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My closing post on Kuching, Sarawak. :)
Guess who came to say hello and welcomed us right after the boat ride?
Yuppie yup!
This shameless dude here did. Wild boar?!?! Methinks he was more like a goat. Really.
The boys got really thrilled at the mere sight of this fella. He was erm.. grazing? Yup, he did the goat/ cow thing actually.
Hmmm.. this is strange.
You know i complained quite a bit on the accommodation at Bako but after looking at the pictures above, somehow it looks a lot better than in reality! Why ah?!?!
But in all honesty, it was bad. I should have taken closer shots instead. :P The beds looked like they were infested with bed bugs and the bedsheets, hmmmm. Then we have bathroom and toilet… let's just say i was really tempted to skip shower altogether despite the fact that i was all sweaty and dirty. I didn't even wanna step in until the very last minute. To do my business, i opted for the deserted public loo instead which was conveniently around the corner. Much cleaner!
Fortunately, the men whom i traveled with are quite something. When they are around, this mother can just sit back and watch. The men, especially the grandfather, will happily take care of everything, including bathing the boys. More often than not, the grandmother and i will stand at the side and watch happily. Naturally, this meant that i only needed to step into the eee eee toilet and bathroom just once. PHEW!
So how did i sleep at night?
I was all covered, from head to toe. Long sleeves top with hoodie (uh huh, i had the hoodie on while sleeping!), long pants and socks! I totally chucked the pillow and made used of my own towel. And nope, none of the accommodation here is air conditioned, which was quite alright. Natural breeze, best!
However, don't let this deter you from going to Bako. From what i saw and what i heard from a gf who was there 2 days earlier, the rest of the accommodation wasn't this bad. It was just our luck to be stuck with Chalet 2, one of the 3 earliest, oldest chalets in Bako. Hmphhh! (if all the accommodation were as such, perhaps i won't be this whiny!)
We only managed to do 2 trails during our time spent in Bako. With the boys around, we had no choice but to pick the shortest trail. Short, yes but never shortchanged. Mosses, i've only seen them in magazines but to feel them for real at Bako? It was quite surreal. For some strange reason, i was more excited to see the moss than the proboscis monkey. Ha!
Even though beep didn't whine or complain, the very fit grandfather insisted on piggyback the boy. I guess it's just the way he expresses his love for his grandchildren. He's not a man with many words, but a lot of times, his actions say quite a bit about him. :)
Our second trail was a little longer and apparently, a little more dangerous. Because of 'danger' part, we chose to leave beep behind, together with the helper.
This time round, it was a lot more educational for the boys. And hey, pitcher plants! My first up close encounter with them.
Alas, proboscis monkeys…
We spotted quite many of them actually, throughout our 2 day stay.
Hermit crab!
Again, another first. Tee hee hee. I know i know! I'm really quite ulu!
Hermit crabs are really quite adorable. We spotted abundance of them, in all shapes and sizes. Do you know that to get them out from their house, all you need to do is to whistle and they'll come crawling out?
Fascinating creature!
All in all, Bako National Park is strictly for nature lovers. Not a place for honeymoon for sure. A good place to go if you want some exposure for the children. But if you intend to stay a night or 2, avoid Chalet 1, 2 & 3. By all means, request for newer lodgings!
And, if you must know, the food sucks. Big time. Both in taste and hygiene. So, bring along cup noodles and whatnots. The effort is worth it, trust me.


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North Central College theatre alumnus Peter Sipla '07 is starring in a production of "Maple and Vine" through Dec. 4 at Next Theatre in Evanston.
Oct. 10, 2011—Lauren Visher, a 2007 alumna of North Central College, is featured in The Naperville Sun for her work with Naper Settlement.
Visher spoke about her work as part of The Sun's "Five Questions" feature. She said as a young girl, she enjoyed volunteering at the Settlement and giving tours. She graduated from Naperville Central High School and majored in musical theatre at North Central.
She enjoys her work as special events coordinator at Naperville's "living history museum."
North Central College presents "Distracted," a thought-provoking comedy about parents and the tough choices they face.
Four performances of "Distracted" will be presented at 7:30 p.m. Oct. 20, 21 and 22 and at 2 p.m. Oct. 23 in North Central College's Madden Theatre at the Fine Arts Center, 171 E. Chicago Ave., Naperville. Tickets are $10/adults and $8/students and senior citizens; call the North Central College Box Office at 630-637-SHOW (7469) or visit northcentralcollege.edu/showtix.
North Central College students will stage six performances of Frank Loesser's Tony Award-winning favorite "Guys and Dolls" in early November.
Oct. 11, 2011—An Oct. 21 performance by members of North Central College's music faculty will include a piece written by a local resident.
The free performance begins at 8 p.m. Friday, Oct. 21, in North Central College's Koten Chapel at Kiekhofer Hall, 329 E. School St. No tickets are required.
Will Allan was named among the seven Hot New Faces of Chicago Theater by Chicago Tribune theater critic Chris Jones. "What do they have in common? Talent. Artistry. And a desire to entertain Chicago and beyond. You'll be seeing them around," Jones writes. He summarizes Allan's career this way: Allan first attracted attention when he appeared in "The History Boys" at TimeLine Theatre, a production that launched several Chicago careers.
Vocal talents of local junior high and high school students, as well as North Central College's choral ensembles, will be showcased Oct. 13 during the annual Choral Invitational Festival at North Central College.
Student groups performing during this year's festival are Naperville's Lincoln Junior High School Eighth Grade Chorus—under the direction of Ross Berkley—and the Rockford Christian Concert Choir. This is the first time a junior high ensemble has been invited to participate in the festival.
North Central College presents the student-written and –directed drama "House of Cards," an innovative play about relationships that unites multiple forms of theatre.
Three performances of "House of Cards" will be presented at 7:30 p.m. Oct. 6, 7 and 8 in North Central College's theatre at Meiley-Swallow Hall, 31 S. Ellsworth St., Naperville. Tickets are $5/adults and $3/students and senior citizens; call the North Central College Box Office at 630-637-SHOW (7469) or visit northcentralcollege.edu/showtix.
July 25, 2011—Chicago Tribune's theater critic Chris Jones unveiled his 2011 list of "The Hot New Faces of Chicago Theater" and featured 2009 alumnus and theatre major Will Allan among his seven top picks of the year.
North Central College prepares students to be informed,
involved, principled and productive citizens and leaders over their lifetime.

subset: commoncrawl


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Southern 6 Trail Race and Kids' K!
Registration opens Jan 1 at 3AM EST.Register Now
Click here for race details!
Southern Adventist University, Wild Trails and Rock/Creek present the Southern Six Trail Race. This is a short, family-friendly event taking place on Southern's beautiful Biology Trail. Everyone ages 12 and above is invited to participate in the 6k trail race and compete for prizes, including ceramic mugs created by local potter Loren Howard. Children ages 12 and under are encouraged to participate in the second-annual Kid K! Door prizes will also be available. Proceeds from the event will go directly to the University's Outdoor Leadership program and will help with development and maintenance of the Biology Trail.
For step-by-step directions with Google Maps, click here >
For complete race details, including directions, click here >


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The Sacred Heart football Gremlins held their first practice session of the 2014 season Monday night at SFCC. According to the school's own website, the Gremlins' first game is scheduled for Aug. 22 at St. Paul's Lutheran.
The online schedule shows only two home games this year — Sept. 20, which is Homecoming for the Gremlins, playing Windsor — and Sept. 26, playing University Academy. All home games start at 7 p.m. A total of nine regular season games are listed.
The football Gremlins are coached by Ben Lyles, Ethan Henson, Len Holland, John Smeltzer and Ron Williams. Photos by Randy Kirby, Sedalia Weekly Observer.


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Microsoft jumping even deeper into the hardware pool with Surface tablet [the competition] 68
Alternate Title: How do you feel about webOS now, HP?
So today out in Los Angeles brought forth an interesting piece of hardware: a tablet running Windows 8 (nothing exciting there, we've seen plenty of those) to be built and sold by Microsoft (wait, what?). Yep, Microsoft is diving headfirst into the hardware pool with their new 10.6-inch Surface tablet. It's a strange entrance, not that the tablet itself is strange, just the way that Microsoft is going about things.
First and foremost is the idea that Microsoft is going from wading into hardware with the successful XBox series and a series of otherwise complete and utter duds (Zune, Kin, et al) to performing a cannonball at the swim meet where it's supposed to be the coach. The introduction of the separete-but-sort-of-the-same-but-still-different Nvidia- and Intel-powered Surface tablets strikes us as not only unusual, but possibly threatening to Microsoft's third party manufacturers.
It's a problem Palm had to deal with when they tried to license Palm OS to Sony and other companies while also making Palm-branded hardware themselves. It led to the company being split into two halves over the concerns of Sony and the like with competing with their supplier.
We can't help but imagine that Asus, Dell, HP, Lenovo, and everybody else who has plans to build or has been considering a Windows 8-based tablet is feeling a little anxious right now. Palm, even when one company, wasn't a particularly powerful firm. Influential and well-known in the mobile space, yes, but not a force to be reckoned with.
Microsoft, on the other hand, is a force with which the companies and many more are going to have to reckon. Microsoft CEO Steve Ballmer says the point of the Surface tablets is to "prime the pump," so to speak, but we can't stop thinking of it as the swim team coach yelling, "Let me show you how it's done!" and jumping into the pool at regionals in front of stunned students and their family.
Will the entry of Microsoft into the tablet hardware space send chills down the spine of their hardware partners? No doubt the idea has been for some time. But will it be enough to send these companies in search of alternatives, and what are they going to find?
If Microsoft's Surface foray proves to be successful, it could both assuage and stoke the fears of hardware partners - simultaneously demonstrating demand for Windows 8 tablet products while delivering that product, brand loyalty, and profit exclusively into Microsoft's lap. If it goes poorly it will assuage and stoke the fears of hardware partners - simultaneously demonstrating that Microsoft can't single-handedly upend the established order of the Windows ecosystem and that customer demand for Windows 8-powered tablets isn't what they had hoped (or is exactly what they feared).
Either way, we have little doubt that Microsoft's Surface is going to send more than a few hardware manufacturers on at least a side quest to see if there's another operating system worth considering. They'll find two: Android, which people know but have been reluctant to buy in expensive tablet form, and webOS, which is in the process of an open source transition with questionable support from the company that's pushing it into open source. It's hard to gauge customer demand for webOS in tablet form, given the incredibly odd and short shelf life of the TouchPad tablet. And with webOS being open source, tablet manufacturers would be able to customize it as do with Android, and much unlike what Microsoft is allowing them to do with Windows 8.
So here's the big question: Does Microsoft Surface provide an opening for webOS? Yes, it does. It's a narrow one and it'll require that HP and the Open webOS team play their cards just right if they have any hope of convincing somebody to make webOS tablets where HP didn't have the will to succeed. Then again, maybe HP could just try again themselves.
68 Comments
It is fun to dream that someone would snap up webOS and re-present it to the world. I was showing my Pre2 to someone yesterday, and he said "Wow...that's pretty slick. How come I've never seen that?" I'm still hoping for open source to where I can boot webOS on a new piece of hardware...to think that it would be released again on new hardware is almost too much to wish for. Would be so nice...
open webOS Ahoy!
HP can't make hardware sexy like that, so they'll have to differentiate with software.
I'm not a MS hater, but just how many platforms can MS support successfully? You talk about Android fragmentation or IOS being a closed system, but MS is all over the map. I like the RT version, but in order to be successful it needs to be sub $300 (pref $250). The upcoming Acer Iconia A210 (with similar specs) is being targeted at this price range. Then there is the x86 tablet. Why do we need a x86 version? I mean really. Most of the x86 legacy software is not optimized for tablet use. Seems like a good idea, just not a practical one. The x86 version will be too expensive, so expensive that you might as well get an ultrabook with an i7 chip. Don't believe me? Read the Windows 8 Pro Tablet specs, "Pricing: To be determined; competitive with ultrabooks". Sounds like upper $700 to me. Instead of increasing manufacturing costs, MS should bombard every platform with their software suites. How cool would it be if MS made a Metro interface (ala HTC Sense) for android or IOS? or Visual Studio for the iPad? People would gladly pay for it. Google just has to drop the Nexus 7 on the market and Apple a 7 inch iPad, that would be enough to flood the market and make it extremely hard for MS to be get a foothold. Stick to what you do best MS, write great software.
Where you see fragmentation, I see unification.
Just imagine being able to use most, if not ALL your desktop apps on your tablet w/o having to pay for them again.... The Pro version with pen input looks promising ...I'll be buying one...The RT version, not so much.
I am still looking for my first occurrence of user-impact fragmentation in Android. One thing is clear, the LG Optimus is a device to avoid. But the Evo line and Galaxy series phones....its one big happy functional phone.
Fragmentation was the WebOS rally cry against Android. In the end, just about every device had their own WebOS required version, and few apps supported more than one of them. You had your Pre/Pre+ fragment, Pre2 fragment, Pixi fragment, Veer fragment, Pre3 fragment, and tablet fragment. Now that is Fragmentation done with Malice Majure!!!
All this fragmentation, clearly Sprints fault. And Obama helped.
Hmm, webOS is "long dead" and your still here bashing? Wow. Anyway if you had an Android device and spent 4.6 seconds in the Play store, you'd see in the reviews the 100's of "Doesn't work on my phone! " and "This is not compatible with my XX phone! ". Heck, you'd even experience it yourself.
So if you're gonna sit here claiming Android fragmentation is a myth... Yeah ok buddy. /smh
WebOS isn't fragmented? The most popular WebOS phones only run188.8.131.52 unless you try and kludge 2.0 in.
@cobrakon, didn't say "long dead", said fragmented. WebOS has become a hobby. Ham Radios and canning vegetables are not dead, neither is WebOS.
And I've always said, stay away from fringe hardware on Android. It's as poorly supported in some cases as WebOS products. Feedback on devices like the Optimus and Replenish are bad, buy an odd bit of hardware, pay the price.
Each device on WebOS pretty much has it's own o/s version that may, and often isn't, supported by apps. Each device, it's own o/s, all devices made bay Palm/HP. That my friend, is the ultimate fragmentation.
And I didn't say fragmentation on Android is a myth, WebOS is a glass house, a total poster child for fragmentation. I noted the irony that our old rally cry bit us on the behind.
Anything else I didn't say you'd like to take issue with?
So in your purview, everything is fragmented, even Apple. iOS 6 can be installed on iPhone 3GS, but it won't work 100% (no Siri). Fragmentation.
Yet unlike Apple, there's plenty of Apps (the majority of the ones I have at all, in fact) that work on both my Touchpad and my Pre 2 (albeit in tiny phone emulator mode, but it's still there.) My Pre 2 isn't running on webOS 3.0.5, yet the app still works.
Different OS versions in webOS is not an absolute qualifier for fragmentation.
What apps are not running other than Siri on an iPhone G3s?
The iPhone G3s was released at the same time as the Pre and the debut of WebOS.
The iPhone G3S is supported with current iOS software. It is slated to get the iOS 6 update as well.
What WebOS version did Palm or HP send out as an update to the Pre?
@rennock, Pre wont run 2 or 3, Pixi can't run all that Pre can because of screen size and subgrade processing, Pre2 can't run 1 or 3 and could not do TTS, Veer could not run 1 or 3, and had an odd screen size the blocked some Pre3 apps, Pre 3 could not run 1 or 3, TP could run some 2 apps but stuggled with 1's despite the higher hardware capacity. Most apps were written for 1, the best apps were written for 3....fragmented.
Unification indeed! Last week while laptop shopping I was dreading the impending Windows 8 drop. Now that I see how they're completely rounding out the ecosystem, I'm actually looking forward to it.
By my math, the bigger 8 version with tactile keyboard is a form factor that just edges out (or close to it) the small MacBook Air. Run tablet and full-fledged desktop apps? Still play nice with all my old stuff? Have it light and sexy at the same time? Yes please!
The smart cover that doubles up as a keyboard is pretty interesting....
Agree...110% and love what they showed.
Some aspects...ran into problems (OS wise and system glitches), but the Smart Cover looked rock solid and I like what they did almost completely hardware wise. Smart Cover, Integrated Stand, Modern Hardware Support, and the lists goes on.
OS wise...hmm...noticed a bunch of hits and misses during the presentation (very much like webOS two years ago when they introduced Palm Pre to the market). WebOS wasn't perfect and it didn't get better till webOS 2.04 almost 6-months after its initial release on Sprint.
That is scary proposition coming from Microsoft and too be honest...what does this mean above Microsoft future plans and hopes when it comes to its new OS. Microsoft is smart...really smart and has a way of pulling out the magic if and when needed, but if HP released webOS today and expected to hit a home-run compared to IOS 6...I just don't know.
Either way, I love what I saw and glad to see MS stepping up to the plate and making their own dream hardware happen. Sorli...
It's freakin' awesome and an answer to old desire. Only bummer is the screen won't stay up right without the kickstand having something to rest on.
Hate to say it, but if all this sexy was wrapped up in an ultra-premium version of the Transformer form factor (self-supporting hinge... extra battery optional in the base), it would be perfection.
Still, this is so svelte while giving me buttons that I likey!
I'm with you @jelvis. All my android friends say that they would use webOS if it was presented/ported to a better hardware (they liked the Pre³ btw) and a "little" more apps wouldn't hurt at all.
And yes, I do think that any well driven/marketed/business_planned company would see that this is the wakeup call to put a 100% effort into their own $ 1.3 bill ($ 3.3 bill w/loses ¿…?) mobile OS which is envied/admired by the most reckoned mobile gurus (Jobs once said he appreciated webOS' influences on the mobile OSes) and if well implemented, presented and executed, could flourish into what its potential has always dictated!
Thank you!
LONG LIVES webOS!!!
In one of the Forum chats, I talked about why people shouldn't be so quick to dismiss webOS. Yes it can fail. But yes it can rise to the challenge. The only certainty in the Tech World, is that it offers no guarantees - the mighty rise, and the mighty fall - ask Nortel. But, it`s also a world where weakness became an art that Apple perfected to success.
What webOS represents to HP, is more than just an operating system. It may occupy a small fraction of net worth, but it also represents an avenue for HP to remain where it has been for so many years - a giant in the computing world.
Assuming that HP Shareholders would like to remain amongst the top tech companies, it`s reasonable to believe that recent circumstances might dictate HP having their own OS.
Google has Android & Motorola
Apple has the Ipick-your-consumer-tech-good, and software iOS
RIM has CrackBerries & Playbooks ..... and BB10 -
And Microsoft, the purveyor of software, and normally not a Competitor for HP in the PC world, about to be like those above - A Windows tablet maker & software guru.
Which leaves HP - a supposedly large dynamic company - now playing with the little guys. And in an area they once dominated - personal computing devices. Mimicking to a certain degree, with how RIM has given way to its competitors. The difference being though, HP has a much stronger stomach to handle the crowd.
How it battles its competition, is how HP's fortunes will be decided. For me, webOS is a no brainer. They've written off the 2 Billion dollar brain fart. And now they have a software that could excel - with Homebrew enthusiasm, and HP support behind it. Add the fact that HP has all kinds of hardware that was almost started, or barely allowed to surface.
That all has to mean some value, should HP make the right decision and play like a Big boy. And not the cowardly lion that was Leo.
> ...a supposedly large dynamic company ...
Supposedly not as bureaucratic paperwork apparently gets in the way to often. Being huge is not necessarily always a benefit — that makes a difference anyway.
This is a game changer no matter how you look at it and WebOS had better be looking like the future when open source is released, not 3 years ago.
Agree, this is a game changer. webOS is really beyond reviving if no hardware is released by either HP or third parties this year. After that everyone will move to another platform. Even I felt teased by this new offer.
So HP it's your move now. Still want to be in the hardware game? Better start competing with a compelling offer. Choose between competing with Microsoft or having your own ecosystem based on webOS? Shouldn't be a hard choice. No money to invest? Better stop and get out of the hardware market and dump webOS (still hope to have an open webOS release for my current devices though).
Maybe Leo's idea to move out of the hardware was not that bad after all.
> ...it'll require that HP and the Open webOS team play their cards just right ...
Can they, or is that gonna put the last nail in the coffin for webOS?!
From previous articles outing inside information from HP, it seems they have huge obstacle in making innovation take years instead of months, making it necessary to predict a fast pacing market years ahead — a tough job if you're not creating the market trends.
So, there are three parts of the population to convince here; the iPad accepting/worshiping mass, the alternative-accepting mass — and the occasional nerds. Neighter will put up with mediocre products/hardware if they are somewhat enlightened about alternatives.
The firs lot will easily be lost to the almighty god-pad (yes, I deliberately omitted the capital G), and the nerds are best served with for instance Samsung/Asus ... stuff made for android, that we can slap our belowed OS onto.
The second lot can be more easily convinced, but not with more mediocre attempts. It will just be another hard earned "lesson". Time is essence, and HP can't afford to play with to much of it unless another failure is the "goal".
HP needs to look towards Asus's innovative pace and learn a lesson. Downsizing the business is hopefully done by removing a lot of middlemen and procedures/protocols that hold innovation back until it's less than mainstream functionality/solutions.
Everything so far has indicated that Open webOS has nothing to do with targeting new, innovative hardware and everything to do with just shoring up the OS for the Open Source community. Which, honestly, sounds like BeOS-level talk.
Unless something is announced soon hardware-wise to complement the Open webOS project, well, there's a level of futility here that I don't think HP's "investment" is going to recover from.
A Microsoft tablet? Meh.
I want my brand new cutting edge wOS tablet, please!
Me too!
This is gorgeous. I hate a 16:9 tablet, so I stay on webOS.
(Win8 for the touchpad? pretty please?)
WebOS, as a mobile and tablet operating system, has always been and still is, amongst us and in the IT press, well received.
What let it down, in terms of not enough apps and insufficient market penetration, was perhaps the lack of hardware quality and choice, as well as poor marketing.
Our best hope may be for an OpenWebOS that can be ported onto a non-webOS mobile/tablet. I hope so.
Then after say a year of hopefully increasing numbers of webOS users and apps, new hardware dedicated to OpenwebOS may follow.
By the way, there is rumour of a Facebook mobile in next 12 months, but what might its platform be, a version of Anrdoid or OpenWebOS or switchable between the two...?
When will you people realize that webOS is dead! Sheesh
The moment you stop coming here to tell us webOS is dead, is the moment we realize it. As long as people have to keep telling us it is dead...
webOS is the zombie that has been modified to have 2 brains and 3 hearts, so there is always a sequel that will have you coming back to see what is going to happen next.
Stay tuned. See you back soon.
Well said my friend...
I hate giving out those +Ones .... but in this case ...... +1
MS is officially in the hardware business now. Please, please, please see the writing on the wall. If you continue to just make hardware for Windows (any version), you need to realize that you are (and will continue to be) competing against any number of other hardware manufacturers; now even MS will compete with any tablet/laptop hardware you make. If you think that your profitability has suffered from the decline in PCs in the last few years, just imagine what that will be like without a real foothold in the growing mobile world.
The success of Surface will be dependent on many factors, but price, cross-platform compatibility and the adoption of the "metro" interface will be some of the primary ones.
Regardless...almost the only way that HP can truly compete now is to harness the power of WebOS and GO ALL IN! Remember the goal of putting WebOS on PC and every other type of device?!? You may want to reconsider doing that...including phones. I'm not saying to stop making hardware for other OSs...hell...keep making Windows-based stuff if you like, but you need to develop WebOS hardware (and software) NOW. In the near future, Android devices may be primarily made by the former-motorola (& Google). We know where iOS is. We now know where MS is going. HP...WHERE THE HELL ARE YOU GOING?!?!?
Good news for WebOS its all I can say...
HP it's your turn now!
I'm chuckling at how you guys see this as an "opportunity" for webOS. Let's see...a compelling new device from Microsoft with a slick OS and top-notch hardware (or so it appears) and plans to actually release a product by end of the year. That's a wonderful opportunity for webOS and HP to, uh...what was the argument again?
It's a great opportunity for HP to make Windows tablet clones (competing with Acer, Levono and the Koreans should give them huge profit margins) and transition their webos team to designing HP crapware for Windows Metro!
This article is fantasy - nobody is going to put out a WebOS tablet to compete against the Windows Tablets - especially the pro one - which enterprise is going to pick a WebOS tablet over a windows one - not even HP will do that.
The real question is whether HP will stick to their dates and deliver a product in September or will they sheepishly scale down (is that even possible?) this endeavor and let it die off.
Only if that is achieved can you consider hardware possibilities and then the answer is still none from HP. They have real problems. They make a lot of mediocre machines ( the Touchpad was oh so mediocre) , a few good ones, that aren't differentiated and probably aren't that profitable. They aren't nimble or quick. Their plans are already laid out, good or bad, and my guess is that no division is including Webos in those plans.
If you're looking for equipment running WebOS, it's going to be homebrew porting it onto someone else's equipment. And it will probably be a tablet. But I want it on a phone. My Veer is nice and all that, but it's not the Pre3 that I was waiting for (Damn you, Leo, forever!) and I'm getting bored with it. I miss the updates and the excitement of something new. I have a pathetic life, I know. But, it is still a capable phone and system, just falling farther behind every day.
whitman told cnbc on May 24 that she is debuting a back to school win8 tablet. So presumably manufacturing already started. What will become of this, can it be repurposed for webOS? It would be great to get some reporting on this subject.
They would come back to the marketplace with open source WebOS 1.0? That would be more stupid than buggy WebOS 3 in last year's launch.
Although it makes for interesting arguments and questions for their hardware partners, the price point for these is at $1000. Yeah, that's already an uphill battle for something that's not even on the market yet. Did they not learn from Xoom and their failures? Price point is almost everything right now.
Also, taking a play out of the handbook that just doesn't work well: Announce a product in June for a release "sometime" in the Fall. Really MS?
Well, I'm glad you were able to report hard numbers on price before anyone else, considering that Microsoft hasn't released any yet!
I guess I was the only one in the nation that read the article on CNN about it's price points according to MS? Windows RT (the first release of this tablet) will run on an ARM chip. This version will be priced "comparably" to other tablets on the market (already two different OS systems). The full versions will be on an Intel chip.
"The full Windows 8 version will be available three months later in 64- and 128-GB versions. Microsoft plans to set its price point in the same zone as ultrabooks, which typically run around $1,000." -CNN
If HP doesn't want to reinvest into building proper hardware to run WebOS, I don't think anyone else will. What's it going to be Meg?
She has already told everyone. Open WebOS with a team that has been gutted.
The only thing that some people do is blame marketing or a CEO for the failure of the Touchpad.
The bad reviews are ignored, the fact that it was late in the game and already outdated seems to be swept under the rug.
There was a huge marketing campaign for the Touchpad. It didn't sell. If anyone believes that HP is going to release a new tablet with open WebOS, they are dreaming.
Ditto all Android tablets. HP was the best seller with the highest name recognition aside from the iPad when it was killed.
In reality, not until it fell to fire sale prices.
The number being used for #2 is the number of units shipped to retail, not the number that were sold to customers.
Best Buy sold around 25K units in the 6 weeks before the fire sale but they ordered 250K units,
At retail it was dubbed the Ouchpad.
You guys are drunk. This will never spark life into WebOS. It died with the Great Pumpkin. Move on already!
But why should we move on, when you havent moved on. WebOS is that ex-girlfriend you always go back to, for whatever reason. She has money, she can cook, you have a baby by her, or whatever. That is webOS, and that is why a lot of people are still here visiting, trying to see how we are doing.
We are ok, Bruh-Man, we are ok.
Can we get a couple of dollars 'til times get better?
Lmao!! ^^^^^ The ex-girlfriend reference was hilarious! Well played 48, well played!
Hahaha well said...
No, webOS is that ex-girlfriend that commands a morbid interest in the increasingly desperate calls for help through stupid meme images until you can't take anymore, because it's become so absurd and insular in its degree of hatred that you can't help but feel sorry for her and her remaining friends that actually remain. It's more of a sideshow people take marginal-but-vested interest in than a legitimate display of actual potential at this point.
Not to say it isn't interesting from a sociological point of view, but it's at best a masochistic display of believers to continue thinking that there's some sort of retail relevance that'll come of Open webOS given how many retailers they burned the second time around. And, believe me, as someone that's been there with a retail webOS device since June 6, 2009, I'd love to believe...The Verge article pretty much validated everything many thought to be true; it was half-baked and the revolving door killed webOS.
How many people leaving does it take to convince people that even those involved in the creation of the OS no longer have faith, really?
So, based on your very own logic, your argument has zero merit, my friend.
After putting down my Google dictionary, I came to the conclusion that I first, can't stand my begging ex-girlfriends, though they did provide a service back then. Second, I can't think of WebOS as an ex, because I am still dating my Pre and my TP, and third, WebOS is still wonderful. Leo, with the help of HP, and whoever else, gave us a what if about WebOS.
I use my Pre to make phone calls, which it is great at. I use my TP or my PC when I want to do anything else. The only thing that I can think of that I dont like about my Pre is the navigation and maps. It could have faster Web browsing, but since I got my TP I dont use it for that enough to worry about it. I still have Free Tether, and all on my Pre-.
Before anyone asks, yes, I have tried others. BB, Evo, I went back to WebOS though.
On another note, I may get an Iphone next if it is bigger. Only if.
Thank you for reading.
Get the paddles. Clear! Whew, that was a close one, I thought we lost her for a minute.
Yeah, most people who have lost a loved one do come back and put flowers on the grave every now and then.
No different. However, ".....The dead are conscious of nothing at all ...." (Ecclesiastes 9:5)
I just want people to quit talking to the grave like someone is listening
RE: Money... Here is $2.00 fuyyah. Go to the pet store, buy a rat, to knaw that thing off of your face....
(Uncle Buck)
What HP needs to do is take their "Back to School" tablet, running Win 8, have the WebOS UI they talked about at "Think Beyond". Continue to make PCs with the WebOS UI available.
This would give people a "Plus 1" device. When solving issues with customers you always go "Plus 1". Give them a bit more than they bargained for.
I am not taking about the OpenSource WebOS, I mean the latest version, all upgrades, etc., including the tap to share for those lucky few that have Pre 3.
Offer a "New" back cover for existing Palm/HP devices, similar to the touch stone back cover that will enable tap to share. That is a "Plus 1". I would get that tomorrow if they offered it.
Continue to make HP hardware with Win 8. Offer the WebOS as an option or free. Imagine it on their touchscreen computers?
Once you get the millions of users with WebOS enabled Win 8 devices, you will have people clammering for more of those WebOS devices. Make calls from your computer (no skype), sync of contacts, calendars, etc. It is a no brainer.
Heck. Roll out a firmware update or software update for current HP computer owners to download and use on their devices.
We can even have OpenSource send to these devices for trial.
HP has so many avenues to chose from but they just don't.
And for those that say WebOS is dead, why are Pre Phones still selling for over 200.00 on ebay and touchpads still selling for more than that? Obviously there is still a demand.
Lots of requests for things for free,
HP should give people things for products off the market? For $99 Touchpads?
Is it free? No. It has been written off. They did the work on it already. What I am saying is include it on their laptops like they said they were. Included it on computers and any new tablets coming out, especially the win 8 tablets. This is an investment.
Get people interested in WebOS. If it is on their computer, they will see it. ALL THE TIME!
HP has an updated manager on their systems. Include on that update manager a listing of available apps for WebOS, user videos, etc. Bombard the consumer of HP products with this great OS.
Hell, include footage from Think Beyond. Phone integrating with tablet. The big seller is synology. If you have a HP laptop that syncs with your HP computer and your HP touchpad, whats not to like?
Recent events with MS EAS failing and changing protocols screwed many people like myself that have their contacts and email pushed to their devices and computer. This would solve that issue.
Good marketing gets the idea out there, implants it in the minds of the consumer with product placement, innovation. HP has the tools and it cost them nothing to do.
So if this thing succeeds, it might push HP to reconsider putting webOS on a tablet,
and if this thing fails..., it might push HP to reconsider putting webOS on a tablet.
Could it be that HP just might want to reconsider putting webOS on a tablet? ;)
P.S. HP may be out of the phone business, but what about at "Pre Touch" (or "Pre Player")?
I have always said that Bill Gate$ and MS were somehow behind the "failure" of webOS. If it had succeeded and HP, the #1 maker of PCs, put it on all of their computers- what would happen to MS and windows? That's a big chunk of change to fall off the table. I have no proof MS did anything to sabotage webOS...just sayin'
Now MS is jumping back in the hardware/tablet game. I'd love to see HP grow a pair and put them on the table. "We are going to continue to make hardware and put webOS on all of them". Going big and taking over...or going home trying. Never going to happen though. Risk/benefit is too risky.
"how did it get like this for webOS "
Hanky panky would be my guess. Not that there's anything wrong with that.
As it was, that single event initiated the rift in wedOS's timeline, that would expand once the ranks were adjusted for the umpteenth time. The new sheriff was in.
To say webOS was hampered by a guy who wasn't enamored with his arranged marriage, would be understating things. He preferred the Bride in the clouds. And then in the interests of breaking-even, he presented a product before its' time .... Literally.
An unpolished piece of tech, with an unpolished piece of software, all for the low price of an ipad
Then he pulls off a brilliant move ... He pumps out a piece of hardware at a "we're new" price point. And it sells. Plentiful. Success at last.
Unfortunately, he did the right thing for the wrong reasons, and then proceeded to take it in another direction. None.
All this by the man whose "actual" success has been fleeting in the business world - which begs the question ... "How did he get this far".
Right now - he's irrelevant. What he did though, shouldn't be to HP. They're in uncharted waters - what some might say is where they've been for the last few years. Whether HP endures themselves to its' adopted child, is now in the hands of Meg and her merry band of Directors.
At least until September.
Regarding the closing sentence: not a chance if Windows 8 pulls off its tablet trick. Firstly because it'll be a third major OS competitor. But more significantly, the level of investment that went into this new hardware combined with Windows Phone 8, Windows 8 and the opportunity Microsoft has with the new kernel changes? That's a significant barrier to make an attack on from any direction (even with a full Linux kernel backing webOS). Especially while still already flanked by the other two mobile OS players.


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Vanessa Cooper Wiki | Vendula Bednarova Wiki
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Since S. cerevisiae undergoes closed mitosis, the nuclear envelope of the daughter nucleus is continuous with that of the maternal nucleus at anaphase. Nevertheless, several constitutents of the maternal nucleus are not present in the daughter nucleus. The present study aims to identify proteins which impact the shape of the yeast nucleus and to learn whether modifications of shape are passed on to the next mitotic generation. The Esc1p protein of S. cerevisiae localizes to the periphery of the nucleoplasm, can anchor chromatin, and has been implicated in targeted silencing both at telomeres and at HMR.
Upon increased Esc1p expression, cell division continues and dramatic elaborations of the nuclear envelope extend into the cytoplasm. These "
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Esc1p is the first non-membrane protein of the nuclear periphery which – like proteins of the nuclear lamina of higher eukaryotes – can modify the shape of the yeast nucleus. The elaborations of the nuclear envelope ("escapades") which appear upon induction of excess Esc1p are not inherited during mitotic growth. The lack of inheritance of such components could help sustain cell growth when parental nuclei have acquired potentially deleterious characteristics.
The position, shape, size and orientation of organelles varies among differentiated cells, thereby allowing distinct cell types to be recognized. The approximately spherical nucleus itself generally is near the center of the cell and in yeasts the spindle pole body (SPB) provides a landmark at one pole of the nucleus, thereby allowing one to witness changes of nuclear orientation during the cell cycle [1,2]. Related to these observations is the question of whether all portions of the nuclear envelope (NE) are necessarily inherited by the daughter nucleus at mitosis – an issue which bears directly on the ability of cells to cope with the impact of damage or change accrued during a single generation. The present investigation shows that a novel landmark of the perimeter of the yeast nucleus is not inherited.
The nuclear lamina in higher eukaryotic cells governs nuclear morphology and serves as a scaffold for the organization of the nucleoplasm, where it anchors heterochromatin and can affect both DNA replication and transcription. The best-characterized proteins of the lamina are the intermediate filament lamins, which self-associate via coiled-coil domains, generating a compact meshwork at the nuclear periphery [3].
Alterations in the structure, organization, and composition of the nuclear lamina are likely to account for aberrant shapes of the nuclei of malignant cells, as well as the distinct nuclear morphology of neutrophils. Moreover, the increased titer of mutant lamins can have major consequences for the shape and integrity of the nuclear surface [3-6]. Lamin orthologs are absent from yeast, and it is not known whether a structural or functional equivalent of the lamina exists in S. cerevisiae. When higher eukaryotic lamin B1 or its receptor is expressed in S. cerevisiae, these proteins concentrate at the periphery of the nucleoplasm [7].
Several non-membrane proteins that normally localize to the periphery of the nucleoplasm in yeast possess coiled-coil domains (Esc1p, Mlp1/2p, Sir4p, Smc5/6p) [8-11]. Esc1p (Establishes Silent Chromatin), contains three equally spaced coiled-coil domains, can function as an anchor for chromatin, interacts with Rap1p and Sir4p, promotes targeted silencing at telomeres and HMR, is needed for proper organization of the "nuclear baskets" of nuclear pores, and promotes nuclear retention of unspliced transcripts [8,12,13]. Esc1p is larger than lamins (187 vs 70 kDa) and lacks the C-terminal isoprenylated CAAX motif present in A and B-type lamins.
The present study demonstrates that accumulation of Esc1p at the nuclear periphery causes dramatic modifications of the nuclear envelope. These modifications are structurally distinct from those which have been reported in a screen of deletion strains [14] or upon deletion of an ER/NE membrane phosphatase [15-17], mutation of components of the nuclear pore complex [18], proteins which function in the early secretory path [19,20], or Acc1p, which reduces very long-chain fatty acids [21]. Interestingly, the titer of Esc1p affects levels of several transcripts, only some of which originate from telomere-proximal loci. The structural abnormalities of the NE which are caused by Esc1p are not passed on to daughter cells, showing that inheritance of components of the nuclear perimeter can be selective.
To learn whether the composition of the nuclear periphery influences nuclear shape, and to extend previous observations on Esc1p [8], we have induced synthesis of GFP-tagged Esc1p in S. cerevisiae. After 3–5 hrs we observe the progressive elaboration of fin- and ring-like "
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Figure 1. Structure of Escapades. (A) (A') Haploid cells expressing GFP-Esc1p from a galactose-inducible promoter (ATY3258) were induced for 5 hrs (left) or overnight (right) with 2% galactose. Escapades are seen as "fins" (F), "rings" (R) or "patches" (P). Phase images are in blue. Induction with 10–100-fold lower concentrations of galactose reduced the number of cells which were fluorescent but did not affect the appearance of their escapades. Note that fewer fins are seen upon overnight induction. The scale bar in this and all figures designates 1 micron. (B) Strain ATY2957 expressing both untagged Esc1p from a galactose-inducible promoter and a GFP-tagged ER marker (a truncated form of HMG-CoA reductase 1-GFP [59]), after overnight induction. Two cells are illustrated – one toward the upper right and one in the lower middle portion of the field. Note the ring-like escapades (e) in both cells. Fin-like escapades are seen after shorter periods of induction and no such structures are seen in the absence of galactose. The peripheral ER is indicated by the arrow. N: The nucleoplasmic volume. (C) Cells which express mRFP-tagged Htb2p and Nup49p-GFP as well as galactose-inducible untagged Esc1p (ATY3156) were induced overnight. Note the presence of the Nup49p-GFP signal (arrow) outside the margin of the red chromatin mass. By contrast, controls in glucose medium show a conventional circumferential distribution of GFP signal (not shown). (D) (D') GFP-Esc1p-expressing cells (ATY1483) were induced overnight, fixed and processed for transmission EM. Note the constant width of the double membranes that extend from the surface of the nucleus, and the presence of nuclear pores in these extensions (arrows). (E) Time-lapse sequence of a GFP-Esc1 expressing strain (ATY2102) after overnight induction. Note the progressive conversion of the fin to a ring-like escapade. Such conversions are seen only infrequently. N: The spherical portion of the nucleus. (F) (F') Htb2p-mRFP-expressing cells induced for 5 hr to express GFP-Esc1p (ATY3281). The square in the upper field is enlarged and illustrated without the green signal below to clarify the distribution of Htb2p-mRFP. Note the absence of tagged chromatin from the escapades.
Images of cells which express Nup49p-GFP (Fig. 1C) show that escapades include nuclear pores (Fig. 1C). Ultrastructural examination also detects nuclear pores in the escapades and demonstrates that escapades are double-membrane sheets (rather than tubules) of constant width (Fig. 1D). Since escapades are limited by two layers of NE it is reasonable that – in cells expressing GFP-Esc or ER membrane proteins – their fluorescent intensity can exceed that of the rest of the NE (Fig. 1A/B). Time-lapse observations show that the position and contour of most escapades remains essentially constant for tens of minutes; however the fin-like structures occasionally fuse back to the nucleus and generate rings (Fig. 1E). Examination of cells which express a tagged histone (Htb2p-mRFP) as well as GFP-Ec1p shows that escapades include little or no chromatin (Fig 1F).
To learn whether excess of other proteins of the nuclear periphery causes similar changes, we have compared the impact of excess Esc1p to that of the non-membrane protein, Mlp1p [See Additional file 1] which colocalizes with Esc1p [9,22], the tail-anchored inner membrane protein, Prm3p [23] [See Additional file 2], and the integral membrane proteins of the inner membrane, Heh1p and Heh2p [24] [See Additional files 3 and 4]. Excess Mlp1p is known to distribute throughout the nucleoplasm [9]. As shown, Mlp1p has no obvious effect and Prm3p causes changes of the NE roughly comparable to those caused by Esc1p. By contrast, the Heh proteins are much more perturbing, with Heh1p not even leaving the quasi-spherical shape of the chromatin mass intact. Induction of either Esc1p or Mlp1p allows continued growth, while induction of Heh1p, Heh2p, or Prm3p is toxic (not shown). We therefore have not pursued these membrane proteins further.
Additional file 1. Impact of excess Mlp1p. Cells expressing the ER/NE membrane protein, Sec61p-GFP (ATY3255), or Htb2p-mRFP (ATY3256) were induced to overexpress Mlp1p for 5 hr by addition of galactose. Note the conventional distribution of GFP signal at the NE and throughout the peripheral ER (left panel) as well as the roughly spherical chromatin mass (right panel).
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Additional file 2. Impact of excess Prm3p. Htb2p-mRFP-expressing cells were induced to overexpress GFP-Prm3p for 5 hr by transfer to methionine-free medium (ATY3244). Note the appearance of GFP-positive ring-like structures and a fin at the margin of the nucleus.
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Additional file 3. Impact of excess Heh1. Htb2p-mRFP-expressing cells were induced galactose to overexpress Heh1p-GFP for 5 hr by addition of 2% (ATY3245). The red and green images have been separated for clarity. Note the massive change of organization of both chromatin and the GFP signal, which often encircles the chromatin.
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Additional file 4. Impact of excess Heh2p. Htb2p-mRFP-expressing cells were induced to overexpress Heh2p-GFP for 5 hr by addition of 2% galactose (ATY3246). The red and green images have been separated for clarity. Note the massive change of organization of both chromatin and the GFP signal, which generally encircles the chromatin.
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Relation of Escapades to Intranuclear Structures
In cells which express the nucleolar marker, Sik1p-mRFP, and can be induced to express GFP-Esc1p, systematic examination of through-focal series after 5 hr induction shows that > 90% of escapades contact the perimeter of the nucleus at or immediately adjacent to the nucleolus (Fig. 2A, and see Additional file 5). The nucleolus is not however present within the escapades themselves. The yeast spindle pole body (SPB) is embedded in the NE at a position which is usually opposite the nucleolus [25,26]. Consistent with the observed nucleolar association, the localization of escapades and karmellae does not coincide with the tagged SPB protein, Spc42p (Fig. 2B and see Additional file 6). Systematic counting of through-focal series of cells which exhibit escapades shows that only ~10% of the tagged SPBs contact escapades. Another modification of the NE, the "karmellae" which result from overexpression of HMG-CoA reductase, also associates with the nucleolus [27] and avoids the SPB (Fig. 2C). A third instance of association with the nucleolus is that of the "flares" which appear upon deletion of the ER/NE membrane proteins, Nem1p or Spo7p. In this case, the flares define a pocket which encloses the nucleolus [15].
Additional file 5. Spatial relation of escapades to the nucleolus. Cells expressing the nucleolar marker, Sik1p-mRFP, were induced to express GFP-Esc1p for 5 hr by addition of 2% galactose (ATY3258). Systematic examination of through-focal series detects association of escapades and Sik1p-mRFP in > 90% of cells which have escapades. Nevertheless, in a given section the extent of association often appears only modest.
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Additional file 6. Spatial relation of escapades to the spindle pole body. Overview comparison of the localization of GFP-tagged Esc1p and the spindle pole body, in a strain (ATY3276) which expresses Spc42p-mRFP and has been induced by addition of 2% galactose for 5 hr. Systematic examination of through-focal series detects association in ~10% of cells which have escapades.
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Figure 2. Relation to Nuclear Structures. (A) GFP-Esc1p was induced for 5 hr to compare the localization of GFP-tagged escapades to the nucleolus, in a strain (ATY2101) which expresses Sik1p-mRFP. Systematic examination of through-focal series shows that > 90% of escapades contact the perimeter of the nucleus at or immediately adjacent to the mRFP-positive nucleolus. See Fig. S5. (B) Comparison of the localization of GFP-tagged escapades with the spindle pole body, in a strain (ATY3276) which expresses Spc42p-mRFP and has been induced for 5 hr. An overview is given in Fig S6. (C) Comparison of the localization of GFP-tagged karmellae with the nucleolus and the spindle pole body, in a strain (ATY1577) which expresses Sik1p-mRFP and Spc42p-GFP and carries plasmid [pGAL-HMG-CoA Reductase I-GFP]. Karmellae are ER membrane stacks associated with the outer nuclear membrane which result from overexpression of HMG-CoA reductase type1. (D) Comparison of the localization of GFP-tagged escapades with a centromere, in a strain (ATY2098) which expresses a GFP-lac repressor fusion and an insertion of lac operator arrays near CENIV and has been induced for 5 hr. An overeview is given in Fig. S7. (E) Comparison of the localization of GFP-tagged escapades with a telomere, in a strain (ATY2097) which expresses a GFP-lac repressor fusion and carries an insertion of lac operator arrays near telomere XIVL and has been induced for 5 hr. An overview is given in Fig. S8. (F) Comparison of the localization of the GFP-tagged telomere-associated protein, Rap1p, and Htb2p-mRFP-tagged chromatin in ATY3275. Induction was for 5 hr. An overview is given in Fig. S9.
Since centromeres are close to the SPB during most of the cell cycle [2], we also studied their relation to escapades, using cells which express lac operator arrays integrated near a centromere and GFP-tagged lac repressor. No obvious association is seen between this centromere and escapades (Fig. 2D and see Additional file 7). Equivalent experiments to localize a telomere again show no obvious association (Fig. 2E and see Additional file 8). In both cases, systematic counting of through-focal series of cells which exhibit escapades shows that only ~10–12% of the tagged loci contact escapades. A functional GFP-tagged form of the telomere-associated protein, Rap1p, which binds Esc1p [8], also is not detected outside the chromatin mass, as would be expected if it were in escapades (Fig. 2F and see Additional file 9). This is also the case for Sir4p [8].
Additional file 7. Spatial relation of escapades to a centromere. Overview comparison of the localization of GFP-tagged Esc1p and a centromere, in a strain (ATY2098) which expresses a GFP-lac repressor fusion, an insertion of lac operator arrays near CENIV and Nup49p-GFP. It has been induced by addition of 2% galactose for 5 hr. Systematic examination of through-focal series detects association in ~10% of cells which have escapades.
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Additional file 8. Spatial relation of escapades to a telomere. Overview comparison of the localization of GFP-tagged escapades and a telomere, in a strain (ATY2097) which expresses a GFP-lac repressor fusion, an insertion of lac operator arrays near telomere XIVL and Nup49p-GFP. It has been induced by addition of 2% galactose for 5 hr. Systematic examination of through-focal series detects association in ~10% of cells which have escapades.
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Additional file 9. Spatial relation of escapades to Rap1p. Overview localization of Rap 1-GFP in a strain (ATY3275) which expresses Htb2p-mRFP and allows galactose induction of untagged Esc1p. It was induced by addition of 2% galactose for 5 hr. Note that the labeled foci often are at the periphery of the chromatin mass, but – unlike escapades – do not extend centrifugally toward the cytoplasm.
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Formation of Escapades-Relation to Cytoplasmic Structures
Following the separation of sister chromatids at the onset of anaphase, the nucleus of the mother cell quickly extends into the bud, generating a dumbbell-shaped structure with a narrow bridge of NE connecting the two nuclei [28]. Fission of the bridge leaves protruding membrane remnants that are normally resorbed by the two resulting nuclei. Escapades superficially resemble these remnants and therefore might be derived from them. Nevertheless, time-lapse microscopy of cells which exit mitosis shows that GFP-Esc1p-positive remnants are efficiently resorbed, as in control cells (see below). Moreover, cells treated with α-factor (3 hr, 1 μg/ml) [See Additional file 10] or hydroxyurea (3 hr, 0.1 M) [See Additional file 11] can generate escapades when Esc1p expression is induced in the absence of cell cycle progression.
Additional file 10. Induction of escapades in cells treated with mating factor. GFP-Esc1p was induced by addition of 2% galactose for 5 hr in cells which had already been treated with 5 μg/ml α-factor for 2 hr (ATY2101). Note that the appearance of escapades is comparable to that illustrated in Figure 1A.
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Additional file 11. Induction of escapades in cells treated with hydroxyurea. GFP-Esc1p was induced by addition of 2% galactose for 5 hr in cells which had already been treated with 0.1 M hydroxyurea for 2 hr (ATY2101). Note that the appearance of escapades is comparable to that illustrated in Figure 1A.
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The formation of escapades does not appear to depend on the integrity of the tubulin or actin cytoskeleton, judging from experiments in which they are induced in the presence of doses of nocodazole or latrunculin A which effectively depolymerize the corresponding cytoskeletal structures [See Additional files 12 and 13].
Additional file 12. Induction of escapades in cells treated with nocodazole. GFP-Esc1p was induced by addition of 2% galactose for 5 hr in cells which had already been treated with 15 μg/ml nocodazole 2 hr (ATY2101). Note that the appearance of escapades is comparable to that illustrated in Figure 1A.
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Additional file 13. Induction of escapades in cells treated with latrunculin A. GFP-Esc1p was induced by addition of 2% galactose for 5 hr in cells which had already been treated with 200 μg/ml latrunculin A 2 hr (ATY2101). Note that the appearance of escapades is comparable to that illustrated in Figure 1A.
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Like the surface of the yeast nucleus [29], escapades are intimately associated with vacuoles, as detected with the membrane dye, FM4-64 (Fig. 3A). Live-cell imaging reveals that escapades can originate at or near the nucleus-vacuole junction (NVJ), prior to extending along the surface of the vacuole (Fig. 3B). Nevertheless, small ring-like escapades and patches of Esc1p can be induced in pep3-Δ cells which lack conventional vacuoles [30] (Fig. 3C).
Figure 3. Escapades Associate with the Vacuole and Originate at the NVJ. (A) Association with the vacuole, as detected by confocal examination of strain ATY2102 stained with FM4-64 (red) after galactose induction for 3 hrs. In each case, the chromatin-containing portion of the nucleus is designated (N) and the escapade is indicated by (*). (B) Time-lapse confocal sequence of strain ATY2513 that expresses Sik1p-mRFP (nucleolus: Nu) and Vac8p-YFP, which marks the vacuole membrane (Vac) and concentrates at the nucleus-vacuole junction (NVJ). GFP-Esc1p expression was induced with galactose 30 minutes prior to and maintained throughout time-lapse imaging, to follow the progression of GFP-Esc1p accumulation. Note the GFP-Esc1p at the periphery of the nucleus (N) and its progressive extension along the surface of the vacuole, generating an escapade (*). The MET25p-VAC8-EYFP plasmid was from D. Goldfarb. (C)pep3Δ cells which allow induction of GFPEsc1p were induced for 5 hr and examined (ATY2103).
Inheritance of Escapades
The distribution of escapades was examined as cells pass through anaphase. Intriguingly, they are excluded from > 95% of daughter nuclei (Fig. 4/Table 1). The asymmetric distribution is especially conspicuous in a mob1-77 mutant which makes it possible to maintain cells in late anaphase for hours (Table 1) [31].
Table 1. Maternal Retention of Escapades
Figure 4. Escapades are Asymmetrically Inherited. GAL-GFP-Esc1p strain ATY2102 was induced for 3 hours at 23°C prior to imaging at 2 min intervals in glucose medium. Boxed regions indicate cells undergoing mitosis and the arrows indicate the direction of elongation of the nucleus during anaphase. Mother (M) and daughter (D) nuclei are indicated in the last panel. Note the retention of escapades by the mother. The color image at the bottom includes a phase image to illustrate the cell outlines just prior to t = 0.
Why are Escapades not Present in Daughter Nuclei ?
Escapades could be actively retained by the maternal nucleus, excluded from the daughter nucleus, or could simply not be able to diffuse in the plane of the membrane. Since septin filaments at the bud neck can restrict transfer of proteins between mother and bud [32-35], we have examined inheritance in strains that carry mutations in septin subunits and disorganize the septin collar at the restrictive temperature (cdc3-3, cdc10-1). In these mutants, escapade exclusion from daughter nuclei is again seen (Table 1). Karmellae are also restricted to the mother in wt and in septin mutants (not shown). Consistent with the suggestion that there is no structural impasse at the bud neck, most escapades which remain in the mother do not accumulate at the neck. Moreover, electron microscopic examination shows that a significant space separates the outer nuclear membrane from the inner aspect of the plasma membrane at the bud neck during anaphase [36].
To evaluate the maternal retention model, we have studied the distribution of escapades in cells for which nuclear division does not require traversal of the bud neck. For this purpose, we have disrupted actin filament integrity with latrunculin A, which allows the spindle to deviate from the mother/bud axis, with the result that nuclear division can occur entirely within the maternal cytoplasm. Strikingly, escapades are restricted to a single nucleus in such binucleates (Fig. 5). This finding led us to investigate whether association with the vacuole could account for maternal retention; however, escapades are overwhelmingly retained in a pep3Δ strain (Table 1).
Figure 5. Distribution of Escapades in Binucleates. Cells expressing GFP-Esc1p and Htb2p-mRFP (ATY2509) were pre-induced, washed and recultured for 8 hrs at 23°C in glucose medium supplemented with latrunculin A to produce binucleate cells, resulting from occasional incorrect orientation of the spindle. A diploid strain was used to facilitate spatial resolution. (A) Confocal z-sections of a binucleate cell show retention of escapades (arrow) in a single nucleus. (B, C) Epifluorescent imaging of binucleate cells. Phase images indicate that the nuclei are contained within the maternal cytoplasm.
To inquire whether escapades and karmellae are intrinsically immobile, we have examined the constancy of their position when the nucleus with which they associate fuses with a conventional nucleus during karyogamy. As shown in Fig. 6, they show no tendency to migrate into the trans nucleus over a period of time greater than that required for anaphase. Nevertheless, tagged nuclear pores are able to access the trans nucleus [37,38]. Note the rapid transit of GFP-Esc1p to the trans nucleus. At least at this level of expression, it has considerable mobility.
Figure 6. Distribution of Escapades and Karmellae upon Karyogamy. Escapades and karmellae remain with the nucleus of their origin upon fusion with nuclei which express Htb2p-mRFP.
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Functional Consequences
Despite dramatic changes in nuclear shape, cells that express excess Esc1p or GFP-Esc1p are not seriously growth impaired in liquid culture or on solid media. Furthermore, they show a distribution of DNA content comparable to wt, and can mate (not shown and Figure 6). The maternal restriction of escapades may direct most physiological consequences of their presence to the mother and thereby contribute to the lack of a major growth phenotype, as each cell division produces progeny which initially lack escapades.
It is therefore of interest that a differential effect at the level of cell cycle progression is evident. Normally, when a mother cell gives rise to a daughter, the mother will rebud quickly, while daughters must grow to a critical size before budding [39]. By contrast, when pre-induced GFP-Esc1p-expressing cells are observed, the bud interval is slowed in mothers (possibly due to interference by the escapades themselves) and accelerated in daughters. As a result, the difference in timing of budding between mothers and daughters is reduced by more than 50% (Fig. 7). The net impact of these events could account for the modest reduction of relative growth rate which can best be detected in mixed cultures in which isogenic wt and GFP-Esc1p-expressing cells are both present. Pure cultures of cells which can be induced to express GFP-Esc1p remain fluoresecent for days of culture in galactose medium, while addition of an equal number of wild type cells prior to culture is followed by a gradual and progressive reduction in the proportion of fluorescent cells.
Figure 7. Kinetics of Bud Formation. GAL-GFP-Esc1p strain ATY2102 and an isogenic wild-type strain (ATY2500) were pre-induced in galactose media for 3 hrs, washed and examined on agarose pads in glucose media over 5 hr, during which time escapades remained visible (ATY2500). DIC images captured every 10 min allowed measurement of the timing of bud initiation for each cell. Time measurements were all relative to the emergence of the first bud by each mother cell, M1. For example, the mother's bud interval, M2-M1 measures the delay prior to the appearance of the second bud on the mother. Correspondingly, D-M1 measures the interval prior to the appearance of the first bud on the daughter. (A) Histogram showing the bud intervals for a single experiment for the wild type strain. Note that there is minimal overlap between mothers and daughters and that mothers re-bud considerably faster than daughter cells. (B) Histogram showing the bud intervals for an isogenic escapade strain. Note, in contrast to (A), that there is considerable overlap between mothers and daughters. The bud interval is slowed in mothers and accelerated in daughters. (C) Histogram illustrating the mother-daughter bud differential calculated as: (D-M2 = bud differential) where D is the time at which a bud appears in the daughter cell and M2 is the time at which a second bud appears in the mother cell. Note that the bud differential is decreased by 50% by the presence of escapades (average bud differential is 65 min. in wild-type strain and 28 min. in escapade strain). Results are cumulative over four separate experiments for each strain.
To further investigate the consequences of excess Esc1p, we initiated microarray analysis of poly(A)+ mRNAs of a pair of isogenic strains grown in galactose medium, only one of which allows Esc1p induction from a GAL1 promoter (Table 2). Upon overnight growth in galactose medium, there is a ~20–30x increase in the signal corresponding to ESC1 mRNA and only 48 other changes which exceed 1.8x fold. Both positive and negative changes are seen. Considering that GFP-Esc1p concentrates at the extreme periphery of the nucleoplasm along with telomeres, it is of interest that both the increases and decreases (> 1.8 fold) are enriched within 40 kb of telomeres, i.e. even though this region accounts for only ~4.1% of the length of the average yeast chromosome, it accounts for ~30% of these changes. Nevertheless, ~70% of the changes are not in this region. No notable changes in mRNA levels were seen for genes whose products associate with Esc1p, such as RAP1 and SIR4. Since deletion of ESC1 does not obviously affect nuclear morphology [See Additional file 14], we have not investigated corresponding transcriptional changes.
Additional file 14. Nuclear shape in the absence of Esc1p. The contour of the NE was defined in cells which lack Esc1p by monitoring the distribution of Sec63p-GFP in a corresponding deletion strain (ATY3278).
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Table 2. Microarray Data
The morphology of organelles is intimately related to their function and deviation from normal morphology can have profound physiological consequences. It is thus plausible that NE shape alterations contribute to events which cause cellular malfunction in disease.
Nevertheless, radical changes of the composition of the nuclear periphery can be compatible with cell survival, as in a variety of yeast deletion strains lacking transmembrane proteins which localize to the NE and ER (Nem1p, Spo7p, Ssh1p), nucleoplasmic proteins (Thp1p), or nucleoporins (Seh1p) [14,17]. None of these proteins normally are restricted to the periphery of the nucleoplasm itself.
Interestingly, the shape of the nucleus is not affected by deletion of Esc1p or deletion of two other proteins which normally localize to the periphery of the nucleoplasm, Mlp1p and Mlp2p [9,22,40]. Moreover, the growth of nem1-Δ, seh1-Δ, spo7-Δ, ssh1-Δ and thp1-Δ [14] is not obviously affected by overexpression of Esc1p (not shown).
Mutations in nuclear lamina constituents, most notably lamin A and C, cause a diverse spectrum of diseases, the laminopathies. Laminopathies caused by excess pre-lamin A at the nuclear periphery are characterized by bleb-like expansions of the nuclear surface [41-44]. The dependence of the shape of the yeast nucleus on both nuclear membrane proteins and proteins that concentrate at the periphery of the nucleoplasm is reminiscent of a distinct laminopathy (Emery-Dreyfuss Muscular Dystrophy), which can result from either mutation of the inner nuclear membrane protein, emerin, or mutation of lamin A [45,46]. Moreover, overexpression of a GFP-tagged form of the inner membrane protein, Prm3p, distorts the shape of the NE in much the same fashion as Esc1p, and excess Heh1p and Heh2p [24] grossly distort the NE and chromatin mass, while Mlp1p induction has no obvious impact. These differential effects may signify that the Heh proteins have a high affinity for chromatin, that Esc1p and Prm3p are more closely linked to the inner nuclear membrane per se than to chromatin, and that Mlp1p is relatively independent.
The characteristic structure of escapades and distribution of excess GFP-Esc1p are compatible with the hypothesis that excess Esc1p forces enlargement of the NE due to end-to-end association of the protein, that chromatin has an intrinsic coherence which tends to preserve a roughly globular shape, and that the inner aspect of the NE (or perhaps Esc1p itself) can also self-associate laterally (Fig. 8). Indeed, although there has never been an experimentally accessible model for investigation of this latter issue, the NE of malignant cells – like escapades – is frequently characterized by focal self-apposition of the lamina and/or the nucleoplasmic surface of the inner nuclear membrane [47,48].
Figure 8. Model of Escapade Structure. Nuclear pore complexes (NPC), outer nuclear membrane (Outer), inner nuclear membrane (Inner), and Esc1p are indicated. The small circles at the surface of the membranes are ribosomes. The model has been drawn to illustrate a tight apposition of Esc1p layers within the escapade, which may account for the constant width that is observed with EM and the exclusion of chromatin.
There is no reason to expect that Esc1p is fully comparable to higher eukaryotic lamins. For example, unlike lamins, tagged Esc1p (or Mlp1p) expressed from its own promoter does not completely encircle the nucleus, being absent from beneath the nucleolus [13,40]. Moreover, the observation of rapid diffusion of GFP-Esc1p upon karyogamy shows that at least the overexpressed protein is mobile.
The transcriptional consequences of overexpressing Esc1p emphasize the importance of this protein (or escapades themselves) for gene expression. Since both negative and positive changes are seen, Esc1p appears to be a complex regulator, not only an enhancer of silencing. In this regard, it resembles many transcriptional regulators including Rap1p [49]. It is also notable that ~70% of the changes which we detect occur at loci which are further than 40 kb from telomeres (which concentrate at the periphery). In addition to their intrinsic interest, these microarray data provide a possible prototype against which to evaluate the transcriptional characteristics of laminopathies, which presumably account for their cell type-specific effects.
Daughter cells differ from mothers in several regards [50,51]. Escapades are not transferred to daughter cells, apparently due to their immobility. These structures – and karmellae – are thus part of a "lagging" domain of the nuclear perimeter. The nucleolus may also be part of this domain, judging from its association with escapades and karmellae, as well as the observation that it is one of the last nuclear components to reach the bud during anaphase [52,53]. Extra rDNA circles [54] and ARS plasmids [55] are also retained.
This study demonstrates the extreme structural plasticity of the yeast nucleus and shows that the unusual "escapades" which can be generated are essentially immobile and therefore are not inherited. Their lack of inheritance provides a striking example of the exclusion of epigenetic change. Such mechanisms – coupled with equivalent normalization of phenotype at the molecular level – are likely to sustain cell identity through mitosis. Higher eukaryotic cells may cope with such issues by extensive disassembly and reassembly of the NE during mitosis, which could provide an opportunity to avoid incorporation of structurally aberrant components.
Yeast Strains and Plasmids
Yeast strains and plasmids used in this study are listed in Table 3 and Table 4. The GFP-tagged form of Esc1p which we employ is known to complement the plasmid partitioning defect of esc1Δ [8]. Strains were constructed using standard methods for transformation, mating, and sporulation. To place ESC1 transcription under control of a galactose-inducible promoter, we used the plasmid, pFA6A-pGAL1 [56], as template to generate PCR products which could be targeted upstream of ESC1. Plasmid pMET25-GFP-ESC1 was constructed by PCR sub-cloning of the full length Esc1p coding sequence from plasmid pEDA129 [8], into pGFP-N-FUS [57]. Restriction endonuclease sites, SpeI and XhoI, present in the polylinker, were utilized such that the ESC1 ORF is in-frame down-stream of GFP and expression is driven by the methionine-repressible MET25 promoter and terminated by the CYC1 terminator. Recombinant clones were identified by restriction endonuclease analysis and DNA sequencing of the 5' cloning junction to verify orientation and reading frame. The level of expression observed in transformed strains cultured under inducing conditions was sufficient to generate NE alterations.
Media and Supplements
Cells were grown in complete synthetic medium or the appropriate dropout medium supplemented with 2% D-glucose, D-raffinose or D-galactose. Generally strains were maintained in mid-log phase in raffinose medium and were induced by addition of galactose. All incubations were at room temperature except when ts conditional strains needed to be shifted to 37°C. As required, media were supplemented with: 200 μM latrunculin A (Sigma) diluted from a 20 mM stock in DMSO, 5 μg/ml α-factor (Sigma) diluted from a 5 mg/ml stock in sterile water, 0.1 M hydroxyurea (Sigma) diluted from a 1 M stock in sterile water, 15 μg/ml nocodazole (Sigma) diluted from a 3 mg/ml stock in DMSO, or 40 μM FM4-64 (Molecular Probes) diluted from a 2 mM stock in DMSO.
GAL-GFP-Esc1p strain ATY2102 and isogenic wild-type ATY2501 were grown overnight to mid-log phase in galactose-containing synthetic medium at room temperature. Total RNA was purified from duplicate 1 ml samples by mechanical disruption with 0.2 micron glass beads in an equal volume of hot acid phenol and used for biotinylated cRNA synthesis according to Affymetrix protocols at the Case Cancer Center Gene Expression Array Facility. Replicate samples were hybridized to Affymetrix GeneChip yeast genome Y98 arrays. Scanned chip images were analyzed with the Affymetrix GeneChip Operating Software (GCOS) using the MAS 5.1 algorithm for fold-change calculations. The results were tabulated with Microsoft Access and annotated using NETAFFX (Affymetrix) and Genespring (Silicon Genetics) software packages. In the tabulation in Table 2, the average values for GFP-Esc1p-overexpressing cells are divided by the averaged data for control cells.
Budding Kinetics
GAL-GFP-Esc1p strain ATY2102 and isogenic wild-type strain ATY2501 were pre-induced in galactose media for 3 hrs at 23°C, washed and examined on agarose pads in glucose medium over 5 hr, during which time fluorescent escapades remained visible. DIC images captured every 10 min allowed measurement of the timing of bud initiation for each cell in four replicate experiments. Time measurements were all relative to the emergence of the first bud by each mother cell, M1. For example, the mother's bud interval, M2-M1 measures the delay prior to the appearance of the second bud on the mother. Correspondingly, D-M1 measures the interval prior to the appearance of the first bud on the daughter.
The time difference of budding between mothers and daughters is referred to as the budding differential and is calculated as: (D-M2 = bud differential) where D is the time at which a bud appears in the daughter cell and M2 is the time at which a second bud appears in the mother cell.
Epifluorescent, phase time-lapse, and DIC microscopy were performed on a Leica DMLB microscope equipped with a SPOT camera (Diagnostic Instruments Inc.) and the SPOT Advanced software package. Confocal microscopy for live cell Z-stack and time-lapse imaging were performed on either a Zeiss LSM510 or a Leica AOBS. Volocity (Improvision) and Metamorph (Molecular Devises) software packages were utilized for image analysis and generation of time-lapse movie sequences. Other z-stack images were collected with a DeltaVision microscope and deconvolved before inspection. For quantitation of association of escapades with various structures, through-focal series were examined systematically and the distribution of nucleoli, etc. was counted in greater than 100 cells.
Vacuole membrane staining with the vital dye FM4-64 was conducted according to [58] with the following modifications. Briefly, log phase galactose-induced cultures were concentrated by centrifugation and resuspended at OD600 = 1 in glucose-containing medium supplemented with 40 μM FM4-64. The cells were then incubated at 30°C with shaking for 30–60 minutes, followed by extensive washing to remove excess dye. Stained vacuoles were imaged at 546 nm with a Zeiss LSM510 confocal microscope.
For time-lapse microscopy, dilute cultures were briefly centrifuged and 2 μl aliquots from the pellet were applied to the middle of 1.5% Agarose pads prepared on microscope slides in glucose- or galactose-containing culture medium, as appropriate. Samples were overlayed with a coverslip and sealed with vaseline before examination.
To learn whether escapades or karmellae are free to move in the plane of the NE, we have crossed cells which express these structures with cells of the opposite mating type and then observed the structure of the nucleus during and after nuclear fusion. For this purpose, pairs of cells grown overnight in galactose-containing medium were mixed, sedimented, and applied to an agarose pad in glucose-containing complete synthetic medium (as above). After approximately 2 hrs, they were observed.
Transmission electron microscopy was performed on samples prepared from cultures grown to mid-log phase at 30°C. Samples were fixed in 2% glutaraldehyde followed by secondary fixation/staining in 4% potassium permanganate and uranyl acetate en bloc. Samples were dehydrated, embedded in Spurr resin, stained with lead citrate and uranyl acetate and examined in a JEOL 1200CX EM.
Authors' contributions
TH conducted most of the experiments as part of his doctoral work.
EA, who made the first observations of deformation of the nuclear envelope/nucleus upon Esc1p induction, provided frequent advice.
AT made the initial observations on escapade induction in this laboratory, coordinated the study, and wrote most of the manuscript.
All authors read and approved the text.
E. Bi, G. Blobel, V. Doye, J. Drazba, E. Fabre, S. Gasser, D. Goldfarb, J. Haber, M. Hitomi, W.-K. Huh, E. Jones, J. Kilmartin, M. Lam, J. Lithgow, M. Luca, P. Lusk, A.G. Matera, D. MacDonald, D. Meyer, F. Najm, D. Narendra, T. Rapoport, M. Rout, K. Runge, D. Shore, R. Sternglanz, P. Tiedrez, E. Townsley, A. Vasanji, K. Weis, H. Worman, R. Wright, D. Wu and Y. Zhang. Confocal Microscopy Core Facility in the Comprehensive Cancer Center of CWRU/UHC (P30 CA43703-12). T.H. was supported by an NIH training grant: "Normal and Abnormal Development" T32-HD7104. A.T.'s contribution was supported by institutional resources.
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