Youtube News: Youtube daily report w Apr 1 2018

This is information about the actual presentation and some information will be made available

at the links that you see at the bottom.

I will give an introduction to the pathogen detection isolates browser.

Some of you may not be familiar with it. I will give you background of the project. It

started five or six years ago. It came about because the food safety agencies in the U.S.,

the FDA and the CDC were thinking about switching from pulsed field gel electrophoreses to whole

genome sequencing. And the FDA started that by forming what they call the genome tracker

network and you see on the left-hand side the number of labs that are part of the genome

tracker network and they wanted to focus on salmonella but the CDC in the summer of 2013

said we should focus on Listeria as the pilot project. All Listeria collected in the U.S.

for both clinical, food and environmental samples would be sequenced and submitted in

real time. We had a meeting where we agreed to contribute to that project.

The reason to focus on Listeria is it has a very low incidence in the population so

it is a tractable problem and all the isolates could be sequenced in real time and it has

a high morbidity and mortality rate. Sequencing these would have a major benefit to the health

of Americans.

This is a slide from one of our collaborators at FDA showing how the network functions now.

Samples from clinical/human, food, animal and environmental are taken by these agencies

as part of the network. That includes the FDA's GenomeTrakr, CDC's PulseNet, State clinical

health labs, and the USDA Food Safety Inspection Service who take samples as well. They submit

the raw genomic sequence data to NCBI and these are typically from Illumina instruments,

either 2x250 or 2x150, and they supply minimum metadata . All the data is publicly available.

Of course they have highly protected metadata that they store locally that we do not see.

So what facility this isolate was taken from and information about the patient.

We built an analysis pipeline that wants to answer two things at this point in time. Are

these isolates clonally related? Is there a point source for a food outbreak for example?

And the second thing I will touch briefly on, what is the set of genes that encode antibiotic

resistance in these isolates?

I won't go into a lot of details of the analysis pipeline. We are making changes to it and

we have publications coming out at some point describing this in detail, but the basic idea

is the data coming from the surveillance network goes into SRA we assemble it and do some quality

filtering. We also pull in assemblies from GenBank for the same organism, let's say Salmonella.

And we cluster them together using single linkage clustering. Right now we use a 50

SNP max breakpoint. We will be switching to multi-locus whole genome sequence typing in

April. The idea is we want to form clonal clusters of closely related isolates and we

are not intending to do the full phylogenetic tree of all salmonella this way. So for salmonella

we have several thousand clusters size 2 through size several thousand. Within each cluster

we do a phylogenetic reconstruction and we make that available and I will show you examples

in a few minutes.

For the antimicrobial resistance we first put together a reference database of acquired

antimicrobial resistance genes and cwe have created software to identify those genes and

we are working on a manuscript to describe it called AMRFinder and we attached antibiograms

to BioSample submissions. So the antibiograms are tabular formats of the antibody susceptibility

tests, either MICs or disc diffusion and we attach those to the BioSample database if

the submitter is willing to supply the data.

We integrate that all into the isolates browser. So the Pathogen Detection Isolates Browser

is using new technologies a NCBI including things like solr but what that means is because

we are trying to do this quickly and get it out to our collaborators as soon as possible,

it's not fully integrated into other resources.

So some of you will be familiar with the drop-down database menu option on the left and you won't

see Pathogen there or in the global database list. It is kind of hidden under Health, which

I'm sure you are all familiar with.

This is a screenshot and then after this I will give you a demo. This is the Pathogen

Detection Isolates Browser in Beta which means it's a work in progress. We are continually

evolving the capabilities and it's not something we would have done in the past where we would

have spent two years building something and release it. We released in 2016 and have been

continually adding features to it. That means there is no help documentation yet but there

is a fact sheet that you see in the upper right menu under learn more. There is a PDF

that gives you basics of how to use the resources. It's similar to what I cover in this webinar.

We will work on help documentation in the future. Part of the reason for that is that

the search capabilities are things we are playing with. We want to make sure the search

is as streamlined as possible for people to use and not document it in an incomplete form

now. What that does mean is that the search syntax is not the same that you would typically

see in Entrez databases like Nucleotide or Genome.

So at this point I will give you a demo.

It's available at the base URL/ pathogens. This is the webinar we are giving now. This

gives you a brief description of what is going on. A couple of example searches and then

I will cover this in a minute. And then you have some basic information including the

fact sheet I told you about and I will open this for a second. It covers a couple basic

examples of how to do searches. Information on antimicrobial resistance and some of the

reference database we put together and how to submit data. If you are interested in submitting

data to us you can contact us and follow these links. For exploration of data we have some

options.

We have all the data available on FTP so if you want to do batch downloads you go there

but I won't cover FTP today. And we have Find Isolates that takes you directly to the browser

and we also break it down by the organism. Here we have the top four foodborne pathogens

in the U.S., we have salmonella, E. coli and Shigella, Listeria, and a Campylobacter. And

we see the total number of isolates. We have over hundred thousand salmonella in the system.

And we have the new isolates. If you recall I said we make single linkage clusters by

50 SNP's. We attempt to do that within every 24 hours. Sometimes it doesn't work. You see

that more fully here on the full list. These are all the organisms we are currently clustering.

Many are not foodborne pathogens so I won't cover those in much detail. Some big examples

you won't find here right now or Staphylococcus aureus. We plan to add those later on this

year we make the switch to whole genome MLST clustering.

Let's look at the top row, salmonella. This is the version that was released on March

16. The latest isolate that was added that was included in this released was from March

14. We have a two day delay and trying to get that down . Basically it means from the

previous release which is version 1147 we had 78 new isolates added to the system. The

breakdown of that is 51 clinical isolates and 27 environmental isolates. All these links

go into the pathogen browser. As well as this link here.

The browser is basically unlike most of the databases at NCBI, it presents information

without having to do a search, in sort of a tabular format.

For the isolate browser every row in the table as an assembled genome. Either assembled through

our system of assembly and annotation or one we collected from GenBank. You have a search

box which I will go into detail. We have some default organism groups and those match the

ones in the table I showed you. We have columns that include metadata supplied by the submitter.

This particular isolate was collected in New York. Some things that we calculate and I

will cover a couple of those, but there are additional columns and you can choose which

columns you want to show with this tool here.

There are a couple of critical columns I want to cover in more detail and that is the minimum

diff. We make clusters of 50 SNPs and we also categorize isolates by two types, environmental

or clinical.

So this column is the minimum SNP distance from this isolate to one of the opposite type.

The first row is not a SNP cluster and not related to anything in our system so it could

be 51 SNPs away or 500. We don't know at this point in time. We are trying to identify clinical

isolates that may be a point source of a foodborne outbreak. The second row is an isolate also

from New York and it is 12 SNPs away from something of the opposite type and it's an

environmental type and it is 12 SNPs away from a clinical type.

If we scroll down here in row 16 we see another isolate and another Listeria is 33 SNPs away

from something of the opposite type. It's clinical and that means it's 33 SNPs away

from something that's a food or environmental source. So if you sort in this column you

can identify things that are clonally related. So I will give you an example. I will do a

search for the new isolates.

And if I sort that and focus on Listeria, there are 22 new isolates for Listeria. Here

is an isolate that is 12 SNPs away and that grows. These SNP distances might be sufficient

for someone in public-health labs to look at this and determine they do not need to

do further investigation.

If I switch to something like salmonella, now we get down to two SNPs and this is an

isolate from Virginia and two SNPs away from a clinical isolate. So not only do we do these

SNP distances but we also provide the SNP phylogentic tree and I will open that up.

This is what we call the SNP tree viewer. We have three panels in this view. We have

a navigation panel and a show you how that works in a few minutes. We have a table with

all the metadata similar to the table on the front page and then we have the tree viewdown

here. This is like Google maps and you can pick it up an drag it and zoom in or out.

You see this isolate is in a SNP tree of 978 salmonella isolates.

This is the section for that. This obviously is a pretty large SNP cluster you can see

parts of the tree are very closely related and others are more distant. The interesting

thing is you can make selections both on the table and the tree. From the search I had

identified this isolate as a new isolate so it came in on the 14th. I can actually highlight

another isolate.

What happens when I have more than one isolate selection is I get the SNP distance measurement

up on the navigation panel. You see there are only two isolate so the minimum, maximum

and average are all the same. 4 SNPs separate these two isolates from each other. From August

last year to March this year. The other interesting thing and I will go back to the search view

is we intersect the searches with a number of isolates.

Here we see when I search for the new isolates, five isolates are in the tree that I was just

looking at. Now this navigation panel becomes useful because not only does it tell you SNP

distances and the breakdown, you can see these are environmental isolates from Maryland,

Virginia and New York and a clinical isolate. When you click on this, it jumps immediately

to where that isolate is on the tree.

We built this part because these large trees were increasingly much more difficult to drag

around like in Google maps. You see how long it takes to get to the next isolate. Whereas

this, you just zoom around landmarks in a map.

We also have a filter and this is the only filtering the metadata that's here so I could

say I only want the ones from Maryland and those come to the top, and you can clear that

filter.

We also have a search box. I can do a search for chicken . Now it will highlight everything

in the tree that is from chicken. I can add that to the selection set. Now you can see

almost everything in this tree comes from chicken. That's a reminder to properly cook

the chicken you are eating. Please.

We also get this breakdown by year. This navigation panel is something we are evolving now and

it will probably change in a few months or sometime in the summer. This search box allows

you to both highlight items in the tree and also to add to the selection set on the left-hand

panel. It will also do subtractions. I see things from Maryland and I think this will

work. I will do a subtraction. And the Maryland isolate's disappear and I think there was

only one so he isolates drop from 539 to 538.

You can clear this and make a selection directly on the tree itself for multiple isolates by

selecting all leaves. So here I have highlighted a small set of isolates, looks like they are

all from the Boston or the U.S. from two different years and I can make a subtree.

If I click on this button now I just have the isolate that I selected by this action

node . And you can collapse it as well and expand it if you want. This allows you to

do an export. We have a warning if you try to export large trees but exporting small

trees is easy and you can dump it in a PNG file. That's not a very good viewer but you

get the idea. You can export in PDF and as a newick tree

if you want to put it in your own tree software do additional work with that. You can share

this view. If you hit this button, you now have a box that you click on and you can copy

and paste that into another viewer or send it by email if you find something interesting

to send to colleagues.

You can just use the share button. When you are in the subtree view it tells you you're

in the subtree view up here and you can go back to the full tree here. What it does is

causes a collapsing view. This is a feature we were testing and it collapses nodes not

selected and when you highlight those nodes you can see the breakdown. You see there are

224 isolates in the subtree mix of various sorts of clinical and environmental.

This is something still being worked on and not fully worked out yet . If you want to

go back to the full tree view just hit hold Treeview when you get back to the original

view we came in on.

This isolate, you can hit the information button and you get an additional information

on metadata but that similar to what you see when you hover over the tree. Some people

might find this annoying you can turn this off. Than that does not happen. You can do

things like control spacing of the tree. So you can separate it for better visibility.

You can collapse that if you want. There are a few other options. You can expand and contract

the tree branches. This is the SNP distances to make it more

spread out if you want.

I should also point out the selections allow you to control what is selected in the table

view. You can see we selected seven isolates. Out of the many we have, and you can download

this. There is a download button here that dumps a TSV file that is just the rows and

isolates suggested. You can choose what columns going the table similar to the one on the

main page and the selection controls which isolates are selected for that tabular download.

You can make selections on the tree, in the table to control the download for whatever

selection of isolates you are interested in.

The only other thing I want to touch on is the antimicrobial resistance. Let's look at

something that is highly antibiotic resistant, Klebsiella. And we make two columns available.

One is the resistance phenotypes as supplied by the submitter, and the genotypes. And you

can highlight that in the filter tab. There are number of filters and two of them are

the phenotypes and genotypes, so I will select the phenotypes, there are 516 of the total

Klebsiella that have supplier-submitted phenotypic data. Now you see this column has information

and you can expand that and you can see the breakdown. Basically in this panel we put

resistance calls, the SIR calls, from whatever interpretation criteria we used. You see that

colistin doesn't have interpretation criteria so it's in the category of other. We also

put the genotype information.

First I want to cover the AST briefly and this is what is stored in the BioSample database.

Now you have this tabular format and it gives you the actual MICs, the interpretations and

you see that many are done by the CLSI standard, actual measurements and some information is

optional and some not supplied. You see things like colistin is not defined because it's

missing and there is no CLSI standard at this point in time.

By adding it to this you can do some interesting searches. I won't cover those in much detail.

I will go back to the main page and see that we have a search for isolates encoding a mobile

colistin resistance gene and a KPC beta-lactamase.

If I click on this I get six isolates that have both MCR and a block KPC allele. They

have Assembly links so they have all been submitted to GenBank and not assembled by

our pipeline. You see they are from Brazil, Portugal and Italy . You see the list of genes

here, we have KPC- three and mcr 1.1, KPC-2 and mcr 1, etc.

You can do searches and there are examples in the fact sheet I pointed out. We are working

on, those of you who know things about aminoglycoside modifying enzymes , you see that the encoding

of that genetic information is troublesome to many computer search programs so we are

working to make the searches for things like aphc3 prime prime 1b easier so you could just

plunk it in to the search box and do the search. It doesn't happen now. So it's a new feature

we will add in the summer.

You may ask I'm not a public health lab so why is this useful for me? Not only could

you get antibiotic resistance or sets of isolates with antibiotic resistance, but we are planning

to expand the sort of genes we make available in the system. So that includes virulence

genes, metal-resistance at biocide resistance we are expanding this tool to incorporate

those other genes of interest to allow you to subset and select the full data set.

So right now we have over 107,000 salmonella and we hope this interface will allow you

to more effectively subset the data rather than doing let's say a blast search with a

gene of interest. You can imagine searching across all 100,007 would be time-consuming.

So we hope these interfaces for large-scale data more effective at NCBI in general.

I think I will stop there and mention these are the people that worked on the project.

You see this email address highlighted in yellow.

pd-help@ncbi.nlm.nih.gov. If you have questions after the seminar, and

this address is linked on our Pathogen page, so just send us an email and we will answer

your questions. I think at this point in time I will take any questions, Peter.

[Peter} I'm afraid I lost power at the beginning of the webinar so I can not participate in

the question part on the phone.

[Bill] Sure.for those of you connecting remotely Washington DC is in a massive snowstorm so

many are losing power. I will go through a couple of questions. What W G MSLT scheme

is used? It was developed in-house at NCBI. We will be making them available at some point.

Basically we developed them for the four foodborn pathogens from TB and I think for C. difficile

and we have been trying to coordinate with CDC on some schemes and we have an upcoming

meeting after the ASM general meeting in Atlanta in June where we are supposed to talk about

that as well.

Another question, urinary tract infection of foodborne disease?

The organisms that causes UTIs like E. coli is often found as foodborne disease. So we

do see a mix of both of those in our system. It's not strictly related to just foodborne.

S. typhi is a hn obligate human pathogen not typically associated with food but we also

see those, so we basically pull in all the isolates under a particular species from our

surveillance network plus k.

Is there any correlation between SNP clustering and classic STCC?

I think the answer is they will highly correlate so the question is are there correlations

between SNP clusters and classic sequence types and clonal complexes? So simply based

on the seven or whatever gene sequence types that we are classically defined before, you

would see a high correlation but you can't guarantee that because of course a small change

in an allele would give you a different allele and possibly sequence type, and different

clonal complex even though it might be part of the same SNP cluster.

So it is not guaranteed and it is something we thought about adding as a feature and doing

the classical sequence typing and adding that as information so you can see ST258 isolates

for example.

Another question, is the number of SNP differences between two isolates the absolute number without

filters, so the SNPs passed filtering or the number of compatible SNPs from the compat

program? The SNP distances now are from the Compat program. I didn't go into details on

this but one of our colleagues at NCBI improved a method that is 30 or 40 years old called

maximum compatibility for the phylogenetic reconstruction. It's now published and that

software is available. It is very useful for highly clonal isolates which is what our system

was developed for. It's not very useful for highly divergent isolates so the maximum compatibility

system looks for columns that are compatible with the phylogenetic reconstructions which

means sometimes it throws away some columns. Why would you want to do that? We found sometimes

when we look at GenBank genomes they have incorrect SNPs with respect to the tree and

we suspect many times it's because of assembly problems. So the system helps filter out some

of the bad data.

Next question how to decide if it's a new isolate? We do the SNP clustering every 24

hours if new data is submitted. So that New is a recency check on whether something has

been submitted since the last time we did the calculation.

Can we use the pipeline to analyze our own gene sets? I didn't touch on this but we built

the system for public health with the idea from colleagues that they would submit data

to us and make it publicly available. That is something we are pushing that people who

want to integrate their isolates into the system are publishing them as part of papers

or research or surveillance networks to make the data publicly available. Right now the

pipeline is not available for download. We will make certain parts of the pipeline available.

I didn't touch on this but we are making a new assembler available. I think the paper

will be submitted by the end of the month. We will put links on the main page at some

point showcasing some features that will be made available.

Is there publication related to this? As I said we will describe that at some point in

the future.

How can I add this as a project to my undergraduate students?

I'm not sure if you are saying how you can get your undergraduate students to use this

project? I'd be happy to touch base with you after this.

Is this connected to Patric? Patric is a NIAID funded system as part of the bioinformatics

resource Center and it is not directly connected to Patric but we certainly coordinate with

them on things like antibiotic resistance.

Would it be possible to add a data column with classic mlst sequence typing? Yes, that

something we are thinking of doing in the future.

One are the future plans of NCBI for GenBank of such disease causing pathogens? Besides

providing a system like this that allows you to easily interrogate for interesting features

it's something we need to talk about because we have 100,000+ salmonella and that will

quickly be 1 million salmonella in a few years so we need to think how to deliver effectively

to researchers when we have such large volumes of data. We cannot expect someone to download

and do this analysis themselves. We are interested in hearing from people on use cases for what

they would use this data for or things their research is interested in investigating across

such large volumes.

Are there better ways of submitting data on a weekly basis other than SRA wizard? SRA

has multiple ways to submit. We have the web-based wizard for submission portal But there is

a completely automated XML-based submission format that they've developed and they are

all completely automated. So I urge you to contact SRA about that. I'm not an expert

on those. If you are interested in submitting data to us, please contact them 1st.

How can I see salmonella serotype on the Treeview? We make the serotype, serovar, available as

a field but that is based on what the submitter sent to us. As you can imagine, well I will

give you an example . Let's get rid of the search and switch to salmonella.

Here is the serovar here and you see it's Saintpaul, Barielly, etc. and if you're talking

about adding the label of serovar onto the tree, that is something not yet built into

the system. I want to point out that we find out very often that this serotype is incorrectly

made and we are thinking of adding additional in silico calculation of the serotype based

on tools available. That will be later on this year.

That's a follow-up on the SRA. If you have problems with SRA submissions , Peter, I think

they can contact the helpdesk and you can help guide them through that stuff. I don't

work for SRA.

Yes I'm here. If you are having trouble with submissions, right to the info address which

is info at ncbi dot nlm dot nih dot gov. We will get that to someone who can help you

with that. And Bill we are out of time so we need to wrap this up.

If there are any other questions we will endeavor to answer the remaining questions in writing

and I will send that to everyone when the recording is available on YouTube and that

is written up.

I think there is just one more question.

What's the difference in the relationship in the following resources, the national sequence

database of resistancd pathogens.

Basically they are all -- the first two are integrated into this system. So saying it's

just a national database of antibiotic resistance pathogens. We say that we are making a database

of pathogens and reporting on antibiotic resistance genotypes reported within those pathogens.

The third one is the Resistance gene database and those are genes that are reference set

of genes and alleles we use to make the calls of the genotypes. I didn't have time to go

into details, I think we can have a separate webinar on anabiotic resistance. I suggest

anyone who has questions to send us an email. And the last question was submission and again

you can always write to the submit-help or the info address for help on submissions.

So I'd like to thank everyone for their time . I will stop the recording now and you can

always send us emails and we will follow up with you individually.

Thank you, Bill.

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15.

PETE DUNNE.

TECHNICALLY WWE UK CHAMPION PETE DUNNE HAS ALREADY MADE HIS DEBUT ON MONDAY NIGHT RAW,

BUT THIS COULD HAVE JUST BEEN WWE'S WAY OF ENSURING THAT "THE BRUISERWEIGHT" RECEIVED

A LOUD REACTION WHEN HE WAS MADE PART OF THE SHOW, SINCE THE SHOW WAS IN MANCHESTER, ENGLAND

JUST DOWN THE ROAD FROM WHERE DUNNE GREW UP.

DUNNE DEFEATED ENZO AMORE BEFORE HE WAS INVITED TO 205 LIVE THE FOLLOWING NIGHT, BUT IT SEEMS

THAT HE HAS SINCE BEEN FORGOTTEN.

AMORE HAS CONTINUED HIS RIVALRY WITH KALISTO AND DUNNE RETURNED TO NXT AHEAD OF THEIR NXT:

TAKEOVER WAR GAMES SHOW, WHERE HE DEFEATED JOHNNY GARGANO.

DUNNE HAS BECOME QUITE A POPULAR STAR WHEN IT COMES TO THE WWE UNIVERSE BECAUSE MANY

FEEL HE IS THIS GENERATION'S ANSWER TO WILLIAM REGAL.

DUNNE WOULD DEFINITELY FORCE CHANGE ON THE MAIN ROSTER, BUT WITH A WWE UK WEEKLY SHOW

BEING RUMOURED, HIS FUTURE ON THE MAIN ROSTER IS CURRENTLY UNCLEAR.

14.

KAIRI SANE.

THE WINNER OF THE INAUGURAL MAE YOUNG CLASSIC TOURNAMENT BACK IN THE SUMMER WAS INSERTED

INTO THE FATAL FOUR-WAY MATCH FOR THE NXT WOMEN'S CHAMPIONSHIP BACK AT NXT TAKEOVER:

WAR GAMES LAST WEEKEND, BUT SHE CAME UP SHORT AS EMBER MOON WAS CROWNED CHAMPION AFTER AUSKA

VACATED THE CHAMPIONSHIP A FEW MONTHS AGO.

THE NXT ROSTER NEEDS A STRONG WOMAN RUNNING THE DIVISION AFTER ASUKA'S LENGTHY REIGN

OVER THE PAST YEAR, WHICH COULD BE WHY THE COMPANY CHOSE EMBER, WHICH MEANS THAT KAIRI

WILL EITHER BE MADE THE MAIN CHALLENGE FOR EMBER MOON MOVING FORWARD, OR SHE COULD BE

THE NEXT FEMALE WRESTLER TO INVADE THE MAIN ROSTER ALONG WITH THE FIVE WOMEN WHO DECIDED

TO MAKE AN IMPACT ON RAW AND SMACKDOWN THIS PAST WEEK.

SOMETHING IS DEFINITELY GOING ON, AND SOMEONE IS DEFINITELY MAKING A STATEMENT WITH THE

WOMEN OF NXT AND THAT COULD MEAN THAT MANY MORE DEBUTS ARE ON THEIR WAY.

13.

TM61.

NICK MILLER AND SHANE THORNE HAVEN'T BEEN SEEN IN NXT FOR A NUMBER OF MONTHS BECAUSE

THORNE SUFFERED A KNEE INJURY LAST YEAR WHICH HAS KEPT THE DUO OFF-SCREEN, EVEN THOUGH THE

AUSTRALIAN DUO DID MAKE THEIR RETURN TO ACTION BACK IN SEPTEMBER.

TM61 WERE ONE OF THE TEAMS IN NXT MUCH LIKE DIY THAT BIG THINGS WERE EXPECTED AND IT WAS

OBVIOUS THAT THE FORMER INDEPENDENT STARS WORKED WELL TOGETHER.

MUCH LIKE THE REVIVAL, EVER SINCE THEIR MAIN ROSTER DEBUT, INJURIES HAVE AFFECTED THEM

OVER THE PAST YEAR, BUT IT IS HOPED THAT WWE OFFICIALS SAW ENOUGH POTENTIAL IN THE TAG

TEAM STARS TO SEE THAT THEY WOULD BE ABLE TO HEAD TO THE MAIN ROSTER SOON.

NXT'S TAG TEAM DIVISION IS STACKED WITH THE LIKES OF SANITY, THE UNDISPUTED ERA, THE

STREET PROFITS AND TINO SABBATELLI AND RIDDICK MOSS SO IT SEEMS THAT THERE IS NOWHERE THAT

TM61 FIT IN ANYMORE.

12.

TYLER BATE.

TYLER BATE WAS CROWNED THE FIRST EVER WWE UNITED KINGDOM CHAMPION BACK IN JANUARY BEFORE

HE WAS DETHRONED BY PETE DUNNE AT NXT TAKEOVER: CHICAGO BACK IN MAY.

SINCE THEN BATE HAS BEEN REDUCED TO BEING PART OF WWE LIVE EVENTS AND PART OF THE DARK

MATCHES ON THE MAIN ROSTER, BUT THE COMPANY WON'T ALLOW HIM TO MAKE A FULL DEBUT.

BATE DID APPEAR ON 205 LIVE FROM THE UK ALONG WITH MANY OTHER UK STARS A FEW WEEKS AGO,

BUT AS ALREADY STATED, WWE HAS ACTED AS THOUGH THIS NEVER HAPPENED OVER THE PAST FEW WEEKS,

SO IT IS UNKNOWN WHERE BATE CURRENTLY STANDS.

THE BRITISH STAR IS REPORTEDLY IN A RELATIONSHIP WITH MAIN ROSTER STAR LIV MORGAN, SO THIS

COULD BE THE PULL THAT WWE NEEDS TO FINALLY PUSH BATE TO WHERE HE BELONGS.

HOWEVER, WITH THE UK SHOW HANGING OVER THE BRITISH ROSTER IN EARLY 2018, THEIR FUTURE

IS HARD TO PREDICT.

11.

LARS SULLIVAN.

LARS SULLIVAN HAS BEEN PAVING OUT HIS OWN DESTRUCTIVE ROUTE ON NXT OVER THE PAST FEW

MONTHS AND IS STILL CURRENTLY UNDEFEATED AS A SINGLES STAR SINCE HE MADE KASSIUS OHNO

HIS LATEST VICTIM BACK AT NXT TAKEOVER: WAR GAMES.

LARS COULD BE THE ANSWER WWE IS LOOKING FOR WHEN IT COMES TO NEUTRALIZING BRAUN STROWMAN

ON THE MAIN ROSTER.

LARS IS ALSO A GIANT WHO HAS BEEN BOOKED LIKE A MONSTER OVER THE PAST FEW WEEKS, WHICH MEANS

THAT IF HE WAS TO CONTINUE THIS WAY IN EARLY 2018 THEN HE WOULD BE A FANTASTIC PICK FOR

A DEBUT ON THE RAW AFTER WRESTLEMANIA NEXT YEAR SO THAT HE COULD STEP UP TO STROWMAN

FOLLOWING WRESTLEMANIA.

WWE HAS STEPPED AWAY FROM THE BIG MAN MATCHES IN RECENT YEARS, BUT THIS WOULD BE A FEUD

THAT WOULD BE WORTH BRINGING BACK BIG MAN MAIN EVENTS FOR, BECAUSE BOTH BRAUN AND LARS

HAVE THE ATHLETICISM OF MEN HALF THEIR SIZE.

10.

KASSIUS OHNO.

KASSIUS OHNO WAS THE SUPERSTAR THAT CM PUNK ORIGINALLY PITCHED TO BE PART OF THE SHIELD

WHEN HE GAVE WWE EXECUTIVES THE IDEA FOR THE THREE-MAN STABLE AHEAD OF THEIR DEBUT AT SURVIVOR

SERIES BACK IN 2012.

KASSIUS HAS MADE A NAME FOR HIMSELF AS CHRIS HERO ON THE INDEPENDENT CIRCUIT OVER THE PAST

FEW YEARS BEFORE HE CAME TO NXT AND IT SEEMS THAT HE HAS VERY LITTLE LEFT TO PROVE ON THE

DEVELOPMENTAL ROSTER.

OHNO HAS EITHER MADE THE RIGHT OR WRONG IMPRESSION AT THIS POINT, WHICH MEANS HE WILL EITHER

BE RELEASED FROM NXT IN THE NEAR FUTURE TO MAKE ROOM FOR NEW TALENT OR HE WILL BE PROMOTED

ALONG WITH A NUMBER OF OTHER STARS.

OHNO HAS GAINED A LOT OF HEAT BACKSTAGE IN RECENT YEARS BECAUSE HE HAS GAINED A LOT OF

WEIGHT, WHICH NOW FORCES HIM TO WRESTLE IN A SHIRT.

THIS COULD WORK AGAINST HIM WHEN IT COMES TO PROGRESSING THROUGH THE RANKS IN WWE.

NIKKI CROSS.

NIKKI CROSS DEBUTED AS A UNIQUE CHARACTER IN NXT WHEN IT WAS REVEALED THAT SHE WAS PART

OF SANITY.

NIKKI HAS SINCE HELPED HER STABLE TO BECOME TAG TEAM CHAMPIONS, EVEN THOUGH SHE FAILED

TO CAPTURE THE NXT WOMEN'S CHAMPIONSHIP AT NXT TAKEOVER: WAR GAMES THIS PAST WEEKEND.

NIKKI HAS BEEN PERFORMING ON THE INDEPENDENT CIRCUIT IN THE UK AND THE US FOR A NUMBER

OF YEARS AND SHOULD BE IN A POSITION NOW WHERE WWE CAN SEE HER WORKING ON THE MAIN ROSTER.

NIKKI WOULD BE THE PERFECT, UNPREDICTABLE ENTITY THAT THE COMPANY NEEDS IF THEY WANTED

SOMEONE TO BE PART OF ANOTHER INVASION ANGLE.

THE ONLY ISSUE HERE WOULD BE THAT NXT WOULD WANT TO ENSURE THAT THEY STILL HAD WOMEN ON

THEIR ROSTER THAT COULD CARRY THE DIVISION OVER THE NEXT FEW MONTHS AND HELP CREATE NEW

STARS.

WHILE NIKKI WOULD BE MOST PEOPLE'S FIRST CHOICE, WWE MIGHT DECIDE TO HOLD HER BACK

TO ENSURE THE FUTURE IS IN GOOD HANDS.

VELVETEEN DREAM.

THE VELVETEEN DREAM HAS JUST BEEN PART OF HIS FIRST MAIN STORYLINE IN NXT AGAINST ALEISTER

BLACK AND HE STOLE THE SHOW AT NXT TAKEOVER: WAR GAMES.

IT SEEMS THAT WWE HAS FOUND A DEFINITE FUTURE STAR WITH VELVETEEN DREAM, EVEN THOUGH HE

WAS ONCE A STAR THAT THE COMPANY THOUGHT ONLY HAD A FANTASTIC ELBOW DROP.

DREAM HAS PROVED THAT HE IS AT A LEVEL ABOVE ALL OTHER STARS ON THE ROSTER RIGHT NOW, SO

IT WILL BE INTERESTING TO SEE IF HIS NEXT FEUD IS AGAINST SOMEONE WHO IS COMPLETELY

DIFFERENT TO BLACK SO THAT THE COMPANY CAN SEE HOW HE ADAPTS.

DREAM COULD BE A STAR LIKE TYLER BREEZE WHO IS LOST AMONGST THE SHUFFLE ON THE MAIN ROSTER,

SO IT IS MUCH BETTER FOR THE COMPANY TO USE HIM ON NXT FOR THE TIME BEING WHILE THE CREATIVE

TEAM COME UP WITH AN INTERESTING ANGLE THAT WILL ALLOW HIM TO MAKE AN INSTANT IMPACT ON

THE MAIN ROSTER.

RODERICK STRONG.

RODERICK STRONG HAS BEEN PART OF NXT FOR A WHILE NOW AND HE DOESN'T SEEM TO BE MOVING

ANY FURTHER FORWARD.

STRONG WAS ONCE PUSHED TOWARDS THE NXT CHAMPIONSHIP BUT CAME UP SHORT IN HIS HUNT, WHICH MANY

OF THE WWE UNIVERSE HOPED WAS BECAUSE HE WAS SET TO MAKE THE SWITCH TO WRESTLING ON THE

MAIN ROSTER INSTEAD.

RODERICK RECENTLY MADE HISTORY WHEN HE WAS PART OF THE FIRST EVER WAR GAMES MATCH AT

THE NXT TAKEOVER SHOW OF THE SAME NAME AND HE MAY NOT WALK OUT OF THE CAGE THE SAME WAY

HE WALKED IN.

NOW THAT THERE IS A HEEL NXT CHAMPION, IT IS ALSO FEASIBLE THAT STRONG COULD BE SEEN

AS THE NEXT CHALLENGER FOR ANDRADE ALMAS.

STRONG IS ALWAYS A SAFE PAIR OF HANDS AND IF NXT GOING ONTO A TRANSITIONAL PERIOD THEN

HE WOULD BE THE PERFECT STAR TO CARRY THE MAIN EVENT PICTURE ALONGSIDE ALMAS IN THE

COMING MONTHS.

NO WAY JOSE.

NO WAY JOSE HAD ONE OF THE MOST INFECTIOUS GIMMICKS ON THE NXT ROSTER UNTIL HE DISAPPEARED

EARLIER THIS YEAR.

THE DANCING STAR ALWAYS ENSURED THAT THE LIVE CROWD HAD A GOOD TIME AND SEEMED TO HAVE THE

RIGHT AMOUNT OF ABILITY IN THE RING AS WELL, BUT HE HAS SEEMINGLY BEEN AWOL FROM NXT FOR

THE PAST FEW MONTHS.

USUALLY, WHEN NXT STARS ARE ABSENT IT IS BECAUSE WWE IS PREPARING THEM FOR THEIR MAIN ROSTER

DEBUT AND ENSURING THAT THEY HAVE THE COVER THEY NEED WHEN THEY ARE GONE.

JOSE'S GIMMICK WAS ALWAYS GOING TO BE A HARD ONE FOR THE MAIN ROSTER AUDIENCE TO COMPREHEND,

BUT IT WILL BE INTERESTING TO SEE IF WWE IS PREPARING FOR THE INFECTIOUS STYLE OF JOSE

TO BE PROMOTED TO THE MAIN ROSTER, OR IF HE IS ON THE LIST OF SUPERSTARS WHO COULD BE

GETTING THE CHOP IN FAVOUR OF THE NEW TALENT COMING THROUGH FROM THE PERFORMANCE CENTRE

RIGHT NOW.

JOHNNY GARGANO.

JOHNNY GARGANO IS ONE OF THE MOST POPULAR STARS ON THE NXT BRAND RIGHT NOW AND WAS RECENTLY

PART OF ONE OF THE BIGGEST MATCHES OF HIS CAREER WHEN HE FACED UK CHAMPION PETE DUNNE

AS PART OF NXT TAKEOVER: WAR GAMES.

GARGANO IS MORE TECHNICALLY ADVANCED THAN ANY OTHER STAR ON THE BRAND AFTER YEARS OF

PERFORMING ALL OVER THE WORLD.

THE FORMER NXT TAG TEAM CHAMPION AND HIS ONE TIME BEST FRIEND TOMASSO CIAMPA STILL NEED

TO HAVE THE FEUD THAT BEGAN BUILDING WHEN CIAMPA ATTACKED GARGANO AT TAKEOVER: CHICAGO

BACK IN MAY.

THE FEUD HAS SINCE BEEN DELAYED SOMEWHAT OVER THE PAST FEW MONTHS BECAUSE CIAMPA HAS SINCE

BEEN OUT WITH A KNEE INJURY, BUT IT SEEMS THAT HE COULD BE BACK IN THE COMING MONTHS

AND THE DUO CAN FINALLY HAVE THEIR SHOW-STEALING FEUD.

THIS WOULD BE THE PERFECT FEUD FOR JOHNNY WRESTLING TO BOW OUT ON BEFORE HE TAKES HIS

RIGHTFUL PLACE ON THE MAIN ROSTER.

PEYTON ROYCE AND BILLIE KAY.

BILLIE KAY AND PEYTON ROYCE WERE VOTED "THE BREAKOUT STARS OF THE YEAR" IN NXT LAST

YEAR, WHICH SHOWS JUST HOW MUCH THE ICONIC DUO HAVE CONNECTED WITH THE NXT UNIVERSE OVER

THE PAST FEW YEARS.

THE AUSTRALIAN DUO HAS BEEN OVERLOOKED IN THE WOMEN'S DIVISION SO MANY TIMES, EVEN

WHEN FANS THOUGHT THAT IT WAS FINALLY PEYTON'S TIME TO BECOME CHAMPION LAST WEEKEND.

BILLIE AND PEYTON SHOULD HAVE BEEN CHOSEN BY PAIGE LAST WEEK TO BE PART OF THE STABLE

THAT INVADED MONDAY NIGHT RAW.

THE WOMEN ARE MUCH MORE CAPABLE IN THE RING AND ON THE MIC THAN SONYA DEVILLE AND MANY

ROSE, BUT ONCE AGAIN THEY HAVE BEEN OVERLOOKED.

IT IS HOPED THAT IF THERE ARE ANY MORE WOMEN SET TO JOIN RAW OR SMACKDOWN LIVE IN THE COMING

WEEKS, THEN BILLIE AND PEYTON WILL DEFINITELY BE THE TWO WOMEN THAT THE WWE UNIVERSE WOULD

LOVE TO SEE ON THE COMPANY'S FLAGSHIP SHOW.

THE AUTHORS OF PAIN.

NO TAG TEAM IN NXT HAS BEEN MORE DOMINANT THAN THE AUTHORS OF PAIN.

THE DUO WASN'T ONE OF THE BEST TEAMS IN NXT WHEN THEY FIRST DEBUTED AND SHOWED THAT

THEY HAD A NUMBER OF FAULTS AND WENT ON TO MAKE MANY MISTAKES, BUT FAST FORWARD MORE

THAN A YEAR AND THEY HAVE WON OVER MANY OF THE WWE UNIVERSE WITH THEIR UNIQUE STYLE.

AKAM AND REZAR HAVE BEEN RUMOURED TO BE SET TO DEBUT ON THE MAIN ROSTER IN THE COMING

MONTHS AHEAD OF WRESTLEMANIA 34 SINCE THEY NOW HAVE NOTHING MOVING FORWARD IN NXT.

THE AUTHORS OF PAIN, WHO ARE MANAGED BY WWE HALL OF FAMER PAUL ELLERING, HAVE ALREADY

LOST THEIR NXT TAG TEAM CHAMPIONSHIPS TO SANITY AND CAME UP SHORT IN THE FIRST EVER WAR GAMES

MATCH LAST WEEKEND, WHICH COULD HAVE BEEN THEIR SWAN SONG.

SMACKDOWN WOULD DEFINITELY BENEFIT FROM THE DOMINANT DUO, SO HOPEFULLY, THEIR PROMOTION

IS SOONER RATHER THAN LATER.

DREW MCINTYRE.

WWE SEEMS TO FOLLOW A SYSTEM WITH NXT CHAMPIONS.

IT USUALLY SEES ALL FORMER CHAMPIONS DROP THE TITLE TO THEIR PREDECESSOR BEFORE THEY

ARE THEN PROMOTED TO THE MAIN ROSTER.

SHINSUKE NAKAMURA, BOBBY ROODE, SAMOA JOE AND A NUMBER OF OTHER STARS HAVE FOLLOWED

THIS TRADITION OVER THE PAST FEW YEARS AND SINCE DREW DROPPED HIS CHAMPIONSHIP LAST WEEKEND

TO ANDRADE ALMAS, IT SEEMS THAT HIS TIME COULD FINALLY BE COMING.

OF COURSE, DREW HAS ALREADY BEEN ON THE MAIN ROSTER AND WAS RELEASED FROM WWE BACK IN 2014

ALONG WITH A NUMBER OF OTHER STARS BEFORE HE WAS RESIGNED EARLIER THIS YEAR.

DREW HAS SINCE PROVED THAT HE HAS RETURNED TO THE COMPANY AS A COMPLETELY DIFFERENT MAN,

BUT HE WAS INJURED IN HIS MATCH AGAINST ALMAS AT TAKEOVER: WAR GAMES, WHICH MEANS THAT HIS

CALL-UP COULD BE DELAYED BECAUSE OF THE INJURY, BUT IT IS DEFINITELY IN THE WORKS.

ALEISTER BLACK.

ALEISTER BLACK HAS CERTAINLY MADE AN IMPRESSION ON THE NXT BRAND OVER THE PAST FEW MONTHS

EVER SINCE HIS DEBUT BACK IN APRIL.

THE UNDEFEATED STAR STOLE THE SHOW LAST WEEKEND AS HE TOOK ON VELVETEEN DREAM AND MANAGED

TO MAINTAIN THAT UNBEATEN RECORD.

BLACK IS A STAR WAITING TO HAPPEN AND WWE NEEDS TO CASH IN ON HIM AS SOON AS POSSIBLE

AND HOPE THAT THE MAIN ROSTER AUDIENCE WILL TAKE TO HIM THE SAME WAY THAT NXT HAS.

BLACK IS AN ENIGMA; A MAN HIS SIZE SHOULD NOT BE ABLE TO MOVE THE WAY THAT HE DOES,

WHICH MAKES HIM EVEN MORE APPEALING TO WWE SINCE THEY DON'T HAVE ANYONE ELSE QUITE

LIKE HIM TO FILL THAT VOID ON RAW OR SMACKDOWN.

BLACK HAS THE ABILITY TO STEAL THE SHOW ON ANY OCCASION AND AGAINST THE STARS THAT WWE

ALREADY HAS ON THEIR MAIN ROSTER, BLACK WILL DEFINITELY BE AT HOME.

For more infomation >> TOP 15 NXT Stars Who Could Be Next In Line For A Main Roster Debut - [Facts Wrack] - Duration: 16:17.

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Tamma Tamma Loge DJ Remix Song || Bollywood Dance DJ || Old Is Gold DJ || Old Hindi DJ Song - Duration: 4:49.

Tamma Tamma Loge

Tamma Tamma Loge DJ Remix Song

Tamma Tamma Loge DJ Remix Song || Bollywood Dance DJ || Old Is Gold DJ || Old Hindi DJ Song

Tamma Tamma Loge DJ Remix Song

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Những Pha Phòng Ngự Hay Nhất Hành Tinh Của Bộ Đôi Quái Vật PEPE & Ramos Ở Real Madrid - Duration: 12:00.

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LINQ e expressões Lambda - Duration: 6:07.

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MBB Gay Games - Duration: 2:01.

My name is Bernardo I'm part of the Meninos Bons de Bola

It's the first amateur soccer team of transsexual men of Brazil

We are a group of boys who believe that trans men can be where they want to be

Including sports courts, playing soccer

Besides athletes we're a quite diverse group

regarding what is being a man

We are white, black, thin, fat, with breasts, without breasts, with beard and without beard

We want to participate in Gay Games because our bodies are bodies in transformation

political and resistance bodies

The Gay Games need to come up with this issue, need to put the question of

trans men in games

We know of the difficulty of the event in sponsoring a team, but because of this

specificity of the team we'd like to make

this request because we are one but we can be many teams of trans men and women around the world.

For more infomation >> MBB Gay Games - Duration: 2:01.

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Renault Clio TCE 90pk [removed]R-link/Airco/Cruise/16''LMV) - Duration: 0:54.

For more infomation >> Renault Clio TCE 90pk [removed]R-link/Airco/Cruise/16''LMV) - Duration: 0:54.

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Volvo V40 2.0 D2 R-DESIGN BUSINESS Navi | Stoel verw. | Dealeronderhouden | LMV - Duration: 0:54.

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Renault Captur 0.9 Tce Dynamique Navi/R-Link/Ecc/Pdc/17inch - Duration: 1:01.

For more infomation >> Renault Captur 0.9 Tce Dynamique Navi/R-Link/Ecc/Pdc/17inch - Duration: 1:01.

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Ford Transit Custom 290 2.0 TDCI 130PK L2H1 Limited Navi/schuifdeur L+ R - Duration: 0:56.

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Ford Transit Custom 290 2.0 TDCI 130PK L2H1 Limited Navi/schuifdeur L+ R - Duration: 1:01.

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Ford Transit Custom 290 2.0 TDCI 130PK L2H1 Limited Navi/schuifdeur L+ R - Duration: 0:59.

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Introducing the NCBI Pathogen Detection Isolates Browser - Duration: 31:24.