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Download the Game Design and Indie Game Marketing Freebook 07/19/17GameDev.net and CRC Press have teamed up to bring a free ebook of content curated from top titles published by CRC Press. The freebook, Practices of Game Design & Indie Game Marketing, includes chapters from The Art of Game Design: A Book of Lenses, A Practical Guide to Indie Game Marketing, and An Architectural Approach to Level Design. The GameDev.net FreeBook is relevant to game designers, developers, and those interested in learning more about the challenges in game development. We know game development can be a tough discipline and business, so we picked several chapters from CRC Press titles that we thought would be of interest to you, the GameDev.net audience, in your journey to design, develop, and market your next game. The free ebook is available through CRC Press by clicking here. The Curated Books The Art of Game Design: A Book of Lenses, Second Edition, by Jesse Schell Presents 100+ sets of questions, or different lenses, for viewing a game's design, encompassing diverse fields such as psychology, architecture, music, film, software engineering, theme park design, mathematics, anthropology, and more. Written by one of the world's top game designers, this book describes the deepest and most fundamental principles of game design, demonstrating how tactics used in board, card, and athletic games also work in video games. It provides practical instruction on creating world-class games that will be played again and again. View it here. A Practical Guide to Indie Game Marketing, by Joel Dreskin Marketing is an essential but too frequently overlooked or minimized component of the release plan for indie games. A Practical Guide to Indie Game Marketing provides you with the tools needed to build visibility and sell your indie games. With special focus on those developers with small budgets and limited staff and resources, this book is packed with tangible recommendations and techniques that you can put to use immediately. As a seasoned professional of the indie game arena, author Joel Dreskin gives you insight into practical, real-world experiences of marketing numerous successful games and also provides stories of the failures. View it here. An Architectural Approach to Level Design This is one of the first books to integrate architectural and spatial design theory with the field of level design. The book presents architectural techniques and theories for level designers to use in their own work. It connects architecture and level design in different ways that address the practical elements of how designers construct space and the experiential elements of how and why humans interact with this space. Throughout the text, readers learn skills for spatial layout, evoking emotion through gamespaces, and creating better levels through architectural theory. View it here. Learn more and download the ebook by clicking here. Did you know? GameDev.net and CRC Press also recently teamed up to bring GDNet+ Members up to a 20% discount on all CRC Press books. Learn more about this and other benefits here.
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beegor posted a topic in Your AnnouncementsHi, We (my company) just published our first 3D game for android. It is running on 3D engine called Rajawali which is relatively new so this was kind of engine testing. Check it out on Google Playstore: https://play.google.com/store/apps/details?id=com.mabuga.gravity.android.before4 I can say that Rajawali is very nice although it still has few major bugs, but community is working actively to fix them. Here are some screenshots...
Hi frob, thanks for your answer! I know there are lots of projects out there, that's why I 'm asking for recommendation (to save some time). I am looking for something that is 1. Stable on all android versions since Android 2.2 2. Cross-platform if possible, but stability is way more important 3. Preferably opensource (but I am willing to pay if I get stability and cross-platform). Somehow I doubt that cross-platform engine would work good on all platforms but I would be happy if somebody proved me that I was wrong.
Thanks, I will check those out, although I prefer opensource projects.
beegor posted a topic in General and Gameplay ProgrammingHi, I recently wrote blog post about developing 3d game for android using Rajawali 3d engine. It is very nice engine with lots of tutorials and examples, but I am wondering if there is more mature (stable) engine out-there. Has anyone tried to develop for android with http://jmonkeyengine.org/ or http://minko.io/ and if so would you recommend it? Thanks!
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Finalé Audio is a new addition to Triode Lab Ltd. family. While Triode Lab specializes in the production of the finest triode and low powered tube amplification, our goal is to appeal to a broader audience of discerning listeners who want the best sound possible at a reasonable cost. To this end, we offer a wide range of high caliber, affordable tube amplifiers, from low powered single-ended designs to 100W powerhouses, many priced around the $2,000 CAD mark (and so you can think of us as offering the Grand Finale!).
Like Triode Lab, all of our products are designed and engineered in Canada, and made to the same exacting standards. Visually, our amplifiers have a clean and simple elegance, but their true sophistication is revealed in their sound. We would rather invest in high quality parts and transformers than in elaborate design features that you can see but not hear, while most other tube amp maker invest the cost into the looks and packaging instead because most customer does not know a fine tube amp MUST come with good quality transformers and parts. Please contact us and let us help you find the right amplifier for your system. We are confident that your search for musical truth will end with Finale.
General Inquiries / Sales and Order : email@example.com
Speakers/ Art Director : Mr . Robert Gaboury (English / Français) - firstname.lastname@example.com
Dealers / Marketing / PR (North America) : Mr . Peter Wolter (English / German) - email@example.com
Production / Brand Support : Mr . Frank Ing (English / Cantonese) - email@example.com
HIRING : Buyers for over sea markets, as well domestic
Starting from 2017, our products will categorized into three lines to couple to the rapid HIFI market to satisfy different customers and markets.
1. FINALE AUDIO - The original brand became the " custom works " department , made to order world class tube amplifiers. (Logo in center, trademarked).
2. New finalé logos- became the Signature / Dealer Line , mainly the speakers to be. (Logos at right two, in the process of registering it's trademark)
3. Pentode 6 (left two) - it will be the direct sale label
And here is a good read about our companies , and our collaborations with our art director / speaker designer Robert Gaboury of Arteluthe Design.
ROBERT GABOURY / Q&A @ 6moons
"I am a designer. If I can focus on designing, I get the opportunity to bring to life new and exciting ideas. We have almost a dozen concepts on the books, some of which are in advanced prototype stages like the Vocé, a bigger version of the Mini and more of a descendant of the Katana than Vivace. The collaboration with Triode Lab has been very stimulating. It is sonic 'haute-couture' but we also share a passion for speakers of the 50s and 60s and we even have a project to bring back value kits based on the JBL and Altec designs of yore. This industry has changed. We need to adapt to survive. I can now pursue projects across multiple brands and not be locked into one single sonic signature which is liberating. "
http://www.6moons.com/audioreviews2/finale/2.html 2016 November
WELCOME TO JOIN OUR TEAM / SELLING NETWORK - as long you like music and our products! you will have your own protected sales territory as well immense support from us. Most sales are made to order other than the bigger sellers. So dealers / re-sellers SHOULD have a reasonable markup for bringing a item in or for demo. The good news are, sometimes we can use customer's preferred parts.
Prices does not include tubes most of the time simply we cannot beat the prices on internet. BUT, if you are ready to purchase a amplifier or speakers from us or our dealers, always ask for discount! or free shipping at least! or next item with deeper discount. We take proud in providing the best customer services while the best pricing.
Subject to price changes of local region due to shipping / currency fluctuations. CONTACT your local re-sellers for the most accurate price and information.
WE LIKE GREEN, and we ONLY use high quality and safe shipping boxes made out of recycle materials. We always use double boxes arrangement with lots of cushioning. Conventional foam/box/plastics are not used across the line due to the wide range of available options and upgrades, so it lower cost on your side too.
Anything higher grade will be custom make tripe layer hard cartoon shipping box with perfect fit high quality foam. Wood crate is option as well insurance, and any heavier item will be shipped on pallets, or by cargo.
Although our amps are known to be reliable and rock solid (not a coincident!). But every amplifier has a 2 years warranty, transferable and from normal use, starting from its manufacture date. That does not including use of wrong/failing tubes, lighting and shipping damage, or any form of modifications.

subset: commoncrawl


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Chubby cutie getting double teamed and l
Date: 2011-11-07 08:24:44
Date: 2011-11-07 08:24:44
Smut Bus - Free Preview What the fuck is the SmutBus? We're just a bunch of horny fucking guys with an arsenal of pimped out rides. We've got a school bus, a van and a bad ass camper van. We drive around all day long looking for hoes that might need a ride (or two, hahahahah). Access all of our exclusive Smutbus adventures right now! Smut Bus Presents: Holly Holly wasnt looking to get laid at all, unfortunately for her she was without a ride and desperate to find one! We offered her a lift, but of course shed have to earn it! Watch the group sex action inside! Start downloading the Full Video Now! When you become a member, you also get free bonus access to over 112 Exclusive Reality Sites, including High Definition sites for only ! So what are you waiting for? Take advantage of the best deal on the internet before it s too late! Get Your Password Now! 18 U.S.C. 2257 Record-Keeping Requirements Compliance Statement


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Robert Twomey (b. 1979, Washington, DC) is an artist exploring the poetic intersection of human and machine perception. Trained as a painter and engineer, he integrates traditional forms with new technologies. He has created a computer simulation of a grandmother with Alzheimer's disease, a body of work exploring the fantasy of an imaginary daughter, and recreated John Searle's Chinese Room as an exchange between synthetic child voice and robotic child drawing.
Twomey has worked in a range of research centers throughout his career, notably the Center for the Study of Learning, the Center for Research in Computing and the Arts, and the Experimental Design Lab with Natalie Jeremijenko. He has used neuroimaging to explore the basis of reading and language perception, built pollution-hunting Feral Robotic Dogs, created experimental game interfaces and 4K stereo animation, and is completing his dissertation on the home, A Machine for Living In, at the Center for Digital Arts and Experimental Media at the University of Washington.
Twomey is currently an Assistant Professor in Digital Media at Youngstown State University. He received his BS from Yale University with majors in Art and Biomedical Engineering, his MFA from the University of California, San Diego, and is a PhD Candidate in Digital Arts and Experimental Media at the University of Washington. He has shown his work in the US and abroad.
rtwomey [AT] ysu [DOT] edu


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Panel / Chassis Mount Sensor Multiplexers Datasheets
Multipoint Paperless Videographic Recorder -- SM3000
from ABB Measurement & Analytics
from ABB Measurement & Analytics
Award-winning technology from ABB. The SM3000 Multipoint Videographic Recorder features state-of-the-art data storage and security technologies. Up to 36 universal analog inputs, communicated inputs or math results can be recorded and displayed in a variety of operator views. 8Mb of onboard flash... [See More]
- Form Factor: Panel / Chassis Mount
- Digital I/O Channels: 6
- Analog Channels: 36
- Accuracy: 0.1000
16 or 32-Channel Relay Multiplexer -- AM16/32B
from Campbell Scientific, Inc.
from Campbell Scientific, Inc.
The AM16/32B has replaced the AM16/32A. The AM16/32B supports two clocking modes: Mode A works like the AM16/32A and sequentially advances through each channel. Mode B uses a relay address to go directly to a specific channel —reducing power consumption and wear on the relay switches. The new... [See More]
- Form Factor: Panel / Chassis Mount
- Excitation: Voltage
- Analog Channels: 32
- User Interface: None
Paperless Videographic Recorder -- DXAdvanced DX1000/DX1000N/DX2000
from Yokogawa Corporation of America
from Yokogawa Corporation of America
Introducing R3, the next stage in the evolution of DXAdvanced, Paperless Videographic Recorders [See More]
- Form Factor: Panel / Chassis Mount; Rack Mount (optional feature); Stand-alone; Bench Top
- DC Voltage: 60 V
- Analog Channels: 48
- DC Current: mA with Resistor


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Vogue 8655; ©1955; ONE PIECE SLACKS SUIT AND CUMMERBUND. Blouse with notched collar joins the slacks with an inset band at waist-line. Buckled front belt is joined at the underarm. Slacks are long and tapered or below the knee length. Fly front closing with zipper. Long sleeves gathered to buttoned band cuffs, short sleeves or sleeveless. Bias cummerbund is gathered at sides.
Links to reviews/blog postsEdit
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Poolenalena Beach, Maui
This beach doesn't tend to get nearly as crowded as others in the area, making it a sunbather's dream - take your pick from endless sunny spots!
If you want an extra secluded piece of sand to yourself, pick your way north along the beach, past the large gravel parking lot, and past a series of rocky outcroppings. You'll pass one mini-beach after another and they are all beautiful!
The further you go, the more secluded it gets. My 5 year old son and I were exploring here to see how far we could walk and ended up getting views that we never anticipated. We soon discovered that it gets so secluded that some folks use these parts for nude sunbathing. Just figured I'd let you in on that little secret so you're not caught off guard as I was!
It's nothing like Little Beach mind you (a nearby unofficial nude beach)... the small number of people who decided to forgo their bathing suits were more discreet here, staying well back from the water line - my son never even noticed them.
If you come in the morning when the water tends to be calm, you can snorkel near the rocky points at the north end of the beach, and also at the south end near the Makena Surf.
Calm waters make for great swimming too!
In the afternoon, the winds picked up and so did the waves. We took full of advantage of this and had a ball boogie boarding for hours! Fun, fun, fun! I know I've said this before, but I just HAVE to say it again: if you haven't tried boogie boarding yet, you really should try it, it's so much fun and it's super easy to learn on your own just by trial and error!
At the south end of the beach, follow the pretty hibiscus-lined path that winds it's way through the Makena Surf resort.
Alongside this walkway you'll find the stone ruins of a fishing heiau that is sacred to the Hawaiians. The area is considered KAPU (forbidden), so you can't walk through the ruins, but it's beautiful to see from afar all the same.
There are no lifeguards here. Signs warn of the potential for strong currents and a dangerous shorebreak.
If you enter the beach from the large gravel parking lot, resist the temptation to walk barefoot from your car to the beach. There are lots of kiawe trees here that you'll have to walk past on your way to the beach, and these trees have a nasty habit of dropping large thorns onto the ground below. The thorns can even pierce through the bottom of your flip flops, so please tread carefully.
There are showers near the Makena Surf. There are port-a-potties in the gravel parking lot at the north end of the beach.
The beach park is open from 7 a.m. until 7 p.m. daily.
To get here from Wailea, head south down Wailea Alanui Drive. It will change names to Makena Alanui Drive as you pass the sign for the Wailea Golf Club. The turn-off for the beach park will be on your right.
There is a large, free gravel parking lot at the north end of the beach with room for tons of cars - look for the yellow gates on the makai (ocean side) of Makena Alanui Road.
Alternatively, you can drive a little further south where you'll find 9 free stalls at the Makena Surf. You'll see a blue beach access sign, yellow gates, and cars parked in the lot parallel with the road. Once you park at the Makena Surf, you'll have to walk through the metal gate at the left side of the lot. (If you've passed the intersection with Makena Road, you went too far.)
A sign on the gate says it will remain unlocked from 7 a.m. until 20 minutes past sunset daily. From here, just follow the paved path to the ocean!
If you stay until the sun goes down, you'll be treated to a gorgeous view. The islands of Kahoolawe and Molokini are silhouetted against the glow, the waves sparkle in the dimming light...
As we watched the sunset, we were even treated to the sight of a couple getting married here - the sound of the conch shell sounded to mark the ceremony, the waves crashed, the palm trees rustled... pure magic!
The Makena Surf Resort is a highly rated resort property that backs onto Poolenalena Beach.
The grounds are lush and beautiful - you can even pick your own bananas and other fruit from gardens maintained by the staff! How cool!
Go to Homepage from Poolenalena Beach, Maui
Please excuse me while I break my site! :)
I'm moving things around a bit, so you may notice some wonky-looking pages until I'm done! Sorry for the mess!
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11 Replies
Jun 19, 2012 at 11:29 UTC
We were recently compromised, not largely, but still compromised. One of our 'managers' who insist on having elevated priviledges, was browsing for a bit of info and downloaded a compromised PDF file. This in turn compromised our mail server and opened up a SPAM session apparently originating from the mail server itself.
Our ISP blocked outgoing emails from our domain until I could get this under control. Fifteen hours of scouring servers (only 7 of them) and cleaning the resultant mess.
We have a UTM firewall and enterprise antivirus and it still got through.
Fortunately it only generated SPAM and did not rob us of anything else apart from email access.
We are a small electrical contracting firm with less than 60 staff.
It does happen. Hopefully not to you.
Best of luck.
Jun 20, 2012 at 12:21 UTC
Thank you for sharing that with us! I can take this to the powers that be, for sure. I could use more like what you wrote up.
I SHOULD say that I have pretty much exhausted Internet searching for "Small business hacked", "small business breach, small business siem, etc. I DID get some fairly good information to add to MY knowledge base, but nothing really convincing to upper management.
What are the chances, they'll argue. What are the risks? Is the price of an appliance worth the the insurance factor?
An article I read called UTM Firewalls and UTM Anti-virus at the desktops, Legacy. And the more I think about it, THEY ARE! How many of us REALLY tighten up the firewall rules to restrict what goes out? How many users acknowledge all updates and changes to applications when the anti virus software asks them to allow, blindly clicking ALLOW?
Without network flows and sampling of packets inbound AND outbound, we never know how the traffic correlates to a new "update". Sure port 113 is "auth", but what if it is REALLY a telnet session or FTP in disguise running on a port that sometimes needs to be open for SMTP to function properly, i.e. Comcast, Verizon, and other e-mail client/server activity.
So, is there a list of companies posted somewhere of recent compromise? ftc.org, Verizon security site, securelist.com, fcc.org? I found an article about a small ma and pop convenience store that got hacked where credit card info was stolen and abused. Another story regarding an engineering and construction company, Golden State Bridge in Calif was robbed of more than $125,000 when cybercriminals hacked into it's bank account. This one I'll use ! There is a real name associated with the breach that lends credibility.
I think we as I.T. pro's in SMB shops need ammo to push forward with our objective to secure the network we've built and defend. I know our shop is tight and there is no way the company wants to hire a dedicated security person. A SIEM, while not a SIO (Security Information Officer) in a box, it sure beets going through logs and trying to correlate activities, which is not possible "real time", SIEM's can do it real time and alert as soon as there is a questionable event!
Done MY research, now to get more compelling evidence for upper management.
Thank you much, if you are still following along...
Jun 20, 2012 at 3:23 UTC
Here's a nice article on security.
Also You can check on stories about security in the spotlight section of community.
Hope that helps.
Jun 20, 2012 at 10:21 UTC
Part of what makes finding more information on this a bit of a challenge is that many smaller companies would not actually know they've been compromised. For other companies that have been compromised, most wouldn't admit to it unless it were required by law.
Jun 20, 2012 at 11:20 UTC
Thank you Riga,
Good basic ground rules we follow and have put into practice. Layered.defense.
Jun 20, 2012 at 11:25 UTC
Good points! Either ignorance is bliss, or "I don't want anyone to know". When I get a chance I will post links to articles I have found that help to show the need for small business to adopt yet another layer to work together with the legacy security tools we've been using for years and the organized criminal groups know all to well how to circumvent.
Thanks again,
Jun 20, 2012 at 11:41 UTC
First thought is "insider threats" from the innocent clicking on an malware link in an email, to a manager bringing in his home wireless router, to a board member siphoning confidential company documents before jumping to a competitor.
What would be great is a list of companies negatively impacted, and how. Companies, of course, do not want such information public. Can think of one egregious case: WikiLeaks.
CERT has a section for the threat vector:
Book on the topic, but may not be readily available:
- Insider Threats in Cyber Security
/ rd.springer.com/ book/ 10.1007/ 978-1-4419-7133-3/ page/ 1#page-1
Jun 20, 2012 at 5:38 UTC
HHS Breach Notification list:
It is a HIPAA list, which requires public notification. State to state notification level varies. Would be a good list to emphasize the enormous security risk.
Jun 22, 2012 at 3:48 UTC
Hello Rogue Packet,
Good stuff! CERT, of course!
HHS Breach Notification List, good info, but definitely a target group. Now, if we could find a repository of SMB's in this same format, that would be cool, but I agree with you that companies do NOT want thier breaches to become public knowledge for fear of negative exposure and customers, partners scrutiny of their ability to take care of their own business.
So, what have I found?
Google "small business breach" brings tons of articles, of course.
The two links below I think are the best resource I found, giving statistics that upper management wants to see and know of.
(Choose to watch "The Triple Threat to Today's Cybercriminals")
(The above Requires registration)
Hackers Not Attracted to Small Businesses? False | Fox Small Business Center
Cyber security not a serious matter for small businesses, according to The Hartford
Why Hackers Target Small Businesses Who Use Macs, iPads & iPhones | Cult of Mac
Cybersecurity for Small Business | FCC.gov
GFI Software Survey: 40% of SMBs Have Suffered a Security Breach Due to Unsafe Web Surfing | EON: Enhanced Online News
Local techie: Small businesses more vulnerable to online hackers -- fox59.com
What's Next For IBM Security? Q1Labs- Security - Security administration/management - Informationweek
SMB Security Monitoring Gaining Sophistication - Security - Security administration/management - Informationweek
SMBs in the Crosshairs - Imperva Data Security Blog
The Top Five Ways Attackers Target Small Businesses -- And What You Can Do About It - Dark Reading
!SMBs may not be as secure as you think : SecurityNewsPortal.com for the latest computer network Internet security and anti virus news
SMBs in the Crosshairs- Computer Network and Internet Security News Resource Portal
Phishing, P2P, Drive-Bys Top SMB Security Risk List - Smb - Business/E-Business - Informationweek
Thanks again Guys. I think I have what I need. I hope this will help others get what they need to get to the third tier of protection; Firewall, Anti-virus and SIEM.
I want to know if the first two are doing their job, As I see it, SIEM is the only way to know for sure.
Jun 22, 2012 at 9:46 UTC
....Now, if we could find a repository of SMB's in this same format, that would be cool....
Thanks for the links!
The topic is a mess legislatively and remains by design at the state level:
/ www.computerworld.com/ s/ article/ 9218554/ House_panel_approves_data_breach_notification_bill
/ community.spiceworks.com/ topic/ 237241-us-senate-proposes-national-data-breach-notification-act
California has SB 1386, but the language is for certain types of breaches ("credit card breach notification" gets attention). Doesn't specific a public list from what can be seen (or, yet). SANS Reading Room could be a resource.
Issue is "risk level" and "financial liability", and associated comfort level. SIEM is aimed as a mitigation factor -- knowing what is happening sooner to be able to react to it. Would be looking for numbers -- frequency, penalties, et al. Loss of reputation is the nebulous one hard to pin down and attribute a cost -- say, Global Payments' breach or Heartland's breach.
If the buy-in isn't what is desired, it can be a report/memo to pass to the decision makers (board?) as a budget item. Refresh it next year, and re-submit. In this way it is an awareness and education item.
Jun 24, 2012 at 7:10 UTC
Rogue Packet,
Thanks again for your input and links. Yes, the issue IS risk and liability. No one/no business wants to spend more than they have to, regarding security, that is a given. I feel that the standard is changing and we DO need to look at something beyond firewalls and anti-virus.
DLP is becoming a strong player in the security market now also, and is more affordable than SIEM. DLP will not tell us what is inside trying to get out as well as SIEM, but it will watch for out bound proprietary information leakage and may cover a good sized portion of what we desire SIEM to do, depending on the rules you create for your product and what you aim to accomplish.
Personally, I like having a second means to determine whether or not my UTM firewall and UTM anti-virus are working as advertised or are they letting bad stuff happen and slip through the cracks. Many times an assortment of anti-virus is used within and organization for this purpose.
I think I will back down now in this thread. I don't suppose it will go much further. I DO thank you all for your participation. It will be a while yet before I know whether or not our company will adopt SIEM. Thought I had it two weeks ago, but found out it was NOT an approved line item in our budget. ARGH!
This discussion has been inactive for over a year.
You may get a better answer to your question by starting a new discussion.


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Hello I have been swarming these forums this whole week after coming back for information and i am very grateful for all the help ^^.
Anyways I have a question about armor( one of the things that ticked me of in beta also )
-where do you purchase armor int his game? All i see are level 1 starting weapons and armor level 1 which you get your own free ones anyways when you begin. This was one of the things that specially ticked me off when i was trying out beta have they added armor and weapon shops in town yet? Such as final fantasy 11 or almost 100% of all rpg/mmorpg which at the very least sells starting equipment? I hope I am wrong about this, and hope you don't have to be forced to make everything for yourself.
-The ability to equip any armor at any level for any class sounds weird as **** also, whats up with this? does this mean i can wear a uber level 50 armor on a level 1 guy @.@ even with penalty this seems flawed.
Thank you ahead of time ^^


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This 10 yard ribbon is a fuzzy pom pom. The wire allows you to bend and shape your creations and decorate with ease. These pom poms would add a fun touch to party décor and holiday DIY project. Try it on an altered gift box or make a fun centerpiece with it.
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The hotel was clean and the breakfast was satisfactory, but our room was only OK. There was no fan in the bathroom and it felt as though there were some ventilation issues in the room generally, which was really stuffy. That said, our beds were very comfortable and the staff was friendly. I wouldn't make this my first choice but for an inexpensive stay in Banff, there are worse properties.
- Official Description (provided by the hotel):
- - Located on Banff Avenue, one short block from downtown Banff- Includes Full buffet breakfast with a made to order omelette station- Indoor hot tubs, sauna, steam and fitness equipment- The Meatball Pizza and Pasta- 134 cozy hotel rooms- The Sundance and Atrium function rooms for small to mid-size - meetings/receptions- Complimentary wireless internet access in all guest rooms- Heated underground parkade with free parking (first come basis)- Complimentary Roam Bus passes for the Banff transit system. ... more less
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Stolen specialized rockhopper
Posted by Stolen Bike Listings on September 21st, 2006 at 3:49 pm
rockhopper written across front bar
Posted by Stolen Bike Listings on September 21st, 2006 at 3:49 pm
rockhopper written across front bar
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It came on the heels of "Hair." For the first four years we were just a summer theater. After the success of "Hair," we asked how we could keep the momentum going. We thought a fall production would be a fun addition. I tried to think of something cool to do at Serenbe. I looked at the stables here. The Headless Horseman came to mind and that is how "Sleepy Hollow" was birthed. We thought we'd do it and see how it went. It literally sold out before we even opened. We knew we had to make it good. We included live horses and all the actors playing instruments. It felt old world meets modern world.
What's new this season?
The stables are great but there are only so many people you can fit in a stable. Last year, we moved it into an open field venue. Adam Koch designed a set in a field and came up with a skeleton version of the town.
This year I am using a new script which I have adapted. The script uses new music and it becomes our version. I have the rights to the script. Now I can tweak it in any way. I am also going to be doing it in Massachusetts, but every version will be unique.
Why has the story held up?
People ask that all the time. There is something about the legend that transcends all ages. It really is the most iconic Halloween story of all time. For visuals, the visual of a Headless Horseman looking for a head is pretty terrifying and iconic. There are so many versions, films and cartoons, a TV show. It's scary but not gory scary. Everyone has their own version. It's become part of Halloween.
'The Sleepy Hollow Experience' What are your crowds like?
Very diverse. Our audiences do skew younger for shows where you are on your feet. This is a mix of theatergoers, people who go to haunted houses and the typical Serenbe audience coming for the experience. People become immersed in the environment and they tend to come every year because it's now part of their tradition. We are seeing new people come as well.
Are most people familiar with the story?
I think people have an understanding of what it is. What's interesting here is that you get some background story. Like, where did Ichabod come from? How did the town get its name? I think people enjoy that.
What is the gay appeal?
I feel like Halloween has almost become a gay holiday. The holidays the gays have embraced. We see so many LGBT audience members, especially for these immersive shows. This show celebrates the fall, and Halloween. There is plenty of great eye candy in the show too. That should get people in the door.
"The Sleep Hollow Experience" is playing at Serenbe Playhouse through Nov. 6. (Publicity photo)


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It takes a very rare skill to transform the beauty of Old Europe into a table as exquisite as this. The Queen Anne Coffee Table excels in showcasing fine details - including its turned, curved legs, its hand-finished surfaces and seamless drawer - and bringing them together into a final design that comes to life with a beauty greater than the sums of its parts. Whether providing the focal point for the living room, the reception, or the office, the Queen Anne coffee table offers decades of true versatility and refinement.
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is it possible to upload previous forms to the dropbox for archive etc.Asked by MJ on April 28, 2015 at 02:25 PM
is it possible to upload previous forms to the dropbox for archive etc...
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Passover Brownie Cake
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I experimented for years to come up with a brownie made with matzoh… this is the best!
1 In a large mixing bowl, beat the eggs and the sugar together. Add the oil. Mix in the cake meal and the cocoa.
2 Add nuts, or use the nuts as a topping. I put nuts on top rather than in this cake for two reasons. They make the top look good without having to frost it. Also, for those who don 1t or can 1t eat nuts, they are easily removed without having to search for them.
3 Pour batter into a greased 9 x 13 inch pan. Bake at 325 degrees F (165 degrees C) for about 35 minutes. The secret to a moist brownie cake, Pesach or otherwise, is to underbake it. Check it at about 30 minutes.


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A gene-rich, transcriptionally active environment and the pre-deposition of repressive marks are predictive of susceptibility to KRAB/KAP1-mediated silencing
KRAB-ZFPs (Krüppel-associated box domain-zinc finger proteins) are vertebrate-restricted transcriptional repressors encoded in the hundreds by the mouse and human genomes. They act via an essential cofactor, KAP1, which recruits effectors responsible for the formation of facultative heterochromatin. We have recently shown that KRAB/KAP1 can mediate long-range transcriptional repression through heterochromatin spreading, but also demonstrated that this process is at times countered by endogenous influences.
To investigate this issue further we used an ectopic KRAB-based repressor. This system allowed us to tether KRAB/KAP1 to hundreds of euchromatic sites within genes, and to record its impact on gene expression. We then correlated this KRAB/KAP1-mediated transcriptional effect to pre-existing genomic and chromatin structures to identify specific characteristics making a gene susceptible to repression.
We found that genes that were susceptible to KRAB/KAP1-mediated silencing carried higher levels of repressive histone marks both at the promoter and over the transcribed region than genes that were insensitive. In parallel, we found a high enrichment in euchromatic marks within both the close and more distant environment of these genes.
Together, these data indicate that high levels of gene activity in the genomic environment and the pre-deposition of repressive histone marks within a gene increase its susceptibility to KRAB/KAP1-mediated repression.
Gene expression is modulated through the alteration of chromatin states by epigenetic regulators. Krüppel-associated box zinc finger proteins (KRAB-ZFPs), which together constitute the single largest group of transcriptional repressors encoded by the human genome, partake in this process
Using chromatin immunoprecipitation (ChIP) and a tiling array, KAP1 has been documented to bind more than 7000 sites in a human testicular embryonal carcinoma cell line
Characterization of thousands of KRAB/KAP1-targeted gene traps
To study the impact of specific genomic features on KRAB/KAP1-induced silencing, we used the recently described trapping/silencing (TrapSil) system
Figure 1
Isolation of thousands of KRAB/KAP1 recruitment sites in genes with variable silencing phenotypes
Isolation of thousands of KRAB/KAP1 recruitment sites in genes with variable silencing phenotypes. (A) Experimental setup used to target tTRKRAB to endogenous genes through the retroviral vector-based promoter trapping/silencing (TrapSil) system.
Since we were interested in elucidating differences between KRAB/KAP1 repressible and non-repressible promoters and genes, we reasoned that "all or none" phenotypes would facilitate subsequent analyses. Therefore, we selected cells in which trapped promoters were highly active at baseline, and either strongly repressed ("repressed clones" containing a "repressing integrant") or almost completely resistant to this process ("non-repressed clones" containing a "non-repressing integrant") when the trans-repressor was allowed to bind its target (Figure
After the isolation of cell populations with differential silencing phenotypes, we mapped proviral integration sites, in order to identify the trapped genes. For this, we combined linker-mediated PCR (LM-PCR) of proviral-genomic junctions with massive parallel DNA pyrosequencing
Additional file 1
List of mapped integrants and integrant gene groups.
Click here for file
Prior to further analysis, we validated our experimental approach by deriving clones from each population. All of the 32 clones analyzed exhibited the expected silencing profile in flow cytometry measurements. Moreover, the clones comprised 10 non-repressed (LI I-X) and 8 repressed (LR I-VIII) LV-TrapSil clones, in addition to 8 non-repressed (MI I-VIII) and 6 repressed (MR I-VI) MLV-TrapSil clones, (Additional File
Additional file 2
KRAB/KAP1 repression profiles from clones isolated from repressible and non-repressible TrapSil populations. Reporter gene activity in the presence and absence of doxycycline (Dox) was monitored in clones derived from repressible and non-repressible TrapSil populations.
Click here for file
Additional file 3
KAP1 and H3K9me3 are present at non-repressible clones in the presence of repressor binding. Chromatin-immunoprecipitation (ChIP) in combination with qPCR was used to verify proper KAP1 recruitment to gene traps in the absence of doxycycline in non-repressible clones. In addition, levels of H3K9me3 monitored the enzymatic activity of the KRAB/KAP1 silencing complex. The relative enrichment was calculated as a percentage of the total input (% of total input).
Click here for file
Genomic environment of repressing and non-repressing gene trap integrants
We characterized the genomic environment of the integrants segregated according to their phenotype by using ROC (Receiver Operator Characteristic) curve analysis
In order to characterize the genomic features surrounding the integrants in our different TrapSil groups, we made use of the same approach. We first calculated the area under the ROC curve, which is a common measure of a predictor variable's ability to discriminate between two classes of events. In our case we compared the average enrichment of a given feature at a set of genomic sites (such as integration sites in our case) relative to that of a set of random matched control sites. The read-out of this comparison is illustrated in color-coded heatmaps, where each rectangle represents the specific enrichment of a feature within the indicated intervals of distance. The relative enrichment between the integration and control site group is scored on a scale from 0 to 1. 1 is scored when a specific feature is enriched in the experimental integrants when compared to matched controls, 0 is scored when the opposite is true. A value of 0.5 indicates no difference between the two groups. The patterns of the genomic features surrounding the TrapSil integrant groups largely reflected the preferential genomic environment associated with either LV or MLV integrations (for values see Additional File
Additional file 4
Values from Receiver Operator Characteristic (ROC) curve analysis.
Click here for file
Figure 2
Characterization of the genomic environment of KRAB/KAP1-docking integrants
Characterization of the genomic environment of KRAB/KAP1-docking integrants. LV and MLV TrapSil integrants were split in repressing (REP) and non-repressing (NREP) groups according to the effect of KRAB/KAP1 recruitment on the trapped promoters. The genomic environment of the different proviral integrant groups was analyzed for the indicated genomic features by ROC curve analysis. This method serves to calculate the relative abundance of a given genomic feature around the integrants of a group for specific intervals. The resulting values are depicted in color-coded heat maps. 1 indicates that the specific feature is enriched in integrants, 0 means that it is depleted. Relative abundance scores of repressing and non-repressing integrants were compared for each trapping vector subtype and the statistical method used included the non-central chi-square test (** p < 0.01; *** p < 0.001). The different genomic feature categories tested were: "gene density", with all of the Refseq annotated genes; the "highly expressed genes" and "expressed genes" group, including genes expressed in the top 1/16th, or the top 1/2 of all genes measured in a transcriptional profiling analysis;
We then compared the ROC values, which are proportional to the levels of genomic features at these sites, between TrapSil groups harboring differential susceptibilities to KRAB/KAP1-silencing. We did this by making relative comparisons between a chosen reference and other gene groups. The reference gene groups are indicated by the symbol "---" within the whole results section. Using this approach we compared the levels of specific genomic features between the respective REP gene group and their corresponding NREP counterpart. When statistical differences were assessed, we found that LV-TrapSil repressing integrants were located within gene-denser genomic regions than non-repressible integrants (Figure
Genomic features of matched repressed and non-repressed transcriptional units
Repressing integrants were on average closer to the transcriptional start site of their targets, compared to non-repressible integrants (Figure
Figure 3
Generation and characterization of matched gene groups displaying differential KRAB/KAP1 silencing phenotypes
Generation and characterization of matched gene groups displaying differential KRAB/KAP1 silencing phenotypes. (A) Matched gene groups with differential KRAB/KAP1-silencing phenotypes. The cumulative histograms illustrate the distribution of repressible (blue) and non-repressible (red) LV-TrapSil and MLV-TrapSil integrants in the transcribed region of genes trapped multiple times. Three groups were distinguished based on the pattern of repressible and non-repressible integrants over 20 kb. Group 1 genes harbored mainly repressing integrants ("long-range repression"), while group 3 contained mostly non-repressible integrants ("absence of repression"). Group 2 genes exhibited an intermediate phenotype, with repressing integrants clustered over the first 10 kb of their transcribed region ("limited repression"). (B) The three gene groups (group 1: "long-range repression", group 2: "limited repression" and group 3: "absence of repression") were analyzed by ROC curves as described in Figure 2 for genomic features over various DNA stretches. (C) Comparison of mean expression levels of the three gene groups based on publicly available microarray data. Group 1: "long-range repression", group 2: "limited repression" and group 3: "absence of repression". Statistical comparisons were made with a non-parametric Wilcoxon test. P-Value Legend: ** p < 0.01.
The genomic context of the three gene groups was reminiscent of observations made in the integrant-centered analysis (Figure
Chromatin features of matched repressed and non-repressed transcriptional units
We first assessed the levels of putative barrier elements such as CTCF, H3.3/H2Az or chromatin modifiers in the different groups
Additional file 5
Heterochromatin barrier elements in the environment of the matched gene groups. The relative level of barrier elements was assayed in the three gene groups (group 1: "long-range repression", group 2: "limited repression" and group 3: "absence of repression"). Calculations were based on ROC curves as described in Figure 3 and included various DNA stretches around the promoters of the different gene groups. Non-central chi square statistical analysis indicated no differences between the groups. The features considered in the analysis were (A) Histone variants associated with active remodeling (H3.3, H2Az), (B) CTCF or (C) chromatin modifiers, such as the histone acetyltransferases p300, TIP60, PCAF and MOF, in addition to the histone deacetylases HDAC1, 2, 3, and 6. The genome-wide binding data for these factors came from published work in HeLa cells (H3.3, H2Az, p300) or CD4 cells (TIP60, PCAF, MOF, HDAC1, 2, 3 and 6).
Click here for file
We then assessed the abundance of a series of histone modifications present at an interval of 1, 10 or 100 kb or 1 Mb around our promoters of interest. We first measured the levels of posttranslational histone modifications correlating with active gene expression, including histone H3 lysine 27 acetylation (H3K27ac), H2BK5 monomethylation (H2BK5me1), H3K4 mono- and trimethylation (H3K4me1, me3), H3K36me3 and H4K20me1. For this we generated genome-wide histone modification maps using a ChIP coupled to deep sequencing (ChIPseq) approach in HeLa cells. Then we used ROC curve-based heatmaps to obtain relative enrichment values for the three groups. When the long-range repressing group 1 was compared to the non-repressing group 3 most of the active histone modifications were enriched in group 1 (Figure
Figure 4
Characterization of the chromatin environments of the matched gene groups with respect to active histone marks
Characterization of the chromatin environments of the matched gene groups with respect to active histone marks. (A) The levels of specific posttranslational histone modifications around the promoters of the different gene groups were calculated over different DNA intervals ranging from 1 kb to 1 Mb. The gene groups included group 1: "long-range repression", group 2: "limited repression" and group 3: "absence of repression" and ROC curve analyses was employed. Briefly, the illustrated heat maps, contain squares giving the relative abundance of the studied histone mark. 1 is scored when the modification is enriched in the gene group when compared to a control group, while 0 indicates depletion of the mark. 0.5 is scored when there is no difference. Non-central chi-square statistical analysis compared differences between the repressible group 1 and the non-repressible group 3. P-Value Legend: * p < 0.05; ** p < 0.01. The histone modifications in the analysis included H3K27ac, H2BK5me1, H3K4me1, H3K4me3, H3K36me3, H4K20me1, which are mostly found within active chromatin. (B) Relevant histograms representing the relative H3K4me3 and H3K36me3 values over 100 kb and 1 Mb, which reached statistically relevant differences when all three gene groups were compared.
We then measured the levels of histone modifications normally present at silent genes, such as H3K9me2/me3, H3K27me3 and H4K20me3. For this we generated histone modification maps by ChIPseq in HeLa cells or relied on a published dataset for the distribution of H3K27me3 in this cell line
Figure 5
Characterization of the chromatin environments of the matched gene groups with respect to silent histone marks
Characterization of the chromatin environments of the matched gene groups with respect to silent histone marks. (A) The levels of histone marks associated with silent chromatin were analyzed at specific intervals around the promoters of the three gene groups (group 1: "long-range repression", group 2: "limited repression" and group 3: "absence of repression"). The calculations were based on ROC curve analysis as described in Figure 4. Non-central chi-square statistical analysis compared differences between group 1 and group 2, and group 1 and group 3. The p-Value Legend is * p < 0.05; ** p < 0.01. The histone modifications in the analysis included H3K9me2, H3K9me3, H3K27me3 and H4K20me3, mainly associated with silent chromatin. (B) Relevant histograms representing relative H3K9me3 and H4K20me3 values over 1 kb and 10 kb, which reached statistically relevant differences when the three groups were compared.
Previous analyses on the mechanisms of KRAB/KAP1-mediated gene regulation have mostly examined the impact of this system on the expression of transfected promoter-reporter units. Here, we investigated KRAB/KAP1-induced changes within the context of endogenous genes. Using a combination of promoter trapping and drug-controllable KRAB/KAP1 recruitment, we previously observed that this complex, when docked to the bodies of transcriptionally active genes, could induce silencing over distances of several tens of kilobases
Comparing KRAB/KAP1-repressed and non-repressed genes gave no indication for a role of putative obstacles to the spread of heterochromatin, such as CTCF binding, accumulation of H3.3/H2Az or recruitment of HATs (reviewed in
Although both repressed and non-repressed genes were situated within euchromatic regions, as expected from the promoter-trapping approach used for their selection, we observed significant differences in both their local and broader chromatin environments. Repressed genes were in regions containing generally higher levels of major euchromatin-associated marks and higher levels of transcription compared with non-repressed genes. Therefore, there is a positive correlation between efficient KRAB/KAP1-silencing and high gene activity. This is suggestive of a model whereby genes situated in more heterochromatic environments can only be highly expressed if endowed with an intrinsic ability to resist repressive influences, while genes located in more euchromatic environments do not need such protective mechanisms
Genes repressed by the TrapSil system also carried higher levels of the repressive marks H4K20me3 and H9K9me3 at baseline at and around their promoters, compared with their repression-resistant counterparts. Noteworthy, these contrasting chromatin configurations were not only observed when comparing a selected set of multiply hit repressed and non-repressed genes (Figure
Additional file 6
Characterization of the chromatin environment of all KRAB/KAP1-docking integrants. LV and MLV TrapSil integrants were split in repressing (REP) and non-repressing (NREP) groups according to the effect of KRAB/KAP1 recruitment on the trapped promoters. The chromatin environment of the different proviral integrant groups was analyzed for the indicated features by ROC curve analysis. (A) The histone modifications in the analysis included H3K27ac, H2BK5me1, H3K4me1, H3K4me3, H3K36me3, H4K20me1, which are mostly found within active chromatin. (B) The histone modifications in this analysis included H3K9me2, H3K9me3, H3K27me3 and H4K20me3, mainly associated with silent chromatin. Non-central chi-square statistical analysis compared differences between the integrant groups with the differential silencing phenotypes. P-Value Legend: * p < 0.05; ** p < 0.01; *** p < 0.001.
Click here for file
Additional file 7
Characterization of the chromatin state of KRAB-ZFP genes. Illustration of H3K9me3 and H3K36me3 enrichments at KRAB-ZFP gene bodies in HeLa cells. KRAB-ZFP gene lengths were equalized by division into 40 bins and included a 2 kb flanking region on both sides of the genes. See Additional File
Click here for file
Additional file 8
List of known and putative KRAB-ZFP genes included in the analysis.
Click here for file
A difference between genes targeted by our TrapSil system and endogenous KRAB-ZFP genes lays in the finding that the latter do not seem susceptible to KRAB/KAP1-mediated long-range repression
Other mechanisms potentially involved in conferring resistance to KRAB/KAP1-mediated silencing are suggested by the analysis of genes that were hotspots of proviral TrapSil targeting and carried both repressible and non-repressible integrants (Additional File
Additional file 9
Genes with multiple TrapSil integrants of different silencing phenotypes. Graphic representation of 6 genes, where multiple TrapSil integrations of different phenotypes have occurred: calnexin precursor (CANX), heterogeneous nuclear ribonucleoprotein A2/B1 (HNRNPA2B1), karyopherin beta 1 (KPNB1), nucleophosmin 1 (NPM1), Y-box binding protein 1 (YBX1), laminin alpha 5 (LAMA5). The integrants with a repressible phenotype are depicted as black lines above the baseline (y = 0), whereas the non-repressible counterparts are depicted as light grey lines below the baseline. Some intragenic sites carried multiple but distinct proviral integrants and subsequently harboured more than 1 integrant count (see y-axis).
Click here for file
In summary, the present work indicates that the impact of KRAB-mediated docking of KAP1 on the expression of targeted genes is more variable than previously suspected. It further reveals reciprocal influences between the functional outcome of KRAB/KAP1 recruitment to DNA and the chromatin features of the involved loci. More broadly, the approach described in the present study, which combined an analysis of the functional consequences of exogenously introduced
pLV-tTR-KRAB-Red was previously described
Additional file 10
List of primers used in this study.
Click here for file
Cell culture and Fluorescence activated cell sorting (FACS)
HeLa cells were grown under standard conditions. Doxycycline (Sigma-Aldrich) was used at a concentration of 1 μg/mL. Clonal tTRKRAB-expressing HeLa cell lines dsRK4 (pLV-tTR-KRAB-Red, LV-backbone) and KiN1.25 (pLTetR-KRAB-NG95, MLV-backbone) were derived after infection with pLV-tTR-KRAB-Red or pTetR-KRAB-NG95, respectively. The LV based HeLa dsRK4 clone contains approx. 15 vector copies as titrated by Taqman and was used for MLV-TRAPSIL assays while the MLV-based KiN1.25 clone contains 10 vector copies and was used for all LV-TRAPSIL assays. In view of this mapping strategy, 2 × 108 dsRK4 or KiN1.25 HeLa cells were infected with 1.6 × 106 MLV-TrapSil or LV-TrapSil infectious particles, respectively, with a multiplicity of infection of 0.04. Cells were sorted based for GFP expression by using the Beckton Dickinson FACSVantage SE turbo Sorter with Diva Option. Flow Cytometry analyses were performed on BD FACScan flow cytometer.
Quantitative PCR (qPCR)
qPCR reactions were carried out with a standard PCR program in ABI PRISM 7900 HT in duplicates or triplicate using either SYBR green detection 1× Power Sybr or 1× Taqman Universal Mix, No AmpErase (Applied Biosystems). Primers were used at a final concentration of 100 nM. When SYBR analysis was performed, cycling reactions were followed by a dissociation curve analysis to validate specificity of amplified products. The increase in fluorescence was analyzed with the SDS software, version 2.2.2 (Applied Biosystems). For all amplification plots the baseline data were set with the automatic cycle threshold function. Primer sequences for all qPCR reactions are listed in Additional File
Linker-mediated PCR (LM-PCR), 454 pyrosequencing and data processing
LM-PCR was used to map integration sites following a previously described protocol
Integration site mapping in genes: Integrant orientation was annotated as determined during sequence processing. UCSC known gene
Receiver Operator Characteristic (ROC) curve analysis
Data analysis was based on a "nested case control" strategy using a collection of TSS characterized by a given behavior with respect to repression along with control sites sampled from the genome to make inferences about the probability of a TSS to display a given response to repression based on genomic/epigenetic features characterizing its environment. More detailed description of statistical basis for this analysis can be found in
Chromatin immunoprecipitation (ChIP) and ChIP followed by sequencing (ChIP-Seq)
ChIP reactions were performed according to published protocols with minor modifications (
Additional file 11
List of antibodies used in this study.
Click here for file
The sequencing libraries from all ChIP products were prepared using the ChIP-seq Sample Preparation Kit (Illumina; San Diego, California, USA; Cat. No. IP-102-1001) according to the protocol supplied with the reagents and using 10 ng of ChIP sample quantified using the Qubit fluorometer (Invitrogen; Carlsbad, California, USA). One lane of each library was sequenced on the Illumina Genome Analyzer IIx using the Single-Read Cluster Generation Kit v2 (Cat. No. FC-103-2001) and 36 Cycle Sequencing Kits v3 and v4 (Cat. Nos. FC-104-3002 and FC-104-4002). Data were processed using the Illumina Pipeline Software v1.5.1. Illumina GAII data were mapped to the genome with Bowtie
Microarray data
HU133a arrays for HeLa cells were downloaded from GEO/NCBI
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
Conceived and designed the experiments: SM, ACG, DT. Performed the experiments and computational analyses: SM, ACG, GA, NM, NZ, AK, AK, JR. Contributed reagents/materials/analysis tools: SQ, AC, FDB, PB. Wrote the paper: SM, ACG, DT. All authors have read and approved the final manuscript.
We thank Joanna Roberts and Miguel Garcia for the FACS analyses, and Keith Harshman and colleagues for help with the DNA sequencing. This work was supported by grants from the Swiss National Science Foundation, the Strauss Foundation, the European Union (PERSIST program) and the Infectigen Association to DT, by an MD-PhD scholarship from the Swiss National Science Foundation to SM, and by NIH grants AI52845 and AI082020, by the University of Pennsylvania Center for AIDS Research and by the Penn Genome Frontiers Institute through a grant from the Pennsylvania Department of Health to F.D.B.
KRAB-containing zinc-finger repressor proteins
SysZNF: the C2H2 zinc finger gene database
Adaptive evolution in zinc finger transcription factors
A comprehensive catalog of human KRAB-associated zinc finger genes: insights into the evolutionary history of a large family of transcriptional repressors
Evolution of C2H2-zinc finger genes and subfamilies in mammals: species-specific duplication and loss of clusters, genes and effector domains
A census of human transcription factors: function, expression and evolution
The ancient mammalian KRAB zinc finger gene cluster on human chromosome 8q24.3 illustrates principles of C2H2 zinc finger evolution associated with unique expression profiles in human tissues
Sequence-specific transcriptional corepressor function for BRCA1 through a novel zinc finger protein, ZBRK1
A maternal-zygotic effect gene, Zfp57, maintains both maternal and paternal imprints
KRAB-type zinc-finger protein Apak specifically regulates p53-dependent apoptosis
Kruppel-associated boxes are potent transcriptional repression domains
KAP-1, a novel corepressor for the highly conserved KRAB repression domain
A novel member of the RING finger family, KRIP-1, associates with the KRAB-A transcriptional repressor domain of zinc finger proteins
Transcriptional repression by RING finger protein TIF1 beta that interacts with the KRAB repressor domain of KOX1
A possible involvement of TIF1 alpha and TIF1 beta in the epigenetic control of transcription by nuclear receptors
KAP-1 corepressor protein interacts and colocalizes with heterochromatic and euchromatic HP1 proteins: a potential role for Kruppel-associated box-zinc finger proteins in heterochromatin-mediated gene silencing
A novel nuclear receptor corepressor complex, N-CoR, contains components of the mammalian SWI/SNF complex and the corepressor KAP-1
Reconstitution of the KRAB-KAP-1 repressor complex: a model system for defining the molecular anatomy of RING-B box-coiled-coil domain-mediated protein-protein interactions
Targeting histone deacetylase complexes via KRAB-zinc finger proteins: the PHD and bromodomains of KAP-1 form a cooperative unit that recruits a novel isoform of the Mi-2alpha subunit of NuRD
SETDB1: a novel KAP-1-associated histone H3, lysine 9-specific methyltransferase that contributes to HP1-mediated silencing of euchromatic genes by KRAB zinc-finger proteins
Association of the transcriptional corepressor TIF1beta with heterochromatin protein 1 (HP1): an essential role for progression through differentiation
The Kruppel-associated box repressor domain can trigger de novo promoter methylation during mouse early embryogenesis
Regulated recruitment of HP1 to a euchromatic gene induces mitotically heritable, epigenetic gene silencing: a mammalian cell culture model of gene variegation
The KAP1 corepressor functions to coordinate the assembly of de novo HP1-demarcated microenvironments of heterochromatin required for KRAB zinc finger protein-mediated transcriptional repression
Genome-wide analysis of KAP1 binding suggests autoregulation of KRAB-ZNFs
Functional analysis of KAP1 genomic recruitment
The HP1alpha-CAF1-SetDB1-containing complex provides H3K9me1 for Suv39-mediated K9me3 in pericentric heterochromatin
KRAB-zinc finger proteins and KAP1 can mediate long-range transcriptional repression through heterochromatin spreading
Tetracycline-reversible silencing of eukaryotic promoters
Silencing of RNA polymerases II and III-dependent transcription by the KRAB protein domain of KOX1, a Kruppel-type zinc finger factor
HIV-1 integration in the human genome favors active genes and local hotspots
Retroviral DNA integration: ASLV, HIV, and MLV show distinct target site preferences
Genome sequencing in microfabricated high-density picolitre reactors
HIV integration site selection: analysis by massively parallel pyrosequencing reveals association with epigenetic modifications
Indexing strategies for rapid searches of short words in genome sequences
Retroviral DNA integration: viral and cellular determinants of target-site selection
Promoter trapping reveals significant differences in integration site selection between MLV and HIV vectors in primary hematopoietic cells
Selection of target sites for mobile DNA integration in the human genome
Global analysis of the insulator binding protein CTCF in chromatin barrier regions reveals demarcation of active and repressive domains
H3.3/H2A.Z double variant-containing nucleosomes mark 'nucleosome-free regions' of active promoters and other regulatory regions
Genome-wide mapping of HATs and HDACs reveals distinct functions in active and inactive genes
Insulators: many functions, many mechanisms
Insulators: exploiting transcriptional and epigenetic mechanisms
Position-effect protection and enhancer blocking by the chicken beta-globin insulator are separable activities
Heterochromatin protein 1: don't judge the book by its cover!
Characterization of the contradictory chromatin signatures at the 3' exons of zinc finger genes
ZNF274 recruits the histone methyltransferase SETDB1 to the 3' ends of ZNF genes
PHF8, a gene associated with cleft lip/palate and mental retardation, encodes for an Nepsilon-dimethyl lysine demethylase
A functional link between the histone demethylase PHF8 and the transcription factor ZNF711 in X-linked mental retardation
Hypomethylation of multiple imprinted loci in individuals with transient neonatal diabetes is associated with mutations in ZFP57
Position-effect variegation after 60 years
Conditional suppression of cellular genes: lentivirus vector-mediated drug-inducible RNA interference
Isolation of a human gene that inhibits HIV-1 infection and is suppressed by the viral Vif protein
Mitotic phosphorylation of histone H3 is governed by Ipl1/aurora kinase and Glc7/PP1 phosphatase in budding yeast and nematodes
Comparing the areas under two or more correlated receiver operating characteristic curves: a nonparametric approach
Pharmacogenomic identification of targets for adjuvant therapy with the topoisomerase poison camptothecin
Integration of retroviral vectors induces minor changes in the transcriptional activity of T cells from ADA-SCID patients treated with gene therapy
Exploration, normalization, and summaries of high density oligonucleotide array probe level data


text:
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The "Stand Up" app is bringing the funniest stand up shows to your mobile phone. We made a list of the funniest comedians and are frequently adding new comedians and stand up shows to this list. For suggestions please send an email to email@example.com
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House Sitter in Birmingham
Hi my name is Maggie Hendon and i would love to take care of your pets! i would also walk them feed them clean up around your house for you or even take the trash out and water any plants that need watering i have pet sat for a few of my family and friends and me and my family have 3 dogs. i would love to help you out with your pets and home!! my email is
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text:
The cocktail I designed for this evening's festivities. Designed to be simple enough for for-hire bar service and the ability to be prepared in punchbowl quantities, if needed.
The Noodle & Boo Cocktail
1 1/2 oz. 80-proof Bourbon
3/4 oz. St. Germain liqueur
scant dash orange flower water
Shake with ice and strain into a cocktail glass.
Similar Posts:
- Recipe: Grand Elderflower Cocktail
- Blood Orange Liqueur Recipes from Jackie Patterson
- Original Recipe: Tuesday Afternoon
- Original Recipe: The Downtown Casino


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|Specific References
|Related Products
|FLJ12815, SEMAF, semF, SEMA5A
|A DNA sequence encoding the amino acids (Met 1-Thr 765) of human SEMA5A (NP_003957.2) extracellular domain was fused with Fc region of human IgG1 at the C-terminus.
|Human Cells
|In general, recombinant proteins are provided as lyophilized powder which are shipped at ambient temperature.
Bulk packages of recombinant proteins are provided as frozen liquid. They are shipped out with blue ice unless customers require otherwise.
|> 90 % as determined by SDS-PAGE
|< 1.0 EU per μg of the protein as determined by the LAL method
|Samples are stable for up to twelve months from date of receipt at -70℃
|Glu 23
|The recombinant human SEMA5A (aa 1-765)/Fc is a disulfide-linked homodimer. The reduced monomer consists of 984 amino acids and predictes a molecular mass of 110.7 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh SEMA5A/Fc monomer is approximately 125-135 kDa due to glycosylation.
|Lyophilized from sterile PBS, pH 7.4
1. Normally 5 % - 8 % trehalose and mannitol are added as protectants before lyophilization. Specific concentrations are included in the hardcopy of COA.
2. Please contact us for any concerns or special requirements.
|Store it under sterile conditions at -20℃ to -80℃. It is recommended that the protein be aliquoted for optimal storage. Avoid repeated freeze-thaw cycles.
|A hardcopy of COA with reconstitution instruction is sent along with the products. Please refer to it for detailed information.
Semaphorins are secreted, transmembrane, and GPI-linked proteins, defined by cysteine-rich semaphorin protein domains, that have important roles in a variety of tissues. Humans have 20 semaphorins, Drosophila has five, and two are known from DNA viruses. Semaphorins are found in nematodes and crustaceans but not in non-animals. They are grouped into eight classes on the basis of phylogenetic tree analyses and the presence of additional protein motifs. Semaphorins have been implicated in diverse developmental processes such as axon guidance during nervous system development and regulation of cell migration. Semaphorin-5A, also known as Semaphorin-F, Sema F, SEMA5A and SEMAF, is a single-pass type I membrane protein which belongs to the semaphorin family. Semaphorin5A / SEMA5A contains one PSI domain, one Sema domain and seven TSP type-1 domains. It may act as positive axonal guidance cues. Semaphorin5A / SEMA5A is an axon regulator molecule and plays major roles during neuronal and vascular development. It plays an essential role in embryonic development. Semaphorin5A / SEMA5A induces endothelial cell migration from pre-existing vessels. It also plays a role in autism, reducing the ability of neurons to form connections with other neurons in certain brain regions.
|Product name
|Product name


text: Are you concerned that your food is lacking in essential vitamins and nutrients? Check out this super simple sprouter. Grow sprouts on your counter for pennies a day. Sprouts contain vitamins A-Z, they are hydrating, alkalizing, biogenic, regenerative, contains fiber and more protein than meat in addition to being a digestic enzyme.


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|Message view
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|"Jean T. Anderson" <...@bristowhill.com>
|Re: [doc] Incubation Policy -> "Acceptance of Proposal by Sponsor"
|Mon, 20 Mar 2006 00:29:05 GMT
Justin Erenkrantz wrote: > On 3/14/06, Jean T. Anderson <email@example.com> wrote: > >>I'm resurrecting the thread that fell off into /dev/null after >>http://email@example.com%3e >> >>So this sentence in >>http://incubator.apache.org/incubation/Incubation_Policy.html#Acceptance+of+Proposal+by+Sponsor >>doesn't match current reality and confuses TLP sponsors who think they >>need to wait for an Incubator PMC vote: >> >> >>>The Incubator PMC MAY immediately accept the Candidate, or may (at the discretion of the Incubator PMC) require a successful VOTE by the Incubator PMC. >> >>Here are a couple replacement suggestions that give the Incubator PMC an >>opportunity to voice any objections: >> [snip] > a+b. =) > > How about following the board's system for acknowledging new PMC members: > 1. PMC *chair* of Sponsor sends in notification to general@incubator. > 1. Any Incubator PMC member can then send an ACK and start a 72hr clock. > 2. If any PMC member says "hold" before 72hrs are up, a formal > discussion/vote is then conducted. > 3. If no PMC member has any negative comment at the end of 72hr clock, > then the PMC chair can commit the status file and 'start' the podling. > > How does that sound? -- justin I committed pretty much what Justin wrote, but worked in some wording from http://www.apache.org/dev/pmc.html#newpmc . Also, instead of "the PMC chair can commit the status file and 'start' the podling', I put "the status file may be committed and the podling started" -- once approved, is there any reason for a mentor on that project to not commit the status file? -jean --------------------------------------------------------------------- To unsubscribe, e-mail: firstname.lastname@example.com For additional commands, e-mail: firstname.lastname@example.com
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- You are here:
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3. You shall have more sex!
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text: Holiday & Seasonal All kinds of ideas and DIY projects to celebrate the holidays and seasons all year long! Rosemary-Infused Fresh Grapefruit Spritzer Beautiful Outdoor Living Spaces Distressed Wood Outdoor Side Table Casual Outdoor Brunch Table Setting Pimm's Cocktail Jug for Your Next Outdoor Party Printed Veggie & Herb Garden Markers "Grow a Tomato Sauce Garden" Gift Basket Backyard Deck Makeover (Part One) DIY Gift Ideas for the Gardener Joyful Spring Decor in Our Kitchen Chocolate Truffle Hearts with Fleur de Sel Valentine's Day Chocolate Chunk Skillet Cookies Canadian White Christmas Home Tour Vintage Winter Wonderland Christmas Mantel Ways to Make Your Christmas Tree Look Fuller Chic Halloween Decorating Ideas Canadian Fall Home Tour Fall Decorating Ideas Using Nature Scents of Fall Simmer Pot Recipe Mini Pavlova with Summer Fresh Fruit Blended Berry & Mint Green Iced Tea Fresh Mint: Garden to Table Recipes Stylish Deck & Patio Makeover Ideas Succulents in Weathered Pots Flea Market Style Outdoor Table Setting Gardening Apron Spring Gardening Gifts for Mom Mini Wreath Gift Tags & More Christmas Wrapping Ideas Holiday Gifts that Give Back Fall Front Porch Decorating Ideas Geraniums for a Pop of Backyard Color Summer Decorating Ideas Easy Breezy Summer Front Porches & Decks Decorating Outdoor Spaces (Best of the OE) Ikea Hack Bar Cart (The Details) DIY Bar Cart for Outdoor Entertaining Lavender & Chai Panna Cotta with Crunchy Maple Topping Country Garden Party Table Centerpieces Vintage Tool Box Planters Homemade Pink Grapefruit Salt Soak Easy Spring Project: Forcing Branches to Bloom Setting the Spring Table: Napkins Pretty Spring Decorating Ideas Beautiful Front Door Paint Colors Ten Minute Project: Crocus Planter Valentine's Day Printable Cards Mini Gingerbread Bundt Cakes Holiday Recipe Ideas (Link Party Features) Vintage Glam Christmas Gift Wrap Take Pause this Christmas Season Holiday Recipes Link Party Festive Pomegranate Mojitos Christmas Inspiration & Holiday Countdown Update No. 7 Trim the Tree Party Vintage Christmas Gift Tags & More Wrapping Ideas Our Favorite Things Giveaway! Gifts for the Holiday Hostess Front Porch Decorating: Winter Swag Woodland Wreath & One Item Project Challenge (Twigs) White Chocolate Pumpkin Pie Steamer Halloween Caramel Apples & More Boo-tiful Ideas Pumpkin Oat Muffins Changing of Seasons: Fall Front Porch Fall Inspired Recipes Rustic Apple Galette Lavender Infused Sugar Summer Dessert Recipes {Be Awesome Features} Watermelon Mint Paletas + More Summerlicious Treats Garden to Table Recipes {Be Awesome Features} Celebrate Summer! {Be Awesome Features} Jar Recipes to Take Along on Your Next Picnic Camping Dessert: Skillet S'more Brownies Summer Entertaining: Outdoor Oasis Party Weathered Bench Container Garden Lounging Around: Our Outdoor Living Space Potting Benches, Succulents & Garden Upcycling Refreshing Basil Cucumber Mojito + More Summertime Drinks Easy Garden Themed Hostess Gift with Jar Printable Cold Tolerant Annuals for Early Spring Planting Spring Front Porch: Adding Cottage Charm Create Your Own Seed Packets Container Herb Garden Ideas Nature Inspired Easter Tablescape Living Room Refresh for Spring Early Spring Gardening: Starting Seeds Indoors Vintage Key Art Valentine's Day Colorful Gifts for Her Keeping the Winter Blues Out of Your Home Valentine's Day Treat Bags with Iron-on Transfer Printables All Things Christmas {December Link Party Features} Holiday Countdown Planner {Finale} Holiday Countdown Link Party Decorating the Christmas Tree {Our House} Hot Chocolate Bar {All the Details} Holiday Countdown Planner {Week Eight} Holiday Recipes Link Party Festive Homemade Marshmallows Thumbprint Cookies {Like Grandma Used to Make} Nostalgic Hot Cocoa Station Holiday Countdown Planner {Week Seven} Handmade Gifts and Gift Wrap Link Party Easy Entertaining Ideas for the Holiday Season West Elm Inspired Drink Coasters Holiday Countdown Planner {Week Six} Burlap Table Favors for Christmas My Version of the Book Page Wreath Holiday Countdown Planner {Week Five} Swoon-worthy Handmade Gift Ideas Log Slice Chalkboard Ornaments Holiday Countdown Planner {Week Four} Painted Glass Canisters {One Item Project Challenge} Holiday Countdown Planner {Week Three} Holiday Countdown Planner {Week Two} Holiday Countdown Planner {Week One} Pumpkin Cake with Chocolate Chips & Pecans DIY Ombre Glitter Pumpkin My Lavender Garden {Oh the Possibilities!} Fresh Peach Crisp in a Jar What I've Learned About Caring for Outdoor Succulents Keeping it Simple: Green and White Plants You are my Sunshine Bench Makeover Outdoor Brunch Tablescape Strawberry Pot Herb Garden Outdoor Decorating Ideas: Lanterns Outdoor Lanterns {A Few of My Favorites} Ways to Use Mirrors in an Outdoor Space Welcoming Spring With Tulip Blooms Freshen Your Space With Spring Decorating Trends Adding Spring Color and Life in the Midst of Winter Christmas House Tour {Countryside} Creative and Personalized Gift Wrapping Ideas Holiday Countdown Series Finale + Link Party Christmas Decorating and Ideas Project E-book Holiday Countdown Series Recap {Week Eight} Cookie Exchange Recipes & Link Party Christmas House Tour {Elma Street} Holiday Countdown Series Recap {Week Seven} Memory Candy Jar & Other Handmade Gifts Jingle Bell Ornaments Holiday Countdown Series Recap {Week Six} Holiday Countdown Series Recap {Week Five} Easy Button Gift Embellishments Holiday Countdown Series Recap {Week Four} Handmade Gift Planning Holiday Countdown Series Recap {Week Three} Planting Bulbs for Christmas Snowflake Printed Tea Towels Holiday Countdown Series Recap {Week Two} Pinterest for Holiday Inspiration Hot Apple Cider for a Cold Hallow's Eve Holiday Countdown Series Recap {Week One} Fall Comfort Food: Coconut Curry Chicken Soup Refreshing Summer Drinks Made With Berries Summer Entertaining Al Fresco Style Outdoor Extravaganza: Plants & Flowers Projects Recap Kitchen Container Potager + Outdoor Extravaganza Link Party Last Minute Easter Ideas Spring Trendspotting: Florals Valentine's Day Color {Just in Time!} Holiday Countdown is On! {Reader Round-up} Holiday Cookie Exchange {Get all the Details} Holiday Countdown is On! {The Finale} Gifts {Almost} Too Pretty to Open Holiday Countdown is On! {Week 8} Holiday Countdown is On! {Week 7} Using Cloches in Holiday Decorating Holiday Countdown is On! {Week 6} Yummy Handmade Gifts {You May Want to Keep} Holiday Gift Guide {For Her} + Giveaway Holiday Countdown is On! {Week 5} Holiday Countdown is On! {Week 4} Holiday Countdown is On! {Week 3} Holiday Countdown is On! {Week 2} Soft and Chewy Ginger Cookies- oh, so good! Holiday Countdown is On! {Week 1} Changing it Up: Fall Decorating


text:
Hindsight is 20/20
I agree with the 4 points but im also satisfied with the job the Gharib is doing with the team. It was a friendly so the coach cared more about getting experience for the players most. I think if this was a tournament game, most of these chances wouldnt have occurred. I would of liked to see what Saleh Gomaa can do in this formation also. Hopefully Ahmed Hegazy regains form soon! Also i think that by playing in Portugal that Shikabala will be able to perform better with the national team. I also think that Zidan has a few more years left in the tank.
Overall not a bad result for a team missing Gedo, Soliman, Shika, Hegazy, Zaki, Gomaa, Trezuguet, Koka etc.


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Previous Parliament blown up (CYOAH
The British government officially surrenders to the German Reich. The former nation of Great Britain is divided. England and Wales are annexed to the German Reich. The Kingdom of Scotland is established as a puppet regime under the former Duke of Windsor as King Edward I, and Northern Ireland went to the Republic of Ireland (the newest member of the Axis). Without England, the rest of the British Empire become independent states, and remain neutral, excluding Australia and Canada, which join the Allies. President Roosevelt, puzzled as to the UK's surrender, but nonetheless furious at the Germans, declares war on Germany on August 1st, 1940. He then issues a draft for fifty thousand troops, which is met surprisingly fast. He then appoints Dwight D. Eisenhower Supreme Commander of the Allied Forces.
What does Eisenhower do with these troops?
Send them to the Eastern front
Yankovic270 17:11, July 27, 2010


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Heath A. DawsonThe Daily JeffersonianTheres no doubt that sophomore Megan Pryors 12-plus pitch at-bat against Indian Valley starter Sarah Schweitzer in the bottom of the fifth inning set off an alarm clock by Barnesville Highs sleeping bats.Pryor eventually reached first base after fouling off several pitches and the Lady Shamrock offense erupted for six runs with two outs en route to a 6-3 victory over Indian Valley in the Division III Sectional Championship on Wednesday evening at the Cambridge City Park.Indian Valley (15-12) took a 1-0 lead in the second inning as Megan Holmes was hit by a Natasha Skinner offering with two outs before Katy Kreinbihl then smoked a double down the left field line to score Holmes.After putting the ball in play over and over again in the first four innings, with only two hits to show for their efforts, the Lady Shamrocks (10-7) finally broke through in the fifth.Bethany Wable started things with an infield single and Kassie Ralston followed with a single to center. After Sapphire Straquedine sacrificed the runners, Pryors key at-bat took place. Pryor eventually reached first as home plate umpire Jerry Williams called catchers interference to load the bases. Amy Hines then hit into a fielders choice as Wable was thrown out at home for the second out.However, Kayla Ralston ripped a two-run single to center field, plating Kassie Ralston and Pryor for a 2-1 edge. Skinner followed with a clutch, RBI single to left, plating Hines for a 3-1 cushion. Senior shortstop Brianna Cook then smacked a two-run single (Kayla Ralston and Skinner scored) off of the glove of an Indian Valley defender, pushing the lead to 5-1. The final run of the inning came on a double, which allowed Cook to score, to right-center by Tiffany Stephen.The Lady Braves managed to score one run in each of the sixth and seventh frames, but it was not enough.BHS first-year head coach Eric Kerns was elated for his squad, which is peaking at the right time of the season.Its been since the mid-1980s since Barnesville last won a sectional championship, Kerns noted. That not only speaks for the girls, but also for former head coach Paul Burkhart. I was left with a very good team and some great leadership in the senior class. We have had some freshman and sophomores step up for us this year. Theyre ripping the ball and theyre not making errors in the field. Ive just been blessed to step in and take over a program that was left to me.Although the Lady Rocks took a while to get going, Kerns said he knew that a big stanza was right around the corner.Every inning, it just seemed like we did something silly to take ourselves out of an inning, Kerns related. I kept telling the girls that we were hitting the ball and putting it in play. I knew they would start falling for hits. Ive been preaching to the girls all year long that they cant leave runners out there. I feel comfortable anywhere in the lineup that if we have runners on base, were going to get them in.I told Megan that she kept getting a piece of it and to stay in there and be positive. That was her best at-bat of the night. When Megan was awarded first base on the play, I saw their teams morale go down, Kerns added.Skinner struck out one and walked two in the win. Kassie Ralston led Barnesville with a single, triple and run. Wable added two singles.Schweitzer recorded one K and walked none in the setback. IVHS was led by Jess Davis and Kreinbihl, who had a double and RBI each.With district play up next for the Lady Shamrocks, Kerns said he feels his team is ready and can play with anybody.Monroe Central is now the No. 1 team in the state in D-III and we outplayed them the entire ball game (in the Ohio Valley Athletic Conference 2-A quarterfinals), but we came up just short, he noted. We know were going to play a very good team in Sandy Valley or Ridgewood. The girls told me that theyre up for the challenge. They know our defense is solid enough and that we can hit well enough to play with anyone right now.NEXTBarnesville will play the winner of the Ridgewood-Sandy Valley game in the D-II Cambridge District Tournament on Tuesday at 5 p.m.