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Hi everyone!! I am officially back from San Francisco! Sorry this post is so late — it took awhile to write!
My mom and I had an absolute blast — not that I'm surprised, we always do! We ate, drank, and pretty much we go, go, go the entire trip!
Where we ate:
I didn't take many food pictures because many of the restaurants were dimly lit – great for ambiance, not so great for photos!
Gary Danko: Absolutely Amazing!! This restaurant is definitely pricey, but the food was incredible. It is price-fixe so my mom and I each ordered separate items and shared. I am pretty sure the menu changes seasonally – but I HIGHLY recommend the lobster and trying the Tete de Moine cheese. The macaron ice cream sandwiches were pretty wonderful too! 🙂
Boulevard: Another win!! This restaurant was incredible as well. My mom had the filet and I had the pork. Both were wonderful. I am also craving another bowl of their amazing cream of artichoke soup— SO GOOD!
Cha Cha Cha: This restaurant is located in Haight-Ashbury and serves the most amazing tapas and Sangria! We tried the wings, creole shrimp, mushrooms, and the pork bbq quesadillas. All of it was delicious, but the Chipotle pepper-guava glaze on the wings was unforgettable. My mom wants me to recreate them — anyone know where to get guava fruit? 🙂
The Stinking Rose: This restaurant is famous for its incorporation of garlic into every dish. Seriously — they even have garlic wine and garlic ice cream! We didn't try the wine or ice cream, but the garlic brie, gnocchi, and 40 clove garlic chicken were really fantastic!! Another great thing about this restaurant — reasonable prices!
Nob Hill Cafe: Mom and I found this cute little restaurant via Yelp reviews and they certainly did not disappoint. We wanted to go to Top of the Mark at the Mark Hopkins Hotel and wanted to find a restaurant nearby. First, I would definitely recommend getting drinks at Top of the Mark, because the views of the city below are incredible! They only serve light fare though so Mom and I wanted something else for dinner. Nob Hill Cafe serves delicious and reasonably priced Italian food – we split an antipasta salad and then each got pasta – mom had a roasted eggplant parmesan and I had a tortellini special with a brie cream sauce and mushrooms. Both were wonderful! The portions were huge and I struggled to leave any delicious tortellini behind!
Scoma's: Mom and I decided to try Scoma's for lunch after taking a bus tour of the city and exploring the Fisherman's Wharf. We each had soup – mom had a butternut and dungenous crab bisque and I tried the lobster bisque – both were good! We also split a really great prime rib sandwich that I think was a special that day. I really loved the interior – you could tell it was a longtime San Francisco tradition!
Boudin Bakery: We decided to try this delicious sourdough bread bakery after our visit to Alcatraz. It is located in the Fisherman's Wharf area and is a huge restaurant — they have a food court area, bakery tours, and a nice restaurant upstairs. The views from the restaurant overlooking the water were gorgeous! Mom and I split a sourdough bread bowl full of dungenous crab and corn chowder — SOO GOOD!!! There was not a bit of this left – except for the outer shell of the bowl. We also split the dungenous crab louie which was shockingly light and the perfect complement to to our heavy soup.
Bottega: Bottega is owned by Michael Chiarello, a food network chef. My mom and I checked out his restaurant when we were in the beautiful town of Yountville, CA during our wine tour. My mom and I split a meat and cheese antipasta as an appetizer – it was fantastic and I really liked that it was served with almonds — they added an unexpected, but welcome twist! I had a really light linguine pasta with artichokes and mushrooms and my mom had their risotto of the day. Both were delicious and a perfect lunch portion for two hungry wine-tasters!
The Spinnaker: The Spinnaker was our last lunch of the vacation and was absolutely perfect. The Spinnaker is located in gorgeous Sausalito and is right on the water – featuring huge windows with views of Marin County, San Francisco, sailboats on the bay, and playful sea lions! This lunch was extra delicious because my mom and I had just finished an 8-mile bike ride across the Golden Gate Bridge! We started our meal with perfectly chilled glasses of white wine and a cheese plate full of delicious creamy cheeses. I had a seafood caesar salad with bay scallops and baby shrimp, and my mom had a spinach salad with baby shrimp and blue cheese. We both really enjoyed our meals at this restaurant and would definitely go back!
Cafe de la Presse: This cute little French bistro was located right next to our hotel. We had two breakfasts here and one dinner. All our meals were fantastic! I tried the spinach and feta omelette on our first visit and the yogurt parfait on the second. When we tried it for dinner we both tried the french onion soup, which was delicious! Our waiter said it was "only a cup" so we both got our own – incorrect! It was definitely a crock like your typical french onion. Next time I think we would just split a bowl, but we weren't complaining about excess french onion soup either! For dinner, I had the ahi tuna special, which was SO good!! The tuna was cooked perfectly and had a wonderful flavor. My mom tried the Beef Bourguignon — her first Beef Bourguignon experience! It was excellent!! It was also served with a delicious macaroni and cheese, which was really great!
Hollywood Cafe: This was our favorite breakfast spot. They served overflowing bowls of gorgeous fruit that were a meal in themselves — not to mention the french toast or pancakes served alongside the bowls! We had breakfast here twice and we had the same things both times. I had the pancakes which were dense and fluffy at the same time! My mom had the french toast and she loved it as well! They also had really thick sliced bacon, which we could resist. The star of the show was definitely the fruit bowl though, with everything from blackberries to kiwi! I need to find a breakfast spot like this in DC!
Wayfare Tavern: This restaurant is owned by Tyler Florence and is known for its fried chicken. Honestly, my mom and I were not overly impressed with the restaurant in general. We were seated directly next to another two-top, which is one of my pet peeves so that wasn't helping the situation. The fried chicken was definitely good, but the portion was so huge that I felt guilty leaving so much! Seriously — five pieces of chicken per one entrée! I left feeling underwhelmed and overstuffed. If you love fried chicken I would definitely check this place out, but I won't likely visit if I find myself in San Francisco again.
Where we stayed:
Hotel Triton: I highly recommend the Hotel Triton. It is a cute and quirky little hotel in the Union Square/Chinatown area. We were able to walk to Chinatown, Union Square's shopping district, the cable cars, North Beach (little Italy), and the Fisherman's Wharf! Basically almost everywhere except for the Haight and Golden Gate Park area.
Hotel Triton's staff was so friendly and incredibly helpful. They made sure all of our tours were arranged properly, and that we either had transportation or knew how to get there if we wanted to walk.
Another bonus is that this is a Kimpton Hotel, which means they have a free wine happy hour from 5-6 PM every day! Mom and I usually missed the first half of happy hour because we were still out and about, but it was always nice to be greeted with a glass of wine before we prepared for the evening!
The hotel is also dog friendly and has a great little workout room where I did my half marathon training! (I was a little terrified to take on San Fran's hills!!)
What we did:
Bus Tour of the City: On our first full day in the city, my mom and I decided to do a hop-on-hop-off bus tour of San Francisco through Big Bus Tours. We ended up on a bus with a live tour guide, which I would recommend as opposed to the earphone option. We stayed on the bus for the entire tour and got to go across the Golden Gate Bridge, and saw city hall, the Haight-Ashbury area, Golden Gate Park, the Fisherman's Wharf while learning fun facts about San Francisco. This was a great way to get acquainted with the area and helped me decide which areas I wanted to go back and visit later in the week.
San Francisco City Hall – the dome has real gold on it!
Alcatraz: We knew we wanted to do a tour of Alcatraz during our stay in San Fran. We also knew that the tours book up quickly so we booked this first thing after we arrived. We did the tour on our second full day in the city and had an absolutely gorgeous day to explore the island. We boarded our ferry around 11:30 AM and left for the island at noon. We watched a short 20ish minute movie first and then began the audio tour. The audio tour was actually really great and I learned lots of fun facts — like that Scarface served time there in the late 1930's.
View of Alcatraz Island from the ferry
Closer view of Alcatraz
View of the Golden Gate Bridge from the ferry to Alcatraz
Cells where the prisoners carved their getaway path from the famous 1962 Alcatraz escape
Wine Tasting: We arranged a small bus tour through our hotel to do a wine tasting. That way our transportation was taken care of and the wineries were predetermined. We had a seriously gorgeous day for our wine tour and it was not surprisingly one of my favorite days of the trip. We learned a lot about California wine in general – like how certain grapes like Chardonnay grow better in Sonoma because of the fog while other grapes like Cabernet Sauvignon grow better in the hotter and dryer Napa climate. In addition we learned that the best Pinot Noir (Chris' favorite!) grow on the Oregon coast. I actually tried an awesome Oregon wine at Gary Danko called "OPP" or "Other People's Pinot." I need to find that again!
The wineries we visited were Viansa, Odette Estate, and Homewood.
All three of these wineries were great, but Viansa really stood out in my mind. It is located on beautiful grounds – in fact a portion of the land was donated to Ducks Unlimited in an effort to conserve the natural wildlife.
Golden Gate Park: After a full day of wine tasting my mom and I decided we needed a day with a lower intensity schedule. I really wanted to see the Japanese Tea Garden and my mom wanted to visit the Georgia O'Keefe exhibit at the de Young Museum.
The Georgia O'Keefe exhibit was really wonderful! It featured paintings that she did while staying at Alfred Stieglitz's family estate on Lake George in upstate New York. I was happy that we decided to do the audio tour because I didn't know a lot about her relationship with Stieglitz prior to the exhibit so it gave me a deeper understanding and helped me explain what I was seeing in the paintings! I would highly recommend visiting this exhibit. We were pretty tired after the exhibit, so we didn't explore the rest of the de Young Museum. Next time! It looked like they had a lot of other exhibits that would be fun to explore.
We end up just enjoying the view of the Japanese Tea Garden from the outside because we were tired and hungry after the Georgia O'Keefe exhibit. It was absolutely gorgeous though and I would definitely go back to explore and sip tea!
Painted Ladies: Before my mom and I headed to Golden Gate Park we stopped by Alamo Square to see the best view of the city and the "Painted Ladies," which were made famous by the show Full House! They were really pretty in person!! It made me want to watch Full House again!
Haight-Ashbury: After the Golden Gate Park, my mom and I walked to Haight-Ashbury, which was a beautiful walk through the park! After stopping for a fabulous lunch at Cha Cha Cha we checked out the surrounding area. It is full of really fun vintage clothing stores and other super funky stores with really fun pieces. I ended up getting a really cute white top and lavender flats and my mom got a fun pair of square toed suede shoes with rhinestone heels! We also saw where Jimmy Hendrix lived and took obligatory Haight-Ashbury photos! I would absolutely go back to Haight-Ashbury to explore more. I didn't expect the shops to be so neat!
At the Haight-Ashbury intersection
Where Jimmy Hendrix lived
Biking the Golden Gate Bridge: On our last day in San Francisco, my mom and I decided to bike the Golden Gate Bridge. We used Blazin' Saddles and had a pretty good experience once we got used to the bikes! The bike ride over to the bridge wasn't too bad – but there were definitely some "hills" that we walked up — they were more like mountains to these two flat land Delaware girls!
The views were absolutely incredible though and I am really glad we decided to bike over the bridge! The only part that was concerning was after we crossed the bridge and continued into Sausalito. There was no bike lane so you were basically just biking on the road with cars. If I was a more experienced biker I am sure I would be fine with this, but my mom and I were both very uncomfortable and ended up walking our bikes from the bridge to Sausalito. Therefore, I would definitely not recommend this as an activity to do with kids! It was a little stressful and we were both happy to park our bikes and head to lunch in downtown Sausalito.
This is definitely a must do, but maybe not a must do again! Next time, I'll walk the bridge if I want to get the view or just take a ferry to Sausalito. 😉
Sausalito: After ditching our bikes, my mom and I set out to explore gorgeous Sausalito. This is definitely an area I would like to come back and visit again. The houses were absolutely gorgeous and all of the sailboats on the water just made it a really pleasant experience in general. We spent most of our time in Sausalito at the Spinnaker, which was great because we could enjoy the view in the air conditioning! We did get to check out a little bit of the town– there were a ton of little jewelry shops and other fun artsy stores that would be fun to visit again!
Overall, we had a really, really wonderful time in San Francisco. We could not have asked for better weather and did pretty much as much as one could do with 5 full days in a city! I hope I am able to go back again someday!


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Locations|Contact Us|Careers
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Horizon Bancorp, Inc. is an independent, commercial bank holding company serving Indiana and Michigan through its commercial banking subsidiary, Horizon Bank. Horizon Bancorp, Inc. may be reached online at www.horizonbank.com. Its common stock is traded on the NASDAQ Global Select Market under the symbol HBNC.
Michigan City, IN and Attica, IN– Horizon Bancorp (NASDAQ GS: HBNC, "Horizon"), the parent of Horizon Bank, N.A., ("Horizon Bank") announces the completion of the acquisition of CNB Bancorp ("CNB"), parent company of The Central National Bank and Trust Company ("Central National Bank & Trust") of Attica, Indiana, effective November 7, 2016. A subsidiary of Horizon was merged into CNB, and then CNB was merged into Horizon. At the same time, Central National Bank & Trust was merged into Horizon Bank. As previously announced, both banks will operate under the name Horizon Bank.
As part of this transaction, Horizon Bank will add two new branches to expand its branch network to fifty-six offices throughout northern and central Indiana and southern Michigan. "We're confident customers will be pleased that the familiar faces they've come to know and trust will remain with Horizon at these branches. Their commitment to their customers shines through and truly shows in all that they do," said Craig M. Dwight, Horizon Bank's Chairman and Chief Executive Officer.
William C. McAdams, former Chief Executive Officer of Central National Bank & Trust stated, "We are pleased to join a company that instills the same values as CNB has throughout the years. Horizon's demonstrated commitment to preserve community banking, retention of local staff, commitment to community involvement, and personal service is why we trust Horizon will serve our customers and local communities just as we have."
Dwight further commented, "Horizon looks forward to the enhancements the Central National Bank & Trust's customer base will experience through our extensive product and service offerings, and on-line banking services, as well as higher lending limits and cash management tools to better serve the business and agricultural communities."
Horizon is pleased to name a life-long resident of the community, Sherri McGraw, as local Market President. "I look forward to all of the new opportunities Horizon will bring to Attica where I can continue to personally serve the banking needs of my local community," commented Ms. McGraw.
The companies are finalizing integration plans that include a systems integration and sign change expected to take place the weekend of December 3,2016. As a result, customer accounts will automatically change to Horizon accounts with no action required on the part of the customer. The terms and conditions of existing deposit and loan accounts are expected to remain unchanged. In nearly all cases customer checks, direct deposits, payments, and account numbers will also remain unchanged in order to make this transition as smooth as possible.
Horizon Bancorp was advised by the law firm of Barnes & Thornburg LLP. CNB Bancorp was advised by the law firm of SmithAmundsen, LLC and the investment banking firm of Renninger & Associates, LLC.
About Horizon Bancorp
Horizon Bancorp is a locally owned, independent, commercial bank holding company serving northern and central Indiana and southwestern and central Michigan through its commercial banking subsidiary Horizon Bank, NA. Horizon also offers mortgage-banking services throughout the Midwest. Horizon Bancorp may be reached online at www.horizonbank.com. Its common stock is traded on the NASDAQ Global Select Market under the symbol HBNC.
Forward Looking Statements
This press release may contain forward-looking statements regarding the financial performance, business prospects, growth and operating strategies of Horizon. For these statements, Horizon claims the protections of the safe harbor for forward-looking statements contained in the Private Securities Litigation Reform Act of 1995. Statements in this press release should be considered in conjunction with the other information available about Horizon, including the information in the filings we make with the Securities and Exchange Commission. Forward-looking statements provide current expectations or forecasts of future events and are not guarantees of future performance. The forward-looking statements are based on management's expectations and are subject to a number of risks and uncertainties. We have tried, wherever possible, to identify such statements by using words such as "anticipate," "estimate," "project," "intend," "plan," "believe," "will" and similar expressions in connection with any discussion of future operating or financial performance.
Although management believes that the expectations reflected in such forward-looking statements are reasonable, actual results may differ materially from those expressed or implied in such statements. Risks and uncertainties that could cause actual results to differ materially include risk factors relating to the banking industry and the other factors detailed from time to time in Horizon's reports filed with the Securities and Exchange Commission, including those described in "Item 1A Risk Factors" of Part I of Horizon's Annual Report on Form 10-K for the fiscal year ended December 31, 2015. Undue reliance should not be placed on the forward-looking statements, which speak only as of the date hereof. Horizon does not undertake, and specifically disclaims any obligation, to publicly release the result of any revisions that may be made to update any forward-looking statement to reflect the events or circumstances after the date on which the forward-looking statement is made, or reflect the occurrence of unanticipated events, except to the extent required by law.
Horizon Contact Information:
Craig M. Dwight
Chairman and Chief Executive Officer
Phone: (219) 873-2725
Mark E. Secor
Chief Financial Officer
Phone: (219) 873-2611
Fax: (219) 874-9280
Sherri L. McGraw
Market President - Attica
Phone: (765) 762-2414
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subset: commoncrawl


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I agree totally that he is capable of manipulating Bella while being immature. But to manipulate him on the level required in this story as a whole? Not a chance.pennybug84 wrote:I have to agree w/Corona on the previous conversation. Jacob is a manipulator. Just because he is immature (one of the reasons I don't like him) doesn't mean he can't manipulate Bella. I know someone who is immature & yet she is a very good manipulator. For me one of the biggest examples of this is the kiss in Eclipse. Jacob manipulates Bella into kissing him.
There are certainly times in the story that he does manipulate (such as in the kiss), but that's not high level manipulation. Saying "I'm going to kill myself!" is pretty basic. But the rest of it is not. To have it as such a plan as you're suggesting here is just ridiculous. He would have to be pure evil for that, and there's no suggestion that is so. When not overwhelmed by jealousy he does do the right thing.
Edward, as I recall, calls him out on it once. I don't recall anyone else saying at any point that they think Jacob is manipulating her. Alice is worried about her spending time with Jacob, but that's more because she can't see her when she's with Jacob. She also comments at the end that it's hard for Bella having to "choose between two good things". Do you honestly think she'd say that if she thought Jacob was masterfully manipulating her for nothing but selfish gain? No way!It's why Edward is always calling him on it. Bella doesn't see his manipulations for what they are. But EVERYONE around her does. Why he does it, ie immaturity, lashing out, overwhelmed by his emotions, whatever, doesn't make it any less manipulative.
When Angela mentions Jacob, she suggests that he is in love with Bella, not manipulating her, and I can't recall another mention. Charlie much prefers Jacob. There is no evidence in the books that everyone around her is warning her to stay away from Jacob.
It's interesting to note that most of the arguments you're using to prove that Jacob is manipulating Bella purely for selfish gain are exactly the same ones I've heard Team Jacob use to prove that Edward is manipulating Bella purely for selfish gain. Perhaps there is some middle ground where both teams can meet ....


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Posted 8 years ago
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Very rare fishing creel made in Portland, Oregon in the early 1900's. Tooled buckskin leather with original strap. McMonies is the correct spelling of the company. A typo was made on a batch of the name badges. Rather than re-order correctly spelled badges, they used up the misspelled badges on creels until they were gone. Thus, making this creel even more rare and unusual!


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Faculty Workshop: Xuan-Thao Nguyen, IU Robert H. McKinney School of Law
UConn Law Faculty and SJD candidates are invited to join Xuan-Thao Nguyen, Gerald L. Bepko Chair; Director, Center for Intellectual Property Law and Innovation, IU Robert H. McKinney School of Law for a faculty workshop, "Banking the Unbanked Innovators."
The UConn School of Law faculty workshop series provides a weekly forum for the presentation of legal scholarship and fosters academic discussion among colleagues and peers. UConn faculty and scholars from around the world present papers --typically works in progress-- and the faculty attendees are given the opportunity to ask questions and provide comments. The workshops are an important focal point for the intellectual life of the school, providing an opportunity for the UConn Law community to meet leading scholars from other institutions. For the full list of faculty workshops, visit the Faculty Workshops page.
The UConn Law faculty workshops series reserves a limited number of seats for UConn Law students. Student attendance is by registration only, and the availability of seats is evaluated on a case-by-case basis. Interested J.D. or LL.M. students should email firstname.lastname@example.com to reserve a seat based on availability. Students must sign up no later than the Monday before the workshop in order to confirm seat availability.


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These sticks, called Wood Stock, are like Pantone chips, but for wood. So if you're clueless about wood like me, you'll be able to tell the difference between beech and birch in no time.
As cool as I think it is, I feel like the type of people who need something like this don't work with wood. And the people who do work with wood don't need something like this. In any case, Wood Stock will teach you the uniqueness of cherry, birch, acacia, ash, beech, alder, oak, larch, maple and chestnut wood. [Abitare via NOTCOT via CrunchGear]


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The Guitron Family has a long tradition of braiding Quality Rawhide and Leather Tack in the Spirit of the Vaqueros for nearly 50 Years. Through the years, our selection of products has grown to include a full array of meticulously hand crafted tack, such as romal reins, bosals, headstalls and cavesons. Made from only the finest leather and rawhide. Our Family hopes this web site will give you a sense of the superior craftsmanship that is evident in our vast product line.
Contact Us | (760) 726-3042 or (800) 252-7913 | P.O. Box 878 | Vista, California 92085
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It's 20 minutes after AC St. Louis falls to the Kansas City Wizards in a preseason friendly, and Steve Ralston is worried the team bus is going to leave without him. From any viewpoint but Ralston's, the idea is absurd.
Forget that he's the most experienced player in the history of professional soccer in the United States with a record 412 appearances. Forget that he's Major League Soccer's all-time leader in assists (135), appearances (378), starts (372) and minutes played (33,143).
This isn't some nameless rookie trying to make the team. Ralston is the face of AC St. Louis, a hometown boy who returned to his roots to help launch professional soccer in a city with a historical appreciation for the beautiful game.
Although he's still six weeks away from playing on his surgically repaired right knee, Ralston is easing into his dual role as the club's marquee signing as well as an assistant coach to manager Claude Anelka.
"I've enjoyed it so far," Ralston said, "but I'm anxious to get back on the field. I'm doing rehab on the knee, and I think first and foremost that I'm here to play."
If his age-defying career in MLS is any indication, Ralston surely has at least a few productive seasons left in the tank for St. Louis. Over 14 seasons with the Tampa Bay Mutiny and New England Revolution, the MLS original scored 76 goals, played in four MLS Cups and was a seven-time All-Star.
And although he can't wait to join his teammates on the field, Ralston is also enjoying transferring the leadership skills he cultivated as a player into his role as assistant coach.
"The coaching bit is new for me so I'm learning as I go here," he said. "I pull guys aside and talk to them about things I have experienced. It's been good. I'm really enjoying that side of it right now."
Not as much as he will be when he gets back to delivering the pinpoint crosses and through-balls that made him one of MLS' most dangerous -- and consistent -- offensive threats for more than a decade.
Since entering the league as a rookie in 1996, Ralston has seen the league progress from barely keeping its head above water to expanding into cities starving for a professional soccer presence. In the process, the infrastructure surrounding the league's clubs has grown as much as the quality of play on the field.
"The things that stick out to me are the stadiums because back then they were all football stadiums and also the age of the players," Ralston said. "I was a young kid. I was 21 when I came in the league. Now they are all 16, 17, 18 years old, making a difference."
With a youthful roster that includes at least seven players from the St. Louis metro area, Ralston will have plenty of opportunities to mentor the game's next generation of talent while setting the table for a future career in coaching.
And what about the recent play of Clint Dempsey, a young player who developed under Ralston's wing in New England? Dempsey scored a delightful, no-look chip that tumbled just under the crossbar to topple Italian giants Juventus last week and send Fulham to the Europa League quarterfinals.
"It was nice," Ralston said with a sly grin. "I wish I could say I taught him that."


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Microsoft plans to release five critical update bulletins next Tuesday, all critical, in the September edition of its regular Patch Tuesday update cycle.
As usual, not much information on the contents of the planned patches is available. All supported versions of Windows (2000, XP, Vista, Server 2003 and Server 2008) will need updating to defend against various, as yet unspecified, critical security risks.
The list of affected software leaves out mention of Microsoft's IIS Web Server software, which is currently the target of exploits capitalising on a zero-day vulnerability. More specifically, the flaw involves problems in the Microsoft FTP services component bundled with IIS 5.0, IIS 5.1, IIS 6.0 or IIS 7.0. In most cases, the bug creates a means to crash a vulnerable webserver, and only in cases where FTP is enabled. Remote code injection only becomes a risk for shops running IIS 5.0 on Windows 2000.
The lack of mention of IIS in Microsoft's pre-alert implies a set of patches for Microsoft's web server software software will have to wait until at least October.
Alongside the critical patches on its menu, Microsoft also plans to update its Malicious Software Removal Tool, as explained in its pre-release notice here. ®


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BISC219/F12: RNAi Lab 8
Lab 8: Series 3- Creating the feeding strain of bacteria for RNAi in C. elegans
To do on the night before this lab:
You and your partner will return to the lab to make an overnight broth culture of one of the colonies that you are sure contains the gene of interest (determined from your visualization of successfully amplified, appropriately sized DNA seen on your gel photo). The sub-culture you will set up tonight will create many identical copies of bacteria that carry the plasmid containing your gene of interest.
- Find your LB+amp plate in the glass front refrigerator in a rack labeled with your lab day. Make sure there is some bacteria remaining on the plate of a colony that you saw successful gene amplification in the PCR product.
- Begin by obtaining two tubes of LB broth (each will have 5 ml of broth) from the refrigerator in the back left hand corner of the room.
- Add 5 microliters of the 50mg/ml ampicillin stock (also found in the refrigerator) to each tube. You may use your P20 micropipet and the middle size tips or a P10 and the smallest tips. (Make sure that you adjust your pipet to the proper setting which will look different on a P10 vs. a P20!)Calculate the effective concentration of ampicillin that you will have in your LB tube (remember V1 x C1= V2 x C2) and record that information in your lab notebook.
- Gently swirl your LB +amp broth to mix the contents. DO NOT invert the culture tubes as the tops are not spill resistant!
- Label the two sterile glass culture tubes with tape in your team color. Label one with "pPD129.36 lsy-2" and your initials. Label the other with your initials only.
- Inoculate the broth with your bacteria by using a sterile disposable loop or a sterile toothpick (both found in the supply area near the door to L304) to scrape your candidate colony off the plate. Be sure not to touch the plate with the loop except on the desired colony. Try to avoid picking up any satellite colonies or another colony nearby! Gently swirl the loop in the LB+amp broth - you should be able to see the colony come off the loop. If you use a sterile toothpick, you can just drop it into the broth so that the tip with the colony is in the broth. Note thatthe part you touched does not come in contact with the medium. (The second tube of broth labeled with just your initials is a control and should not be inoculated with bacteria as it is your control for contamination.)
- Balance the 2 tubes across from each other on the rotating wheel in the 37C incubator at the front of the room when you come in the door. DO NOT USE THE wheel that is at Room TEMP!
- Incubate these broth cultures at 37°C overnight. Do not forget to make sure the wheel is rotating when you leave!
Plasmid Isolation and Transformation
Last week, you tested two colonies of previously transformed bacteria in order to find one that contains a plasmid with the C. elegans lsy-2 gene. You have confirmed that the colony you have sub-cultured last night contains E. coli bacteria-- not just any pPD129.36 plasmid but a copy of pPD129.36+lsy-2. Now we can isolate the plasmid DNA from these cells and transform these plasmids into a different special strain of E. coli bacteria, HT115(DE3). We will use this strain because we can induce the HT115(DE3) strain to overexpress the plasmid genes, resulting in the presence of a lot of double stranded RNA specific to lsy-2,our gene of interest in these bacteria.
To recap: Yesterday you inoculated a few milliliters of LB broth containing E. coli that we are sure have maintained our genetically engineered plasmid, pPD129.36, modified to contain an antibiotic resistance gene to ampicillin and all or part of the C. elegans lsy-2 gene that you want to investigate. You added ampicillin to the broth to ensure that the plasmid DNA would be maintained by the cells. Overnight the bacteria have grown to high density and the plasmid DNA has undergone many replications. However since you started with a single colony of bacteria and that colony grew from a single transformed cell, all the copies of the plasmid DNA in your overnight culture should be identical ("clones" of one another). To isolate the plasmid DNA from this bacterial strain that expresses C. elegans lsy-2 , you will perform what is commonly called a "mini-prep". This term distinguishes the procedure from a "maxi-" or "large scale-prep" which involves a larger volume of cells and additional steps of purification. The overall goal of each "prep" is the same--to separate the plasmid DNA from the chromosomal DNA so that a certain gene on the plasmid DNA can be studied further.
To isolate plasmid DNA from an overnight culture of cells, the media is removed from the cells by centrifugation. The cells are resuspended in "Solution I" which contains Tris to buffer the cells and EDTA to bind divalent cations in the lipid bilayer, thereby weakening the cell envelope. Upon the addition of "Solution II," the chromosomal DNA and the plasmid DNA are denatured by the sodium hydroxide, and the cellular proteins and lipids are dissolved by the detergent, sodium dodecyl sulfate (SDS). The pH of the solution is returned to neutral by the potassium acetate in "Solution III." At neutral pH the SDS precipitates from solution, carrying with it the dissolved proteins and lipids. In addition, the DNA strands renature at neutral pH. The chromosomal DNA, which is much longer than the plasmid DNA, renatures as a tangle that gets trapped in the SDS precipitate. The plasmid DNA renatures normally and stays in a water based solution. In this way the plasmid DNA is separated from the chromosomal DNA, the proteins and the lipids of the cell. Plasmid DNA can be precipitated out of solution in absolute ethanol and then put back into solution (in an appropriate concentration) in water. The ingredients and concentrations of a stock solution of all reagents (such as Solutions I, II, and III) can be found in the Media Recipes section of this section of the wiki.
Today you will transform the isolated plasmid DNA, pPD129.36+lsy-2, into an IPTG inducible strain of E. coli called HT115(DE3), and spread the transformed bacteria onto LB agar media supplemented with ampicillin (resistance conferred by a gene expressed from the plasmid).
Part 1: Plasmid DNA Isolation
Only one of your two overnight cultures should appear cloudy with bacterial growth. If your control is cloudy, please inform your instructor. (You can find the ingredients and concentrations for ingredients of Solutions I, II, and III in the Series 3 Media Recipes page.
- To start the plasmid DNA isolation, label 2 microfuge tubes with pPD129.36+lsy-2 (our plasmid name + our C. elegans gene), plus your initials and team color.
- Check to be sure that your cells have not settled. If they have, mix the cultures but do NOT invert them because they will leak. Then pour some of the overnight culture so that both microfuge tubes are almost full. If you are nervous about pouring the bacteria, you can pipet 750 microliters into each tube twice, so that there is a total of 1.5 ml in each tube. The exact volume doesn't matter but the tubes should be quite full when you close the cap.
- Place the tubes in the room temperature microfuge so that the hinges of each cap is facing out. Paying attention to this small detail will help you know where in the tube to find your pellets. While it is not essential to do this in this step, it is a good habit to get into since sometimes pellets are very small and hard to see. Be sure your tubes are balanced, then spin the tubes for two minutes at 8,000 rpm. Check the rcf speed of the centrifuge while it is spinning (by hitting the toggle that changes the readout) and record the rcf's in your lab notebook. Relative centrifugal force (rcf) is what should be used in M&M to describe your speed since it is universal and rpm is rotor and centrifuge dependent.
- If the supernatant is not clear, recentrifuge until it is. When the supernatant is clear, pour it into the waste beaker that is on your bench, then flick the tube with the cap open to remove the last few drops of liquid. The cell pellet will not fall out.
- Resuspend your cell pellets COMPLETELY in 100 microliters of solution I. Pipet up and down until ALL of the cells are uniformly suspended in Solution I. Be sure to change tips between samples. Leave the cells at room temperature as you prepare solution II.
- To make solution II, mix 500 microliters of 2% SDS with 500 microliters of 0.4M NaOH in a microfuge tube. Close the cap and invert the tube several times to mix the contents. Add 200 microliters of solution II to each miniprep and invert the tubes five or six times to mix. In some cases the minipreps may appear to "clear" but don't worry if you don't see a big change in yours. Place the tubes on ice for five minutes.
- Add 150 microliters of solution III to each miniprep and immediately vortex each tube for 10 seconds with your vortex set at the highest setting. White clumps should appear in the solution after you vortex it. Place the tubes in the room temperature microfuge and spin for 3 minutes at the HIGHEST speed.
- While the minipreps are spinning, label another set of microfuge tubes with the name of the plasmid (pL4440+C. elegans gene name)and your initials or team color. The new tubes can be left at room temperature.
- A white pellet should be visible when you remove your minipreps from the microfuge. Use your P1000 to transfer 400 microliters of each supernatant to the appropriate clean microfuge tube. It's OK to leave some of the supernatant behind. Try to avoid transferring the white pellet.
- Add 1 ml of room temperature 100% ethanol to each new tube. The tubes will be quite full. Close the caps and invert the tubes at least five times to thoroughly mix the contents.
- Spin the tubes in the room temperature microfuge for 2 minutes at HIGHEST speed. It is important to orient your tubes so that the hinges are up this time, as the pellets are expected to be barely visible.
- Pour off the supernatants into the waste beaker on your bench then flick out the last few drops. The tubes do not have to be completely dry.
- Carefully, wash the pellets with 500 microliters of 80% ethanol. (You may need to make a few mls of 80% ETOH from the 100% ethanol stock provided). When you wash the pellets, add the 80% ethanol slowly and carefully, so that the liquid flows down the side of the tube away from the pellet. After the ethanol has been dispensed, immediately remove it with the same tip, making sure to keep the tip on the side of the tube that doesn't have your pellet. Some liquid will remain in the tube, and it can and must be removed using your P200, set to 100 microliters. Try to remove as much of the ethanol as you can (without removing your pellet!). The pelleted DNA will have to be completely dry before you add water or your DNA will not go into solution.
- After all the DNA pellets have been washed, put your tubes with their caps open in the SpeedVac that is at the back of the lab. If you were able to remove most of the ethanol with the P200, then after two or three minutes the last drops of ethanol will have evaporated and the pellet will be barely visible. If the DNA pellet is invisible please see your instructor to determine how much water to use in the next step.
- If you have a visible pellet of DNA, add 30 microliters of sterile water to each tube and vortex each tube enough for all of the DNA to be off the wall of the tube and into solution. The pelleted plasmid DNA must dissolve completely in the water. The liquid can be brought back to the bottom of the tubes by spinning for a few seconds in the microfuge. You can then combine the dissolved plasmid DNA into one of the two tubes so that you have approximately 60 microliters of plasmid DNA as one aliquot. Store the DNA on ice until you are ready to use part of it and then give the rest to your instructor to freeze. DO NOT DISCARD ANY OF YOUR PLASMID ISOLATE!
Measuring the concentration of plasmid DNA using the NanoDropper
Use the ThermoScientific NanoDrop Spectrophotometer in L308 to measure DNA by taking Absorbance at A260nm. This spectrophotometer uses only 1 microliter of sample and does not require cuvettes. The sample is held in place by fiber optic technology and surface tension that holds the sample in place between two optical surfaces that define the pathlength vertically and dynamically. Measurement can be assessed in a range of 2 to 3700nm/microliter dsDNA. These are expensive machines so make sure you follow the directions carefully and ask your instructor for guidance as needed.
More information is available from the manufacturer's website at: | http://www.nanodrop.com/HowItWorks.aspx
Using the Nanodropper
1. Clean the upper and lower optical surfaces of the sample retension device by pipetting 2 microliters of clean deionized water onto the lower optical surface. Close the lever arm and tap it a few times to bathe the upper optical surface. Lift the lever arm and wipe off both optical surfaces with a Kimwipe.
2. Open the NanoDrop software from the Desktop of the computer and select the nucleic acids module.
3. Initialize the machine by placing 1 microliter of clean deionized water onto the lower optic surface, lower the lever arm, and select initialize from the NanoDrop software. Once initialization is complete (~10sec.), clean both optical surfaces with a Kimwipe.
4. Perform a blank measurement by loading 1 microliter of purified deionized water.
Note that as in traditional spectroscopy, the blank will be subtracted from subsequent measurements. If you want to determine the contribution of a specific buffer or diluent, measure the buffer first using distilled water as a blank. If the buffer does not contribute to the A 260nm reading, then deionized water is fine to use as the blank. The water or buffer should always be measured to be sure that the instrument has been zeroed properly. The measurement of water or buffer should be zero or very close. All measurements are automatically normalized to 340nm.
5. Measure the nucleic acid sample by loading 1microliter of sample and selecting "measure". Record your DNA concentration in your lab notebook and on the tube of plamid DNA. Once the measurement is complete. Clean both optical surfaces with a Kimwipe and the machine is ready for the next sample.
You should ensure that the appropriate constant (50 for dsDNA or 40 for RNA) has been chosen. The software automatically calculates the nucleic acid concentration. If the calculation is done by hand, the A260nm is represented as a 1cm path for convenience, even though 1-nm and 0.2nm paths are actually used during the measurement cycle.
Clean Up
When the last sample was been measured, clean the sampling device by repeating step 1.
Part 3: Transformation of isolated plasmid DNA into E. coli strain HT115(DE3)
The competent HT115(DE3) bacterial cells are on the instructor's bench You will transform some of your plasmid DNA into these bacteria. The cells are very fragile, so treat them gently.
The prep staff prepared these competent HT115(DE3) cells for you using the Inoue Method Media:Inoue bacterial transformation.doc. Reference: Inoue H., Nojima H., and Okayama H. 1990. High efficiency transformation of Escherichia coli with plasmids. Gene 96: 23-28. The cells were made competent to take up free plasmid DNA by this treatment. This treatment makes the cell wall and membrane more permeable and our transformation efficiency much greater but it weakens the cells; therefore you must keep them cold and mix them gently throughout the transformation (no vortexing!).
- Obtain a tube containing 50 μL of competent cells from your instructor.
- Label the top or the side of the tube with HT115(DE3), pPD12936+lsy-2, and your initials or team color.
- Consult with your instructor to determine how much of your plasmid DNA you should add to the cells to start the transformation. When you have determined the correct vol., pipet between 1μL and 10μL of your plasmid DNA to the tube. (The volume you should use is dependent on the conc. you achieved in your mini-prep.) Pipet up and down once to mix the DNA and the cells. Close the cap and let the transformation mixture sit on ice for 10 minutes.
- Heat shock by incubating the transformation mix at 42°C for 90 seconds, exactly. This step must be timed exactly. Remove the tube at the end of 90 seconds to your ice bucket while you get your LB ready.
- Consult your instructor to determine the volume of warm LB broth you should add (usually between 250-500 microliters) to the transformation mix. LB aliquots should be found in the 37°C mixer/incubator on the green/blue team bench. When pipetting the media, remember to release your thumb on your micropipet slowly, to avoid splashing the liquid on the end of the barrel. The barrel is not sterile and if you see the liquid touch it, then discard the media in the waste beaker and try again with a new tip.
- Once you have added the LB, close the cap and invert the tube once or twice to mix the contents. Incubate at 37°C for 30-45 minutes in the bench top incubator/ mixer with occasional gentle mixing.
- While the plasmid DNA is being taken up by the competent cells and the new genes provided by the plasmid are being expressed by the bacteria, label two LB + amp agar plates. Label the bottom of the plated with the strain's identity (HT115(DE3)), the plasmid used, the date, your initials and team color. You must label the bottom of the plated since the tops are easily switched. Differentiate them by putting 50μL on one and 200μL on the other. Put these plates in the hood with the blower on and with the lid ajar to dry the surface of the agar for about 10 minutes or until the surface looks dry but is not badly dehydrated.
- Once the transformation mix has incubated at 37°C for 30-45 minutes, invert it to mix the contents and pipet 50 microliters of transformed cells onto the center of a labeled slightly dehydrated LB + amp plate prepared in the previous step. Pipet 200μL on the other plate. Pour 5-10 glass beads onto the plates. Put the lid back on and gently swirl the beads all over the plates to spread the transformed bacteria around. When you are done - pour the beads into the beaker with disinfectant near the sink.
- Replace the lids and leave the agar plates undisturbed for a few minutes.
- Once they have dried enough that the surface doesn't appear wet, invert the plates and incubate them at 37°C for 24-48 hours. The plates should be incubated with the agar side up so that condensation will not drip onto the surface of the agar and smear the colonies that will be growing there.
- Save the remaining transformation mix for 24 hours or until we are sure that there is at least one colony growing on each of your plates.
What would it mean if you had no colonies on your plate? Normally, you would expect to have around 100 pale color colonies on each plate. If you have at least one well isolated colony on the plate, you're all set. After the 24 hour growth period the plate should be placed in the rack in the refrigerator labeled with your lab day. You will use a single colony from the plate to make an overnight broth culture on the day before next lab. If you have no colonies on one or more of your plates, please notify your instructor right away.
Before leaving lab today, give the rest of your isolated plasmid DNA to your instructor in a labeled microfuge tube. Make sure your tube is labeled with your name, lab day, plasmid name, DNA concentration and color coded with a piece of tape in your team color.
To do on the day before the next lab:
You and your partner will return to the lab to make an overnight broth culture of your selected colony as described below. This process will create a sub-culture of many identical copies of the bacteria containing the plasmid carrying the construct to RNAi the gene that you want to study.
- Find your plate in the glass front refrigerator in a rack labeled with your lab day. Select a colony to start your overnight culture. At this point, ALL of your colonies should contain your plasmid of interest.
- Begin by obtaining two tubes of LB broth (each will have 5 ml of broth) from the refrigerator in the back left hand corner of the room.
- Add 5 microliters of the 50mg/ml ampicillin stock (also found in the refrigerator with the broth) to each tube. Calculate the effective concentration of ampicillin that you have in your LB tube (remember V1 x C1= V2 x C2) and record that information in your lab notebook.
- Add 5 microliters of the 12.5mg/ml tetracycline stock (also found in the refrigerator with the broth). Calculate the effective concentration of tetracycline that you havein your LB broth tube. Record that info in your lab notebook.
- Gently swirl your LB +amp+tet broth to mix the contents.
- Label the two sterile glass culture tubes with tape in your team color. Label one with "pPD129.36 lsy-2" and your initials. Label the other with your initials only.
- Inoculate the broth with your bacteria by using a sterile disposable loop to scrape your candidate colony off the plate. Be sure not to touch the plate with the loop except on the desired colony and don't pick up any satellite colonies! Gently swirl the loop in the LB+amp+tet broth - you should be able to see the colony come off the loop. (The second tube of broth labeled with just your initials is a control and should not be inoculated with bacteria as it is your control for contamination.) If you prefer to use a sterile toothpick rather than a loop, you may pick up the colony with the sterile toothpick and drop the toothpick into the broth culture. Note that the tip with the colony is in the broth and the contaminated part you touched with your fingers does not touch the sterile medium.
- Balance the 2 tubes across from each other on the rotating wheel in the 37C incubator at the front of the room when you come in the door. DO NOT USE THE ROOM TEMP WHEEL!
- Incubate these broth cultures at 37°C overnight. Do not forget to make sure the wheel is rotating when you leave!


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brazil nut pie
- 3 eggs
- ½ cup (90g) rapadura sugar or brown sugar
- 1 tablespoon honey
- 2 tablespoons vanilla extract
- 1½ cups (240g) brazil nuts, roughly chopped
- natural Greek-style (thick) yoghurt and honey, to serve
brazil nut pastry
- 1 cup (160g) brazil nuts, ground
- 2 teaspoons vanilla extract
- ½ cup (70g) buckwheat flour (see note, page 102)
- 1 egg
- ½ cup (50g) LSA+
- 2 tablespoons rapadura sugar or brown sugar
- Preheat oven to 160°C (325°F).
- To make the pastry, place the ground brazil nut, vanilla, flour, egg, LSA and sugar in a bowl and stir to combine. Turn out onto a work surface and bring together to form a smooth dough.
- Press the dough into a lightly greased, 22cm loose-based fluted pie tin, trimming the edges.
- Place the eggs, sugar, honey and vanilla in a bowl and whisk to combine. Add the nuts and mix to combine. Pour the mixture into the tart shell and cook for 30 minutes or until just set. Refrigerate until cold.
- To serve, slice and top with yoghurt and honey. Serves 6–8.
+ LSA is a mixture of ground linseeds, sunflower seeds and almonds. Find it in the health food aisle of the supermarket.
+ Buckwheat flour is rich, nutty and gluten free. It's available from health food stores.
Bea Long
I LOVE this pie! The crust just sends it over the top. WOW, it's delicious.


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Josh Powell's Siblings Try to Claim Life Insurance
Josh Powell changed beneficiaries of $2.5 million life insurance policy.
March 8, 2012 -- In the months before he killed himself and his two children in a fiery explosion, Josh Powell altered the terms of a $2.5 million life insurance policy on himself, his sons, and his missing wife, Susan Cox Powell.
According to court documents filed by the New York Life Insurance Company in U.S. District Court, two of Powell's siblings have tried to make claims on the life insurance in the wake of their brother's murder-suicide.
Powell, who was called a "person of interest" in his wife's 2009 disappearance, killed himself and sons Braden, 7, and Charlie, 5, during a supervised-custody visit in February.
The insurance company is petitioning the court to decide who should receive the $2.5 million payout, since Powell is considered a "slayer" of his children and "person of interest" in his wife's disappearance, terms that could disqualify him and his family as beneficiaries, they argue.
According to the lawsuit, Josh Powell changed the terms of his family's life insurance to designate his brother, Michael Powell, as the main beneficiary, with his sister and father to receive smaller percentages of the payout.
The insurance was first taken out during the summer of 2007, when Josh and Susan Powell took out $1 million policies each on themselves, and $250,000 riders on each of their children. At the time, they designated one another as the beneficiaries. The couple later set up a family trust, designating each of their fathers as beneficiaries of the trust, and naming the trust the beneficiary of the life insurance.
Then, two years later, in December 2009, Susan disappeared. Police have never located her body and have made no arrests in connection with her disappearance, though Josh Powell was the only named person of interest in the case.
In October, 2011, just weeks after his father was arrested on charges of child porn and voyeurism, Josh Powell changed the beneficiary of the life insurance. No longer would the family trust receive the life insurance payouts; instead, Josh Powell's siblings would become the beneficiaries, according to the court documents.
The changes resulted in Susan Powell's father, Chuck Cox, losing his claim on the life insurance.
Two months later, Powell once again changed the beneficiary information, designating his brother, Michael Powell, as the beneficiary who would receive 93 percent of the pay out, while his sister Alina would receive 4 percent, and a third sibling would receive three percent. He designated his father Stephen as a secondary beneficiary.
Powell made the final beneficiary change on Dec. 3, less than two months before he killed himself and his children on Feb. 5, 2012.
Nine days later, both Michael and Alina Powell contacted New York Life Insurance Co. to make a claim on the policies.
The insurance company is asking the U.S. district court to decide whether the Powells have a rightful claim to the money, despite Josh Powell's status as a "slayer" of the two boys and a "person of interest" in Susan's disappearance, or whether Susan's parents or other interested parties have a claim on the payout.
The filing also draws Josh Powell's competency into question, asking the court to determine whether the murder-suicide affects the beneficiary changes.


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One-Pot Wonders: Chicken-Sausage Frittata With Corn and Feta
Get the Recipe
Frittatas are the perfect vehicle for clearing out extra ingredients in the fridge while feeding a hungry group of people quickly and easily. At a bachelorette weekend I was at not too long ago, a friend of mine took our leftovers of chicken sausage, grilled corn, grilled asparagus, and parmesan, tossed them all together in a pan, added beaten eggs and threw it in the oven. Did I mention she had seven hungry and slightly hungover girls waiting at the table? Genius.
Inspired by her resourceful frittata, I recently decided to make my own version, using chicken sausage, corn, and feta cheese. While there are many methods for making a frittata, I turned to our easy method, which uses a scrambled-egg technique to create light and fluffy eggs, while taking advantage of both the stove-top burner and the oven's broiler to cut down on cooking time.
I first beat the feta with the eggs and a generous pinch of salt, then set it aside while I soften and brown the peppers, sweet kernels of corn, onions, and pre-cooked pieces of chicken sausage. I then pour the eggs over the mixture and gently stir, so that the fillings are evenly distributed throughout. With a quick blast under the broiler, the frittata is cooked. To finish it off (and make it slightly more of an indulgence) I like to top the dish with creamy diced avocado, and serve a simple green salad on the side to round out my meal.
Get the Recipe
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How to Turn Your Handset Into a Facebook Phone
Facebook unveiled "Home" this morning, the company's modified version of the Android software able to be installed on Android devices.
How to get it? On April 12, head on over to Google's Play store and search for "Facebook Home." Download the software — it's apparently a quick install, and should load on your phone immediately. You'll also have to download Facebook's other apps — Messenger and the like — and also register for the "Facebook Intent," an under-the-hood mechanism that makes Android work.
From there, you'll be able to access all of the key features that come with Facebook's new Android skin, including Cover Feed and Chat Heads, the persistent messaging application that flows throughout apps across the entire phone.
Sadly, only a handful of devices will be supported at launch. The HTC One, HTC One X, Samsung Galaxy S3, Samsung Galaxy S4 (in the future) and the Galaxy Note 2. Facebook plans to have more devices supported, but for now only these few are capable of being upgraded.
Home is designed to run on Ice Cream Sandwich and Jelly Bean versions of Android, Facebook said.
A separate program will allow device makers to incorporate Facebook Home right out of the box.
The first two partners are HTC (as we reported last year), as well as carrier AT&T. Their collaboration — a phone called the HTC First — comes in four colors (ooh, aah). It'll be released on April 12 for $100.
The company plans to update "Home" on a monthly release cycle, adding new features to install regularly.
So, it's not ready for all Android lovers, but in Facebook's version of the mobile future, it will be.
- Ten Minutes With Adam Mosseri, the Guy in Charge of Facebook Home
- If Facebook Made a Real Facebook Home (Comic)
- The Buffy Story: Facebook's Long Road Home to an Android Phone
- Facebook's Phone Also Coming to France Telecom-Orange, but First to AT&T
- Content, Content Everywhere In Facebook's Ideal Mobile World
- Can Facebook Build an Android-Style Home on the iPhone? Probably Not.
- How to Turn Your Handset Into a Facebook Phone
- HTC First: The Hardware Side to Facebook Home
- Buffy Has Landed: Facebook Launches Home, Its Android Phone Project


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Double click on above image to view full picture
Powerful and versatile, FUJFILM EF-42 flashguns provides complete TTL performance. For precise lighting, the flash can be moved 90° upward, 180° to the left and 120° to the right, flexibly supporting bounce shooting and other operations. Allowing reference by different values, the focal length display can be switched between the digital display (APS-C) and the 35-mm film size converted display on the LCD while the wide range of functions and user-friendly controls make the EF-42 easy to use. The guide number of 24 to 42 changes automatically to the lens focal length while the flash will also zoom automatically between 24mm and 105mm (up to 20mm with the wide panel). Allowing full manual control, the EV can be adjusted in 7 steps (-1.5, -1, -0.5, 0, +0.5, +1, +1.5) while the power ratio can be adjusted in 7 steps (FULL,1/2,1/4,1/8,1/16,1/32,1/64). When shooting in the dark or dimly lit places, the near infrared assist light is automatically illuminated to enable automatic focusing.
Full TTL Compatibility
HS50EXR / HS30EXR / HS33EXR / HS25EXR / HS28EXR / HS22EXR / HS20EXR / SL240 / SL260 / SL280 / SL300 / SL1000 / FUJIFILM X100 / FUJIFILM X20 / FUJIFILM X10 / FUJIFILM X-S1 / FUJIFILM X-Pro1 / FUJIFILM X-E1 / FUJIFILM X-M1 / FUJIFILM X-A1
Additional Information
|Manufacturer Part Number


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Where Zombie Bosses Come From
Where zombie bosses come from is a mystery rarely explained in most game and movies these days. Usually starting out as your common run of the mill zombie something sets them apart for greatness and soon their tongue starts to lengthen or perhaps they grow to prodigious size and ferocity. One thing is for certain, we need them.
You can only mow down legion after legion of common zombie cattle fodder for so long before you get bored. Video game logic needs… No it demands that there must be a greater challenge to be overcome in order to make your apocalypse experience greatness. Enter the zombie bosses. He's there to ratchet up the tension after the players have sated their blood lust on the minor zombie hordes.
Logically you may say why would there be zombie bosses? That makes no sense. Why would people be turned into zombies, but then a select few, marked for greatness are allowed to become boss zombies.
That's when I say to you, "Look pal you are willing to accept the dead rising from their graves to attack the living, but you can't handle some of them changing into something a little tougher….. Really? This is where your suspension of disbelief craps out on you? Not with the first zombie? Stop being so damned picky."
Do any of you remember when Carrot Top didn't look like a giant bee stung Chucky Doll? No? Just me… alrighty then.
In addition I'm not sure why the doctor has a poster hanging up of Carrot Top, but I'm sure it's something sinister.


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Posts Tagged 'Technical Writing'
Academic Advice Collection
মে 28, 2012 এখানে আপনার মন্তব্য রেখে যান
[Source: http://people.engr.ncsu.edu/txie/advice.htm]
Ph.D. dissertation/research advice:
- Advice by Tao Xie
- Imagining Research & Development: R&D student internships/student job advice
- Getting a Ph.D. at the University of Southern California by Yuriy Brun
- Pithy advice for graduate school by Rachel Pottinger and Kathryn. S. McKinley
- Some grad school advice by Noah Smith
- Advice by Frédo Durand
- How to choose an advisor and a topic by Vibha Sazawal
- Advice on Getting a Start into Research and Advice on Writing Research Papers by Tao Xie
- Writing a Research Paper by Jim Reggia
- Networking by Mary Jean Harrold and Kathryn. S. McKinley
- The Researcher's Bible by Alan Bundy et al.
- Advice for Prospective Research Students on Contacting Potential Advisors by David Evans
- Suggested guidelines for finding "related work" for conference papers. by Gail Kaiser.
- A course on Preparation for Statistical Research offered at NCSU Stat Dept
- Principles of Effective Research by Michael A. Nielsen
- The Ph.D Experience by Mihir Bellare
- Tom Henzinger's advice on doing research
- Citations: Why do we cite papers? by Jeff Offutt
- The scientific method by Jeff Offutt
- What is a PhD? by Jeff Offutt


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2016 Outstanding Young Virginian Awards
The Virginia Jaycees will honor outstanding young people between the ages of 21 and 40 through the Outstanding Young Virginians (OYV), Outstanding Young Farmer (OYF), Outstanding Future Leaders (OYFL), Outstanding Young First Responder (OYFR), and Outstanding Future Leader (OFL) programs. The awards will be presented during the banquet of the November Annual Conference. In addition to the presentation of the awards, there will be a guest speaker who is yet to be determined. The Virginia Jaycees have been dedicated to publicly recognizing Outstanding Young Virginians since 1942. Modeled on the United States Junior Chamber of Commerce's Ten Outstanding Young Americans program, this program gives the state organization the opportunity to recognize young Virginians who best exemplify through their activities the ideals expressed in the Jaycee Creed.
Since 1954, the Virginia Jaycees have honored an Outstanding Young Farmer. This program recognizes young farmers for achievements in agri-business as well as contributions made through the involvement in community, agriculture and other organizations. The purpose of the award is to foster better urban-rural relations by contact between Jaycees and farmers to create a greater public interest in, and understanding of today's farmer and their contribution to society.
In 1992, the Virginia Jaycees established the Outstanding Young First Responder Award to honor unselfish, life-saving contributions of pre-hospital providers. This award not only recognizes the contributions of the recipients, but honors the world's first rescue squad, established in Roanoke, Virginia by Julian S. Wise.
In 2003, the Outstanding Future Leaders category was created to recognize young people below the age of 21 for their accomplishments and leadership in the community. The Virginia Jaycees may consider scholarships for Future Leader recipients.
In 2013, the Virginia Jaycees unveiled a new award, Outstanding Local Business, to honor businesses in Virginia that have significantly supported our local chapters and in that way helped the Jaycee mission succeed and flourish.
Jaycee chapters, community organizations, businesses, and corporations throughout Virginia will submit the nominations. All nominations must be postmarked by October 5, 2016 (new deadline). A prestigious panel of judges, which will include Jaycee members, community leaders and representatives from the programs' sponsors, will review and select this year's recipients. Recipients will be recognized through a certificate and will be provided lodging, meals and admittance to Jaycee board meeting activities while attending the awards program. In addition, the Virginia Jaycees will select one recipient in the OYV category to nominate for the USJCC Ten Outstanding Young Americans award.
For questions or to submit your forms, please contact email@example.com.


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TROUT LAKE, Klickitat County — A former forest-ranger station popular with overnight visitors to Gifford Pinchot National Forest until it burned down last year is being replaced with a former bunk house and tool shed located 25 miles away.
The Willard Tool House, built by the Civilian Conservation Corps in 1940 and scheduled for demolition, will be moved to Peterson Prairie to replace the historic Peterson Prairie Guard station.
The Gifford Pinchot National Forest hopes to have the structure moved from Willard in eastern Skamania County to the Peterson Prairie location northwest of Trout Lake and available for rental by 2015.
Peterson Prairie Guard Station was built in 1926 to house U.S. Forest Service rangers. For many years, it was part of the recreation rental program and popular for overnight stays. The cabin was only 2.5 miles from Atkisson Sno-Park, which made it accessible to skiers and snowshoers during the winter. The cabin rented for $50 a night.
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In September, while the huge Cascade Creek forest fire raged on the south slope of Mount Adams, the Peterson Prairie Guard Station burned down one night in an unrelated fire. The guard station was many miles away from the Cascade Creek fire.
"When we heard about Peterson Prairie Guard Station burning down that morning we couldn't really believe it with everything else we had going on," said David Wickwire, Mount Adams District recreation program manager. "We went right out to see, and I don't have to tell most who loved this place about the empty sinking feeling when I first saw it."
The guard station was a total loss with only the chimney remaining.
"We received an outpouring of interest from local communities and numerous partners and volunteers expressing willingness to help up rebuild a cabin at the site," said Nancy Ryke, Mount Adams District ranger.
Willard is a tiny community along the upper Little White Salmon River on the southern edge of the Gifford Pinchot National Forest. It once was its own ranger district.
Willard Tool House was a tool shed and bunk house, said Rick McClure, heritage and tribal programs officer for the Gifford Pinchot.
"It has been sitting empty for 12 years, but it's in good shape structurally," McClure said.
The tool house is larger than Peterson Prairie Guard Station.
It has one large room on the ground floor that can be framed to make two rooms. Upstairs also is one large room that can be divided into two.
McClure said the tool house could be configured to have two private bedrooms upstairs and a living room and kitchen downstairs. The ground floor can be wheelchair-accessible.
Six to eight persons could stay in the tool house once it is refinished, he said.
The Forest Service has the money to move the tool house to Peterson Prairie. McClure said he envisions that happening this year or 2014.
Volunteers in the agency's Passport in Time program will be enlisted to help renovate and refurbish the tool house.
McClure said some of the work might be done before the tool house leaves Willard because there is electricity available there but not at Peterson Prairie.
The Mount Adams Institute is trying to raise $75,000 to finance the tool house restoration. The nonprofit has about one-third of the money secured.


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Fast Food: Taco Bell Triple Steak Stack
Would you like fries (or onion rings or chicken tenders) with that?
Taco Bell began the year defending themselves against allegations of false beefertizing, and although the lawsuit filed by an Alabama firm was eventually dropped, you never fully win a war that compels your website to add a tab titled "What Are These 'Other' 12% Ingredients?"
At the risk of sounding like an apologist for the beef extension industry, I'm on Taco Bell's side here. Their taco filling, which they call "seasoned beef," is 88 percent cheap beef and 12 percent the other kinda crap anyone with his head in the game might expect to find in a fast-food taco.
My favorite "seasoned beef" guest stars are oatmeal (to retain moisture) and cocoa powder (to make the oatmeal beef-colored). Maybe you're partial to soy lecithin or trehalose. Either way, it's not as if these crazy-named adjuncts are the ground-up bits of your favorite childhood pet; they're just standard elements of the fast food hustle. Anyone who rings the Taco Bell ought to have surmised beforehand that something a little untenderloin could be involved with a beef taco that nets you change from a Washington.
But if my defense of Taco Bell is based on the notion that a sub-buck burrito is going to come with a certain degree of mystery and modified corn starch, what's to be made of the new Triple Steak Stack? This baby costs $4.99, making it one of the more expensive standalone items on the fast food market.
The triple refers to the portion of steak: It's 4.5 ounces of "100% USDA Select marinated, grilled steak," which is three standard tacos' worth. In a bold departure for the chain, the steak stands mostly alone. It's augmented by a three-cheese blend (standard mozzarella, cheddar, and AmeriJack) and stuffed inside what they're calling a bolillo flatbread.
That's it. No onions or peppers, no lettuce or tomato, no pico de this or guaca-that. You are free to adulterate your Triple Steak Stack with the usual array of Taco Bell sauce, but for research purposes I went missionary.
In-store advertising proudly points out that the TTS is nine inches long. It makes sense that they'd appropriate Subway's ruler to describe what is essentially a steak and cheese sandwich, but the only fast food weight or measure that matters to me is gross tonnage, and the Triple checks out OK in that department. It's a bit thinner than most top-shelf Taco Bell offerings due to the lack of vegetables, beans, or rice, but it's still a fair portion of beef and cheese.
The bread was served medium-soggy and got damper as it cooled. It's thick for a folded sandwich (about ¼ inch), slightly sweet, and mostly bland. A short stretch on a grill or in a press could have smartened it right up, but as served, the limpness obscures the passable flavor.
If you eat it as the taco gods intended, the next layer you hit is the steak, but if you unfold it for research purposes, your immediate post-bread encounter is with an overgenerous spread of melted three-cheese. There's too much of the stuff, which would be a blessing if it were any good, but it's devoid of flavor other than extreme saltiness.
My three-ingredient lunch was off to a fatal two-strike start before I had a chance to consider the steak, which was a pleasant surprise if not a saving grace. It was cut to the ideal sandwich size: strips of about an inch by a half-inch by an eighth. It was tender without being mushy, and the marinade was peppery and reminiscent of a better brand of quickie mart beef jerky.
In light of the high price and troubles with bread and cheese, I can't recommend the Triple Steak Stack, but I will say that Taco Bell is ending the year in good beef standing.
All products linked here have been independently selected by our editors. We may earn a commission on purchases, as described in our affiliate policy.


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Table of Contents :
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Ready to Race
The Kawasaki Jet Ski Ultra 310R personal watercraft (PWC) is the perfect prescription for adventurous riders seeking a pure adrenaline rush on the water. Based on Kawasaki's popular Jet Ski Ultra 310X, R-model draws from decades of Kawasaki's championship-winning PWC racing experience to create the ultimate "sport" personal watercraft that's ideal for hard core recreation and racing. The Ultra 310R is equipped with an 18 position handle bar and a sports seat.
Key Features
- The Supercharged DOHC inline four-cylinder engine produces a staggering 310 horsepower. It's the most powerful production PWC in history
- KX motocross-style electro-polished stainless steel handlebar enhances the 310R's sport riding capabilities while also providing excellent all-day comfort
- 12-position handlebar mount can be adjusted for reach and height to allow operation while standing or sitting and to suit a wide range of riders and conditions
- The Sportseat offers a special support bolster and a grippy KX-motocross-like seat cover for hard sport riding
- Aggressive graphics reinforce the hard-core sport capabilities of the Ultra 310R
- Strong and light, the 301R's deep-V hull offers superb straight-line tracking and precise cornering performance
- Industry-leading, 212L storage capacity lets you bring your gear with you
- 78L fuel tank is the largest in its class, providing tremendous range for long rides
- Separate keys for full power operation and Smart Learning Operation
Made to Perform
The post style handlebar is similar to the kind found on Kawasaki's KX450F motocross bike.....although this bar offers even greater adjustability. Handle position is adjustable both for height and forward / rearward position. A total of 12 positions are available. Riders can choose from three height positions (STD / 5mm up / 10mm up) which can be adjusted using 5mm washers. Two sets of handle bar slots and reversible handlebar holders offers four forward / rearward positions. Optional handlebar clamps add another six positions. The stainless steel handle features and electropolish finish for a high quality appearance and the handlebar comes equipped with a racy bar pad.
Sport Seat
The Ultra 310R is equipped with a stylish sport seat specifically designed for hard sport riding. The moderate contour of the seat supports the rider's hips, offering enhanced lower body support for hard sport riding. The seat shape flares less at the thighs than the standard seat of the Ultra 310X. This makes it easier to grip the seat with the knees when stand-up riding in rough water. Textured material, similar to that used on the KX motocross bikes contributes to the racy appearance and the ability to grip the seat in all situations. Adding to the quality image, having the hand strap start at the pattern line created a more integrated design. A JET SKI logo and black stitching are some other quality touches. An ergonomic deck design gives plenty of legroom and the left-side reverse lever is ergonomically designed for easy operation. At the end of the day on the water, a quick and easy hose connection allows a simple method to flush sea water out of the engine.
310 Horsepower Supercharged Engine
The 1,498cc, liquid-cooled inline four-cylinder engine makes 310 horsepower via a low mechanical loss design and improved efficiency. The Eaton TVS constant-displacement supercharger and liquid-cooled intercooler provide instant and consistent boost for outstanding performance throughout the rev range.
- Heat-resistant plastic intake manifold is shaped to improved low-mid range power
- Cast pistons feature additional ring land V-grooves for better oil retention
- Hardened nickel 33.4mm intake and 28.3mm exhaust valves offer superior reliability
- Two-layer oil cooler offers efficient cooling
- Large cam chain increases reliability in a supercharged engine application
- The reinforced crankcase adds to engine's durability and reliability
- Fuel pump provides a high pressure, high flow fuel for increased power and better fuel efficiency.
- Thick water jackets improve cooling performance
- Check valve system between the cylinder jacket and water muffler that shuts water flow off above 2,000 rpm, to keep the cooling water inside the cylinder jacket for optimum cooling performance
- Piston cooling oil jets increased from one jet per piston to two jets per piston for increased piston cooling
- Oil pan features additional baffle plates to prevent oil slosh during tight turns or extreme acceleration/deceleration
- Crankcase oil return holes are designed for quicker oil return to the oil pan
Fuel Injected
High-flow fuel injectors with 4 orifices are fitted to complement the increased power output and to improve combustion efficiency. A single throttle body is used with a bore diameter of 60 mm.
Air-to-Water Intercooler
Efficiently cools high-pressure intake air – cooler air is denser and makes more power. Liquid-cooled intercooler design maintains consistent thermal performance.
Large Impellor
To suit the 160 mm jet pump, the Ultra 310R gets an impeller designed to efficiently convert all the extra power into forward thrust. The pump impeller features a super-efficient 3-blade design that gives the 310R great initial acceleration.
Rear Mounting
The wide rear deck behind the seat offers plenty of room to set up gear for water sports and facilitates getting back on board from the water. A long rear boarding step makes it easy to climb on board, and the step retracts inside the bumper to prevent damage.
Quattro KSD
The 310R's Kawasaki Splash Deflector (KSD) uses three bottom-mounted ridges and an additional deflector mounted at the top to deflect water splashing forward from under the hull. Whether turning or straight-line riding at any speed the Quattro KSD minimizes spray, allowing greater rider concentration.
Ride Plate
A groove running from the centre to the rear of the ride plate makes it easier to hold a straight course when running straight ahead.
Twin Vortices Supercharger
Four-lobe, constant-displacement design offers significant performance increase over standard roots-type unit. The system force-feeds the engine two liters of air with every revolution, providing a constant supply of high-pressure intake air at all rpm. The compressed air is delivered continuously, not in waves, for smoother power delivery.
Strong, Light Hull Design - Superb in Calm and Rough Water
Designed to emulate the performance of Kawasaki's IJSBA championship-winning race machines the reinforced inner bow is extremely light with ample strength to handle the engine's power output. Exemplary straight-line tracking and responsive handling in both calm and rough-water conditions set performance benchmarks for the class. The lower hull is specially designed to distribute the engine and power delivery loads over a wide area. Deep-V hull design enables a steep bank angle for motorcycle-like turning performance. This reduces the lateral G-loads imposed on riders during turns, allowing them to better concentrate on riding. The 22.5° V-angle at the bottom of the hull helps reduce unwanted spinouts during sharp turns. This all assist in delivering the neutral handling and turning characteristics for which Kawasaki JET SKI watercraft are famous The bottom of the hull slants upward steeply from the center section to the bow, so the 310R can penetrate swells and waves with less shock, pushing through waves rather than bouncing over them at high speeds. Integrated sponsons extend from the hull, providing wallow-free straight-line performance and contributing significantly to the 310's excellent maneuverability while helping to retain the narrow hull design.
Jet Pump
To effectively convert the 310 PS output into thrust, the Ultra 310X utilises a large 160mm jet pump, designed using eight guide vanes. The pump delivers superior "traction" in rough water conditions and significantly contributes to the Ultra 310X's mind-boggling acceleration.
The Ultra 300X features a 4-into-2-into-1 powerboat-style exhaust system, which routes the exhaust pipe upwards. This allows the smoothest flow for exhaust gases, maximising exhaust efficiency.
Electronic Trim Control System
Electronic Trim Control System allows the rider to control the angle of the jet pump nozzle to suit water surface conditions and riding style. Adjustment range is plus or minus 8 degrees from standard.
Electronic Cruise Control
IMPROVED Fuel Economy Assistance mode revised for significant fuel savings and increased range. Convenient electronic cruise control smoothly holds a set speed from 8 kph up to top speed. One-touch, push-button 8 kph mode allows hassle-free no-wake zone riding and the economical Riding Indicator mode can be viewed on a graph on the LCD info screen.
Large Capacity Storage
The huge front storage compartment comes with removable tray combined with a centre console storage area.
- Large-capacity, fully sealed storage area with detachable storage tray offers ample room for recreational equipment
- Compact integrated underseat storage area is ideal for small items like a ski rope
Multiple Riding Modes
The ECU controlled ETV allows multiple riding mode options to be activated with the touch of a button. 8 kph mode conveniently regulates rpm to maintain constant speed in no-wake zones and cruise control fixes speed at a specified level for convenient long distance touring.
Comfort and Convenience
Roomy three-passenger seat offers ample support plus all-day comfort whether riding stand-up solo, seated or with passengers. The passenger handrail on the Ultra 310X is located to a more ergo-friendly location to help reduce passenger fatigue and features a redesigned handle for better grip. The optimally curved mirrors have been designed for a wider rear view and are mounted to the upper deck via new brackets An ergonomic deck design gives plenty of legroom and the left-side reverse lever is ergonomically designed for easy operation. At the end of the day on the water a quick and easy hose connection allows a simple method to flush sea water out of the engine.
Instrument Panel
LCD screen features a face design that's easy to read. The instrumentation includes a fuel consumption function, a boost meter scale, riding modes, external water and air temperature, clock, time, trip and hour meters, tachometer (in digital or graph displays), speedometer and max speed functions. Eco Indicator shows the rider when the speed and throttle opening is in the 310LX's most efficient range. The Kawasaki Eco Mode reduces and smoothes engine performance while conserving fuel.
Beginner Friendly
The Ultra 310LX features Kawasaki's innovative electronic Smart Learning Operation (SLO) system which can be programmed to speed-limit the engine to assist novice riders in mastering their riding technique without worrying about excessive speeds.
|Engine Type
|Supercharged Intercooled, 4-stroke In-Line Four
|1498 cm³
|Bore X Stroke
|83 x 69.2 mm
|Compression Ratio
|8.2 : 1
|Valve System
|DOHC 4 valve per cylinder
|Fuel System
|Electronic Fuel Injection: ø60 mm x 1
|Battery Coil (Transistorised Ignition)
|Inducted water
|Drive System
|Direct drive from engine
|Axial flow, single stage
|8,407 N {857kgf}
|Impeller Diameter
|160 mm
|Steerable Nozzle
|Maximum Power
|228 kW {310 PS} / 8,000 rpm
|Maximum Torque
|269 N-m {29.2 kgf-m} / 6,000 rpm
|Overall Length
|3,370 mm
|Overall Width
|1,195 mm
|Overall Height
|1,155 mm
|Fuel Capacity
|78 litres
|Curb Mass
|475 kg includes full full tank, tool kit, and all fluids at optimum levels


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Blumenthal, Murphy hold panel, talk gun violence
HARTFORD >> Connecticut's U.S. senators see the national attention generated by the Newtown shootings as an opportunity to address gun violence. To begin that process they heard Friday from a community that's no stranger to the subject: Hartford's North End.
U.S. Sens. Richard Blumenthal and Chris Murphy, both D_Conn., held a roundtable discussion Friday afternoon at the Parker Memorial Community Center in Hartford. The panel included elected leaders as well as law enforcement officials, mental health professionals, and faith leaders.
The senators also heard from members of the community, many of whom had lost family members to gun violence in Hartford.
For the rest of the story, please click here.


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NETTUTS.com recently posted a handy tutorial for those wanting to use the font of their choice on their site using sIFR3.
Last week, Philo showed you how to implement sIFR3. This time, I'm going to show you how to implement Facelift Image Replacement (or FLIR), an alternative to sIFR that does not require Flash.
This second part shows you how to install/setup the tool, configure it correctly for your setup and includes some of the pros and cons of using the software.


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Download our detailed WakeUP user guide,
titled "Improving Crop Performance with WakeUP."
You'll see 8 color pages of research and farmer reports showing how WakeUP Spring and WakeUP Summer can improve yields and quality for corn, soybeans, wheat, forages and other crops. The report is in PDF format (Portable Document File). If you don't have Adobe Acrobat Reader to open the file on your computer, contact us. We'll mail you a copy.