norway Spruce project, CBVK.

This is a very large project with over 1 million contigs.

Because of the size it was not assigned GI numbers.

It has one G.I. number that represents the master record.

So CBVK and all those zeroes, that single representative record, which has no data in

it except metadata, that one record is indexed in Entrez.

If you search for Norway Spruce in Nucleotide you would find that record but all of the

million contigs are not available to you from entrée and have not been able to be available

to from the API e-utilities.

They are now.

One question that folks might have is how do I check if a sequence has a G.I. number

or not?

The easy way to do this programmatically is to use ESummary because it will immediately

return to you in error tag telling you that this accession does not have an identifier,

a gi identifier.

And you can do that with a mixed bag of input and esummary will very politely tell you which

ones work and which ones don't.

At this point that's our recommendation as to the best way to check.

Esummary is fast and it can process thousands of records in a given URL, so it's a good

way to parse through a big list if you want to find out what has a G.I. number or not.

There are other approaches if you have massive projects and you are concerned about using

ESummary, write to us, talk to us, we can give you some additional help.

What I'm going to do now before I go to this slide is do some demo.

We will go through a few calls so you can see how they work.

Also this little Perl script, an incredibly simple little script, I've put on the ftp

site in case any of you want to play with it.

I'm just going to explore the calls a little bit.

I'm going to actually show you what the Perl script looks like.

That's it.

All it does is post a URL and I have an array here of eight sample URLs.

You can explore this and edit as you want and play with this functionality.

The first URL is an ESummary and that will get back two sequence records specified by

an accession.

This is something we haven't been able to do in the past, so to run this little script,

I just say perl geturl.pl with index one.

I review that and I get some data.

What you see here are two docsum records.

The first one here.

And the nice thing is it works.

In the past this would have given an error.

I get my accession back and get my record back.

For those of you that are parsing docsums you might want to be aware we have a new tag

within the docsums called Accession.version.

That thing is brand new and it has the accession.version in it.

That is something you can parse out directly and that's where the identifier is now.

Of course the GI numbers going to be there also.

And again, if it doesn't have a G.I. number, ESummary can't find it.

That's how esummary's going to work.

Now esearch.

Two ESearch calls, one will look for foxp2[title] and the other one has human[organism] without

$idtype=acc.

The other one as $idtype=acc appended.

So let's do the first one.

As I was showing in the slides, this is what I get back; I get 48 things back.

Here is the top 20; I get gi numbers.

All I have to do is add that ID type equals acc in that URL and i get back the accessions.

It is quick and simple.

All right, going on, ELink.

We can do elink just as we did esearch, linking from a nuccore record to a protein record.

Here is the id.

It's also NM_001126, so we will do it both ways.

Just know you have to include idtype=acc at the end, if I'm going to do the accession.

So my first elink call, again, has the G.I. number as input and gives me the gi number

for the output for the protein.

If I do idtype=acc, here is the NM_ as the input, the NP_ is the output.

You can do this if you are linking across from non-sequence to sequence databases.

So if I'm going from pubmed to nuccore or pubmed into protein, you will use idtype=acc.

And, of course, PubMed will not have an accesion number.

That's fine.

You'll use pmid's as your input to ELink but the output from ELink will be accession numbers

in protein space.

That will all work just fine.

All right.

Let's take a look at the last ones here.

Here is EFetch.

Here is some of that Norway Spruce data.

This is WGS data that we've not been able to give you before with EFetch.

That URL, here is the ID, that long accession, the accession for the contig.

And I'm going to get FASTA back.

So I do that.

Boom, there is the FASTA for that wgs contig and if I scroll up to the top, there it is,

Picea abies.

There is the data, it has a contig name.

All of this little FASTA definition line is something we are providing.

It doesn't exist in Entrez, but we are providing one to you.

And so here it is, you can get those data.

It's very likely if you want the entire project, ftp might be a better method.

But, this is a way you can get contig by contig if you want to get a set of contigs down from

one of these sets.

Last but not least, ESummary as a check.

How do I check to see whether my sequence has a G.I. number?

I'm going to try ESummary with the CBVK.

The same accession of just retrieved with efetch.

Let me see what happens.

But I'm going to try it in a mixed list of other things; an NM_ record and a GenBank

record.

How would it look if I had a mixed list and a bomb in their that doesn't have a G.I. number,

what is esummary going to do?

ESummary in the case where I just give it that Norway Spruce wgs contig, it very politely

tells me, ERROR Invalid uid at this position.

I know that is the contig that had no UID.

If I give it the three identifiers, two of which have GI numbers and one doesn't.

This is the data I get back.

At the top you see, ERROR Invalid uid at this position.

And then I get the two valid docsums back as expected.

So this should be a fairly rapid way to quickly check a list of data to say what has gi's

and what doesn't.

This little Perl script, this almost brainlessly simple Perl script.

It is on the FTP site and you can play with it if you want.

If you have questions about any of that, we will be taking them in a couple moments or

if you have questions later, write to us.

That will be my last statement before I be quiet and we start discussing.

Continue to follow our social media posts, blog, news, Facebook and Twitter accounts

for more updates and announcements about things that will be happening with these sequence

resources, the changes with sequence identifier space going forward.

The NCBI Learn page is a great place to come for events like this if you want to have more

awareness of the webinars and other courses that we are offering.

If you want to find out about the e-utility updates, it's a great idea if you are using

them, please subscribe to utilities-announce@ncbi.

That's where we send out all of the announcements about these upcoming changes, how you can

prepare for them.

Where we announce in advance that changes changes are happening.

So it's a great place to look for those kinds of things and keep yourself aware and not

be surprised.

Finally, info@ncbi is our general helpdesk.

Please don't hesitate to write to us if you have questions or concerns.

All right I'm going to be quiet and open the floor to questions.

This is Peter Cooper.

We do have a couple questions.

Eric.

Several of them I answered and you sort of answered as you went through, so I won't go

over those again.

Those are the ones I answered in the questions pod.

Interestingly people asked those questions then immediately you said the answers to them.

Here's a few others.

The first one is about downloading WGS sequences from the FTP site.

Given a top-level WGS project ID, so that's essentially that prefix, what's the best way

to programmatically get the ftp location for the entire assembly?

One way you can do that is to use the BioProject database.

That has a good place to search for these data sets.

That has metadata in it that will allow you to find the project that is the source of

the WGS data.

You can link from BioProject into nuccore and that will give you the master record,

if it's not in the BioProject data itself.

That's one avenue.

You then would be able to construct an FTP URL down to the FTP site to get that data

from Genbank.

Another way you could approach that task of getting the appropriate accession is through

nuccore itself.

At this point my understanding is that all of our WGS projects -- or at least the vast

majority -- will be assigned a G.I. number for their master record.

And that master record would be indexed with some metadata.

So if you're trying to search organism or a title or something like that, you may be

able to find it that way.

Then you can construct an FTP URL into Genbank.

I hope that helps.

If you want more detail on that, let us know and we can provide you with some additional

suggestions.

I think the structure of the FTP site is a formal way of doing that based on those accessions.

What I will do is put that -- I'll have to look at it so I can describe it properly -- I

will put that in the Q&A document so everybody can take a look at that.

The other question that I see here is can esummary maintain the order of the input UID

even if there is a GI-less accession.version in there?

That's a good question and one that we're thinking about ourselves.

At the moment the order should be maintained for those records that do have GIs.

It appears what's happening is we are writing the errors at the top of the output screen.

So there's a disruption in the current order of the input.

That's a good suggestion.

We will take that up with the development staff and see if we can do something to address

that.

At the moment you're going to be stuck with finding the errors at the top and the rest

should be in the order in which you submitted them.

All right.

Thank you all very much for your time and for attending.

Good luck out there and let us know if you run into problems or have concerns or suggestions

about these new functions.

Thank you very much and have a great day.

For more infomation >> NCBI Minute: New Version of E-utilities Supports Accession.version Identifiers - Duration: 28:00.

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For more infomation >> Margaret's Weather Picture for February 1, 2017 - Duration: 0:30.

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Legacy Challenge Kickoff – A campaign for UF College of Medicine scholarships - Duration: 2:21.

DUKE: Hello!

Welcome to Gainesville, Florida.

I'm Jim Duke from the class of '85 and president of the UF Medical Alumni Board.

I want to let you know that the UF College of Medicine is working very hard to fill some of its crucial needs

much like college football recruiting.

And through our brand-new Legacy Challenge campaign

we are looking for 100 commits to support our ambitious scholarship fundraising goal.

With me today is Dr. Jason Rosenberg from the class of '95

who, along with his wife, Denise, has a very important decision to make.

Jason, what's it going to be?

ROSENBERG: Denise and I thought long and hard on how best to support our medical school

and those students who have a dream to help others through a career in medicine.

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(Cheering and Band Playing Music)

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Well, I hadn't planned on this

But my wife Pam and I want to sponsor a medical student

and we're going to join the legacy challenge.

We're in!

ROSENBERG: Awesome

(Cheering and Band Playing Music)

For more infomation >> Legacy Challenge Kickoff – A campaign for UF College of Medicine scholarships - Duration: 2:21.

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Daily Tarot Reading for 2 February 2017 - Duration: 16:50.

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Team Bonding is Key for Slovenia's Women's Team | Patagonia Dreaming - Duration: 8:22.

I really like the place here.

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Also if it's not so crowded

there are not so many things to do in the afternoon,

but we like it here.

(PATAGONIA DREAMING)

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And it's good to be here.

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it's a part of... how can I say? World.

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In Ushuaia, we like winter conditions -

the weather, the low temperatures, the snow.

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The main thing why we came here

is to train a lot,

to train on different slopes

because here the ski area offers really different slopes

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because the base is here.

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to prepare ourselves for the season.

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In Slovenia, it is summer.

Training lasts, approximately, until 12 or 12.30pm.

Then they have lunch and some free time.

At 2.30pm, we arrive at the hotel.

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At 5.30pm, they have physical training.

At some point, we analyse the videos.

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We just started with our trainings.

First trainings are a little bit easier

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Yeah, I'm putting my protectors.

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Winter here is pretty cold.

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The high level is not so big, so we can train a lot

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If we're in Europe, we train mostly on glaciers.

The snow is so-so.

The weather is always changing.

So here it's mostly the same every day.

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I'm preparing for the first run

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Yeah, Ana was sick for two days.

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with the Skidoos and walking,

and some lunch in the nature.

For more infomation >> Team Bonding is Key for Slovenia's Women's Team | Patagonia Dreaming - Duration: 8:22.

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BofAML provides a forum for tech leaders to discuss the latest trends - Duration: 1:05.

This is the eighth time we've run this summit, and in the seven prior years what we've

seen is that in the over 270 young and emerging technology companies that we've seen at the summit,

over 16% of those companies have gone on to be vendors of the bank.

Companies that have helped to grow our mobile capability, helped grow our data and analytics

capability and now increasingly this year, we see the trend of robotics and artificial

intelligence beginning to come through.

What is unique about this event is the commitment that we have as Bank of America to the community

here in the valley.

This place is unique.

It is the cradle of innovation.

Many places around the world have tried to copy it, nobody's been able to achieve that

and it's why we come back and we re-double our commitment every year.

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