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70-544 TS: Ms Virtual Earth 6.0, Application Development

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70-544 exam Dumps Source : TS: Ms Virtual Earth 6.0, Application Development

Test Code : 70-544
Test Name : TS: Ms Virtual Earth 6.0, Application Development
Vendor Name : Microsoft
: 134 Real Questions

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Microsoft TS: Ms Virtual Earth

On the brand new Microsoft virtual Lab Certification gadgets | Real Questions and Pass4sure dumps

For several years now, Microsoft has covered simulation gadgets in some of their IT certification assessments. Simulation gadgets are meant to check the candidate’s arms-on skills with the Microsoft product/know-how beneath consideration. a good deal has took place within the virtualization space in ensuing years; the latest innovation is what's common because the digital Lab-based mostly exam merchandise.

What’s the change Between a Simulation and a virtual Lab?

A simulation examination merchandise, they ought to be aware, is a mock-up of an operating equipment and, optionally, different business utility. The simulation merchandise developer contains extra functionality into the mockup with the intention to throw off candidates who are in doubt exactly a way to accomplish the given task.

however, since the simulation is a completely ‘canned’ object, the user interface includes many, many ‘dead’ components and is, at most suitable, a mediocre formulation for assessing a candidate’s useful skillability with a seller’s know-how.

To get a suppose for the way Microsoft simulation items work, please try the following online technology demo from Microsoft (requires information superhighway Explorer):

Microsoft Simulation item on-line Demo

here's a downloadable edition of the identical: Microsoft licensed skilled (MCP) exam Demos

nevertheless, a virtual Lab places the candidate in a are living, Terminal features session with one or extra home windows-primarily based digital machines. sure, that’s relevant: the certification candidate has free reign over an entire working gadget. What this capability, very nearly talking, is that if the candidate does not recognize a way to operate the stated task, then he or she is, to use the IT parlance, “hosed,” so far as that item is concerned.

check out right here hyperlink that demonstrates how Microsoft is enforcing the digital Lab format in its present technology of IT certification exams:

Amila online: Microsoft’s digital Lab assessments: Going inside the Black container

this is a different cool link without delay from the Microsoft studying individuals that additionally showcases the prior to now linked demo, but also does a much better job than me at describing the Microsoft virtual Lab know-how:

Microsoft Born to be taught weblog: Microsoft digital Lab assessments: Going interior the Black box

Which Microsoft checks Are Going digital?

As of this writing (September 2009), only 1 exam has crossed over from typical simulation to virtual Lab structure:

examination 083-640: TS: windows Server 2008 lively listing, Configuring

these of you who had been actively preparing on your windows Server 2008 MCITP credentials could know that the 083-640 examination is the replacement for exam 70-640 TS: windows Server 2008 active directory, Configuring. The unhealthy news is that the 70-640 examination become a walk in the half in comparison to the 083-640 exam. The good news is that those that flow the new 083-640 examination can hang their heads excessive, having correctly verified not handiest their skillability with theoretical windows Server 2008 ideas, but also their competency with useful application of those concepts.

This text comes from the above-referenced prep page for 083-640:

This know-how professional (TS) examination, exam 83-640: TS: windows Server 2008 energetic listing, Configuring, became obtainable in November 2008. This examination maps to the equal objectives as 70-640, but this version (the place available) incorporates a performance primarily based part in addition to the general exam merchandise types. This examination is attainable in English only.

other Avenues for digital Lab Experimentation

I’ve all the time informed my college students that the Microsoft TechNet and MSDN virtual Labs are considered one of Microsoft’s most effective-saved secrets for IT certification candidates. bound, Microsoft doubtless intends these virtual labs basically for enterprise consumers to get a think for brand new Microsoft products and technologies with no need to exert the crucial blood, sweat, and tears of deploying the software on one or more look at various packing containers.

in spite of this, I humbly put up that IT cert candidates can profit helpful hands-on adventure with their MS know-how of choice by using spending time in these free (!), prosperous, and mighty virtual Lab environments. The labs provide 90-minute time blocks by which you're free to work through a structured lab (you even get to down load the absolutely formatted lab e-book).

What’s greater, is all Microsoft skill sets are coated: IT professional, Database, and Developer. check ‘em out:

related Pearson items

Aches & Pains Map indicates the place It Hurts: Yahoo-Microsoft and Google | Real Questions and Pass4sure dumps


The listing of engines like google is shrinking. Yahoo-Microsoft? The most well liked theme.

Search engine market share? The fulcrum of the opposed MS-Yahoo bid — and one of the crucial common searches in Google, Yahoo, MSN, and Ask.

On super Tuesday, they found a way to monitor Presidential primaries on a search engine: a mashup of Google Earth and twitter / twittervision, with a Google gadget and iGoogle thrown in for decent measure. still, key phrase Meta Tag, meta tags, and meta keywords remain their stock in alternate here at Search Engine Watch.

So when Gary price, director of on-line information substances and editor, ResourceShelf and DocuTicker, pinged us with a weather Channel-MSN virtual Earth mashup, they couldn’t face up to featuring a fresh attitude on the Microsoft-Yahoo adversarial bid.

The Aches and Pains Map on suggests Google feeling no pain from the MicroHoo hoo-hah.

Yahoo traders and employees — in the yellow area – don’t look like suffering in Silicon Valley.

In contrast, Microsoft – rebuffed by using Yahoo – seems to affected by high degrees of aches and pains. The Microsoft virtual Earth map of aches and pains (a Google Earth competitor) indicates Redmond almost in the red zone.

The Aches and Pains Index map graphically depicts areas of bigger or lessen ranges of weather-linked pain. areas within the orange areas can predict above regular degrees of aches and pains. Areas shaded in yellow will have normal stages, and gray regions will be below commonplace.

So in case you don’t like the climate, wait until the next day. It’ll exchange, identical to Google’s share of searches – and there’s still nothing you can do about it.

are looking to reside on true of the newest search developments? Get excellent insights and information from their search specialists.

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14 stellar examples of fantastically link-useful digital PR campaigns, damaged into five certain categories of content codecs.

Why do they buy luxury manufacturers? a glance into the psychology using their purchase choices -- and how search marketers should still for this reason consider about method.

It’s been very nearly three½ years in view that Google announced RankBrain, however they now have little element about the way it works. right here's what they really comprehend--and what is fable.

Mojeek's elevator pitch: "independent and unbiased search results and not using a consumer tracking." Deep dive into how they stack up as an alternative search engine.

Watch game of Thrones online: how to move season 8 from anywhere | Real Questions and Pass4sure dumps

well, after well-nigh two years they ultimately bought a brand new episode of video game of Thrones and it definitely lived up to the hype. One hour of pure action, reuniting characters, constructing plot traces and the expected game of Thrones violence - they can say for definite that you simply should watch online game of Thrones. Episode 1 become a should-watch and optimistically, the rest of season eight will continue on this fashion.

video game of Thrones season 8: when and the place to monitor?

the first episode has now aired which potential it's now attainable for capture up. reckoning on the place you are living, there's a special streaming or tv provider to observe online game of Thrones on-line - that you may see a listing of English-talking international locations under.

The relaxation of the episodes can be streaming each Sunday, are living at 9pm ET, 6pm PT within the US, which ability it could possibly grow to be airing early within the morning based where you live (2am within the UK, for example). So it might possibly be rather a late evening for some.

the first episode aired on Sunday evening (or very early morning for some unfortunate time zones) however do not worry in case you missed it, which you can still without problems watch online game of Thrones on-line. There are a number of of distinct streaming features showing the first episode on trap up and even alternatives for international locations now not airing the exhibit - you can find all of your alternatives under.

With six new episodes and a total of just about seven hours of viewing to be consumed, the 8th and last season is one now not to leave out. Rumours are now suggesting the greatest fight in the heritage of television and film, so that you simply know here is going to be huge.

So with the primary episode now obtainable to monitor, all of the first seven seasons effectively obtainable and the rest of season eight not distant - here is the area to be for all your how to watch online game of Thrones online awareness. they have tracked down the locations to watch every and every episode, no be counted the place you are living - that you may discover them all down beneath. and it's even viable to do so at no cost in some areas.

Watch video game of Thrones on-line: movement outside your country

are looking to watch game of Thrones online within the US, UK, Canada, Australia, New Zealand or India? we've got discovered all of your top of the line options and listed them below.

however, in case you find yourself in the unfortunate circumstances the place you've booked a vacation over the period of game of Thrones season eight, then issues get a little extra problematic. For those unlucky enthusiasts, all hope isn't lost yet. you'll locate that looking at online game of Thrones may not be an alternative due to geo-blockading. This capability that broadcasters do are likely to are trying and forestall you from staring at in different nations, putting up some virtual roadblocks.

don't worry, you will nevertheless be able to watch the exhibit, it simply gets a bit bit more durable. by using a VPN, you will nonetheless be capable of capture the display by using changing your IP address to a location back the place you are living, enabling you to observe it all (assuming that does not breach any TS&Cs, of path). 

we've got demonstrated hundreds of VPNs and might suggest these because the three optimal VPN alternatives at the moment attainable: 

1. categorical VPN (comes with a 30 day cash returned guarantee)this is the #1 rated choicest VPN on the earth at this time because of its speed, security and ease-of-use. that you may watch on many contraptions without delay including wise TVs, fire tv Stick, notebook, Mac, iPhone, Android cellphone, iPads, drugs and so on. check out express VPN and get 3 months FREE with an annual plan2. IPVanish helps up to 10 instruments, so first rate on the go and with a spotlight on security3. NordVPN: SmartPlay tech makes NordVPN an excellent, most economical choice for streaming

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Watch game of Thrones on-line within the u . s .

well there actually is not any shock here, if you are within the US then HBO might be the go-to region for every thing online game of Thrones. unfortunately, it's additionally no surprise that this skill you will should sign in to a HBO subscription to monitor the display.

HBO presents just a few programs, all of which are pretty bendy. That potential which you can sign in for one month, finish the exhibit after which cancel if needs be. now not to mention, HBO offers free trials on all of its packages, which helps soften the blow. costs beginning at $14.ninety nine a month with HBO and go up counting on which equipment you choose.

the first episode is now attainable for trap up within the US. The rest of the episodes can be airing each Sunday at 9pm ET, 6pm PT, giving US enthusiasts one of the most superior streaming times out there.

Watch online game of Thrones on-line in the UK FREE

if you are within the UK you get two diverse alternate options when it comes to watching the show: either via Sky Atlantic or by the use of a Now television leisure flow.  

the united kingdom failed to precisely have the easiest live streaming slot (2am is only a little bit too early) so they absolutely have in mind in case you overlooked it. The decent news is that the first episode is now purchasable for trap-up on both NowTV and Sky Go or you can watch it on tv tonight (April 15) at 9pm. 

all of us be aware of that Sky may also be costly, so make certain to take a look at their dedicated Sky television offers and applications guide to be certain you are getting modern-day most suitable expenditures and presents.

NowTV then again has a 7-day FREE trial, that means that you may catch the primary episode without paying a single penny! And, just in time for online game of Thrones, you can get three months of NowTV enjoyment for just £17.ninety nine (25% discount)

And if you're out of the uk and nonetheless want to watch suggests from your Sky or Now subscriptions, then you definately'll should download and install a VPN as described above.

Watch video game of Thrones on-line in Canada

if you are in Canada making an attempt to watch online game of Thrones on-line then you definitely're in success as Crave could be showing the whole lot of the new season on its release. That capacity the same air time as the US, each week on Sunday at 9pm ET, 6pm PT.

And in case you have not yet been in a position to watch the first episode, it is now attainable to watch on seize-up with Crave. 

You can pay for Crave on a monthly basis or get an annual subscription. not like HBO, Crave does not appear to do trials but if you do pay for a subscription, they'll throw in one free month.

fees range for Crave but which you can get applications together with HBO from round $20 for the month.

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Watch video game of Thrones online in Australia

if you're trying to capture the primary episode now its aired (and even the complete reveal) in Australia, there are only two alternate options for you, and that's the reason either signing up to Foxtel's cable provider or the business's streaming carrier, Foxtel Now. Episode 2 could be airing at 11am on Monday April 22, a somewhat extra low cost time compared to the uk's 2am!

Like the entire streaming capabilities above, you will ought to pay to get a subscription and there are a bunch of distinctive packages purchasable, but Foxtel Now offers a free 10-day trial if you haven't already signed up.

Watch online game of Thrones on-line in New Zealand

For these in New Zealand there in fact simplest is one alternative for season 8 and that's the reason SoHo. The Sky top rate leisure channel may be the area to locate season 8, airing at 1pm after which repeating at 8.30pm each and every Monday. the first episode is now obtainable for catch up on the website.

sadly, SoHo doesn't do trial subscriptions. So in case you do not have already got it you are going to need to dive on in on a tv package or improve your present one to a plan with SoHo.

packages together with SoHo beginning as low as $24.ninety one a month and go up from there reckoning on how a good deal you decide to consist of. 

Watch game of Thrones online in India

For enthusiasts in India you might be in good fortune, when it comes to looking at game of Thrones online, India has the most cost-effective choice. quite simply log onto Hotstar and sign up for a premium account. this may cost you both ₹207.39 ($three) for a month or ₹967.eighty four ($14) for a whole 12 months. 

the primary episode is now attainable to observe and Hotstar might be displaying each other episode because it airs. The time to tune in is each Monday at 6.30am. yes that does mean an early morning for Indian fans however they believe like its a small rate to pay to capture this a great deal-expected exhibit's first episode.

the way to watch seasons 1-7 of online game of Thrones on-line

if you wish to do some super speedy re-watching of the historical seasons, they will inform you the top of the line places to observe seasons 1-7 of video game of Thrones however be warned, there is a lot of content to trap up on. 

fortuitously, most of the streaming functions above present seasons 1-7 of the display, that means which you could watch all the historical episodes after which smoothly movement into the brand new season. under now they have listed each country with all of the diverse viewing systems for the old seasons:

US: the USA has a tonne of alternatives when it involves the historical seasons with: HBO GO, HBO Amazon channel, Amazon leading, iTunes, Google Play store, Microsoft save.

UK: With simply as a whole lot option, within the UK which you can watch ancient episodes on. Your best bets are both NowTV or Sky Go. different options encompass Amazon major, iTunes, Sky shop, PlayStation, Microsoft store, Chili, TalkTalk television or the Google Play keep.

Canada: The alternate options in Canada are just a little extra limited, choose between: Crave Plus, iTunes, PlayStation, Microsoft save or Google Play store.

Australia: fairly corresponding to Canada, choose from Foxtel Now, iTunes, Microsoft shop, psor the Google Play keep.

New Zealand: SoHo...yep it truly is pretty lots your only option in New Zealand. but if you need to watch the brand new season you could as neatly watch the complete exhibit appropriate?

India: simply Hotstar, however when its so cheap, do you really want to shop round?

in different places: reside somewhere not on this list? try giving a VPN a are trying and connecting to one of the crucial countries beneath with a variety of alternate options for looking at the show. 

video game of Thrones Season 8 trailer - get yourself excited for the subsequent episode

Episode 1 got you craving some extra motion? Watch the trailer above to help cling you over until the first episode airs. With the entire Starks, dragons and White Walkers, they feel like season eight goes to be greater than value the wait. 

Scroll up to look where to observe the primary episode the place you are living.

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TS: Ms Virtual Earth 6.0, Application Development

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Identification of the Beer Component Hordenine as Food-Derived Dopamine D2 Receptor Agonist by Virtual Screening a 3D Compound Database | real questions and Pass4sure dumps

Molecular properties of FCDB and PhyDB compared to other VS databases

Our first aim was the generation of an in silico 3D food-compound database. Besides this database (FCDB; 12,579 compounds), which they constructed by selecting natural food constituents from the Dictionary of Food Compounds14, they also generated a small natural products database (PhyDB; 987 compounds) based on the catalogue from the vendor PhytoLab Vestenbergsgreuth, Germany (available at The databases are part of the Supplementary information.

For the characterisation of their newly generated databases, they calculated molecular property distributions for FCDB and PhyDB and for samples containing 10,000 randomly selected compounds from the UNPD database15, the ZINC biogenic compounds subset (ZBC) and the ZINC all purchasable subset (ZAP)16. Thus, they could compare their databases with established freely available VS databases that contain natural products (UNPD database and ZBC) and drug-like compounds (ZAP). Detailed data are available in the Supplementary information, Fig. S1.

The calculations revealed that the FCDB and PhyDB databases are substantially different to a typical drug-like library (ZAP). While the compounds in drug-like libraries tend to comply with Lipinski’s rule-of-five17 resulting in Gaussian-like molecular property distributions with distinct maxima, the molecules in FCDB, PhyDB, and UNPD exhibit very broad molecular-property distributions. Hence, the compounds in these databases tend to have higher structural diversity, which is typical for natural compound libraries18. Despite their distinct similarity, differences between FCDB/PhyDB and UNPD still exist, especially in terms of molecular weight. The natural product library ZBC turned out rather to possess drug-like than natural-product-like properties, probably because the authors of ZINC ( took their information about the natural character of a compound from vendor catalogues. The vendors often also include synthetic derivatives of natural products in their catalogues to make them more attractive for medicinal chemists, as also observed by Manallack et al.19. In summary, it turned out that their generated screening databases represent a new type of screening library, which is much more similar to databases containing natural products than drug-like compounds.

Virtual screening process

The next goal was the search for nutritive or natural D2R-ligands in the generated VS databases. D2R is a target of great interest for the pharmaceutical industry, for which agonists have been developed as drugs for treating Parkinson’s disease and antagonists for treating schizophrenia20,21. Although no X-ray structure of D2R has been published yet, a wide variety of ligands has been synthesised that facilitates the effective use of ligand-based VS methods such as pharmacophore searching11. The GPCR Ligand Library22 provides a comprehensive collection of known D2R-ligands compiled from the GLIDA database23. However, since the majority of D2R-ligands are synthetic in origin, the ligand collections contain primarily drug-like compounds, in contrast to those contained in FCDB and PhyDB. Therefore, the selected VS methods needed to be capable of finding VS hits in FCDB and PhyDB that are structurally different to the input ligands. Therefore, they did not only apply VS methods that rely on molecular similarity but rather used pharmacophore searching in combination with hit-list clustering and molecular docking. The overall VS workflow is summarised in Fig. 1.

Figure 1: Schematic representation of the VS process from database generation to experimental testing. Figure 1

The numbers on the left and on the right side represent the number of compounds that passed through the respective VS step depicted in the centre. Due to 230 compounds that are present in both FCDB and PhyDB, the absolute number of different compounds is smaller than the sum of the compounds in the two databases.

Pharmacophore modelling

By selecting relatively large and structurally diverse training sets for both D2R-agonists and -antagonists, they ensured that the resulting pharmacophore models only retain the most critical features for binding. The training sets are depicted in the Supplementary information, Figs S2 and S3. Training sets containing only large and structurally similar compounds would probably have resulted in very specific pharmacophore models with a large number of features, which would not be able to find any hits in FCDB and PhyDB. Additionally, they did not consider excluded volumes in the pharmacophore models to avoid obtaining small hit lists.

It is a prerequisite for an alignment of the ligands to be a good approximation to their binding mode that the template compound for the alignment provides an orientation close to its bioactive conformer. In addition to molecular docking and subsequent molecular-dynamics refinement of the ligand conformers as techniques for approaching the bioactive conformation, they selected some ligands, namely # 21 and # 27 from the agonist training set, Fig. S2, and # 32 from the antagonist training set, Fig. S3. They used the conformers that are stored in the GPCR Ligand Library directly, because the conformational space in low-energy regions is sparsely occupied for structurally restrained ligands. The bioactive conformer has been shown to be often located near the global minimum24 and a low-energy conformer for a restrained ligand can thus be a good approximation. The training set ligand conformers that possess the maximum similarity compared to the template were then determined by ParaAlign (see Methods and Supplementary information). For each ligand, this conformer was extracted and used as input for the HipHop25 pharmacophore model generation algorithm. Since they generated up to 25 pharmacophore models per run, the variation of the conformer types and the templates that were used for ParaAlign resulted in a large number of models, which had to be validated subsequently.

The validation process involved large test databases compiled from the GPCR Ligand Library and GPCR Decoy Database22 in order to determine the overall best pharmacophore models. They found two very similar pharmacophore models with almost identical excellent VS performance in model validation for the D2R-agonists. Both models contained hydrogen-bond donor, aromatic ring and positive ionisable features. They therefore decided to use only the model that gave a slightly better receiver operating characteristic (ROC) curve (Fig. 2b).

Figure 2 Figure 2

The overall best D2R-agonist pharmacophore and its inter-feature distances (a), depicted together with its ROC curve that results from VS of the test database (b). The features are hydrogen bond donor (purple, HBD), aromatic ring (orange, R) and positive ionisable (red, P). The dashed red line in the ROC plot indicates a random selection.

A comparison between this D2R-agonist pharmacophore model (Fig. 2a) and models reported in the literature revealed a high degree of similarity. Both Malo et al.26 and Chidester et al.27 reported D2R-agonist pharmacophore models that also contain hydrogen-bond donor, aromatic ring and positive ionisable features with similar relative orientations. The only difference is an additional hydrophobic feature close to the positive ionisable feature present in their models. However, since not all their training set ligands contain such a feature, they conclude that the hydrophobic feature is able to increase binding affinity but that it is not critical for binding.

We also obtained two pharmacophore models for the D2R-antagonists that exhibited the best VS performance in the validation step (Fig. 3c). In contrast to the agonists, these two models did not show a high degree of similarity concerning shape and features (Fig. 3a,b). While the first D2R-antagonist model contained a hydrogen-bond acceptor, an aromatic ring and a positive ionisable feature, the other model possessed a hydrophobic feature together with the aromatic ring and positive ionisable feature. The relative orientation of the features was also different. However, this observation is not surprising, since their D2R-antagonist training set covered ligands with considerably different shapes (e.g. ligands # 35 and 39, Fig. S3). Such diverse ligands probably adopt different binding modes and, hence, it is not surprising that they obtained two diverse pharmacophore models.

Figure 3 Figure 3

The overall best D2R-antagonist pharmacophore models and their inter-feature distances (a and b) depicted together with the corresponding ROC curves that result from VS of the test database (c). The features are hydrogen bond acceptor (green, HBA), aromatic ring (orange, R), positive ionisable (red, P) and hydrophobic (blue, H). The dashed red line in the ROC plot indicates a random selection. The blue curve corresponds to model a) and the green curve to model b).

The models further resemble a ligand classification for pharmacophore model generation reported by Klabunde and Evers28. They divided their D2R-antagonist training set into two different classes and built different pharmacophore models from the two subsets. Their first D2R-antagonist pharmacophore model (Fig. 3a) resembles their class I model, whereas their second model (Fig. 3b) shows some similarity to their class II model. As their validation database contains a substantial number of actives from both ligand classes, similar VS performance of the pharmacophore models was not surprising. The validation performance is not as good as for the D2R-agonist models, probably because, on the one hand, either model is recognising class I-like actives or class II-like actives better and, on the other hand, hydrophobic and hydrogen-bond acceptor features are not as selective as a hydrogen-bond donor feature. For this study, they decided to use both models for database searching in FCDB and PhyDB and, subsequently, to combine the two hit lists to avoid missing any type of antagonist.

The 3D database-searching step using the selected D2R-agonist pharmacophore model selected 1,441 compounds from FCDB and PhyDB. In contrast, the two selected D2R-antagonist pharmacophore models gave a combined hit list with 1,866 compounds. Note that they treated tautomeric compounds and different ionisation states as different database entries in this step, which led to a lower number of chemically different hit compounds.

Hit-list clustering and compound prioritisation

The next goal of their study was to select structurally diverse compounds for experimental testing. Therefore, they performed a clustering analysis of the hit lists in Discovery Studio 3.1 (Accelrys Inc., San Diego, CA) to divide them into groups that contain structurally different classes (Fig. 1). Moreover, the possible corresponding tautomers and ionisation states for a given hit compound could be pooled as they were assigned to the same compound cluster. After variation of the number of generated clusters, they found that values of 80 clusters for the D2R-agonist hits and 100 clusters for the D2R-antagonist hits represent good values for characterising the hit-list compounds.

Selecting compounds from different clusters ensured that they chose structurally diverse molecules for the following molecular-docking step. The choice was further based on the fit value to the pharmacophore models and on visual inspection. For both D2R-agonist and -antagonist screening, they derived a cut-off limit for the fit value from the pharmacophore validation step, in which databases consisting of known actives and putative inactives were mapped to the models. From the actives mapped to the D2R-agonist model, all but one gave a fit value higher than 2.2. Hence, they used this value as a cut-off limit in compound prioritisation. By contrast, most of the actives mapped to the D2R-antagonist models showed fit values of 1.9 and 2.3 for model A and model B, respectively. They retained pharmacophore searching hits with larger fit values than these cut-off limits for at least one of the models.

Hit-list clustering and compound prioritisation reduced the number of VS hits to 125 and 69 different compounds for the D2R-agonist and -antagonist screening, respectively. Thus, they selected less compounds from the D2R-antagonist screening for the molecular docking step, although the pharmacophore hit list and the number of clusters was larger than for the agonists. However, many compounds in the antagonist hit-list did not pass the fit-value cut-off. Additionally, visual inspection of the hit-list molecules revealed that large and bulky compounds tended to have very high fit values. These molecules were rejected because they are unlikely to fit into the binding pocket.

Molecular docking and final compound selection

The combination of pharmacophore searching with molecular docking enabled an initial check of their database molecules for the required molecular features and their relative orientations. Then they examined more thoroughly, whether the candidates could fit well into the binding pocket and establish important protein-ligand interactions. A D2R inactive-state homology model used successfully in a previous docking study29 served as docking target. The compounds that passed the clustering and prioritisation step were docked together with their corresponding tautomers and ionisation states using AutoDockVina30 with high exhaustiveness and maximum conformer values for detailed examination. For each compound, they carefully inspected both the value of the scoring function and the resulting receptor-ligand complexes. An interaction between a positively charged moiety and the carboxylate group of Asp1143.32 was mandatory for a compound to be regarded as hit in the docking step.

Eventually, they obtained 53 hit compounds and searched for corresponding possible activity data concerning dopamine receptors in the literature. They excluded hits with known bioactivity at D2R and checked the commercial availability of the remaining compounds. Finally, 17 compounds of different origin (Fig. 4a,b, Table 1) were purchased and prepared for experimental testing.

Figure 4 Figure 4

Structures of the experimentally tested D2R-agonist (a) and -antagonist (b) VS hits. The 17 compounds can be classified into natural food constituents (delphinidin, hordenine, kukoamine A, pyrraline, salsolinol, roquefortine C), further compounds from natural sources (fumigaclavine A, leonurine, muscimol, ajmalicine, dihydroberberine, emetine), and food contaminants (clenbuterol, fenpropimorph, halofuginone, robenidine, sarafloxacin).

Table 1: Compound sources, pharmacophore searching fit values and radioligand binding data for the tested VS hits compared to the reference compound dopamine, employing D2L receptors expressed in CHO cells.

We also examined, whether a 2D molecular similarity search would have been sufficient to extract these molecules from the VS databases. Therefore, they calculated Extended Connectivity Fingerprint 4 (ECFP4)-based Tanimoto coefficients (Tc) for the nine D2R-agonist screening hits and the eight D2R-antagonist screening hits compared to the respective training set ligands. For the D2R-agonist hits, the Tc values varied from 0.094 (pyrraline) to 0.324 (salsolinol), while they obtained Tc values between 0.110 (fenpropimorph) and 0.345 (ajmalicine) for the D2R-antagonist hits. When comparing all their screening database compounds to the training set ligands, the top 100 hits using the D2R-agonist training set as a reference gave Tc values of at least 0.234, while the Tc values of the top 100 hits using the D2R-antagonist training set as a reference were at least 0.243. Thus, when applying the 2D molecular similarity search, only three of their selected compounds, ajmalicine (Tc = 0.345), salsolinol (Tc = 0.324) and fumigaclavine A (Tc = 0.288) would have been present among the top-ranked compounds. For details, please refer to the Supplementary information, Table S1.

Experimental evaluation

To confirm the VS predictions, they conducted radioligand binding assays to estimate the binding affinity of their hit compounds at D2R. As they performed VS to identify both nutritive D2R-agonists and -antagonists, they were interested in the ability of the VS hits to activate the receptor or to antagonise the effects of dopamine. Hence, functional assays were carried out to evaluate both G protein- and β-arrestin-mediated signalling pathways.

Ligand binding experiments

The affinity profiles of their VS hits towards the human dopamine D2L receptor isoform were compared with those of the reference compound dopamine by radioligand binding experiments (Table 1).

The binding data revealed Ki values ≤ 50 μM for five of nine tested D2R-agonist VS hits, representing a hit rate of 56%. Two out of eight tested D2R-antagonist VS hits gave Ki values ≤ 50 μM corresponding to a 25% hit rate. All observed binding affinities were in the micromolar range, which is typical for active compounds found by VS31. Fumigaclavine A, an alkaloid with an ergoline-like structure produced by different Aspergillus species32, showed the highest binding affinity with a Ki value that was only seven times higher than for the endogenous ligand dopamine. Furthermore, the coccidiostat and possible food contaminant robenidine33, as well as salsolinol and hordenine bind in the low micromolar range. Salsolinol, for which controversial activity profiles for D2 receptors have been reported34,35,36, is a constituent of cocoa and chocolate37, whereas hordenine is present in barley38 and beer39. The anthocyanidin delphinidin, which has a considerably different molecular scaffold to the remaining VS hits, gave a Ki value of 26 μM. Most active compounds contain an aliphatic amine as a positively charged moiety at physiological pH to enable a salt bridge to Asp1143.32. Only robenidine, which contains a guanidinium group, and delphinidin, which features cationic oxygen, are different.

Functional experiments

Finally, they examined the ability of their VS workflow not only to identify compounds that bind to D2R, but also to predict the compound efficacy. Hence, they performed functional experiments for the five compounds that showed the best binding affinities, namely fumigaclavine A, robenidine, salsolinol, hordenine, and delphinidin. As it has been shown that ligands can induce G protein-mediated signalling and β-arrestin recruitment independently40,41, the individual pathways were investigated separately (Table 2).

Table 2: Intrinsic activities and potencies of the five most active VS hits determined at the dopamine receptor subtype D2S by measuring the inhibition of forskolin-stimulated cAMP accumulation and recruitment of β-arrestin-2 after stimulation of the D2S receptor expressed in HEK cellsa.

Among the five investigated VS hits, three compounds showed significant intrinsic activity in the cAMP accumulation assay. The EC50 values of fumigaclavine A and hordenine are both in the extended range of the Ki values determined in the radioligand binding experiments. Conversely, probably because of a very flat dose-response curve, the EC50 value of salsolinol is considerably lower than its Ki value. For hordenine, they could observe almost full activation (Emax = 76%) compared to the reference compound quinpirole and to dopamine (Fig. 5a). This is particularly interesting, because hordenine did not show significant β-arrestin recruitment (Fig. 5b). In fact, titration curves using a constant concentration of 300 nM of the D2R-agonist quinpirole with hordenine confirm its antagonistic activity (Fig. 5c). Fumigaclavine A and salsolinol also did not show β-arrestin recruitment, but the maximum response of these compounds in the cAMP accumulation assay was well below 50%, categorising them as weak partial agonists.

Figure 5: Receptor activation properties of hordenine. Figure 5

In comparison to dopamine, hordenine shows agonist properties in the inhibition of forskolin-stimulated cAMP accumulation (a). While no β-arrestin-2 recruitment was determined for hordenine (b), the test substance completely antagonised quinpirole-stimulated recruitment (c).

Although the G protein-biased D2R-agonist hordenine shares structural similarities to the balanced agonist dopamine, receptor–ligand interactions obtained after docking and energy minimisation in presence of a D2R homology model appear to be different. Hordenine lacks a meta-hydroxyl group compared to dopamine, rendering it unable to form hydrogen bonds to both residues Ser1935.42 and Ser1975.46 as dopamine does42 (Fig. 6). The receptor–ligand complex that results from docking and energy minimisation cannot explain whether double methylation of the nitrogen atom enables hordenine to form additional hydrophobic interactions compared to dopamine.

Figure 6: Conformation of hordenine and its receptor-ligand interactions obtained after docking and energy minimisation. Figure 6

We used an active-state homology model of D2R and performed MD simulations with the endogenous ligand dopamine42. Dopamine was removed from the model, hordenine was docked into the binding pocket and the resulting receptor-ligand complex was subjected to energy minimisation in a water box. Whereas dopamine is able to form two hydrogen bonds with both Ser1935.42 and Ser1975.46 in the D2UpR model42, their VS hit hordenine forms only a single hydrogen bond to Ser1975.46 due to the lack of a second hydroxyl group.

Taken together, they could observe partial or almost full agonist activity in the cAMP inhibition assays for three of four tested D2R-agonist VS hits. For the D2R-antagonist VS hit robenidine, they could not observe any activation in either assay. These results confirm the pharmacophore models and VS workflow.

Thus, the VS workflow proved to be able to identify hordenine as almost full D2R-agonist in Gi/o activation and simultaneous antagonism in β-arrestin recruitment, indicating that this substance acts as a G protein-biased agonist at D2R. Additionally, two VS hits with partial agonism in Gi/o activation, fumigaclavine A and salsolinol, were identified. The D2R-antagonist robenidine did not show any activation in either assay. The VS method based on the newly generated VS food database can now be applied to other targets for unbiased identification of novel bioactive compounds in food.

Hordenine is a natural constituent of barley and, in particular, beer, a food that is often linked to alcohol abuse. In the present study, hordenine was able to bind to D2R with a Ki value in the low micromolar range. The activation profile of hordenine showed significant bias for G protein-promoted activation over β-arrestin recruitment compared to functionally balanced D2R-agonists like the endogenous ligand dopamine. Given that β-arrestin recruitment leads to desensitisation and receptor internalisation, hordenine-promoted D2R activation may be stronger and more sustainable than activation with the balanced neurotransmitter dopamine. Further studies are necessary to clarify the possible influence of hordenine on the dopaminergic system and food reward.

Cisco firewalls under attack – and there's no patch: Too many SIPs and they drown in data | real questions and Pass4sure dumps

Cisco says miscreants are actively exploiting a SIP vulnerability in its networking gear that it disclosed on Wednesday.

The bug, CVE-2018-15454, lies within code in some Adaptive Security Appliances, and its Firepower Threat Defense software, that handles Session Initiation Protocol (SIP) packets. SIP is the signalling protocol used in IP telephony.

The advisory warns that an attacker can hose a vulnerable system offline “by sending SIP requests designed to specifically trigger this issue at a high rate.”

If your network is being attacked this way, you'll see a large number of incomplete SIP connections per second, viewed with the show conn port 5060 command. Meanwhile, show processes cpu-usage non-zero sorted will show high CPU utilization. If an affected unit crashes, there will be an “unknown abort of the DATAPATH thread” error message, the advisory added.

Since Switchzilla doesn't have a patch ready yet, sysadmins are advised to implement a variety of mitigations:

  • Disable SIP inspection if it's not needed;
  • Block hosts seen sending attack traffic;
  • Filter the “sent-by” address, because Cisco says the attacks it observed used that (invalid) address as the source;
  • Rate-limit SIP traffic.
  • Vulnerable systems use Cisco ASA software 9.4 and later, and FTD software 6.0 and later, on a number of different hardware platforms: the 3000 Series Industrial Security Appliance, ASA 5500-X firewalls, the ASA Services Module in Catalyst 6500 switches and Cisco 7600 routers, the Firepower 2100 and 4100 appliances, the Firepower 9300 ASA security module, and two virtualised products, ASAv and FTDv. ®

    Sponsored: Becoming a Pragmatic Security Leader

    Global PDB File Layout | real questions and Pass4sure dumps

    In my Windows 2000 book [2], detailed information about the extraction of public symbols is found, along with plenty of reusable source code. In this article, I will cover the more general task of parsing a PDB file and dissecting it into its parts without regarding the contents of its various streams. The sample application presented here is a simple Win32 console mode program that expects the path of a PDB file, and stores its constituents in separate files. Depending on the option switches specified on the command line, the following information can be extracted:

  • The PDB file header, containing general information about the file

  • The allocation bit table, which specifies what parts of the file are in use

  • The root stream, containing specific information about all data streams

  • All data streams listed in the root stream

  • The most basic structural property of a PDB file is its subdivision into pages of equal size. The most frequently used page size is 1KB (1,024 bytes), but my research has revealed that 2K and 4K pages are legal as well. (You can verify this by examining how Microsoft's dbghelp.dll processes PDB files.) A PDB stream is a sequence of file pages that contains coherent information. The most essential property of a stream is that its pages can be located anywhere in the file in arbitrary order.

    When a stream is read or written, a stream directory is responsible for telling the application which pages need to be accessed in which order. This directory is itself stored in a stream called the root stream. Additionally, an embedded Allocation Bit Table keeps track of used and unused pages. This table is indispensable as soon as "holes" appear in the PDB file due to the rearrangement of streams. If a stream is rewritten to the end of the PDB file, releasing the pages it occupied before, the allocation bit table reflects that the previous pages are free while the new pages are in use. This scheme is borrowed from simple operating systems such as MS-DOS with its FAT file system, in which a similar table specifies which disk sectors are allocated to files.

    Figure 1 shows the typical basic layouts of a PDB file in 1-KB, 2-KB, and 4-KB mode. Which page size should be used depends on the data to be stored in the streams. If the page size increases, the allocation bit table and the root stream become smaller. On the other hand, a larger page size results in more page overhang; that is, more file bytes are wasted if the stream size isn't an exact multiple of the page size. The same problem occurs in file systems, in which the disk sector size must be chosen properly to avoid excessive sector overhang. Most PDB files, such as the Windows 2000 symbol files and the debugging information generated by Microsoft Visual C/C++ 6.0, employ the 1-KB scheme, as depicted on the left side of Figure 1.

    Figure 1 Typical PDB file layouts.

    In 1-KB page mode, a maximum of 64MB can be stored in the PDB file, which simply results from multiplying the number of allocation bits by the page size. As I will show in a moment, PDB page numbers are stored as zero-based, 16-bit quantities. Therefore, the allocation bit table will never comprise more than 0[ts]10000 bits (8KB).

    The PDB header always occupies the first file page, and is followed by one or more pages containing the allocation bits. The structural definition of the header is shown in Listing 1. The first 44 bytes make up the ID string PDB_SIGNATURE_200, specifying the file type and version. At the time of this writing, the most recent version is 2.00, and this is the version used by the Windows 2000 symbol files and the Visual C/C++ debugging information. The dPageSize member indicates the page size applying to all pages in the file, and wStartPage is the zero-based page number of the first data page following the allocation bits. The size of the allocation bit table can always be computed by subtracting 1 from wStartPage (for the header page), and multiplying the result by dPageSize. The wFilePages member specifies the number of pages stored in the PDB file, and should always match the file size in bytes divided by the page size.

    Listing 1PDB File Header #define PDB_SIGNATURE_200 "Microsoft C/C++ program database 2.00\r\n\x1AJG\0" #define PDB_SIGNATURE_ 44 // size of signature (bytes) // ----------------------------------------------------------------- typedef struct _PDB_HEADER { BYTE abSignature [PDB_SIGNATURE_]; // PDB_SIGNATURE_200 DWORD dPageSize; // 0x0400, 0x0800, 0x1000 WORD wStartPage; // 0x0009, 0x0005, 0x0002 WORD wFilePages; // file size / dPageSize PDB_STREAM RootStream; // stream directory WORD awRootPages []; // pages containing PDB_ROOT } PDB_HEADER, *PPDB_HEADER, **PPPDB_HEADER;

    The RootStream member is another undocumented structure of type PDB_STREAM, as defined in Listing 2. This structure appears wherever a stream is defined. Here, it refers to the root stream that contains the sizes and locations of the data streams within the file. In a moment, they will revisit it when walking down the list of data streams in the root stream. Only the dStreamSize member of the PDB_STREAM structures is of interest when dealing with a PDB file on disk. The pwStreamPages member can obviously be used as a scratch pad by PDB read/write utilities that handle PDB information in virtual memory. Simply ignore this value because it might be a bogus address that was valid once upon a time.

    The RootStream structure is immediately followed by an array of 16-bit page numbers used by the root stream. Most PDB root streams I have seen so far don't exceed one page, so the awRootPages[] array usually contains a single entry only. One exception is the extraordinary large symbol file of ntoskrnl.exe, which has root stream that spans two pages.

    Listing 2 The Basic PDB Stream Structure typedef struct _PDB_STREAM { DWORD dStreamSize; // in bytes, -1 = free stream PWORD pwStreamPages; // array of page numbers } PDB_STREAM, *PPDB_STREAM, **PPPDB_STREAM;

    The allocation bits don't require much explanation. Each bit is associated with an individual page, and a value of 1 means that the corresponding page is currently available. The bits within a byte are ordered from least-significant to most-significant. That is, bit #0 of byte #0 refers to page #0, bit #1 of byte #0 to page #1, and so on. Note that the allocation bits of an on-disk PDB image do not necessarily reflect the status of the stored data stream pages. If you examine a couple of Windows 2000 symbol files, you will find out quickly that some data stream pages are located in pages that are marked free, and some pages marked in-use are not part of any stream. So, the allocation table is probably rebuilt by PDB readers/writers from the data in the root stream, and has meaningful content only while loaded into virtual memory.

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