Split (2)

validation · 215k rows

text stringlengths 1 3.78M	meta dict
Removal of dining club licenses shakes up Park City establishments From left, Jeff Jacobs, Kaleb Doilney, Max Doilney and Joanna Doilney are frustrated at the law requiring them to give up their dining club license. Max Doilney and Jacobs, who own and operate the Corner Store Pub and Grill near Park City Mountain Resort, say that whatever they choose, they will have losses. It's no secret that Utah has strict liquor laws. During the 2017 legislative session, some regulations were loosened while new ones restricted establishments even further. One of the biggest changes, included in H.B. 442, made it so dining club licenses, which allowed places to be both a restaurant and a bar, would be eliminated this year. After failed attempts from dining club owners — including many in Park City — to lobby for a change to or delay of that part of the law during this year's legislative session, the elimination of dining club licenses is expected to go into effect on July 1. Businesses operating under a dining club license must decide by the end of next month whether to become a bar or a restaurant, designations that both carry various requirements. Those in the restaurant industry say the situation has left almost two dozen establishments around Park City struggling to determine their next steps. If businesses choose the restaurant route, they must follow the 70/30 rule, meaning that the ratio of food to alcohol gross sales must be at least 70 percent to 30 percent. Max Doilney, the owner of the Corner Store Pub and Grill, said that is especially difficult because Utah sets the wholesale cost of liquor by requiring restaurants to purchase liquor from the state. For restaurants with high-priced food, hitting the ratio is not a problem, but at places like the Corner Store, where the average cost of a meal is $15, he said it is an obstacle. Recommended Stories For You "The state is essentially dictating our entire food and beverage market by setting their price," Doilney said. "We are in a pickle because we can't hit 70/30 or we'll have to raise our food prices, which alienates families." The alternative, dropping the cost of alcoholic beverages, would reduce the restaurant's net profit, he said. If the Corner Store and other businesses around Park City choose to be licensed as a restaurant, they also must require guests who drink an alcoholic beverage to order food with their drink. With dining club licenses, that was not the case. David Wakeling, the owner of Collie's Sports Bar, said that requiring guests to order food is not good for business. "It's giving our state even more of a bad reputation," he said. Plus, if Collie's is a restaurant, it would have to close at 11 p.m. rather than 1 a.m. But even with those consequences, Wakeling does not want to lose the families that visit his restaurant, which would happen if he were to become a bar. People under 21 are not allowed in bars. "Park City in particular has so few options for families as it is, and this is taking away more of that," he added. Elected officials did pass a law this year, H.B. 456, that allows businesses to hold both a restaurant and a bar license. But the restaurant and bar areas must be in separate rooms with distinct entrances and with shared walls being at least 8 feet high. For many, that would require construction that they do not have the money or space for. Shantel Stoff, sales and marketing director of Red Rock Brewery and co-president of the Park City Area Restaurant Association, said that Red Rock Brewery in Kimball Junction could go that route. There is a room in the back that Red Rock would be able to convert into a bar, but it is not ideal. "The portion of the restaurant that has to become a restaurant versus the bar is not how we would prefer to continue, but it is what we are faced with given the legislation that passed," she said. She said that Red Rock has not yet decided what it will choose to do. It was so difficult to get the dining club license that it had, it is hard to give it up, she said. She served as general manager of Red Rock Brewery for 10 years and, during that time, spent 27 months meeting with the Utah Department of Alcoholic Beverage Control to be permitted a dining club license. Doilney worries what would happen if the majority of the establishments choose to opt for a bar license, since it seems to be easier to comply with. "You might as well have an ID station at the Junction saying, 'Don't come into town with your kids because there is no place for them to eat,'" he said. Doilney predicts that it will balance out, but there does not seem to be a perfect solution for most of the Park City dining clubs. "It's not an easy task for any of the business owners in Park City," he said. Doilney said that he built his business around the previous liquor laws, and now he will need to do construction if he wants to keep the guests, and lifestyle, he has. If he is licensed as a bar, his nephew, who is 15-years-old, will no longer be able to work at the establishment or even step foot in the building. His young son will not be able to either. "They are not just taking away a little bit of our revenue and a little bit of our livelihood, they are taking away our lifestyle," he said. He said that part of his frustration is that he and several other business owners spent months lobbying to get the elimination of the dining club license delayed or changed during this year's legislative session. He said that senators and representatives toured establishments with dining club licenses in Park City and restaurant owners shared with them a list of their concerns. "They gave us assurances that our concerns would be addressed," he said. "In my opinion, they look us in the eye and say they are going to do something and they don't do it." Stoff said that the restaurant association hired someone to help guide it through the legislative session so it could voice its worries about the law, but no progress was made. She said that many businesses were blindsided by some elements of the law last year. Doilney said it seems they are being punished. "The law is incredibly shortsighted," he said. "It puts stresses on small businesses that otherwise have been abiding by the law better than other liquor outlets."	{ "pile_set_name": "Pile-CC" }
An evaluation of non-invasive blood pressure (NIBP) monitoring on the wrist: comparison with upper arm NIBP measurement. The arm is the traditional site for application of an oscillometric non-invasive blood pressure (NIBP) cuff This study, which compares upper arm NIBP to wrist NIBP, involved 510 same arm sequential paired blood pressure (BP) measurements in 85 volunteers. Wrist NIBP consistently overestimated mean arterial, systolic and diastolic pressure by approximately 10 mmHg. Ninety per cent of the mean arterial pressure differences at the wrist were within a range of +/- 9 mmHg around a mean difference of 10.6 mmHg. The systolic pressure difference was 11.2 mmHg with 90% of differences between +/- 12 mmHg. The diastolic pressure difference was 10.2 mmHg with 90% of differences between +/- 9 mmHg. With the device used and within the normal blood pressure range, compensation can be performed by subtracting 10 mmHg from the measured values or simply by elevating the wrist about 15 cm and taking the BP at face value. Wrist NIBP may be a viable clinical alternative in situations where difficulty occurs with upper arm NIBP measurement.	{ "pile_set_name": "PubMed Abstracts" }
It used to be that craft beer nerds living in St. Paul had to cross the river for some variety — there just weren’t enough breweries in the Capital City to satisfy us. But now some of the metro’s best breweries reside squarely in the 651 area code. We officially have 12 breweries and one brewpub in St. Paul proper. Five more will open soon. That doesn’t include the east metro suburbs, which are home to some really great breweries and brewpubs. In case you’re not aware, the difference between brewpubs and breweries is that brewpubs can serve wine, beer or liquor made outside the premises, but they cannot distribute the beer they make. Brewpubs almost always serve food. Breweries can serve food (most choose not to), but they cannot serve outside wine, liquor or beer. They can distribute what they make to liquor stores and restaurants. Hours vary at many breweries. Some are open just a few days a week, so be sure to call or check their website before heading out. Here is our east-metro guide to craft beer drinking at the source, where a glass of suds tastes best. COMING SOON Clutch Brewing Company This brewery will be located in the mezzanine level of the Keg and Case development in the old Schmidt Brewery on West Seventh Street. Owners Jordan Standish and Max Boeke are longtime homebrewers ready to live the craft brewery dream. Expected opening is April 2018. Clutch Brewing: 928 W. Seventh St., St. Paul; clutchbeer.com Waldmann Brewing Owner Tom Schroeder took on a daunting project in renovating the oldest surviving commercial building in the Twin Cities. The saloon was originally built in 1857, and the restaurant and taproom portion of the brewery will reside in the original building. There’s an addition off the back that will house the kitchen and the brewing equipment. The beers here will be classic German styles, and more than 80 percent of what they serve will be lagers. Waldmann will also serve a menu that mostly consists of sausage and sides, including a currywurst, weisswurst and a hot dog from Red Table Meats. Expected opening is Oct. 1. Waldmann Brewing: 445 Smith Ave., St. Paul; 651-222-1857; waldmannbrewery.com Culhane Brewing When Sidhe Brewing on Payne Avenue closed, owner Kathleen Culhane immediately launched a new project, Culhane Brewing. To be located in the former Station 4 building in Lowertown, the brewery will be another super-small batch operation, but this time with a kitchen. Opening date has yet to be determined. Culhane Brewing: 201 E. Fourth St., St. Paul; culhanebrewing.com Stacked Deck Brewing Company Formerly known as Looney Bin Brewing, this brewery, which will be in the Treasure Island Center in the former Macy’s downtown (which will also house a training facility for the Minnesota Wild), will serve a wide variety of beers, according to co-owner Andy Erickson. He and his partners, Brad Randall and Scott Wege, are former homebrewers taking the next step. Expected to open in February or March 2018. Stacked Deck Brewing: 421 Cedar St., St. Paul; stackeddeckbrewing.com Yoerg Brewing Another historical revival, but of a different sort, Yoerg has already brought back beer originally brewed in Minnesota in 1848, but they’ve been brewing it in Wisconsin. The new taproom they’re working on, in the former Wabasha Bar on St. Paul’s West Side, will also serve food, mostly bar food, including a currywurst, pizzas from Giovanni’s in Brainerd, Minn., pickled eggs and pate. They’ll have wine, too. Expected opening is Oct. 1. RELATED: Map of breweries in St. Paul and east metro suburbs Yoerg Brewing: 427 S. Wabasha St., St. Paul; yoergbeer.com ST. PAUL 12welve Eyes Brewing This downtown St. Paul brewery quietly opened in July with a whopping 12 beers on tap. They’re making really small batches, so don’t get too attached to any one beer because they go fast. That said, if something is popular, they’ll probably bring it back around. Related Articles Lunds & Byerlys to stay closed on day after Thanksgiving Surdyk’s adds bar and eatery called Sidebar Neighborhood girl finds and returns chef Justin Sutherland’s stolen knife roll 8 ways to make school-at-home lunches easier Game on! Tailgating takeout ideas for game day The space, in the historic Pioneer Endicott building, is on the garden level, but still manages to get plenty of light. There’s ample seating around the bar and at tables to bring a group or spread out and play games — there was a group playing poker on one of our visits. They have a space in their alley for food trucks that they’ll use eventually, but for now you’ll have to bring your own. What to drink: They haven’t been open long enough to have a flagship beer, but we’ve enjoyed their IPAs. 12welve Eyes Brewing: 141 E. Fourth St., Suite LL2, St. Paul; 651-493-8106; 12welveeyes.com Bad Weather Brewing Bad Weather has quickly evolved into a must-visit spot for most of my beer-nerd friends, and I’m no exception. There is always a huge variety of beer on tap — I can’t remember ever visiting and not finding something new. For all the variety, each one is made with so much care that I never hesitate to order something I’ve never tried. The staff is friendly and efficient, and the space is big enough that you can usually find a table, even when it is busiest. It can be tough to score a spot on the patio, but at least the garage doors that open to the outside offer some fresh air. There’s usually a food truck at Bad Weather, and because they’re usually so busy, they get some of the best street food in the cities. What to drink: Whatever they’re pouring, but our favorite seasonals are Tippin’ It Down (an ESB brewed with Earl Grey Tea) and Firefly Rye (a pale ale made with rye malt). Bad Weather Brewing: 414 W. Seventh St., St. Paul; 651-207-6627; badweatherbrewery.com Bang Brewing You have to admire the moxie of Jay and Sandy Boss Febbo, the owners of Bang. They are making 100 percent organic beer, and that’s not easy to source. In addition, the brewery, which is housed in a slick, stainless steel grain bin, is 100 percent wind-powered. All their good intentions would be for nothing if the beer wasn’t good, but it is. Some of my favorite in the state, as a matter of fact. Add to that the charm of one of the two of them always being behind the bar and the fact that they brew every batch without help, and you’ve got a great story and reasons to come back again and again. Oh, and there’s a sweet patio, too. There is often a food truck or sausage cart, but check their website if you’re counting on it. What to drink: Neat (a sparkling bitter) or Nice (a dark ale that’s great with a shot of coffee in it, which the brewery also offers). Bang Brewing: 2320 Capp Road, St. Paul; 651-243-2264; bangbrewing.com Barrel Theory Squarely at the center of exciting things happening in St. Paul brewing, Barrel Theory is the result of two former Surly employees spinning off on their own. The brewery is churning out expensive, labor-intensive beers that are available only in their sleek Lowertown taproom. Beer enthusiasts have been packing the place, so much so that they tend to run out of popular brews quickly. If you’re a fan of sours, or even if you’re not, try any of their Berliner Weisse variations — they’re best in class. They’re also brewing some really nice, hazy IPAs and currently have my favorite pale ale. No food trucks, but you can order from Dark Horse Bar and Eatery, which is next door and has great bar food, including some of the most underrated pizza in town. What to drink: Everything, but especially Falkor (an American pale) and Key Sublime (a Berliner Weisse). Barrel Theory Beer Company: 248 E. Seventh St., St. Paul; 651-600-3422; barreltheory.com BlackStack Brewing This spacious warehouse brewery shares a building with Can Can Wonderland, the crazy artist-created mini-golf course/eatery/entertainment space in the industrial area off University Avenue near the Minneapolis border. Their lineup of beer might be smaller than some other breweries, but the beers they do have are solid, creative and easy to drink. The taproom is sizable, so feel free to bring a group, and even though Can Can gets crowded, there’s ample parking available. There are food trucks some days but not others — check the calendar on their website. What to drink: Ends Meet (an amber-colored Belgian ale) or whatever northeast-style IPA they’re pouring. Black Stack Brewing: 755 N. Prior Ave., St. Paul; 612-369-2934; blackstackbrewing.com Burning Brothers Brewing This brewery’s tagline, “Don’t fear the beer,” says it all — co-founder Dane Breimhorst has celiac disease but loves beer. So he decided to make something he could drink. The brewery boasts a variety of styles, including a lime shandy in the summer, that are popular among the gluten-free crowd and even those who can have regular beer. The brewery is in the Hamline-Midway neighborhood, nestled in a residential area, and though it’s a cozy spot, we haven’t had trouble getting a seat. They even host gluten-free food trucks pretty regularly — there’s a calendar on the website. What to drink: Parched (a lime shandy), or Roasted (a coffee strong ale) Burning Brothers Brewing: 1750 W. Thomas Ave., St. Paul; 651-444-8882; burnbrosbrew.com Flat Earth Brewing Company Housed in the former Hamm’s brewery facility on the East Side, Flat Earth has a giant bar, a separate sampling room, plenty of seating and a patio for outside imbibing. The brewery also has lots of events, including trivia, live music and quirky Hot Wheels races for kids and adults alike. Flat Earth makes a lot of Belgian ales, but also plenty of American standards and fun seasonals. Food trucks are hit or miss here, so it’s best to eat before you come. What to drink: Belgian-style pale ale or Black Helicopter coffee stout Flat Earth Brewing: 688 E. Minnehaha Ave., St. Paul; 651.698.1945; flatearthbrewing.com Great Waters The only brewpub in St. Paul, Great Waters has occupied the corner of St. Peter Street and Seventh Place in downtown St. Paul for 20 years — long before the current craft-beer boom. They’ve been brewing some award-winning ales during that time and always offer an array of regular and cask-conditioned beers. The restaurant menu is huge, and includes a wide array of bar snacks, sandwiches, entrees and salads. It’s a great place to meet friends before a downtown event or unwind after work. What to drink: Saint Peter Pale Ale or Brown Trout Brown Ale Great Waters Brewing Company: 426 St. Peter St., St. Paul; 651-224-2739; greatwatersbc.com Lake Monster Brewing Housed in the gorgeous old King Koil Mattress Factory just off the Cretin-Vandalia exit of Interstate 94, Lake Monster’s spacious taproom means you can bring all your friends to hang out. The brewery’s beers are generally pretty standard, but solid, but there’s usually one on the list that is at least somewhat experimental. There’s also a pretty sprawling patio out front, and most nights, there’s a food truck. What to drink: Empty Rowboat IPA or India Pale Gose Lake Monster Brewing: 550 Vandalia St., St. Paul; 612-964-6288; lakemonsterbrewing.com Summit Brewing The oldest brewery on this list, Summit Brewing has been making craft beer since before it was cool — founder Mark Stutrud first brewed the brewery’s flagship Extra Pale Ale in 1986. The brewery’s stable of year-round beers are always a solid choice, but in recent years, they’ve really let their brewers get creative, to great effect, with their Unchained series. The beers are limited-release, and they are distributed, but also served in the brewery’s Beer Hall, a community gathering room that basically turned into a taproom on the weekends after the Surly Bill was passed in 2011. What to drink: The beer that started it all, Extra Pale Ale, Oatmeal Stout (which isn’t sold in retail outlets) or any of the Unchained varieties. Summit Brewing: 910 Montreal Circle, St. Paul; 651-265-7800; summitbrewing.com Tin Whiskers Started by three electrical engineers, Tin Whiskers was the first brewery to locate downtown. Their signature Wheatstone Bridge, a wheat beer brewed with chamomile tea and honey, might be the most ubiquitous tap beer in St. Paul restaurants. The taproom is a modern space in the Rossmor Building. Given that there are three restaurants in the building (Black Sheep Pizza, Keys Cafe and Sawatdee), the brewery does not partner with food trucks but encourages patrons to bring in food from its neighbors. What to drink: Wheatstone Bridge or Flip Switch IPA. Tin Whiskers Brewing: 125 E. Ninth St., St. Paul; 651-330-4734; twbrewing.com Urban Growler The only full-fledged brewery in St. Paul that has a kitchen (for now), Urban Growler has recently hired a new chef that has kicked the food up a notch to really good bar food. The brewery is owned by two women, which makes it unusual in the heavily bearded hipster-heavy craft beer scene. The beer here is great, too. Though their flagship beers are firmly in the classic category, their Plow to Pint series, in which they partner with local farmers to add farm-fresh ingredients, and their seasonals offer some seriously fun variety. What to drink: Big Boot Rye IPA or any of the Plow to Pint beers. Urban Growler Brewing Company: 2325 Endicott St., St. Paul; 651-340-5793; urbangrowlerbrewing.com Wabasha Brewing This brewery on St. Paul’s West Side has a pretty, historic taproom, with the brewery operation up above. It’s usually full of neighborhood regulars, which gives it a charming dive-bar feel, but with artwork for sale on the walls. There’s also a small patio adjacent to the taproom. The brewery is probably best known for its West Side Popper, a jalapeno cream ale that is heavy on the fresh, green pepper flavor and pretty light on the spice. What to drink: West Side Popper or Lawnmower Porter. Wabasha Brewing Company: 429 S. Wabasha St., St. Paul; 651-224-2102; wabashabrewing.com EAST METRO SUBURBS Angry Inch Who knew downtown Lakeville was so charming? This storefront brewery is on Holyoke Avenue, the main drag of the historic district. It has a family vibe — there were moms, dads, kids and even grandparents there when we visited. They have a wide variety of beer, with a lot of styles on the sweet, dark and high-alcohol side of things. What to drink: Daddy’s Honeypot (a French saison) or Samoan Kisses (a milk stout with chocolate and coconut notes) Angry Inch Brewing: 20841 Holyoke Ave., Lakeville; angryinchbrewing.com Bald Man Brewing Housed in an industrial strip mall just across from the Eagan Outlet Mall, this brewery is the perfect place to hide while your friends and family who love to shop are across the street. The taproom here is sizable, and there are seven solid beers on tap at any given time. Bald Man has a food truck almost every night, too. What to drink: Tupelo Honey Brown Ale or Young American Pale Ale. Bald Man Brewing: 2020 Silver Bell Road, Suite 28, Eagan; 651-600-3164; baldmanbrewing.com Bent Brewstillery This combo brewery/distillery is the only one of its kind in the Twin Cities. Owner Bartley Blume is one part mad scientist, one part activist, as the state’s liquor laws don’t allow him to serve both beer and cocktails. The beer here, which they are allowed to serve, is creative and the atmosphere cool. Be sure to check out the bathrooms, which are some of our favorites in the Twin Cities. What to drink: Nordic Blonde or any of the beers in their Funked Up series of sour beers. Bent Brewstillery: 1744 Terrace Drive, Roseville; 844-879-2368; bentbrewstillery.com Barley John’s This roadside brewpub in New Brighton has a great patio with a soaring fire pit and hops growing up trellises surrounding it. If you’re lucky, there might be live music, too. They have a full bar, wine and a fairly large menu that includes sandwiches, salads, burgers, pizzas and entrees. What to drink: Stockyard IPA or Old 8 Porter Barley John’s Brewpub: 781 Old Highway 8, New Brighton; 651-636-4670; barleyjohns.com Big Wood Brewing This downtown White Bear Lake brewery embraces its suggestive moniker in more ways than one, first by outfitting its taproom with lots of wood, including heavy wood-carved chairs (short shorts are a no-no here, you could end up with a splinter) and beers with cheeky names like Morning Wood, Bad Axe and Bark Bite. Ignore the names if they are off-putting to you, though, because the beer here is good. What to drink: Morning Wood (a coffee stout) or Amigo Grande (a Mexican lager) Big Wood Brewing: 2222 Fourth St., White Bear Lake; bigwoodbrewery.com HammerHeart It’s the farthest flung of the suburban breweries on this list, but HammerHeart makes some road-trip worthy beers. The brewery has a dark, Nordic theme that is very masculine, but very cool, and the brewery has a tasty niche: smoked beer. There are often some delicious non-smoked varieties on tap if that doesn’t float your boat. Smoke is applied to the grains here with an expert hand and it’s really worth a try. What to drink: Thor’s Hot Pepper Porter or Bog Burial (Scotch barrel-aged peat-smoked old ale) HammerHeart Brewing Company: 7785 Lake Drive, Lino Lakes; 651-348-2654; hammerheartbrewing.com Lift Bridge Brewing A true community gathering spot, Lift Bridge hosts something called Townie Tuesdays, in which the brewery donates to a local charity. They also host a holiday cookie bake-off, an annual hop-picking event and live, local music. It doesn’t hurt that the beers here are some of the best in the entire metro area — good enough to travel for, even if you don’t live in the Stillwater area. What to drink: Anything, really, but Hop Dish, Mango Blonde and The Warden (a rich milk stout) are our favorites. Lift Bridge Brewing Company: 1900 Tower Drive W., Stillwater; 888-430-2337; liftbridgebrewery.com Maple Island Brewing This charming brewery is in historic downtown Stillwater, and the best place to be is the little patio in the back of the shop, with views of the St. Croix River. The beers here trend toward the gimmicky, including one brewed with ice cream from the legendary Stillwater shop Nelson’s, and one named White Butt IPA. Just go with it. What to drink: Maple Island Bock or Burlesque Kolsch Maple Island Brewing: 225 Main St. N., Stillwater; 651-430-0044; mapleislandbrewing.com Nutmeg Brewhouse Related Articles Game on! Tailgating takeout ideas for game day The brave, the few, the takeout only: Booya in 2020 More than a dozen walk-up window ideas for your outdoor dining pleasure Here’s your list of 20+ places offering State Fair food & drinks in lieu of the Fair We haven’t yet been to the original Burnsville location of this suburban brewpub, but the beer is the same in Arden Hills, and it’s good. The food menus are different at the different locations, too, and the Arden Hills spot, which is located squarely in a nondescript strip mall, changes its menu fairly frequently. As with all brewpubs, you can get outside booze here — beer, wine and cider in Arden Hills, and a full bar in Burnsville. What to drink: Any of the rye IPAs Nutmeg Brewhouse: 1905 County Road 42W, Burnsville; 952-892-1438 or 3673 N. Lexington Ave., Arden Hills; 651-340-8812; nutmegbrewhouse.com	{ "pile_set_name": "OpenWebText2" }
1. Field of Invention Relates to a tubular key controlled operation of a terminal based on the coded interruption of a light stream therein as read by a counter adapted therewith to make operational a computer program in circuit therewith, the program being identified by said code as read by said counter. 2. Description of the Previous Art There is not known presently to be any terminal providing access to secured data in a computer by a coded interruption of a light stream generated within the terminal and read by a counter having access to a program in a computer to make the same operational. The applicant in his U.S. Pat. No. 4,322,719 disclosed a coded solid state entry device by the use of a tubular coded key engaging a plurality of switches to energize a circuit to unlatch tumblers of which a code is read which functions to match with an electronic circuit to allow access or entry. Known also are terminals which are operated by coded plastic cards which require card reading machines which in turn are adapted to put a computer into an operational mode to perform a specified task.	{ "pile_set_name": "USPTO Backgrounds" }
With the advent of transmission of multimedia content such as video over the Internet and other communications networks, there is any ever-increasing need for increased data rate capacity on communication networks. Often, optical communication signals are used in backbones of communications network, where the increased traffic at the edges of the network aggregates to several gigabit of network traffic. Therefore, there is an increasing need for optical communication techniques to meet the demand on communications network data capacity. However, laying down additional optical transmission media such as fiber optics often requires large amount of capital expenditure and may not always be a suitable options due to the expenses involved and other regulatory issues. Techniques for improved optical transceiver operation are needed.	{ "pile_set_name": "USPTO Backgrounds" }
Arbitration Arbitration falls under the category of Alternative Dispute Resolution, acronym ADR. Arbitration offers an alternative path for disputing parties to gain a binding decision in a private, confidential process held outside of the normal legal channels, such as filing a lawsuit in court. Arbitration is most commonly selected by signing a contract with an arbitration provision before a dispute arises. It is a very common dispute resolution process in commercial disputes; it is almost non-existant in criminal matters, as the courts generally retain jurisdiction in criminal matters. Arbitration is different than the court system. It is not a judicial process, even though there is sometimes pressure to treat it as such. Arbitration sprang from a premise that business and industry disputes may be best resolved outside of a criminal court system. Arbitration is common in business and commercial contracts; these are the cases that it was originally designed to address, and they represent the bulk of the cases today. Private and Binding: Arbitration is a private process that results in a binding decision, called an award. Arbitration offers a confidential hearing before a neutral decision-maker, often an industry expert, who delivers a binding award with limited appeal after providing the parties with a full and fair hearing. Confidentiality is perceived as a tremendous benefit, protecting the reputations of those involved in the dispute. This is in stark contrast to courts of law, where decisions are published and referred to for years to come. Arbitration is primarily used in business and commercial disputes, and also in family matters. Arbitration is not generally a venue for criminal cases. Parties may agree on many aspects of arbitration either ahead of time, by including an arbitration clause in a contract, or at the time a dispute arises. Agreements about how arbitration is conducted includes what forum will administer the claim, where the hearing is conducted, what type of hearing, whether the award should be a simple award or fully supported (a reasoned award), and other procedural matters. Many arbitrators are appointed or selected based on their expert knowledge and experience within an industry; others are chosen for their understanding of the legal process, such as former administrative law judges or seasoned, reputable attorneys. Arbitrator's Role: An arbitrator is a private judge; a finder of fact in a hearing of equity. Arbitrators must disclose any known conflict of interest and sign an oath that they have no known bias and can be fair in dealing with the parties and the subject matter in dispute prior to accepting appointment to a case. Parties may not contact or communicate with the arbitrator outside the presence of all parties to the dispute. Rules of Procedure: The arbitration process is governed by rules of procedure specified in the arbitration provision of a signed contract. When a contract provides that any disputes will be settled by binding arbitration using specific rules of procedure, the arbitrator conducts the case according to the time frames, document and exhibit provisions, and hearing parameters noted in the rules. Often the arbitration provisions is vague, with the only requirement being arbitration. Other times the provision can be quite specific. The parties may jointly agree to utilize Colorado Mediators & Arbitrator's administrative services rather than the forum specified in the contractual arbitration provision. By joint agreement, the arbitrator may conduct the arbitration utilizing the rules specified in the contract, or that the parties unanimously agree to use Colorado Mediators & Arbitrators Arbitration Rules of Procedure. Size of Claim Determines Procedure: Colorado Mediators & Arbitrator’s rules provide that smaller claims have a less formal hearing than larger claims in order to provide reduced cost and compressed time frames for the benefit of the parties. Small Claims ($14,999 or less) have a flat fee and are decided on the written evidence submitted. Expedited Claims ($15,000 - $74,999) have an administrative fee and a flat hearing fee if conducted in four (4) hours or under. Large Claims ($75,000 and over) consist of one or more pre-hearing teleconferences followed by a hearing attended by the parties and their attorneys, when represented. The necessary hearing time is determined by the parties. Arbitration is Less Formal than Litigation Using the court system to solve problems is often overkill. The length of time necessary to get to hearing is often prohibitive. The legal process is punctuated by formality and is often confusing to non-attorneys. A business or layperson’s understanding of the Rules of Evidence often prohibits submitting evidence in litigation unless it is done in a narrow, specific way. Legal formality fosters the need to hire an attorney, which adds to the cost of resolving the dispute, and the dispute itself is usually costly. It is not uncommon for litigation to take years! Additionally, the element of fairness may not be addressed by a judge because judges are bound by both the written law and an extensive body of case law, created over the years by a myriad of past cases that may or may not seemingly apply to your business case. Arbitration is Not Mediation Mediation also falls under the category of ADR, and often these terms are used interchangeably due to confusion. To clarify, mediation is a formal negotiation of the matter and does not have a guaranteed conclusion. Just like when you offer to buy a car, if the seller doesn’t like the deal, they can walk away. If the seller likes the deal, they sign a contract and there is a legal consequence – the car’s ownership changes hands. However, arbitration results in a decision, like it or not. Arbitration offers a full and fair hearing of each party’s presentation of the evidence and testimony of the parties. Big difference, even though both mediation and arbitration fall under the category of Alternative Dispute Resolution, or ADR for short. Arbitration Testimonials "Used you in 2007 and was very pleased with the outcome. Business dispute."Gail Bryan (Business Arbitration) “Judy Larkins has built a very deserving reputation for ADR excellence. She is skilled as a mediator/arbitrator and also as program manager. She has both vision and drive to increase the use of ADR and to arrive at creative solutions that satisfy parties.” Deborah Schick Laufer, Director, Federal ADR Network (FAN) "I have known Judy Larkins for the past 10 years, and during that time participated in many mediations at Colorado Mediators & Arbitrators and referred many friends and attorneys to Judy. From the business side of the process, Judy is a pleasure to work with. The fees are fair and there are no surprises. She also organizes the mediation process to be cost efficient. From the procedural process, Judy does an excellent job of helping the parties work through their disputes and reach agreements." Von Limbaugh, Attorney "I am an independent arbitrator/mediator on the panel at Colorado Mediators and Arbitrators. Judy asked me to join the panel and I have enjoyed working with her -- both on specific cases and in relation to her efforts to continually improve CoMA's rules and services. She is easy to work with and is very competent and professional. CoMA is her business and she works hard to make it successful by recruiting good arbitrators and mediators to serve on her panels and by providing excellent service to her clients."John Sass, Attorney/Arbitrator/Mediator	{ "pile_set_name": "Pile-CC" }
/* * ==================================================================== * Licensed to the Apache Software Foundation (ASF) under one * or more contributor license agreements. See the NOTICE file * distributed with this work for additional information * regarding copyright ownership. The ASF licenses this file * to you under the Apache License, Version 2.0 (the * "License"); you may not use this file except in compliance * with the License. You may obtain a copy of the License at * * http://www.apache.org/licenses/LICENSE-2.0 * * Unless required by applicable law or agreed to in writing, * software distributed under the License is distributed on an * "AS IS" BASIS, WITHOUT WARRANTIES OR CONDITIONS OF ANY * KIND, either express or implied. See the License for the * specific language governing permissions and limitations * under the License. * ==================================================================== * * This software consists of voluntary contributions made by many * individuals on behalf of the Apache Software Foundation. For more * information on the Apache Software Foundation, please see * <http://www.apache.org/>. * */ package org.apache.hc.core5.http.impl.io; import java.io.ByteArrayInputStream; import java.io.ByteArrayOutputStream; import java.io.InputStream; import java.net.Socket; import java.nio.charset.StandardCharsets; import org.apache.hc.core5.http.ClassicHttpRequest; import org.apache.hc.core5.http.ClassicHttpResponse; import org.apache.hc.core5.http.ContentType; import org.apache.hc.core5.http.HttpEntity; import org.apache.hc.core5.http.Method; import org.apache.hc.core5.http.NotImplementedException; import org.apache.hc.core5.http.ProtocolException; import org.apache.hc.core5.http.config.Http1Config; import org.apache.hc.core5.http.impl.DefaultContentLengthStrategy; import org.apache.hc.core5.http.io.entity.StringEntity; import org.apache.hc.core5.http.message.BasicClassicHttpResponse; import org.junit.Assert; import org.junit.Before; import org.junit.Test; import org.mockito.Mock; import org.mockito.Mockito; import org.mockito.MockitoAnnotations; public class TestDefaultBHttpServerConnection { @Mock private Socket socket; private DefaultBHttpServerConnection conn; @Before public void setUp() throws Exception { MockitoAnnotations.initMocks(this); conn = new DefaultBHttpServerConnection("http", Http1Config.DEFAULT, null, null, DefaultContentLengthStrategy.INSTANCE, DefaultContentLengthStrategy.INSTANCE, DefaultHttpRequestParserFactory.INSTANCE, DefaultHttpResponseWriterFactory.INSTANCE); } @Test public void testBasics() throws Exception { Assert.assertFalse(conn.isOpen()); Assert.assertEquals("[Not bound]", conn.toString()); } @Test public void testReadRequestHead() throws Exception { final String s = "GET / HTTP/1.1\r\nUser-Agent: test\r\n\r\n"; final ByteArrayInputStream inStream = new ByteArrayInputStream(s.getBytes(StandardCharsets.US_ASCII)); Mockito.when(socket.getInputStream()).thenReturn(inStream); conn.bind(socket); Assert.assertEquals(0, conn.getEndpointDetails().getRequestCount()); final ClassicHttpRequest request = conn.receiveRequestHeader(); Assert.assertNotNull(request); Assert.assertEquals("/", request.getPath()); Assert.assertEquals(Method.GET.name(), request.getMethod()); Assert.assertTrue(request.containsHeader("User-Agent")); Assert.assertEquals(1, conn.getEndpointDetails().getRequestCount()); } @Test public void testReadRequestEntityWithContentLength() throws Exception { final String s = "POST / HTTP/1.1\r\nUser-Agent: test\r\nContent-Length: 3\r\n\r\n123"; final ByteArrayInputStream inStream = new ByteArrayInputStream(s.getBytes(StandardCharsets.US_ASCII)); Mockito.when(socket.getInputStream()).thenReturn(inStream); conn.bind(socket); Assert.assertEquals(0, conn.getEndpointDetails().getRequestCount()); final ClassicHttpRequest request = conn.receiveRequestHeader(); Assert.assertNotNull(request); Assert.assertEquals("/", request.getPath()); Assert.assertEquals(Method.POST.name(), request.getMethod()); Assert.assertTrue(request.containsHeader("User-Agent")); Assert.assertNull(request.getEntity()); Assert.assertEquals(1, conn.getEndpointDetails().getRequestCount()); conn.receiveRequestEntity(request); final HttpEntity entity = request.getEntity(); Assert.assertNotNull(entity); Assert.assertEquals(3, entity.getContentLength()); Assert.assertEquals(1, conn.getEndpointDetails().getRequestCount()); final InputStream content = entity.getContent(); Assert.assertNotNull(content); Assert.assertTrue(content instanceof ContentLengthInputStream); } @Test public void testReadRequestEntityChunckCoded() throws Exception { final String s = "POST /stuff HTTP/1.1\r\nUser-Agent: test\r\nTransfer-Encoding: " + "chunked\r\n\r\n3\r\n123\r\n0\r\n\r\n"; final ByteArrayInputStream inStream = new ByteArrayInputStream(s.getBytes(StandardCharsets.US_ASCII)); Mockito.when(socket.getInputStream()).thenReturn(inStream); conn.bind(socket); Assert.assertEquals(0, conn.getEndpointDetails().getRequestCount()); final ClassicHttpRequest request = conn.receiveRequestHeader(); Assert.assertNotNull(request); Assert.assertEquals("/stuff", request.getPath()); Assert.assertEquals(Method.POST.name(), request.getMethod()); Assert.assertTrue(request.containsHeader("User-Agent")); Assert.assertNull(request.getEntity()); Assert.assertEquals(1, conn.getEndpointDetails().getRequestCount()); conn.receiveRequestEntity(request); final HttpEntity entity = request.getEntity(); Assert.assertNotNull(entity); Assert.assertEquals(-1, entity.getContentLength()); Assert.assertEquals(true, entity.isChunked()); Assert.assertEquals(1, conn.getEndpointDetails().getRequestCount()); final InputStream content = entity.getContent(); Assert.assertNotNull(content); Assert.assertTrue(content instanceof ChunkedInputStream); } @Test(expected = ProtocolException.class) public void testReadRequestEntityIdentity() throws Exception { final String s = "POST /stuff HTTP/1.1\r\nUser-Agent: test\r\nTransfer-Encoding: " + "identity\r\n\r\n123"; final ByteArrayInputStream inStream = new ByteArrayInputStream(s.getBytes(StandardCharsets.US_ASCII)); Mockito.when(socket.getInputStream()).thenReturn(inStream); conn.bind(socket); Assert.assertEquals(0, conn.getEndpointDetails().getRequestCount()); final ClassicHttpRequest request = conn.receiveRequestHeader(); Assert.assertNotNull(request); Assert.assertEquals("/stuff", request.getPath()); Assert.assertEquals(Method.POST.name(), request.getMethod()); Assert.assertTrue(request.containsHeader("User-Agent")); Assert.assertNull(request.getEntity()); Assert.assertEquals(1, conn.getEndpointDetails().getRequestCount()); conn.receiveRequestEntity(request); } @Test public void testReadRequestNoEntity() throws Exception { final String s = "POST /stuff HTTP/1.1\r\nUser-Agent: test\r\n\r\n"; final ByteArrayInputStream inStream = new ByteArrayInputStream(s.getBytes(StandardCharsets.US_ASCII)); Mockito.when(socket.getInputStream()).thenReturn(inStream); conn.bind(socket); Assert.assertEquals(0, conn.getEndpointDetails().getRequestCount()); final ClassicHttpRequest request = conn.receiveRequestHeader(); Assert.assertNotNull(request); Assert.assertEquals("/stuff", request.getPath()); Assert.assertEquals(Method.POST.name(), request.getMethod()); Assert.assertTrue(request.containsHeader("User-Agent")); Assert.assertNull(request.getEntity()); Assert.assertEquals(1, conn.getEndpointDetails().getRequestCount()); conn.receiveRequestEntity(request); final HttpEntity entity = request.getEntity(); Assert.assertNull(entity); } @Test public void testWriteResponseHead() throws Exception { final ByteArrayOutputStream outStream = new ByteArrayOutputStream(); Mockito.when(socket.getOutputStream()).thenReturn(outStream); conn.bind(socket); Assert.assertEquals(0, conn.getEndpointDetails().getResponseCount()); final ClassicHttpResponse response = new BasicClassicHttpResponse(200, "OK"); response.addHeader("User-Agent", "test"); conn.sendResponseHeader(response); conn.flush(); Assert.assertEquals(1, conn.getEndpointDetails().getResponseCount()); final String s = new String(outStream.toByteArray(), "ASCII"); Assert.assertEquals("HTTP/1.1 200 OK\r\nUser-Agent: test\r\n\r\n", s); } @Test public void testWriteResponse100Head() throws Exception { final ByteArrayOutputStream outStream = new ByteArrayOutputStream(); Mockito.when(socket.getOutputStream()).thenReturn(outStream); conn.bind(socket); Assert.assertEquals(0, conn.getEndpointDetails().getResponseCount()); final ClassicHttpResponse response = new BasicClassicHttpResponse(100, "Go on"); conn.sendResponseHeader(response); conn.flush(); Assert.assertEquals(0, conn.getEndpointDetails().getResponseCount()); final String s = new String(outStream.toByteArray(), "ASCII"); Assert.assertEquals("HTTP/1.1 100 Go on\r\n\r\n", s); } @Test public void testWriteResponseEntityWithContentLength() throws Exception { final ByteArrayOutputStream outStream = new ByteArrayOutputStream(); Mockito.when(socket.getOutputStream()).thenReturn(outStream); conn.bind(socket); Assert.assertEquals(0, conn.getEndpointDetails().getResponseCount()); final ClassicHttpResponse response = new BasicClassicHttpResponse(200, "OK"); response.addHeader("Server", "test"); response.addHeader("Content-Length", "3"); response.setEntity(new StringEntity("123", ContentType.TEXT_PLAIN)); conn.sendResponseHeader(response); conn.sendResponseEntity(response); conn.flush(); Assert.assertEquals(1, conn.getEndpointDetails().getResponseCount()); final String s = new String(outStream.toByteArray(), "ASCII"); Assert.assertEquals("HTTP/1.1 200 OK\r\nServer: test\r\nContent-Length: 3\r\n\r\n123", s); } @Test public void testWriteResponseEntityChunkCoded() throws Exception { final ByteArrayOutputStream outStream = new ByteArrayOutputStream(); Mockito.when(socket.getOutputStream()).thenReturn(outStream); conn.bind(socket); Assert.assertEquals(0, conn.getEndpointDetails().getResponseCount()); final ClassicHttpResponse response = new BasicClassicHttpResponse(200, "OK"); response.addHeader("Server", "test"); response.addHeader("Transfer-Encoding", "chunked"); response.setEntity(new StringEntity("123", ContentType.TEXT_PLAIN)); conn.sendResponseHeader(response); conn.sendResponseEntity(response); conn.flush(); Assert.assertEquals(1, conn.getEndpointDetails().getResponseCount()); final String s = new String(outStream.toByteArray(), "ASCII"); Assert.assertEquals("HTTP/1.1 200 OK\r\nServer: test\r\nTransfer-Encoding: " + "chunked\r\n\r\n3\r\n123\r\n0\r\n\r\n", s); } @Test(expected = NotImplementedException.class) public void testWriteResponseEntityIdentity() throws Exception { final ByteArrayOutputStream outStream = new ByteArrayOutputStream(); Mockito.when(socket.getOutputStream()).thenReturn(outStream); conn.bind(socket); Assert.assertEquals(0, conn.getEndpointDetails().getResponseCount()); final ClassicHttpResponse response = new BasicClassicHttpResponse(200, "OK"); response.addHeader("Server", "test"); response.addHeader("Transfer-Encoding", "identity"); response.setEntity(new StringEntity("123", ContentType.TEXT_PLAIN)); conn.sendResponseHeader(response); conn.sendResponseEntity(response); conn.flush(); } @Test public void testWriteResponseNoEntity() throws Exception { final ByteArrayOutputStream outStream = new ByteArrayOutputStream(); Mockito.when(socket.getOutputStream()).thenReturn(outStream); conn.bind(socket); Assert.assertEquals(0, conn.getEndpointDetails().getResponseCount()); final ClassicHttpResponse response = new BasicClassicHttpResponse(200, "OK"); response.addHeader("Server", "test"); conn.sendResponseHeader(response); conn.sendResponseEntity(response); conn.flush(); Assert.assertEquals(1, conn.getEndpointDetails().getResponseCount()); final String s = new String(outStream.toByteArray(), "ASCII"); Assert.assertEquals("HTTP/1.1 200 OK\r\nServer: test\r\n\r\n", s); } }	{ "pile_set_name": "Github" }
Order filed June 26, 2015. In The Fourteenth Court of Appeals ____________ NO. 14-15-00440-CV ____________ IN THE INTEREST OF N.N.F., A CHILD On Appeal from the 315th District Court Harris County, Texas Trial Court Cause No. 2013-06707J ORDER This is an accelerated appeal from a judgment in a suit in which the termination of the parent-child relationship is at issue. Appellant's brief was due June 22, 2015. No brief has been filed. Appeals in parental termination cases and child protection cases are to be brought to final disposition within 180 days of the date the notice of appeal is filed. See Tex. R. Jud. Admin. 6.2(a) (effective May 1, 2012). This accelerated schedule requires greater compliance with briefing deadlines. Therefore we order appellant’s appointed counsel Lana Shadwick, to file appellant’s brief no later than July 9, 2015. If the brief is not filed by that date, counsel may be required to show cause why he should not be held in contempt of court. In addition, the court may require appointment of new counsel due to the failure to timely file appellant’s brief. PER CURIAM	{ "pile_set_name": "FreeLaw" }
Introduction ============ Children are exposed to diverse types of evaluative feedback regarding their behavior, intelligence, and performance in their social lives. Parents and teachers often provide children with positive feedback to enhance their intrinsic motivation and boost their self-esteem ([@B16]; [@B5]). However, an increasing body of evidence suggests that praise is not always beneficial for children (see [@B9]; [@B16] for a review). For example, unlike process-related feedback (e.g., "you worked hard"), receiving personal feedback (e.g., "you are smart") following success, when children's positive performance is attributed to their personal traits, could foster resistance to subsequent mistakes ([@B30]; [@B19]). Notably, [@B16] suggested that the effects of praise on motivation depend on not only the praise content (e.g., process-related feedback or personal feedback) but also the recipients' characteristics and interpretation of the praise. Evidence accumulated from research conducted thus far has revealed that some variables, such as perceived autonomy and performance standards, affect children's motivation (e.g., [@B7]; see [@B16] for a review). However, few studies have examined insincerity, another key variable, or have assessed the manner in which children interpret and respond to praise that leaves room for doubt. Developmental studies have shown that young children can produce and understand insincere utterances from around the age of 4 or 5 years ([@B1]; [@B3]). Although previous literature has not directly examined the sensitivity of children to insincerity in praise, this evidence leads us to speculate that children around the age of four or five come to notice insincerity in praise that is given in an inappropriate situation (i.e., failure situation), and begin to display negative responses to insincere praise. The question as to how and when children should be praised is one of the main concerns of adults engaged in parenting and education; however, very few attempts have been made to examine the impact of praise in situations where children fail and evaluate their own performance as poor. Indeed, evaluations received from others are not always congruent with children's own self-evaluation; for example, parents and teachers sometimes praise children when the children themselves feel that they did not perform well on a task (i.e., when they have failed). A recent study that examined children's responses to inflated praise found that school-aged children with low self-esteem sought fewer challenges after they received inflated praise ([@B5]). Crucially, this finding indicated that children with low self-esteem might evaluate their own performance as poor compared to children with high self-esteem, and the disagreement between internal low evaluation and external high evaluation (i.e., inflated praise) might cause the negative response to the praise, because they might recognize the praise as insincere. This study also focused on children's theory of mind that could be one of the key factors that serve to constrain their interpretation of and responses to insincere praise. Theory of mind is defined as the ability to attribute mental states to the self and others to interpret behavior ([@B31]). False-belief tasks are commonly used to measure young children's theory of mind development, and a large body of evidence has shown that children pass first-order false-belief tasks between the ages of 3 and 5 years ([@B39]). Individual differences in theory of mind in early childhood influence children's social lives in diverse ways ([@B17]). Theory of mind allows children to comprehend non-literal speech, such as irony (e.g., [@B11]), white lies ([@B4]), and antisocial lies ([@B34]), by understanding the intention behind the words that are spoken. That is, as theory of mind develops, children can infer the intention underlying non-literal speech and detect its true meaning. Children need to interpret the meaning of praise and compliments in situations where they receive insincere praise. Therefore, in situations where children are praised following failure, they may use theory of mind to determine the reason for the praise. Ambiguity in praise, which is characteristic of feedback in Japanese culture, is another key topic that has been overlooked in previous research. The Japanese language is highly contextualized, and the content of the speaker's discourse is ambiguous, rather than explicit ([@B26]). In Japan, ambiguity is observed not only in language but also in parenting and education. Japanese mothers adopt an "absorbent childrearing" approach, where they do not teach children explicitly but encourage them to understand situations and behave autonomously ([@B2]). In addition, Japanese preschool and kindergarten educators use the machi no hoiku (childrearing of waiting) strategy, whereby they hold back and wait, allowing children the opportunity to handle problems on their own with minimal assistance from teachers or adults ([@B35]; [@B13]), and this strategy might support both positive and negative self-reflection. Against a cultural background involving this language system and these educational beliefs, Japanese children might be more likely to receive ambiguous evaluative feedback without an apparent reason (e.g., "great," "good," or "okay") relative to Western children. To my knowledge, [@B20] conducted the first study to examine ambiguous praise, where the experimenter suddenly approached 2- to 4-year-old children while they were engaged in free play at preschool and provided them with praise with no indication of the reason for which praise had been provided ("great!" or "sugoi!" in Japanese). Children received this feedback in situations where they had not performed activities that was appropriate for evaluation (e.g., success, failure), and their facial and verbal reactions were observed. The results showed that most 2-year-old children and some 3-year-old children smiled and showed agreement with the sudden ambiguous praise, while some 3-year-old children and most 4-year-old children exhibited confusion at the sudden ambiguous praise, with dubiety in their facial expressions, and half of them asked about the reason for the praise. These findings suggest that at approximately 4 years of age, children begin to consider not only praise itself but also the reason for praise. Although [@B20] observation indicated that 4-year-old children were confused by ambiguous praise and cared about the reason for which praise had been offered, children's responses to and interpretation of ambiguous praise in situations involving activities that evaluation is appropriate for (e.g., success, failure) remained unclear. The overall aim of the current study was to examine young Japanese children's interpretations of and responses to insincere praise, which involves ambiguity in the reason of the praise, and explore the association between these responses and the maturity of their theory of mind. The first hypothesis was that participants would display a negative emotional response, low performance rating, and low motivation to insincere praise, similar to that observed in the failure situation without feedback, since they notice that the praise is insincere. The second hypothesis was that most participants would attribute the receipt of insincere praise following failure to their efforts rather than outcomes or their personal attributes. This hypothesis was proposed based upon the Japanese cultural belief system, which emphasizes the importance of hard work, effort, and persistence in self-improvement (e.g., [@B36]; [@B15]). Their tendency to attribute the insincere praise to their efforts may be the basis for low motivation after receiving insincere praise, as this includes the message that "they had already made a sufficient effort." The third hypothesis concerned the relationship between theory of mind and the child's responses to insincere praise. It was hypothesized that children with mature theory of mind would be more likely to report negative responses (i.e., negative emotion, low levels of self-rated performance, and weaker motivation for perseverance) after receiving insincere praise following failure, relative to children with less mature theory of mind. The proposed reason for this was that, whereas children without mature theory of mind could consider the praise as sincere if they take it at face value, children who possess mature theory of mind could interpret the hidden message behind the praise and consider it invalid. Therefore, when children possess mature theory of mind, their positive emotion, performance rating, and motivation for further challenge could be reduced because they speculate the reason of the praise in relation to the failure situation. Methods {#s1} ======= Participants ------------ Seventy-two Japanese children aged 4 years and 7 months to 6 years and 9 months participated (mean age = 5.70 years, SD = 0.61; 24 boys, 48 girls). All participants were native Japanese speakers attending one of two preschools. An ethics approval for psychological research was not required at the time the research was planned and conducted as per the author's institution guidelines (Meiji Gakuin University) as well as national regulations. Though there was no opportunity to submit an ethics application at that time, the author carried out this study in accordance with the recommendations of the Japanese Psychological Association Ethical Principles of Psychologists with careful attention to ethical considerations to uphold high ethical standards. All participants' parents gave the author written informed consent in accordance with the Declaration of Helsinki. Although detailed demographic information regarding the children could not be obtained because of school privacy policies, the sample was recruited predominantly from middle-class families. Materials and Procedure ----------------------- The children were tested individually in quiet rooms at the preschools. They performed a failure-praise task, theory of mind tasks, and a verbal ability test. The order of the tasks was counterbalanced. ### Failure-Praise Task A new task designed to measure individual differences in responses to praise following failure was developed for this study. Children's responses were examined by using stories in which they appear as a protagonist puppet; then, they were asked how they felt in the stories. The task structure and procedure were based on sensitivity-to-criticism tasks that have been used to assess children's responses to criticism in both Eastern and Western countries (e.g., [@B6]; [@B21], [@B22]; [@B27], [@B28]; [@B29]). The stories that were used in the tasks are shown in Appendix [1](#A1){ref-type="app"}. In the failure-praise task, the experimenter read two puppet-based stories (a "no-feedback story" and a "praise story") aloud and directed the children to act out the role of the main character simultaneously, using a puppet. The task flow is shown in Figure [1](#F1){ref-type="fig"}. Before the experimenter read the stories, the children were instructed to choose one of four puppets to represent themselves and introduced it to another puppet that represented the teacher. The children were told that the puppet child (representing the participant) and the puppet teacher were going to play a block design game that involved the use of four bicolored blocks from the Wechsler Preschool and Primary Scale of Intelligence Block Design test ([@B38]). In the block design game, the puppet teacher showed the puppet child a design on a picture card. The puppet child was asked to recreate the design using the blocks, within a limited amount of time. ![Flow of the procedure for the failure-praise task.](fpsyg-09-01684-g001){#F1} To ensure that the children understood the rules of the game, they were provided with similar, but easier, examples and allowed to practice. Upon completion of the two practice trials, they were told the no-feedback story (control story) first, followed by the praise story (test story). This was to avoid the effect of praise on the response in the no-feedback story (control story) and to confirm how they evaluate their failure itself. The two stories contained descriptions of very similar events. In both stories, the child failed to recreate a design, which was shown on a picture design card, within a limited amount of time. The difference between the two stories was that praise was provided in the praise story but not the no-feedback story. The no-feedback story was used as the control story, mainly to confirm whether children were dissatisfied with their performance when they failed. In the no-feedback story, the children were told that the main character (i.e., the child) played a block design game and tried hard to recreate a complex design with the blocks but failed to do so within the time limit. In the praise story, the main character played another block design game with a similar degree of difficulty and tried hard to recreate the design but failed to do so. However, in the praise story, the teacher offered the main character insincere praise without a reason (i.e., "sugoi-ne!" in Japanese that means "great!" in English), in response to the work. The two picture design cards used in the stories were counterbalanced. The following three questions were posed after each of the stories: (Q1) Emotional response: "I want to know how you feel about what happened in this block design game" followed by, "Do you feel happy or not? Do you feel sad or not? Do you feel angry or not?" They answered each question after being shown illustrations of each emotional facial expression. To create an index of a general positive emotional response, one point was awarded for each positive emotion (i.e., happy, not sad, and not angry), and a summed score was calculated (scores ranged from 0 to 3). (Q2) Performance rating: "Think again about everything that happened in this block design game. Should you get a circle (good) or a cross (not good) for what you did?" Children were asked to assign a grade to their block design by pointing to a circle (which means "good" in Japan) or a cross. One point was awarded for a positive evaluation (good), and no points were awarded for a negative evaluation (not good). (Q3) Motivation: "Think again about everything that happened with this block design game. If the teacher asked you to try the game again, would you like to play the difficult game or the easier game?" The children were asked this question to measure their motivation for attempting to perform challenging activities in the stories. They were asked to choose between a card depicting a complex design and a card depicting a straightforward design. One point was awarded for choice of the challenging activity, and no points were awarded for choice of the easy activity. The children were asked another question about interpretation of praise in the praise story, as the praise provided in the story was ambiguous, and they could have interpreted it in diverse ways: (Q4) Interpretation of praise: "Why did the teacher say, 'great!' (sugoi-ne!)?" Children's responses were classified into three categories: "outcome," "effort," and "other" (i.e., other interpretation, do not know, and no response). If the children did not answer, they were asked the following forced-choice question: "Did she offer the praise because you did good work (outcome) or because you tried hard (effort)?" Six of the 72 children (8.33%) were asked the forced-choice question. The author and a second coder coded participants' interpretation of the praise. Interrater agreement, established using Cohen's kappa, was high (K = 0.85). Disagreements were resolved via discussion. In the debriefing, the children and the experimenter played a block design game without a puppet, if the children wanted to try to recreate the design themselves, and they talked about successful experiences in their daily lives. ### Theory of Mind Tasks The children completed theory of mind tasks that included two first-order false-belief tasks ([@B12]) and two second-order false-belief tasks ([@B33]). The example of stories and questions used in the theory of mind tasks are shown in Appendix [2](#A2){ref-type="app"}. These tasks have demonstrated good test-related reliability ([@B18]). Each of the first-order false-belief tasks included two memory control questions, a first-order false-belief question, and a reality question. If the children answered all four questions correctly, they received one point. Each of the second-order false-belief tasks included a first-order false-belief question, a reality question, a second-order false-belief question, and two memory control questions. The children answered the second-order false-belief question and memory control questions only if they had answered the first two questions (the first-order false-belief and reality questions) correctly. If children answered both the first-order and reality questions correctly, they received one point for understanding first-order false beliefs, and if they answered the second-order false-belief and memory control questions correctly, they received one additional point for understanding second-order false beliefs. Therefore, overall theory of mind scores ranged from 0 to 6 (i.e., four points for understanding first-order false beliefs and two points for understanding second-order false beliefs). ### Verbal Ability Test Children's vocabulary was assessed to control for verbal ability in the analysis, as this ability is typically positively related to their performance in theory of mind tasks ([@B25]). The Picture Vocabulary Test-Revised (PVT-R; [@B37]), which was standardized for use with Japanese samples, was used to measure children's receptive vocabulary. The PVT-R is the Japanese version of the Peabody Picture Vocabulary Test ([@B8]). In the task, children were shown an array of four pictures on each page of a stimulus book and asked to select an appropriate picture, which was named by the experimenter, from the four pictures. The standardized score was used for the following analyses. Results ======= Data from 52 children (18 boys and 34 girls) who evaluated their work in the no-feedback story as "not good" (low performance rating after failure without feedback) were included in the main analysis examining their responses to the praise in the praise story. This data selection process was essential because young children tend to be unrealistically overconfident of their ability (e.g., [@B32]) and could be unable to recognize their failure in the story. Table [1](#T1){ref-type="table"} shows the means and standard deviations for data regarding age, PVT-R scores, theory of mind scores, and scores for each measure in the failure-praise tasks, for the 52 children. ###### Descriptive statistics. Measure M (SD) Range ----------------------- --------------- -------- Age in months 68.67 (7.42) 55--81 Verbal ability 26.31 (10.52) 6--46 Theory of mind 3.38 (1.92) 0--6 No-feedback story Emotional response 0.92 (0.65) 0--3 Performance rating 0.00 (0.00) 0 Motivation 0.73 (0.45) 0--1 Praise story Emotional response 2.33 (0.96) 0--3 Performance rating 0.58 (0.50) 0--1 Motivation 0.58 (0.50) 0--1 Note. n = 52. Concerning an overview of the analysis, first, children's responses to the praise story were compared with their responses to the no-feedback story by using Wilcoxon signed-rank tests to test the first hypothesis. To test the second hypothesis, their interpretation of the praise following failure was coded. Lastly, a logistic regression analysis concerning their responses to praise following failure per their theory of mind development was conducted to test the third hypothesis. Children's Response to Insincere Praise --------------------------------------- To test the first hypothesis, which expected that young children would show a negative response to insincere praise, Wilcoxon signed-rank tests for positive emotion, self-rated performance, and motivation were performed. Children's levels of positive emotion (Z = −5.53, p \< 0.001), and self-rated performance (Z = −5.48, p \< 0.001) were significantly higher relative to those observed in the no-feedback story. Their motivation was significantly lower relative to those observed in the no-feedback story (Z = −2.53, p = 0.01). Children's Interpretation of Insincere Praise --------------------------------------------- To test the second hypothesis, which predicted that most participants would attribute the receipt of insincere and ambiguous praise following failure to their own effort in a story, children's interpretation of the praise was coded. The interpretation provided by seven of the 52 children was categorized as "other," and the remaining 45 children were classified into one of two groups based on their attribution of the praise: 27 were classified into the outcome-interpretation group (e.g., children who believed that the teacher had offered praise because the recreated puzzle had been good or they had been able to recreate half of the puzzle) and 18 were classified into the effort-interpretation group (e.g., children who believed that the teacher had offered praise because they had worked hard or done their best to complete the challenging puzzle). Age, t(43) = −0.74, ns; PVT-R scores, t(43) = 1.24, ns; and theory of mind scores t(43) = 0.13, ns did not differ significantly between outcome- and effort-interpretation groups. Theory of Mind and Responses to Insincere Praise ------------------------------------------------ A logistic regression analysis was performed to test the third hypothesis, which predicted that children with mature theory of mind would respond more negatively to the praise following failure, relative to children with less mature theory of mind. Coding of the interpretation of insincere praise following failure revealed that there were two major types of interpretation (i.e., effort and outcome)^[1](#fn01){ref-type="fn"}^. Thus, whether the interaction between theory of mind and the interpretation were significantly associated with their response to insincere praise was also analyzed. First, theory of mind scores were centered, and interpretation of praise was coded into effort (+1) and outcome (−1). To conduct the logistic regression analysis, children's scores for the emotional response question were divided into two categories reflecting higher (2 and 3 points) and lower (0 and 1 points) levels of positive emotion. As dependent variables, children's responses to praise (emotional response, performance rating, and motivation) were examined in each separate analysis. In each analysis, age and PVT-R scores were entered in the first step, theory of mind scores and interpretation of the praise were entered in the second step, and the interaction terms of theory of mind scores and interpretation of the praise were entered in the final step. Table [2](#T2){ref-type="table"} shows the results of the logistic regression analysis for the three dependent variables. For emotional response, the model was not significant, χ^2^(5) = 5.00, p = 0.42. For performance rating, the model was significant, χ^2^(5) = 14.56, p = 0.01, and the main effect of theory of mind was significant, B = −0.87, Wald = 6.90, p = 0.01, indicating that mature theory of mind was related to relatively lower performance ratings after insincere praise. Although the model for motivation did not reach significance, χ^2^(5) = 9.11, p = 0.11, the interaction between theory of mind and interpretation of the praise approached significance, B = −0.38, Wald = 3.80, p = 0.05. A simple slope analysis revealed that theory of mind score was negatively associated with motivation after praise following failure in the effort-interpretation group, B = −0.60, Wald = 3.11, p = 0.08, indicating that there was a tendency that children with mature theory of mind displayed lower motivation after insincere praise, relative to children with less mature theory of mind in this group. On the other hand, for children in the outcome-interpretation group, theory of mind was not significantly related to motivation, B = 0.15, Wald = 0.22, p = 0.64. ###### Results of the logistic regression analysis (final model). Variable B SE Wald df Odds ratio %ΔOdds 95% CI ---------------------------------- ------- ------ ---------- ------ ------------ -------- ------------ Emotional response Age 0.05 0.08 0.37 1 1.05 +5 0.90--1.23 Verbal ability 0.07 0.06 1.40 1 1.08 +8 0.95--1.22 ToM −0.10 0.29 0.11 1 0.91 −9 0.51--1.61 Interpretation (effort, outcome) −0.04 0.44 0.01 1 0.96 −4 0.41--2.26 ToM × Interpretation −0.07 0.23 0.10 1 0.93 −7 0.60--1.45 Performance rating Age −0.12 0.09 1.86 1 0.89 −11 0.75--1.05 Verbal ability 0.17 0.07 6.68^∗∗^ 1 1.19 +19 1.04--1.35 ToM −0.87 0.33 6.90^∗∗^ 1 0.42 −58 0.22--0.80 Interpretation (effort, outcome) 0.50 0.44 1.28 1 1.64 +64 0.70--3.88 ToM × Interpretation −0.26 0.24 1.24 1 0.77 −23 0.49--1.22 Motivation Age 0.07 0.07 1.05 1 1.08 +8 0.94--1.24 Verbal ability 0.05 0.05 0.89 1 1.05 +5 0.95--1.16 ToM −0.23 0.27 0.74 1 0.80 −20 0.47--1.34 Interpretation (effort, outcome) −0.33 0.37 0.82 1 0.72 −28 0.35--1.47 ToM × Interpretation −0.38 0.20 3.80^†^ 1 0.68 −32 0.47--1.00 Note 1. n = 45, † p \< 0.10, ∗ p \< 0.05; SE, standard error; CI, confidence interval; ToM, Theory of mind. Note 2. Emotional response: model fit, χ 2 $5$ = 5.00, p = 0.42, -2 Log likelihood = 40.04, Nagelkerke R 2 = 0.17, goodness-of-fit test (Hosmer--Lemeshow); χ 2 $7$ = 9.88, p = 0.20. Note 3. Performance rating: model fit, χ 2 $5$ = 14.56, p = 0.01, -2 Log likelihood = 46.01, Nagelkerke R 2 = 0.37, goodness-of-fit test (Hosmer--Lemeshow); χ 2 $7$ = 6.13, p = 0.53. Note 4. Motivation: model fit, χ 2 $5$ = 9.11, p = 0.11, -2 Log likelihood = 52.18, Nagelkerke R 2 = 0.25, goodness-of-fit test (Hosmer--Lemeshow); χ 2 $7$ = 11.96, p = 0.10. Discussion ========== This study examined young Japanese children's interpretations of and responses to insincere praise following failure and explored the association between these responses and the maturity of their theory of mind. Children heard stories involving the receipt of insincere praise from a teacher following failure and answered questions regarding their interpretation of the praise, emotion, performance rating, and motivation. Regarding children's responses to the praise, the results showed that the children were generally happy to receive insincere praise following failure. Inconsistent with the first hypothesis, they exhibited positive emotions and their self-rated performance levels increased after the praise. The findings indicate that some of the children increased the positivity of their own evaluations to match those of the teacher. However, motivation to accept the challenge of completing a more demanding task was lower in the situation involving failure followed by praise relative to that observed in the situation involving failure without feedback, suggesting that the receipt of insincere praise from a teacher following failure could negatively affect young children's motivation. This finding suggests that young children began to notice insincerity in praise in failure situations to some extent, but they were still affected by its positive face value. Regarding children's interpretation of insincere praise without an apparent reason (i.e., "great!" or "sugoi-ne!" in Japanese), the results did not support the second hypothesis, which predicted that most of the children would attribute the praise to their effort rather than outcomes or their personal attributes. Approximately 52% of the children attributed the praise to outcome, and approximately 34% attributed the praise to their effort. It should be noted that theory of mind scores did not differ significantly between the outcome-interpretation and effort-interpretation groups, confirming that the difference in interpretation did not reflect children's ability to understand others' mind; rather, it may reflect one's attribution tendencies. Although there are no empirical data to explain the reason that about half of the children attributed the praise to outcome even in the failure situation, one interpretation was that some young children still have a tendency to interpret their work positively and with optimism; specifically, they focus on the beauty of their work rather than the components that require improvement ([@B23]). The main topic of interest in this study was the association between individual differences in the maturity of children's theory of mind and their responses to insincere praise. The results provided partial support for the third hypothesis, which predicted that children with mature theory of mind would receive and respond to the praise negatively. Children with mature theory of mind reported lower levels of self-rated performance, relative to those reported by children with less mature theory of mind after praise in both interpretation groups. One possible explanation for this finding is that children who attributed the praise to their outcome might have questioned what was in the teacher's mind in a failure situation; for example, "She praised me; however, did she really think I gave a great performance?," and this could have led them to give themselves a low performance rating. Children who attributed the praise to their effort might have interpreted that the teacher had no choice but to give praise for their effort because their performance was unsatisfactory, and this could have led them to give themselves a low performance rating. In the effort-interpretation group, children with mature theory of mind also exhibited lower levels of motivation for perseverance relative to those observed for children with less mature theory of mind, following praise. They might have doubted the teacher's praise; e.g., "She praised me; however, did she really think I tried well?" and might have answered, "Yes, I trust her. She thinks I try hard even if I have failed; therefore, I cannot be expected to do more than this." This kind of thought might have decreased their motivation to perform more challenging tasks. The findings revealed that children's interpretation of insincere praise differed depending on the maturity of their theory of mind. Specifically, although theory of mind did not account for the children's positive emotional responses, it explained the observation of lower levels of self-rated performance following the praise in those with mature theory of mind in both groups. Furthermore, theory of mind tended to be associated with weaker motivation for meeting the challenge of the task following the praise in the effort-interpretation group. It could be that young children with mature theory of mind recognize the uncertainty in the teacher's insincere praise and therefore considered the feedback as unreliable. One could also consider the idea that some children interpreted "great (sugoi-ne)" as a sarcastic remark. However, it is unlikely that the participants interpreted the praise as sarcasm. This is because most children attributed the praise to their effort or outcome and did not reported sarcastic intentions, and most of them exhibited positive emotion following the praise. The remark would be more likely be interpreted as sarcastic at the age of approximately 9 years, when children understand irony ([@B10]; [@B24]). Most previous studies examining children's responses to praise have focused on positive feedback following success ([@B30]; [@B19]; [@B16]; see also [@B5]). Within this body of literature, the main strength of this study is its novel focus on insincere praise following failure, and the new finding that process-related praise, which was believed to have positive effect on children's self-evaluation and motivation (cf. [@B19]), has a negative effect in some cases. This implies that praise for effort in a failure situation sometimes conveys the message that adults are sufficiently satisfied with children's effort, thus decreasing children's motivation for challenges. The evidence of the current study indicated that the effects of insincere praise could differ depending on the maturity of children's theory of mind. The findings also have practical implications that there is a need for fine-grained observation of individual children's understanding of mental states prior to the provision of evaluative feedback. As theory of mind dramatically develops in young children, parents and teachers should avoid using thoughtless positive feedback and assess how children evaluate their own work. This study was the first to perform an empirical examination of insincere praise and revealed the association between young children's negative responses to the praise following failure and the development of theory of mind. Although the study demonstrated strengths and important implications, it was subject to some limitations. For example, children's responses to the praise following failure were examined within the context of the scenarios, and the ways that children respond to this type of praise in their everyday lives remains unclear. Another limitation involves the nature of the sample, which included a relatively small sample size and an unbalanced sex distribution. It should also be noted that Japanese children could be motivated by failure, as they have been shown to work harder following failure than they do following success ([@B14]). Additional studies specifically designed to examine the effects of praise in diverse settings and cultures are required to provide evidence for or against the generalization of the current findings. Author Contributions ==================== The author confirms being the sole contributor of this work and approved it for publication. Conflict of Interest Statement ============================== The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Funding. The research was supported by KAKENHI (Grant No. 15K17283). The author thanks the children who participated in this study. Data from seven children whose interpretation was classified as "other" were eliminated from analyses. Instruction =========== You are playing a block design game with Hanako-sensei (teacher Hanako). Hanako-sensei shows you four blocks; one side of these blocks has two colors---white and red---and the other side is colored red. No-Feedback Story ================= Hanako-sensei shows you a picture of a model and says, "Look at this picture and recreate the design yourself using these blocks. Start!" You try hard to recreate the design but fail to do so. Praise Story ============ Hanako-sensei shows you a picture of another model and says, "Look at this picture and recreate the design yourself using these blocks. Start!" You try hard to recreate the design but fail to do so. Hanako-sensei comes to your side and says, "\[Child's name\], sugoi-ne (great)!" First-Order False-Belief Task ============================= I'm going to tell you a little story about Saru-san (Monty) and his lunch box. Look! Here's Saru-san. He wants Kaeru-san (Freddie) to put an apple in his lunch box to take to school, but Kaeru-san says there are no apples left, so he'll have to take a pear instead. Saru-san doesn't like pears at all. He really wanted an apple! He's so cross about the pear that he stamps his feet all the way up the stairs. 1. Q1. How does Saru-san feel when he gets a pear? (Memory control 1) 2. Q2. How does Saru-san feel when he gets an apple? (Memory control 2) But, look! While Saru-san is out of the kitchen, Kaeru-san finds one apple left in the cupboard. He decides to give Saru-san a nice surprise; so, he takes the pear out of Saru-san's lunchbox and puts the apple in there instead. He puts the lunchbox in Saru-san's bag. Saru-san comes back, picks up the bag, and hurries off to school. Saru-san doesn't see what's inside his lunchbox. Now it's lunchtime. Saru-san takes out his lunchbox. 1. Q3. What does Saru-san think is in the box, an apple or a pear? (First-order false-belief) 2. Q4. What is really in the box, an apple or a pear? (Reality) Second-Order False-Belief Task ============================== This is Kouta. Today is his birthday, and Kouta's mom is going to surprise him by giving him a puppy. She has hidden the puppy in the shed until it's time for Kouta's birthday party. Kouta says "I really hope you've got me a puppy for my birthday, Mom"; but, remember, Mom wants to surprise Kouta; so, instead of telling Kouta she got him a puppy, Mom says, "Sorry, I didn't get you a puppy, Kouta. Actually, I've got you a really good toy for your birthday." 1. Q1. What did Kouta think he was getting for his birthday? (First-order false-belief) 2. Q2. What was his mom really giving him? (Reality) \[If the child answers the two questions correctly, continue with the following story\]. Now Kouta decides to go outside to play. On his way out, he goes into the shed to get his bike, and he finds the birthday puppy! Kouta says to himself, "Wow! Mom didn't get me a toy; she really got me a puppy for my birthday!" Mom didn't see Kouta go to the shed; so, she doesn't know he found the birthday puppy. Inside, the telephone rings. It's Kouta's granny, calling to find out what time the party is. Granny says to Mom, "What does Kouta think you've gotten him for his birthday?" 1. Q3. What does Mom say to Granny? (Second-order false-belief) 2. Q4. Did Mom see Kouta go into the shed? (Memory control 1) 3. Q5. What did Mom really get Kouta for his birthday? (Memory control 2) [^1]: Edited by: Ann Dowker, University of Oxford, United Kingdom [^2]: Reviewed by: Virginia Slaughter, The University of Queensland, Australia; Elisabetta Conte, Università degli Studi di Milano Bicocca, Italy [^3]: This article was submitted to Developmental Psychology, a section of the journal Frontiers in Psychology	{ "pile_set_name": "PubMed Central" }
653 F.2d 99 Kim Lee HUBBARD, Appellant,v.Glen R. JEFFES, Superintendent of S.C.I. at Dallas andAttorney General of Commonwealth of Pa., Appellees. No. 80-1369. United States Court of Appeals,Third Circuit. Argued Oct. 17, 1980.Decided July 7, 1981. Richard S. Watt, Haverford, Pa., (Argued), Gerber, Gerber & Shields, Norristown, Pa., Peter T. Campana, Williamsport, Pa., for appellant. Robert F. Banks (Argued), First Asst. Dist. Atty., Kenneth D. Brown, Dist. Atty., Williamsport, Pa., for appellees. Before SEITZ, Chief Judge, ALDISERT, ADAMS, GIBBONS, HUNTER, WEIS, GARTH, HIGGINBOTHAM and SLOVITER, Circuit Judges. OPINION OF THE COURT ADAMS, Circuit Judge. 1 Kim Lee Hubbard appeals from a denial of his petition for federal habeas corpus. He was convicted in the Lycoming County Court of Common Pleas, Criminal Division, of second degree murder. The Pennsylvania Supreme Court twice upheld the conviction, after direct appeals in which Hubbard contended that the admission of certain evidence transgressed his Fourth Amendment rights, that the evidence at trial was insufficient to support the conviction, and that his trial and post-trial counsel had been ineffective.1 Hubbard once again advances these contentions in the current petition. Specifically, he claims that because the police neglected to read him the Miranda warnings before requesting the items, he did not voluntarily consent to the seizure of his boots and car. Thus, he argues, the admission of evidence obtained from this "seizure" violated the standards of voluntariness required by the Fourth Amendment.2 Additionally, Hubbard asserts that his trial counsel was ineffective in failing to make a timely suppression motion and in not interposing objections to allegedly inflammatory closing remarks by the prosecutor. Finally, Hubbard challenges the assistance rendered by post-trial counsel on the ground that he did not raise in post-trial motions the issue of trial counsel's performance. 2 We conclude that the state court afforded Hubbard an adequate opportunity to present his Fourth Amendment claim, and that his other contentions are without merit. Accordingly, we affirm the district court's denial of the habeas petition. I. 3 The following facts were adduced in the state court. Jennifer Hill, a twelve-year-old friend of Hubbard's sister, spent the night of October 18, 1973 at the Hubbard home. She was last seen alive at approximately 4:30 on the afternoon of October 19, shortly after leaving the Hubbard residence, entering a metallic green car. Her body was found ten days later in a nearby cornfield. An autopsy identified the cause of death as manual strangulation, and placed the time of death between 4:30 and 8:00 p. m. on October 19. A search of the cornfield revealed a boot-heel mark under the victim's body, and two tire marks embedded in a mound of clay near the entrance to the field. 4 On October 31, two police officers and the district attorney went to the Hubbard residence to question members of the family. No family member was a suspect at the time. Mrs. Hubbard called the school where her son was a student and asked that he return home. When he arrived at the Hubbard home, the officers requested him to wait in another room until they finished speaking with his parents. According to petitioner's testimony, the police then questioned him for approximately thirty to forty-five minutes out of his parents' presence. Hubbard testified that he was not fearful during this questioning, and that he voluntarily cooperated with the police. He also stated at trial that the officers told him he was free to leave the room or to end the questioning at any time. 5 The officers then asked Hubbard if he owned a pair of boots and a car. Hubbard responded affirmatively and the police asked permission to inspect and to retain possession of these items. The state court found that "it was the uncontradicted testimony of the officers that they twice advised Hubbard that he had a right to refuse to consent to the requested inspection." 472 Pa. 259, 372 A.2d 687, 694 (1977). Hubbard nevertheless turned over his boots and drove one officer back to Borough Hall and surrendered his car to him. 6 Subsequent police investigation revealed that the heel mark under Jennifer Hill's body was made by one of the boots given to the police by Hubbard, and that the tire marks near the scene matched the tires on Hubbard's car. These two items of information became a principal element in the case against Hubbard. 7 At trial, one police officer asserted that Hubbard had been given the Miranda warnings prior to being questioned in his home. However, the other officer, Sgt. Peterson, denied that they read the Miranda rights, because Hubbard "was not a suspect" at the time. Upon hearing Sgt. Peterson's recollection, Hubbard's counsel moved to suppress the physical evidence turned over during the interview at the Hubbard home, arguing that the absence of Miranda warnings had rendered Hubbard's consent involuntary under the Fourth Amendment. In the alternative he moved for a suppression hearing. The trial court denied the motion, ruling that it was not timely under Pa.R.Crim.P. 323(b), which requires motions to suppress evidence to be made before trial. 8 In a post-trial opinion, the trial judge considered the merits of the Fourth Amendment claim. He made a factual finding that Hubbard had "voluntarily surrendered his boots and car for examination." The trial judge then ruled that "Miranda warnings are not a prerequisite to a voluntary surrender of evidence," citing Schneckloth v. Bustamonte, 412 U.S. 218, 93 S.Ct. 2041, 36 L.Ed.2d 854 (1973). Schneckloth holds that, in distinction to the waiver standards that govern custodial interrogations, a knowing and intelligent waiver of the right to refuse a request to search is not a prerequisite to finding a voluntary consent to a search or seizure. The case indicates, however, that knowledge of the right to refuse consent bears on the question of voluntariness. 9 The Pennsylvania Supreme Court affirmed the denial of the suppression motion. It reiterated that the pre-trial filing requirement of Pa.R.Crim.P. 323(b) permits only two exceptions. Untimely suppression motions are allowed 1) where the opportunity to make an application to suppress did not previously exist, or 2) where "the interests of justice require." 372 A.2d at 693. 10 The Pennsylvania Supreme Court rejected Hubbard's argument that the opportunity to make an application did not exist until his lawyer learned, at trial, that no Miranda warnings had been given at the home. Counsel was aware of all the facts pertaining to the interview, the court reasoned, and he therefore had the opportunity before trial to determine whether a suppression motion was advisable. 11 Reaching the merits, the Pennsylvania Supreme Court then concluded that the interests of justice did not require relaxation of the pre-trial rule in this situation, because Hubbard's constitutional objection was not well-founded. Determining that Hubbard was not in custody at the time, that he was aware of his right to withhold consent, and that his consent was voluntary, the court held that under Schneckloth v. Bustamonte, 412 U.S. 218, 93 S.Ct. 2041, 36 L.Ed.2d 854 (1973), the Fourth Amendment had not been violated. II. 12 Our latitude in reviewing the state court proceedings is limited. 28 U.S.C. § 2254(d) directs that the factual findings of both the trial court and the Pennsylvania Supreme Court "shall be presumed to be correct." See Sumner v. Mata, --- U.S. ---- 101 S.Ct. 764, 67 L.Ed.2d ---- (U.S.1981). Sumner holds that a habeas petitioner, in order to overcome state court factual determinations, must demonstrate "by convincing evidence" that the state proceeding was inadequate or the determinations clearly erroneous. We may also be precluded by the Supreme Court's decision in Stone v. Powell, 428 U.S. 465, 96 S.Ct. 3037, 49 L.Ed.2d 1067 (1976), from reexamining the merits of Hubbard's Fourth Amendment claim. In Stone v. Powell the Court held that when a state prisoner raises a Fourth Amendment violation in a habeas petition, a federal court may not consider the merits of the claim if the state tribunal had afforded the petitioner "an opportunity for a full and fair litigation" of his claim. Id. at 494, 96 S.Ct. 3052, 49 L.Ed.2d 1067. 13 We agree with the Pennsylvania Supreme Court that Hubbard had a full and fair opportunity to present his claim to the state courts. His counsel was aware of the interview and of the need to make suppression motions before the commencement of trial. He could have reviewed statements Hubbard made to the police that might have acknowledged receipt of the warnings. He could have questioned Hubbard or his parents about whether Miranda warnings were given. Counsel had all the resources at his command to investigate and present a Fourth Amendment claim at the proper time. The failure to do so was not brought about by any restriction of the opportunity by the state courts. 14 An additional reason supports the conclusion that Hubbard enjoyed the opportunity contemplated in Stone v. Powell. Despite denying the request to conduct a suppression hearing in the middle of the trial, the trial court permitted counsel to explore at trial the questions of Hubbard's custody and the voluntariness of his consent to the seizure. Moreover, the Supreme Court in Schneckloth v. Bustamonte, supra, declared that the voluntariness of consent to a search is a factual question to be determined from the particular circumstances. The trial judge specifically found that Hubbard had voluntarily relinquished his boots and car. Since Hubbard has offered no evidence to demonstrate that the findings were clearly erroneous or that the state proceedings amounted to a denial of due process, the Supreme Court's decision in Sumner v. Mata, supra, makes it clear that we are bound by the finding of voluntariness. 15 In the course of deciding whether the circumstances fell within an exception to Pa.R.Crim.P. 323(b), the Pennsylvania Supreme Court considered the merits of Hubbard's suppression motion,3 and also held that the failure to give Miranda warnings did not nullify the voluntariness of Hubbard's consent. Under these circumstances, where Hubbard had ample occasion to develop relevant evidence and received a ruling on the merits, he had a full and fair opportunity to advance his Fourth Amendment claim in the state court system. 16 Furthermore, on the merits, Hubbard's Fourth Amendment argument rests on a flawed premise. The absence of Miranda warnings does not vitiate consent to a seizure of personal property, because the Miranda protections are addressed to constitutional rights that are distinct from Fourth Amendment rights. Solicitude for individual privacy is the central thrust of the Fourth Amendment. Katz v. United States, 389 U.S. 347, 88 S.Ct. 507, 19 L.Ed.2d 576 (1967). Privacy rights must be balanced, however, against the interest of the community "in encouraging consent (to a search), for the resulting search may yield necessary evidence of the solution and prosecution of crime, evidence that may insure that a wholly innocent person is not wrongly charged with a criminal offense." Schneckloth, supra, 412 U.S. at 243, 93 S.Ct. at 2056. Inasmuch as the integrity of the fact-finding process may be enhanced by admitting evidence recovered from a search or seizure, the Supreme Court has not required a stringent standard for measuring voluntariness in the Fourth Amendment context. As the Court noted in Schneckloth, in refusing to impose the "knowing and intelligent waiver" standard, "almost without exception, the requirement of a knowing and intelligent waiver has been applied only to those rights which the Constitution guarantees to a criminal defendant in order to preserve a fair trial." 412 U.S. at 237, 93 S.Ct. at 2052. The Miranda rights implicate the reliability of the truth determining process, because they apply in an inherently coercive situation and they protect the right to counsel and the privilege against self-incrimination. Knowledge of the right to be assisted by counsel and to remain silent, however, has little bearing on the voluntariness of a consent to a search. So long as the "waiver" of the right to refuse consent need not be knowing and intelligent, the person whose consent is sought need not be made aware of the full panoply of constitutional protections available to a criminal suspect who is undergoing custodial interrogation. Consequently, the absence of Miranda warnings is not dispositive of whether a person voluntarily consented to a search. III. 17 Hubbard's challenges to the effectiveness of counsel are without merit. The Pennsylvania Supreme Court concluded that trial counsel might have been amiss in not ascertaining the facts that would have formed a predicate for the suppression motion. However, the court held that this did not constitute ineffective assistance, because a timely motion would have been unavailing for the reason that the Fourth Amendment claim was without merit. 372 A.2d at 698. We concur in this conclusion. It follows that post-trial counsel was not ineffective in neglecting to assert trial counsel's alleged mishandling of the suppression motion. IV. 18 We also accept the Pennsylvania court's finding that trial counsel's failure to object to allegedly improper closing comments by the prosecutor "was born of a reasonable, calculated, and apparently successful trial strategy." 402 A.2d at 1001. Trial counsel testified that he anticipated that the prosecutor would become shrill and alienate the jury, thereby diminishing the likelihood that the government would gain a conviction for first degree murder. The course chosen by counsel was reasonably designed to effectuate his client's interests, and indeed, the jury failed to return the verdict urged by the prosecution, and convicted Hubbard only of murder in the second degree. Since trial counsel was not ineffective, a fortiori post-trial counsel did not render inadequate assistance when he declined to raise trial counsel's failure to object to the prosecutor's summation as a ground for post-trial relief. V. 19 For the foregoing reasons, the judgment of the district court dismissing the petition for a writ of habeas corpus will be affirmed. 20 GIBBONS, Circuit Judge, concurring. 21 I would hold that the fifth amendment issue involving the testimonial nature of Kim Lee Hubbard's acknowledgment that the disputed items belonged to him was presented in the state courts and thus proper for our consideration. However, because the majority holds that the fifth amendment issue was not clearly raised in state court, I, too, do not reach this issue at this time. I agree with the majority's disposition of Hubbard's fourth amendment claim, but anticipate seeing this case again. 1 In the first appeal, the Supreme Court vacated the judgment of sentence and remanded for consideration of the effectiveness of appellant's post-trial counsel. 472 Pa. 259, 372 A.2d 687 (1977). Following an evidentiary hearing, the lower court held that post-trial counsel had rendered effective assistance, and reinstated the judgment of sentence. In his second appeal, Hubbard challenged the trial court's finding that counsel had not been ineffective, and the Supreme Court affirmed. 485 Pa. 353, 402 A.2d 999 (1979) 2 Although the alleged failure to give Miranda warnings is an element of Hubbard's argument, this appeal presents only a Fourth Amendment issue concerning the voluntariness of Hubbard's consent. We do not address any Fifth Amendment Miranda problems that may arguably lurk in the record. Hubbard asserted no Fifth Amendment claims in the state court, and the parties did not press such claims in the district court or on this appeal. Moreover, had Hubbard raised a Fifth Amendment claim for the first time in his habeas petition, we could not consider it without a demonstration that he exhausted state court remedies, or in the absence of a showing of cause for the failure to raise the claim below and prejudice. Wainwright v. Sykes, 433 U.S. 72, 97 S.Ct. 2497, 53 L.Ed.2d 594 (1977) 3 The "lack of opportunity" exception to Rule 323(b) is similar to the Stone v. Powell standard permitting federal courts to review Fourth Amendment claims when the petitioner lacked a full, fair opportunity to air these claims before state tribunals. This parallel substantiates our conclusion that Hubbard had an opportunity, within the meaning of Stone v. Powell, to present his Fourth Amendment claim to the Pennsylvania trial court	{ "pile_set_name": "FreeLaw" }
The legend continues! The 1972 Chevy Vega #10 is currently in the final stages of restoration. The infamous Willliam “Grumpy” Jenkins Vega #10 has been surrounded by Championship titles, Mystery, and Controversy. In August at the AHRA Gateway Nationals in St. Louis the Vega #10 broke the record by qualifying at 8.97 sec doing 153.05 mph breaking the AHRA record. Shortly after the competition the Vega #10 was stolen in southern Illinois while William “Grumpy” Jenkins was having breakfast at a local eatery. This was an effort by the competitor to obviously find out what kind of technology was used on the engine to make the Vega part of the 8 seconds club. Later on the authorities received an anonymous tip with the location of the Vega #10, However, the engine and transmission were completely pulled out of it. Grumpy being the prepared man that he is could have replaced the motor and trans but opted to wait a couple weeks before bringing the Vega #10 back out to “smack” the competition. William “grumpy” Jenkins finally sold the Vega #10 in 1975. The new owner raced it under a different name and different paint job, everything that made it a Grumpy toy was change to conceal the fact it was the #10. Certain individuals in the racing circle knew Mark Pappas was interested in purchasing an original Grumpy Toy. After extensive negotiations Mark “Grumpy Jr” Pappas Finally had an authentic Grumpy Toy of his own. A lot of skeptics doubted this was the original Vega #10 until William “Grumpy” Jenkins confirmed the secrets of authenticity he implemented when it was originally built by himself. There was a mounted stick in the car known as the Grumpy stick, This over 30 year old broom stick was use to hold up the hatch so rear support modification could be done in the trunk. All the other hidden secrets were revealed to confirm its originality such as: webbing in the dyna 60 had holes drilled in it for weight saving but not compromising the strength. Another confirmation sign was the Grumpy signature on the block plate made with a diamond rotary engraver in 1973, which Grumpy says can not be duplicated. Now begins the rebuilding of a legend. Windy City Rods and Restorations located at*************, is where the renovations will be completed to restore this iconic car back to its original form when Grumpy built it. Windy City Rods and Restorations had there work cut out for them. They had to fix 37 years of stress cracks due to it being acid dipped in 1973, along with the wear and tear of pro-stock racing up until 2008 under a different name. Windy City Rods and Restorations repaired all damaged panels, doors, lower quarter panels and rear tail panel. All the stress cracks around the windows had to be extensively repaired. The Vega was then sanded down vigorously from top to bottom. It was then hand blocked, sealed, and painted to perfection. All floors tins and engine compartment tins where completely cleaned and polished. The roll cage which was painted blue had to be stripped down, primed and painted. The extra bracing that was added to the roll cage was then removed to restore the Vega back to 1973 compliance. The front end suspensions components where media blasted and then painted Grumpy Grey. All wiring was redone to 1973 standards. Windy City Rods and Restorations also installed a new fuse block that was given to them personally from William “Grumpy” Jenkins. The car was then thoroughly wet sanded then buffed to give it that glass looking shine. All period correct stickers were placed on the vehicle. The Vega was then hand pinned stripped and lettered. New seat and bracket were installed in the car. New rear tubs were fabricated, and all glass was installed including the original rear glass. Windy City Rods & Restorations would like to thank the late William “Grumpy” Jenkins for supplying all the original spare parts for the Chevy Vega #10 as well as Jake and the rest of the Jenkins Competition team. Windy City Rods & Restorations reports that they have been receiving surprise parts every week from Grumpy's shop, to ensure its being restored to its original form. Currently Windy City Rods & Restorations is anticipating the arrival of the period correct 331 motor, they are also waiting on the period correct Lenco transmission. The 1972 Vega #10 is a legendary car full of mystery, mystic, and controversy. This car was ahead of it time, and with its championship history it will continue to intrigue people for years to come 8.97 at 153.05 MPG (which wer new AHRA records) Did B.J. come up with the idea of using a tube frame inside a stock car's body? (Note: that i understand that the factory experimental cars had this kind of setup, but this is not the exact same thing---i don't think so at least because the F/X cars had extensive modifications to the wheelbases/lengths and body mods.) b) And therefore the idea of narrowing the rear axle and rear suspension so as to be able to run very wide tires which wouldn't be possible with a stock suspension? BTW, i noticed that BJ's chevy racing engine book is available somewhat cheap at amazon. i would say my favorite prostocks are 74ish to about 81. This is with the really low in the front end with the tucked in front wheels, but the rear was still stock height so you had that interesting rake. Register Now In order to be able to post messages on the Hot Rod Forum : Hotrodders Bulletin Board forums, you must first register. Please enter your desired user name (usually not your first and last name), your email address and other required details in the form below. User Name: Password Please enter a password for your user account. Note that passwords are case-sensitive. Password: Confirm Password: Email Address Please enter a valid email address for yourself. Email Address: Insurance Please select your insurance company (Optional) Log-in User Name Remember Me? Password Human Verification In order to verify that you are a human and not a spam bot, please enter the answer into the following box below based on the instructions contained in the graphic.	{ "pile_set_name": "Pile-CC" }
The balance simplex in non-competitive 2-species scaled Lotka-Volterra systems. Explicit expressions in terms of Gaussian Hypergeometric functions are found for a 'balance' manifold that connects the non-zero steady states of a 2-species, non-competitive, scaled Lotka-Volterra system by the unique heteroclinic orbits. In this model, the parameters are the interspecific interaction coefficients which affects the form of the solution used. Similar to the carrying simplex of the competitive model, this balance simplex is the common boundary of the basin of repulsion of the origin and infinity, and is smooth except possibly at steady states.	{ "pile_set_name": "PubMed Abstracts" }
Q: Does 4-wheel alignment involve taking wheels off? I was wondering if a 4-wheel alignment involves taking off all the wheels. I called NTB to check whether their wheel alignment includes tire rotation and they said it didn't. But if they have to take all the wheels off, why would it matter which wheel they put on which rotor, i.e. doesn't make any difference to them? If they can do it without taking the wheels off, then I would understand. A: An alignment does not require removing the wheels. The equipment is attached to the wheels while they are in place. I often wondered about the shops that will do a free brake inspection but then charge $20 for tire rotation.	{ "pile_set_name": "StackExchange" }
you weren't on there. if you were i am sure you would be a 10.	{ "pile_set_name": "Enron Emails" }
"You little pickaninny." "You'll kill yourself flying like that." "Yes, you are." " Windy?" " You bet." " I thought you was deaf, pops." " I'm sorry, I didn't hear you speak." "Didn't say nothing." "Didn't you?" " Mr. David Burke." " 607, sir." "Excuse me." " Floor, mister?" " Six." "You hear that wind?" "I'm going to get myself grounded out of the Air Force." "That's what I'm going to do." "Ain't going to fly this kind of crate anymore." "Can you blame me?" "Right on time, Slater." "Right on time." "All right, there, Yuley." "Come in." "Here, let me help you off with your things." "What are you doing with such a big old dog in New York?" "I never had a wife." "Right on the button." "That's what I'm looking for." "Someone serious, with a head for business." "All right, here." "Come on over here." "Up." " Whiskey?" " I don't mind." " Straight or with water?" " With water." " I got ice, you want ice?" " No, just water." "Make yourself comfortable, Slater." "We got to talk." "Like the view?" "You got a whole heap of it." "I used to be able to afford a little space inside, too." "We call this dump home, don't we, old boy?" "Not bad." "Well, here's to our mutual benefit." "May it be considerable!" "I guess you know all you have to know about me, Slater." "I know something." "All right, you tell me what you know, go on." "You said you wanted to talk to me." "Since we knew a few people in common, I wanted to come and find out." "I mean the dirt." "I know you were on the police force... had a session with the State Crime Committee... and you got a year for contempt." "Because I wouldn't talk." "I was on the force 30 years." "I had my own squad, and I knew everybody." "Everybody was my friend until they needed a patsy." "I know about you, Earle." "Two stretches:" "One for assault with a deadly weapon, one for manslaughter." " Every time you get a decent job..." " Knock it off." "What's so big about you, Burke?" "How come you make so much noise?" "You've been sniffing around... trying to find a hole in the fence just like everybody else." "What makes you so big, you can call me up to this dump..." " and shoot off your mouth?" " I got an idea, that's why." "How would you like to pick up $50,000, all in small bills, just for yourself?" "How would you?" "Hello." "Johnny?" "No." "When?" "Sure, that's fine." "Well?" " What would I have to do?" " Just walk into a bank and take it." "Maybe you got me wrong, Burke." "I never stolen nothing in my life." "Except maybe a watermelon when I was a boy on the farm." "I don't want a Big Joe with more decorations than Dillinger." "I want a safe thing." "I want a man who needs some money to set himself up." "I want a man with guts." "I want a serious guy in trouble." "And that's you." "That's me." "That's the both of us." "This is a one-time job." "One roll of the dice, and then we're through forever." "Come on, Slater, what do you say?" " Who'd like to make themselves a fortune?" " I would!" " Everybody stays off the car." "Is it a deal?" " Yes." "I'll watch them." " And who'll watch you?" " I don't know." "Six, please." "Do you hear that wind?" "You sure we're going to make it?" "Always made it before." "With my luck, daddy, this could be it." " Maybe on the way down." " I'm walking down." "Shut up, you mutt." " Johnny, come in." " Hello, Dave." "Scram." "He'll get hair all over you." "Take off." " Here, let me help you." " I can't stay long." "Well, it takes some explaining." " How about a drink?" " No, thanks." "You got the floor." "Here." "He keeps shedding." "All right, there." "Come on, get up there." " Those blue coats." " That's all right, don't worry about it." "You don't look like I could sell anything to you." " You look like you have it all, right now." " Come on, fill me in, Dave." " You're into Bacco for too much dough." " Now tell me something I don't know." " You owe him about $6,000, right?" " Wrong, $7,500." "It was $6,000 last week." "The horses are still running, and I'm still losing." " And Bacco's willing to wait?" " What's the pitch, Dave?" "I know how you can pay it off." "And I know what's coming next." "The fix is in." "It's a sure thing, right?" " Now, wait a minute." " No thanks, Davey boy." "I'll go down the drain on my own." "You're crazy." "All right, so I'm crazy." "You're counting your fingers and toes." "I'm talking about $50,000." "I'll take that drink." " Straight." " Just tell me one thing, Johnny." "How come Bacco hasn't knocked your teeth in for not paying off?" "'Cause I'm paying $100-a-week interest." " Well, that can't leave you much." " Just my teeth." " You ever going to pay it off?" " Lf I can ever wear out this streak." "No use stopping now." "I'm way past the life ropes." "I'm praying for a miracle." "What do you mean, $50,000?" "Just what I said, $50,000 to $75,000, like that." " All in small bills." " And you need Johnny Ingram?" "I'm just a bone picker in a four-man graveyard." "It's a bank job." " Are we social climbing, David?" " This is easy money." " Man, you're drifting." " I'm serious." "That's not your line, Dave." "That's the firing squad for you." "That's for junkies and joyboys." "We're people." "Okay, what's my line?" " I'll forget you asked me." " You don't even want to hear?" "I did all my dreaming on my mother's knee." "All right, forget it." " You riding?" " Downtown." "I'll ride along with you." "A man like you thinking thoughts like that." "You're in trouble." "I've got to get out of this trap." "They've kicked my head in." " What can I do?" " Find a hobby, man." "Anything." "They sure changed your color when they rehabilitated you at Sing Sing." "$50,000 can change it back." "Come back here, you." "Get back in here." "You heard me." "Mind the store." " How are those two little women of yours?" " The kid's fine." "That ex-wife of mine, man, she's worse than Bacco." "If that alimony isn't there on the first, her lawyer's there on the second." "Thanks for the lift." "If you change your mind..." "I think I can get you the $7,500 right away to pay off Bacco." "I've got $500 on the nose of Lady Care today." "I can't lose forever." "You'd be surprised." "Just tell me, how come it isn't safe to walk in the park at night in a big city like this?" "It's these wild kids." "Well, if it isn't the old clam himself." "This fresh air will kill you, Coco." "I know it." "But it's pigeon time for the little king." "What's this, a press interview or something?" "Go on, talk to the old man if you want to." "He wants to do you a favor." ""What's the matter with this Davey Burke," he says. "I want to pay him off."" " Hi, Dave." " Hello, Bacco." "Join me?" "Want me to open up something for you, in the operation?" " I told you, whenever you're ready." " You're outside the law." "Well, a little inside, a little outside." "More or less." "Everything is more or less." "Just name it." " A fellow by the name of Ingram owes you." " Sure." "A very entertaining boy at Connoy's place." "What time is it?" "I'm on my way, stupid." "Go back to sleep." " What time?" " 7:30." " Why so early?" " Burke is waiting." "Can you trust this Burke?" "Not like I can trust you, but enough." "Come here, you big clown." " Now you go back to sleep." " Let him wait." "I'm off to make my fortune." "What kind of a fortune?" "Just one of those fortune fortunes." "I told you about it." "Burke got hold of some kind of concession." "He's a friend of that Lefty Gowers, isn't he?" "Yeah, he's the one who told Burke about me." "You knew Gowers in jail, didn't you?" "No, at the Millionaires' Club." "Where else?" " Earle, what does he want with you?" " I'll tell you after I see the proposition." "Do you have to stay overnight?" "Or you just want to stay overnight?" "I have to see the concession." " You got enough money, sweetie?" " Plenty." "How much is plenty?" "About $15." " You better take some more." " Burke is paying." "What do I care if he's paying?" "You take some more and then if you feel like paying, you just pay." "You don't have to take this deal if you don't like it." "If this isn't what you want, you don't take it." "There's no hurry." "There is a hurry." "I have to make it, Lorry, and I have to make it now." "It wasn't too bad when I was grubbing along by myself." "It was always too tough, too greedy, but now, because of you..." "I have to make it on my own because of you, any way I can." "No, you don't." "Not just any way." "You mustn't even try." "I have to." "Earle, listen..." "I have you, right?" "You have me." "What difference does it make where the money comes from?" "They're not going to junk me like an old car." "Now, don't worry." "If it isn't all right, I won't do it." "It's about 100 miles up the Hudson." "I've got it marked there." "Melton." "Now watch the waiter." "Over here." "Now here's the setup." "That side door's the key to the whole job." "The bank stays open Thursdays till 6:00." "Most of the factories pay on Friday." "So the bank is loaded with payroll cash and deposits from the stores." "Every Thursday night, there's close to $200,000 in untraceable cash sitting in there." "A half dozen clerks stick around for an hour totaling and straightening out the books." "The Assistant Manager has a bad heart." "Joe Foss, the guard, is about to retire, has glasses and arthritis." "That's it." "That colored waiter from the drugstore brings coffee and sandwiches after 6:00." "The rest of the town's home at supper." "You could take it with a water pistol." "Well, what do you think?" " There's just one thing wrong with it." " What?" "You didn't say nothing about the third man being a nigger." "At night, I tell you people" "When that cold, cold sun goes down" "At night, I tell you people" "When that cold, cold sun goes down" "I cry, I sigh, I wanna die" "'Cause my baby's not around" "What's the matter, pretty baby?" "Tell me, what's your daddy done?" "Won't you tell me, pretty mama what's your daddy done?" "You've got to come and hold me" "Before the morning sun" "Hi, baby, what's shaking?" "Bacco wants to buy you a drink." "And I want to buy you a shiny new car." "Too bad about Lady Care." "Yeah." "They bobbed her nose." "Believe me, pretty mama" "It's not just me, I know" "Believe me, pretty mama" "It's not just me, I know" "I just can't make that jungle" "Outside of my front door" "Well, at last, the invisible man." " I had a heart attack." " For a couple of horses." "Don't be like that, baby." "I don't care if you want to drop me, Johnny, but send back the key." " Who said anything about dropping you?" " Then you know where to find me." "That's good." "But it was better when you wanted it." "I told you Bacco has a message for you." "It's a long night, pretty boy." "I'm not going anywhere." "Maybe I should lend you a piece of iron." " Maybe." " I can let you have this juvenile delinquent." "I'm only playing with kids." " Johnny?" "Come in." " Hi, Garry." "Hello, Johnny." " Can I see you, Ed?" " Why, sure." " Do you want me to cut out?" " No, not for me, Garry." "Well, Johnny, what do you think of these?" " Who are they for?" " My oldest." "Her sweet sixteenth." " Those are real cultured pearls." " Sixteen, already?" "Yeah, they don't stand still like we do." "Well, what can I do for you?" "Bacco just blew in." "He called me earlier today." "What do you owe him?" "$7,500." " Hello, Ed." " Bacco." " I come down for my money, Johnny." " I don't have it." "Maybe you're not looking hard enough." "Maybe you think I'm a jerk who can wait forever?" "Look, I lost again today." "I've been losing steadily for a month." "I just got to break this streak I'm in, Bacco." "I can't let it knock me out." "Suit yourself." "Just get the money to me by tomorrow night." "All of it." " Suppose I can't?" " There is no "can't."" "I don't like the word "can't." I say have it." "Can you bail me out, Ed?" "I'm sorry, Johnny." "I can't help you." "You already owe me a couple of thou, anyway." "I asked you to stop with the horses." "I told you once and for all to stop." "I can't get it for you tomorrow." " You're saying what to me?" " Look, man, I'm telling you in front." "I don't have it now, and I won't be able to get it by tomorrow." "I'm not dancing with you, Moriarity." "You mean you come here with a gun when I personally call you to talk to you?" "When I stretch you six months with a debt I would kill for?" " You come and pull a gun on me?" " It's in my pocket." "The gun's at my head, that's where it is." "I tell you, Ingram, I want you to know." "Have that dough at my place tomorrow night... or I'll collect it from you, or that ex-wife of yours, or your kid." " You'll do what?" " All right, Johnny, drop that gun." "Tomorrow night at 8:00." "Or I'll kill you and everything you own." "Well, it tells you in the good book" "And they teach the same in school" "Let a man get his hands on you" "And he'll use you for a mule" "My mama gave me warning And now I know it's true" "She said all men are evil And Daddy, that's you" " All men are evil" " All men are evil" "All men are evil" " She said all men are evil" " All men are evil" " All men are evil" " My mama told me" "My mama gave me warning And now I know it's true" " Lady Care in the fifth." " She said that all men are evil" " They bobbed my nose." " My mama used to tell me" " Annie, they bobbed my nose." " Don't ever love no man" "Johnny, please." " He'll use you and abuse you" " Don't ever love nobody." "And that's something I can't stand" "My mama gave me warning And now I know it's true" " Talk that song." "Tell 'em." " She said all men are evil" "And Daddy, that's you All men are evil" "He's been making crazy like that for hours... like someone slipped the leash, or something." "All of them cats is calloused!" "And that's you!" "That little boy's in big trouble." " Is that your last word?" " I'll keep my mouth shut." " Think it over, Earle." " Nothing to think over." "Just the idea of it makes me nervous." "I wouldn't trust my own self on a deal like this with a colored boy." "Okay." "Hi." "You're late." "I'm always afraid to wake up on this once a week father's day." " What time will you get back?" " Why?" "I've got a P.T.A. Meeting tonight." "Should I get a sitter?" " You got your sitter." " Daddy." " How are you?" " Fine." " What do you see up there?" " The park, the lake." " What else?" " I see a merry-go-round." " Is that all?" " I see a red balloon." "I see people skating." "Give me a kiss." " It sounds like Central Park, today." " I'll get my things!" "Mrs. Anker, this is Eadie's father." " How do you do, Mr. Ingram?" " Hello." "We're having a meeting of the P.T.A. Steering Committee here." "Some of my friends." "This is Eadie's father, Mr. Ingram." " How do you do, Mr. Ingram?" " We're glad to know you." "I hope you can steer your way out of it." "Very glad to have met you." "I'm ready, Daddy." "Let's go." "I'll get the elevator." "What did I ever see in you?" " It's here, Daddy." "It's here." " That." " Here's another ticket, Eadie." " How about you?" " It makes me dizzy." " You're too old." " What's the big idea?" " Bacco's afraid you might blow town." "You tell that peckerwood boss of yours... if he looks hard at my wife or baby I'm blowing him a new one." "I don't want to see you here." "You gum up the scenery." "We like it here." "I'm telling you to fade or I'm turning you right in... now, to those cops." "That'll be the day." " Officer." " Yes." "Can I help you?" "Where can I find the zoo?" "Just go through that tunnel and follow the signs." "You stay right here, Eadie." "Daddy's got to make a phone call." " I'm staying." "I'm eating." " Don't you budge." " I'll see that she stays here if you like." " Thank you." " What's your name, little girl?" " Eadie." "Eadie Ingram." "Mr. Burke, please." "Dave?" "Johnny." "Yeah, I know you've been out." "Listen, Dave..." "I'll take out that deal." "Now, wait." "I need $7,500 right away to pay off Bacco." "Yeah." "All right, I'll call back in 15 minutes." "Goodbye." "Hello, Bacco?" "Burke." "Fine." "That's what I'm calling about." "Call those babies off." "I told you, Davey, I got a problem." "It got a little rough, and now he has to make a deal." "It was a public thing." "He has to make a deal." "That's what I'm telling you, he's got a deal." "I'm making it for him." "I'll give you the dough in two weeks." "You know what he did to me?" "He pulled a gun." " He pulled a gun?" " After all I did." "I treated him like a son." "Look." "It can still be straightened out." "You think I have no feelings?" "Babysitter." "What's going on in there, an orgy?" "What's on your mind, Helen?" " Can't I flirt with you a little?" " Some other day." " Are you staying home tonight?" " Why?" "I can't get a babysitter." "Then stay home and take care of him yourself." "Well, if you're busy..." "You see, Sam's boss gave him two tickets to this musical." " I'm to meet him in front of the theatre." " Take the baby with you." "What's the matter with you?" "Am I bothering you, or something?" "It's just because Lorry said that you would." "Just keep taking her around." "I have to make a phone call." "Mr. Burke, please." "Yeah, I'm listening, Dave." "Bacco is paid off." "Start celebrating." "We could be on to something really big." "Stop crowing, man." "There's nothing to celebrate." "That little crumb threatened my kid and my ex-wife." "Yeah, I know I got rid of a headache." "Now I got cancer." "Yeah, I'll see you tonight." "Wake up, Dave." "We're committing suicide." "This is 3:00 in the morning." " Come on." "Just throw anything." " Sure." "Left." " You're out." " Hey, that's pretty neat." "It's all this new atom war stuff." "It's the first thing they teach you now." "I sure could use that technique from time to time." " The same, Earle?" " Hit me with a couple, Mac." "You know, I'd like to teach you sometime." " You mean a girl could do that?" " Why, sure." "Doesn't matter how big the guy is." "Here, I'll show you." "What's this, the kiddie hour?" "It's make-believe war time, Earle." "The hour of sweet romance." "Okay, Georgie, rape me." "I'm sorry, buddy." "I hope I didn't get any on you." " Here, the drink's on me." " I'll buy my own drinks." "Look, kiddo, let's just take it easy." " Show me." " Sure." "What do you do when a guy grabs you this way?" "I give in." " Relax, baby." "Don't fight me." " I thought I was supposed to do that." " Not yet." " Not yet?" "In a minute it'll be too late." "Come on, grab my wrist." "Now don't try to break the hold." "Put your right foot over in front of mine." "Now throw your hips out, bend forward, and throw me." "Honey... if you're going to throw that bum, you throw him the other way." "All right, why don't we all just drink up and quit fooling around?" "Did you say something to me, bud?" "That stuff belongs to my war." "Take her to Canaveral and launch her." "Maybe you know something better." "Now look, fellows, let's just settle down, shall we?" " I'm going." " Maybe you'd like to try?" "You better go back and play with the girls." "Tell them all about Sputnik." "I thought you were an expert, or something." "Come on, throw a punch." "I'll show you if it's bull." " All right, soldier, let's break it up." " It's not a fight." "I just want to show this old veteran how this thing works." "Now, come on." "Don't you want to throw something?" "Get lost." "Just a scientific experiment." "We're just a couple of scientists." "Now come on, throw a punch." "Try it." "All right, Earle." "Let me take care of this." "Now look, soldier..." "Go on, try it." " Any particular hand?" " Any one you like, pop." "The kid was only trying to show off, Earle." "I didn't mean to hurt him." "You slob, what did you do?" "You feeling any better now, honey?" "It's all right, it's just the wind." "Take it easy." " Okay." " Stretch out now." "You'll be all right." "Are you sure he's not there?" "Well, did you look in the back booth?" "There you are." "Sweetheart, I've been calling all over." "Where were you?" "Sweetie, I knew your deal went wrong." "I knew it went wrong the minute... that Helen phoned and she said you insulted her." "I apologized for you, and I said I was sure you could baby-sit for her later." "Honey, I want to hear everything that happened when I get back." "But right now, I'm so late I got to rush right back downtown." "Guess what?" "My boss is going to buy a new place, and I'm going to manage it." "How about that?" "We're going to have dinner with the owner, and he wants me there..." "Maybe I better call and say I can't make it." " Maybe I better stay home tonight?" " You don't have to hold my hand." "All right." "Your dinner's on the stove... and there's some good programs on tonight." "Sweetie?" " You'll be here when I get home, won't you?" " Where else would I go?" "I wish you wouldn't make such a big thing out of it." "Listen, with that deal, it's just as well." "I told you, I don't care how long it takes for you to find the right thing to do." "Sweetie, we're doing fine." "And if my boss does take on this new shop..." "I'll be making much more money." "You might at least say that I look good." "If you're going, why in the hell don't you go?" "You know, I knew you were in trouble when I fell in love with you." "I knew it." "I knew it would be rough for us, honey, and it would take time." "But I didn't care." "You don't have to be the great big man with me, Earle." "I don't care about things like that." " There's only one thing I care about." " I know." "But what happens when I get old?" "You are old now." "You can go straight to hell!" "Hello." "Dave Burke." "Yeah, Burke." "You mustn't do that." "Sorry." "I was dreaming." "A man always dreams about what he wants, or what he's afraid of." "I'm going away for a couple of days." "Maybe you want to take Eadie for a drive or something." "Thanks." "Maybe I will." "No." " How did all this happen to us?" " You know, Johnny..." "I didn't mind what you did to me." "I minded, but I would've gone on." "Are you saying I can come back?" "The door's never been locked against you, not for my sake." "But I couldn't do it to Eadie." "A child can't have a father who lives your life." "Except on visiting days." "Not even on visiting days, but that's the law." " You're tough." " Not tough enough to change you." "For what?" "To hold hands with those ofay friends of yours?" "I'm trying to make a world fit for Eadie to live in." "It's a cinch you're not gonna do it with a deck of cards and a racing form." "But you are?" "You and your big white brothers." "Drink tea with them and stay out of the watermelon patch." "Maybe our little colored girl will be Miss America, is that it?" "I won't listen when you talk like that." "Why don't you wise up?" "It's their world and we're just living in it." "Let go of me!" " Don't let me catch you teaching Eadie..." " Daddy!" "You woke me up." "Baby, I'm sorry." "Sweetheart, listen." "I was just telling your mommy how much I loved you." "And you must never forget it." "We had such a nice time today." "I'm telling you how much I love your mama." "You always mind her, and be good." "We're counting on you a lot in this family." " I want to see Lorry." " Lorry's out." "Come on, now." "Don't be mad." "I'm sorry about this afternoon." "I really am." "Just me and my miseries you walked in on." "Oh, you..." "I'm ready to kiss, make up, and say I'm sorry." "Do you know what that louse of a husband of mine did?" "When I told him that I couldn't get a sitter... he just decided to go there with one of the boys." "How do you like it?" "I don't mind." "The kid's sleeping upstairs." "Come on in, have a drink." "I have to go by 11:00." "I should have told you that anyway." "Was I really mean?" "Don't you remember what you said?" "I'm scared to." "What was it?" "I forget." "To a much more affectionate future." "Come on in and have a visit." "I've got to keep an ear open for the baby." "You can hear the baby plenty, down here." "Come on." " Would you do a thing like that?" " Like what?" "Like he did." "Leave me alone this way." "Not for a minute." "Can you really hear my kid down here?" "Honey, I spent more sleepless nights with you than you know." "I'd like to ask you something." "But you must promise not to be angry." "I promise." " I mean, seriously." " All right." "I promise." "How did it feel when you killed that man?" "I'm sorry." "I'm stupid." " You want me to make your flesh creep?" " No." "Forget it." "I enjoyed it." "It scared me, but I enjoyed it." "I hated that man so, I could've killed him all over again... even though I didn't mean to." " What did he do to you?" " He called me." "He insulted me." "He was a very smart-talking character, and then he called me." "What do you mean?" "He dared me." "Like you are now." "Just this once." "He comes out of the drugstore, and he crosses over with the light." "Then he goes down the side street to the side door of the bank." "It's a regular-sized door." "Like this." "No different." "Except that in the top half, there's a small glass panel for observation." "The guard opens the door." "This is the heart of it, Johnny." "That door is on a chain." "It holds the door open so much." "The waiter hands the guard the sandwiches." " Hello, Burke." " Come in." "I want you to meet Johnny Ingram." "This is Earle Slater." "Sorry I'm a little late, but I had girlie trouble." "Where were you, Dave?" "I was telling him about the chain." "The guard locks the door again without unhooking the chain..." " and the waiter leaves." " That's right." "The chippie is the chain." "We got to figure out an answer on the chain." "Maybe you could have him put a gun on the guard... and just barrel through the door?" "What do you think, Johnny?" "I don't know." "Tell him about the car." "I got a beauty." "A hopped-up motor with dual carburetion and a beat-up station-wagon body." "I bought two stolen plates, and the car can't be traced." "It's a remade job that was used in smuggling." "We got four police specials that have no history... and a couple of shotguns." "I thought this was an easy job." "It sounds like D-day." "Don't you give those guns a thought." "I'll take care of them." "I'm not thinking about the guns." "I'm thinking about the chain." "Don't worry about it, boy." "We'll be right there with you." "All you have to do is carry the sandwiches in a white monkey jacket." "And give him a big smile." "And say, "Yes, sir."" "You don't have to worry, and you don't have to think." "We'll take care of you." " Then you'll have to start right now." " Don't beat out that Civil War jazz here." "We're in this together, each man equal." "We'll take care of each other." "It's one big play." "Our only chance to grab stakes forever." "I don't want to hear... what your grandpappy thought on the old farm in Oklahoma." "You got it?" "I'm with you, Dave." "Like you say, it's just one roll of the dice." "It doesn't matter what color they are." "So as they come up seven." "It's all right, Dave." "I've got an idea about the chain." "Let's say the chain on the door is eight inches." "Or make it 10 for good measure." "If the box with the sandwiches and the coffee is big enough... the guard just naturally has to unlatch that door." "He can't expect me to turn it sideways because of the coffee, right?" "So the box we use has just got to be big enough." "And he just naturally has to open that chain." "You got it, Johnny." "It'll work." "I know it'll work." "Earle, don't put on the light." "I've been crying." "Earle, I'm like all the rest of them." "I keep telling you how to live and not letting you be what you are." "Never mind about me." "I spoil everything." "I can't help it." "I just have to spoil it." "Not for me, darling." "You know, I just kept waiting here, and I thought if you would come home... nothing would ever spoil it for us." "I thought of how I won't let myself see the way you feel about the money." "How it has to be your own, or we just can't last." "And you're right." "That's the way it is." "Only don't leave me, Earle." "Please, darling, don't leave me." "I'll never leave you." "I've been leaving all my life... since when I can first remember." "When the wind blew us off the land in Oklahoma... we left." "After that, I never stayed." "Not in the Army, not in Detroit, not anyplace." "I'd start something, if it didn't work right away, I'd blow it." "It was always something." "A lousy captain, or a Polack foreman in the auto works... or it'd be too slow." "I'm getting too old to take things slow." "If I don't make it now, I never will." "I mean with you, too." "It's now or never." " Aren't things ever easy for you, Earle?" " Only when I get mad." "Then they get too easy." "I think that's why I get mad, to make it easy." "But I got something now... and I'm going to stick with it." " Okay, what'll it be, mister?" " Just gas." "Fill it up." "Can you do 150 on a straightaway?" " I said just gas." " Sorry, mister." "She's strictly 65 on the outside." "I'm a nut on motors." "All set?" "Gassed up, parked, and ready for the switch." "If we get to it." " How you doing, Earle?" " Just waiting for the whistle." "Remember, I'm the one who blows it." "Right on the nose." " Take it easy." "We got to give him time." " I sure hope he doesn't screw it up." " Did you go over the roads?" " Twice." " You kept your gloves on all the time?" " Quit mothering me, Dave." "Doing all right." "I don't mind the action, it's the waiting." "I wasn't made to wait." "I've been waiting all my life." " It won't be long now." " The sooner the better." "When I gassed up, that kid in the station opened the hood and saw the motor." " What of it?" " He surely admired it and me." " You worried about it?" " Not yet, but I'm working on it." " I hope that business with the chain works." " The hell with that chain!" "No fireworks." "Sure." "Just get this in your head, Earle." "Ingram isn't a powder puff." "I know you were a tough cop, Dave... but a sudden noise in the night is like to frighten brother bones." "Johnny was in the same war as you." "Sure, and he had the big white master to hold his hand." " He better not crap out." " He won't." "If he does, I'll hold his hand." "Attention, please." "Bus 120, now arriving from New York... will depart for Albany in five minutes." "Bus 120 will depart for Albany in five minutes." "Thank you." "Hey, there, you." "You with the cap." "Just a minute." "You see the accident?" "No, I just heard the crash and there it was." " You see him?" " I see him." " What's that clown trying to do?" " Keep your shirt on." "All right, thanks." "Could you wait a minute, sir?" " You crazy?" " Shut up." "What did the cop want?" "A bit of bad luck." "He asked if I saw the accident." "So you gave your fingerprints." " Did he see your license?" " How stupid can you get?" "I got nothing in my wallet but a bus ticket and $15." " That's all?" " That's all." "But he got a good look at me, right up close." " You were wearing the glasses." " We were face to face." "Your mother wouldn't know you in those dark glasses." "She would if I held up a bank." "Get wise to yourself." "You're another black spot on Main Street." "Shut your ugly mouth." "Get in the car." "Some day I'm going to snap off your poisoned head." "Listen to me, Johnny." "That cop wouldn't recognize you in 100 years." "We have to take some chances." "You're a gambling man." "Gamble." "It depends on the odds." "Back up." "The odds will never be right." "I know how to handle him." "I've been handling them all my life." "He's no different." " All right, Slater, handle me." " Johnny!" "It's now or never." "We're right up to the line." "We'll be done and away by 6:15." "You're sure you want to go through with this?" "It's going to let us live again." " All right." " Good boy." "We keep apart till 6:00." "Everything like we planned." "Let's move." "The next time you call me, I'm going to see you." "I'll be waiting." "You're not just another white spot to me." "All right, let's go." "Take it easy, pal." "Don't worry about Slater." "He's a hard nose, but he's dependable in the clutch." "It's going to work." " What?" " I don't know." "Just something." "Maybe drive around, something like that." "I don't know which is worse, the atom bomb or you kids in your do-it-yourself cars." "You should've seen the job I gassed up this afternoon." "Just an old beat-up station wagon." "But the motor inside..." "The carton?" " You'll come over and baby-sit with us?" " Sure." "Wait here." "No nonsense with those guns." "Do you hear me?" " There, I got you right in the mouth." " I got you in the eye." " Right in the nose." " You missed me that time." "Let's have the key." " Did you get anything?" " Not a shot." " We jumped a couple of rabbits." " Good." " There, I got you." " I got you again." "All right." "Come on, let's get them girls." "We deserved it." "We got them." "Right in the bellybutton." "Cut it out." " I got them right in the mouth." " I got them on the lip." "Watch out, here's..." "Brats." "We're sorry." "We'll help you." "You're late, Charlie." "They must be using bigger boxes." "Just a second." "There, let's see." "Darn chain, there." "Don't move." "Joe." "Don't shoot." "Shut up." "Now just sit still, everyone, and behave." "Hey, Charlie, got an extra?" "Man, I got a dozen of them all over the sidewalk, back there." "The waiter." "Go ahead." "Let's cut out before he kills somebody." "Give me the keys." " Dave, you get the car." " But Johnny's supposed to get the car." " We got it made." "Come on." " Give me the keys." "Come on!" "Keep the room covered." "Pete, can I see you a minute?" " Sure thing." " I need another book of raffle tickets." "Well, I sure have..." "That's funny." "Hold it, mister." "You there, the hunter." "Stay there." "A stickup." "Get in the car." "He's got the key." "To the car, Dave." "Keep these people back out of danger." "My God!" "My car!" "Cover me, I'll get Dave." "Throw the keys, Burke." "Give up." "You don't have a chance." "The keys, Burke." "You there, in the alley." "Come out with your hands up." "Burke, give us the keys!" "There are your keys." "You still got it made." "Run, Johnny." "I'm sorry." " Let's clear out." " We can't leave Burke here." "Watch it." "Look out." "Drop that gun!" "What do you know?" "He sure ain't going to talk now." "You screwed it up." "You couldn't trust." "You killed him." "No, hold it!" "Well, these are the two that did it." "Which is which?" "Take your pick."	{ "pile_set_name": "OpenSubtitles" }
There are two types of frames: Futon lounger and bi-fold. The Bi-fold frame is made up of two parts, the backrest, and the sitting part. In spite of this, some manufacturers offer matching ottoman instead that can also be used as a bench. Moreover, they also offer twin futon mattress which is suitable for single use. Futon mattresses were first used in Japan but have since spread throughout the globe due to their comfort factor. Futons are made up of two parts: the frame and mattress. Futon can be in different sizes, where the frame and mattress determine the size. But just how big is a futon mattress ? How Big Is A Futon Mattress? Their depth usually varies from 5 to 20 inches depending on the materials used to make the mattress. So finally purchase the standard and best futon for your family and our buying guide must be very much helpful for you. Mostly, cotton is used for basic futon mattresses though; a foam layer may be used to maintain the rigidity. Polyester and pocket coils or foam can also be used with innerspring. While the Futon loungers, also known as loveseats, have an additional lounge extension that folds and hides between the seat deck and futon base. This enables it to give the most sitting and sleeping space in small rooms. The mattresses have also been improved and can be found in both custom and standard sizes which answer the question, how big are a futon mattress? The standard sizes are in the main four groups: chair, full, twin and queen which allows them to fit on standard beds. Types of Futon Mattresses In answering the question of how big is a futon mattress, we will look at the different types present. Futon mattresses act as bendable beds with frames that can be folded, but since modern futon mattresses have western dimensions, some of them cannot fold to 180 degrees. Nevertheless, futons offer maximum utilization of spaces as they can give room for other functions, working as both beds and chairs making them popular for their flexibility. Futon mattresses come in different sizes and designs. The different types of futon mattresses include: The Chair It is a classy chair that can be used as a single person chair by day and a single person bed at night. It usually measures 28 inches wide by 54 inches long when opened to a bed position and can be utilized with an elective ottoman that uses 28 inches wide by 21 inches long mattress. The Twin It is a durable futon mattress that is reversible and can be folded into a coach or opened into a bed. There are two types of twin futons: ·The Twin Split: Dimensions for the twin split are 39 inches wide by 75 inches long, and it needs an non-compulsory stand-alone ottoman along with a compatible 39 inches wide by 21 inches long mattress to be used as a bed. ·The Twin Lounger: The dimensions for the twin lounger is 39 inches wide by 80 inches long and will also need a twin size mattress to be used as a bed. However, they can both fit on twin beds. The Full Futon Mattress Among the best futon mattress , it is considered as a luxurious type of futon mattress that comes in three styles: The Loveseat: This measures 54 inches wide by 54 inches long but requires an optional stand-alone ottoman part and a love ottoman mattress to be used as a bed. The Full: That measures 75 inches wide by 54 inches long and can be used with a full-size futon mattress. The Full Adjustable Seat: which includes an ottoman that slides into the frame and a love ottoman mattress when used as a bed. Nonetheless, all three can open to a full sized bed. The Queen Futon Mattress They are found in three styles: The Queen: It measures 80 inches wide by 60 inches long when opened and uses regular queen mattresses. The Queen Split: Whose dimensions are 60 inches wide by 54 inches long when open conversely; an optional stand-alone ottoman and compatible mattress 60 inches wide by 26 inches long are required for it to be used as a bed. The Queen Lounger: It has the same body as the Queen Split the only difference is that the ottoman part slides into the frame and it needs a Queen Ottoman mattress to be used as a bed. Conclusion Futon mattresses come in different categories allowing the buyer to choose from a variety of alternatives depending on their tastes and preferences. The fact that they can be easily folded and stored elsewhere gives them an advantage over normal mattresses. The problem of estimating how big is a full size futon is quite important in case you want to find out a suitable one for yourself and your family. Chair or twin sizes can be used when one wants to utilize the least floor space possible, but they will need an alternative when they get guests. In such a case they should consider purchasing bigger futons such as Queen Split. Then again, if one requires more sitting space, a Queen Size or full futon would be a great fit. They allow maximum utilization of space as the can serve multiple purposes such as floor couches, play area or even outdoor rest areas. This makes them a must-have home accessory both because of its aesthetics and comfort. Having stated all this, you must now have realized the answer to how big is a futon mattress. Related Posts: How To Fold Up A Futon Mattress How To Put A Futon Cover On What Size Sheets For A Futon	{ "pile_set_name": "OpenWebText2" }
Bullina Bullina is a genus of sea snails or bubble snails, marine gastropod molluscs in the family Aplustridae. Species Species within the genus Bullina include: Bullina callizona Sakurai & Habe, 1961 Distribution : Philippines Description : white ovate shell with four narrow pink spiraling bands. Bullina exquisita McGinty, 1955, the exquisite bubble Distribution : Indian Ocean Length : 7.8 mm Description : found at depths of 90 to 110 m Bullina lineata Gray 1825, the red-lined bubble Distribution : Indo-Pacific, Japan to Australia. Bullina nobilis Habe, 1950 Distribution : Japan, Philippines Length : 10–21 mm Description : ovate shell with wide aperture, narrowing at the top, and white outer lip; shell with horizontally spiraling red-brown bands, crossing wavy vertical bands in the same color, over a white background. Twisted columella. Bullina oblonga Sowerby, 1893 Distribution : South Africa, Réunion. Length : 8-11.5 mm Description : rather rare; almost white shell, crossed by numerous red-brown fine spiraling bands that end at the columella. Bullina torrei (Aguayo & Rehder, 1936) Distribution : Cuba, Virgin Islands, Indian Ocean, Arabian Sea, Gulf of Bengal. Length : 8.5 mm Description : found at depths of 15 to 27 m Bullina virgo Habe, 1950 Distribution : Philippines Length : 8 mm Description : white shell crossed by two pale pink spiraling bands. Bullina vitrea Pease, 1860 Distribution : Australia, New Caledonia, Hawaii, Japan Description : found at depths of 12–15 m; shell with two gray or black spiraling bands and a yellowish color at the anterior and posterior; the snail is translucent clear with white pigmentation. References Category:Aplustridae Category:Taxa named by André Étienne d'Audebert de Férussac	{ "pile_set_name": "Wikipedia (en)" }
Chuhuiv Raion Chuhuiv Raion () is a raion (district) in Kharkiv Oblast of Ukraine. Its administrative center is the town of Chuhuiv which is incorporated separately as a city of oblast significance and does not administratively belong to the raion. References Category:Raions of Kharkiv Oblast	{ "pile_set_name": "Wikipedia (en)" }
ORIGI[{At llntbt@nitt! 9itatts @ourt otftlprul @lufins No. 15-388G Fifed: December2,2015 FILED DEC - 2 ?015 :* * * * ,1. '* :1. * * * * :i * * * * * * i U,S. COURT OF MESSIAH MIZEL, EX REL. KAMAL- FEDERAL CLAIMS JALAL: JAMES, Plaintiff, Pro Se Plaintiff; In Forma PauPeris; v, Lack of Subject Matter Jurisdiction. UNITED STATES, Defendant. * * :1. ,1. * * :l * * :i * * , * * Messiah Aziz El, Ex Rel' Kamal'Jalal: James, pro se, Freehold, NJ' Albert s. larossi, Trial Attorney, commercial Litigation Branch, civil Division, united States Department of Justice, washington, D.C., for defendant. with him were Robert E. Kirschman, Jr., Director, commercial Litigation Branch, and Benjamin c. Mizer, Principal Deputy Assistant Attorney General, civil Division, washington, D.C. ORDER HORN. J. FINDINGS OF FACT filed a complaint2 PIq se plaintiff Messiah Aziz El, "Ex. Rel Kamal-Jalal: James"r in Court of Federal Claims alleging "deprivation of Petitioner's the -UniiJd States 1 The caption for the complaint filed in this court on April 17,2015lists "Messiah Aziz El' Ex. Rel. Kamal-Jalal: James" as the plaintiff. In the first line of the complaint, however, ;Crystat Gabri El, Ex. Rel Crystal-Gabrielle: Hawkins'' is also indicated as a plaintiff' The United onty signature on the complaintwas Messiah Aziz El. Rule 11(a) of the Rules of.the StaleJCourt of Federat Claims (RCFC) (2015) requires every pleading be signed by a farty personally if the party is unrepresented, G1ve1 that neither "Kamal James" nor lCry"tbf GaOri El," if they are two additional individuals, signed the.complaint, this court does not consider eithei as a plaintiff in the above captioned case because they did not comply with RCFC 1 1 (a). Accordingly, Mr. El is considered the only plaintiff in this action. ln its submissions to the court, defe-ndant refers to plaintiff as "Kamal James," which may be his legal name. 2 Plaintiff has filed at least two other complaints in federal court. In March 2013, Mr' El fileJ a complaint in the United States District court for the District of New Jersey, which Constitutional and Internationally secured rights without due process of law" by "the United States through action by the UNITED STATES DISTRICT COURT under U.S.C. 18(United States Code), prohibiting, life, liberty and property of the Petitioner."3 In his complaint, plaintiff alleges that he "is a natural person, in propria persona sui juris, under the laws of the indigenous community Washitaw Dedugdahmoundya Nation State and under the laws of the United States of America Republic," and thus he is "entitled to all protections enumerated within the United States Constitution. Specifically the Fifth Amendment right of due process, to be heard before a court of competent jurisdiction. . . ." As relief, plaintiff seeks "an award of due process under the Fifth Amendment to the Constitution of the United States for the rights reserved by the Petitioner" and "an award of monetary damages under the Fifth Amendment of the Constitution of the United States." Defendant asserts that, in February 2015, plaintiff was convicted of conspiracy to defraud the United States in violation of 18 U.S.C. S 286 (2012), multiple counts of submitting false, fictitious, and fraudulent claims to the United States, and three counts of mail fraud in violation of 18 U.S.C. S 1341 (2012) in the United States District Courtfor the District of New Jersey. Plaintiff does not contest these assertions. The allegations contained in the complaint currently before this court appear to arise from the criminal proceedings involving plaintiffs arrest and eventual conviction. Plaintiff is currently incarcerated in the Monmouth County Correctional Institution in Freehold, New Jersey. Plaintiff's confused and disorganized complaint alleges that the United States violated his Fifth Amendment right to due process by improperly exercising personal and subject matter jurisdiction over him. Plaintiff argues that the United States District Court of the District of New Jersey did not have jurisdiction for several reasons, including that the judges were "defacto" judges "operating under color of law," the United States Supreme Court is the only court with original ju risd iction when the United States is a party, and, "[n]o injured party has been produced and therefore no lawful claim under common- law exists." Plaintiff further argues that his due process rights were violated when he was arrested by agents "acting on a warrant issued by an un-constitutional Judge Defacto. . ."4 Plaintiff argues that the 'UNITED STATES DISTRICT COURT has entered was dismissed for failure to state a claim upon which relief may be granted. See El v. New Jersev, 2:13-cv-01431 (D.N.J. 2013). Shortly thereafter, Mr. El filed another complaint in the same court, which also was dismissed for failure to state a claim upon which relief may be granted. See El v. New Jersev, 2:13-cv-03481 (D.N.J. 2013). 3 Capitalization, grammar, punctuation, emphasis, and other errors are quoted in this Order as they appear in plaintiffs submissions. a To the extent that plaintiff is asserting a claim for false arrest, such a claim alleges a tort, and this court lacks jurisdiction over such claims. See Keene Coro. v. United States, 5OB U.S. 200,214 (1993); Rick's Mushroom Serv., Inc. v. United States,521 F.3d 1338, 1343 (Fed. Cir. 2008) reh'o and reh'q en banc denied (Fed. Cir. 2004), cert. denied' 545 U.S. 1127 (2005); Gadd v. United States,232F.3d 915 (Fed. Cir. 2000); Brown v' United sham pleas of not guilty on behalf of the Petitioner without authorization, trial has commenced and Petitioner has been found guilty through colorable proceedings." Plaintiff contends that he, objected to the Personal and Subject Matter jurisdiction of the UNITED STATES DISTRICT COURT on several occasions throughout these colorable proceedings and all constitutional claims, statements and arguments have been ignored or denied by (defacto) Judges. . . Plaintiff contends that "[t]he UNITED STATES DISTRICT COURT officers, have all intentions on proceeding under color law and disregarding their obligations to the United States Constitution of the Republic and the Petitioner's rights that are secured therein." Plaintiff also alleges that "Defendant has failed to provide the requested oaths of office" that plaintiff requested defendant produce. As relief, plaintiff seeks a judgment from this court: 1. Declaring that the actions of the Defendant in exercising authority under 28 U.S.C. 3231 to prohibit all beneficial uses of the "personal rights" constitute a taking of private property for public use in violation of Petitioner's rights under the Fifth Amendment to the United States Constitution of the Republic. 2. Declaring that the foregoing actions of Defendant under 28 U.S.C. 3231 constitute a deprivation of Petitioner's property without due process of law in violation of Petitioner's rights under the Fifth Amendment to the United States Constitution; 3. awarding Petitioner the amount of $250,000 (silver specie or currency of the UNITED STATES), representing just compensation for these actions of Defendant, as well as reasonable attorney fees, expenses, and all cost ofthese proceedings... Defendant filed a motion to dismiss for lack of subject matter jurisdiction on June 26,2015, to which Plaintiff responded, followed by Defendant filing a reply in support of its motion and a further response by plaintiff. DlscussloN The court recognizes that plaintiff is proceeding pro se, without the assistance of counsel. When determining whether a complaint filed by a pro se plaintiff is sufficient to invoke review by a court, pro se plaintiffs are entitled to liberal construction of their pleadings. See Haines v. Kerner, 404 U.S. 5'19, 520-21 (requiring that allegations contained in a pro se complaint be held to "less stringent standards than formal pleadings States, 105 F.3d 621, 623 (Fed. Cir.) reh'q denied (Fed. Cir. 1997). The Tucker Act specifically excludes actions sounding in tort from this court's jurisdiction. See 28 U.S.C. S 14e1(aX1) (2012). drafted by lawyers"), reh'q denied, 405 U.S. 948 (1972); see also Erickson v. Pardus, 551 U.S. 89, 94 (2007); Huqhes v. Rowe,449 U.S. 5, 9-10 (1980); Estelle v. Gamble,429 U.S. 97, 106 (1976), reh'q denied,429 U.S. 1066 (1977); Mafthews v. United States, 750 F.3d 1320, 1322 (Fed. Cir.2014); Diamond v. United States, 115 Fed. Cl. 516, 524, aff d, 603 F. App'x 947 (Fed. Cir.), cert. denied, 135 S. Ct. 1909 (2015). "However, "'[t]here is no duty on the part of the trial court to create a claim which [the plaintiffl has not spelled out in his [or her] pleading.""'Lenqen v. United States, 100 Fed. Cl. 317, 328 (2011) (alterations in original) (quoting Scoqin v. United States,33 Fed. Cl.2B5,293 (1995) (quoting Clark v. Nat'l Travelers Life lns. Co. , 5'18 F .2d 1 167, 1 169 (6th Cir. 1975))); see also Bussie v. United States, 96 Fed. Cl. 89, 94, aff d,443 F. App'x 542 (Fed. Cir.2011); Minehan v. United States, 75 Fed. C\|.249,253 (2007). "While a pro se plaintiff is held to a less stringent standard than that of a plaintiff represented by an attorney, the pro se plaintiff, nevertheless, bears the burden of establishing the Court's jurisdiction by a preponderance of the evidence." Riles v. United States,93 Fed. Cl. 163, 165 (2010) (citing Huohes v. Rowe, 449 U.S. at 9 and Taylor v. United States, 303 F.3d 1357, 1359 (Fed. Cir.) ("Plaintiff bears the burden of showing jurisdiction by a preponderance of the evidence."), reh'q and reh'q en banc denied (Fed. Cir. 2002)); see also Shelkofskv v. United States, 119 Fed. Cl. 133, 139 (2014) ("lwlhile the court may excuse ambiguities in a pro se plaintiffls complaint, the court 'does not excuse [a complaint's] failures."' (quoting Henke v. United States,60 F.3d 795,799 (Fed. Cir. 1995)); Harris v. United States, 113 Fed. Cl. 290,292 (2013) ("Although plaintiff's pleadings are held to a less stringent standard, such leniency 'with respect to mere formalities does not relieve the burden to meet jurisdictional requirements."' (quoting Minehan v. United States, 75 Fed. Cl. at 253)). It is well established that "'subject-matter jurisdiction, because it involves a court's power to hear a case, can never be forfeited or waived."' Arbauqh v. Y & H Corp., 546 U.S. 500, 514 (2006) (quoting United States v. Cotton, 535 U.S. 625, 630 (2002)). "[F]ederal courts have an independent obligation to ensure that they do not exceed the scope of their jurisdiction, and therefore they must raise and decide jurisdictional questions that the parties either overlook or elect not to press." Henderson ex rel. Henderson v. Shinseki, 562 U.S. 428 (2011l'; see also Gonzalez v. Thaler, 132 S. Ct. 641, 648 (2012) ("When a requirement goes to subject-matter jurisdiction, courts are obligated to consider sua sponte issues that the parties have disclaimed or have not presented."); Hertz Corp. v. Friend, 559 U.S. 77,94 (2010) ("Courts have an independent obligation to determine whether subjeclmatter jurisdiction exists, even when no party challenges it." (citing &Ug-hr-Y-&-H-Q.eIS., 546 U.S. at 514)); Special Devices. Inc. v. OEA. lnc., 269 F.3d 1340, 1342 (Fed. Cir. 2001) ("[A] court has a duty to inquire into its jurisdiction to hear and decide a case." (citing Johannsen v. Pay Less Druq Stores N.W.. Inc., 918 F 2d 160, 161 (Fed. Cir. 1990)); View Enq'o. lnc. v. RoboticVision Svs.. lnc., 115 F.3d 962' 963 (Fed. Cir. 1997) ("[C]ourts must always look to their jurisdiction, whether the parties raise the issue or not."). "Objections to a tribunal's jurisdiction can be raised at any time, even by a party that once conceded the tribunal's subject-matter jurisdiction over the controversy." Sebelius v. Auburn Reo'l Med. Ctr., 133 S. Ct.817,824 (2013); see also Arbauqh v. Y & H Corp., 546 U.S. at 506 ("The objection that a federal court lacks subjecf matter jurisdiction . . . may be raised by a party, or by a court on its own initiative, at any stage in the litigation, even after trial and the entry of judgment."); Cent. Pines Land Co., L.L.C. v. United States, 697 F.3d 1360, 1364 n.1 (Fed. Cr.2012\ ("An objection to a court's subject matter jurisdiction can be raised by any party or the court at any stage of litigation, including after trial and the entry of judgment." (citing Arbauqh v. Y & H Corp., 546 U.S. at 506-07)); Rick's Mushroom Serv.. lnc. v. United States, 521 F.3d at 1346 ("[A]ny party may challenge, orthe court may raise sua sponte, subject matter jurisdiction at any time." (citing Arbauqh v. Y & H Corp., 546 U.S. at 506; &,lde[y.,--U-nj!edsta!S, 379 F.3d 1344, 1354 (Fed. Cir.); and Fannino. Phillips & Molnar v. West, 160 F.3d 717, 720 (Fed. Cir. 1998))); Pikulin v. United States, 97 Fed. CL71,76, appeal dismissed,425 F. App'x 902 (Fed. Cr. 2011). ln fact, "[s]ubject matter jurisdiction is an inquiry that this court must raise sua sponte, even where . neither party has raised this issue." Metabolite Labs., lnc. v. Lab. Corp. of Am. Holdinqs,370 F.3d 1354, 1369 (Fed. Cir.) (citing Textile Prods.. lnc. v. Mead Corp., 134 F.3d 1481,1485 (Fed. Cir.), reh'o denied and en banc suqqestion declined (Fed. Cir.), cert. denied, 525 U.S. 826 (1998)), reh'q and reh'q en banc denied (Fed. Cir. 2004), cert. qranted in pa1! sub. nom Lab. Coro. of Am. Holdinqs v. Metabolite Labs., lnc., 546 U.S. 975 (2005), cert dismissed as improvidentlv qranted, 548 U.S. 124 (2006); see also Avid ldentification Svs., Inc. v. Crvstal lmport Corp.,603 F.3d 967,971 (Fed. Cir.) ("This court must always determine for itself whether it has jurisdiction to hear the case before it, even when the parties do not raise or contest the issue."), reh'q and reh'q en banc denied, 6'\|4 F.3d 1330 (Fed' Cir. 2010), cert. denied, 131 S. Ct.909 (2011). Pursuantto RCFC 8(aX1) and the Federal Rules of civil Procedure, a plaintiff need only state in the complaint "a short and plain statement of the grounds for the court's jurisdiction," and "a short and plain statement of the claim showing that the pleader is entitled to relief." RCFC 8(aX1), (2) (2015): Fed. R. Civ. P. B(a)(1)' (2) (2015); see also Ashcroftv. lqbal,556 U.S. 662,677-78 (2009) (citing Bell Atl. Corp. v. Twomblv' 550 U.S. sn+, sss-sl ,570 (2007)). "Determination of jurisdiction starts with the complaint, which must be well-pleaded in that it must state the necessary elements of the plaintiffs claim, independent of any defense that may be interposed." Hollev v. United states, 124 F.3d 1462, 1465 (Fed. Cir.) (citing Franchise Tax Bd. v. constr. Laborers Vacation Trust,463 U.S. 1 (1983)), reh'o denied (Fed. Cir. 1997); see also Klamath Tribe Claims Comm. v. United States,97 Fed. C;.203,2O8(2011); Gonzalez-McCaullev lnv. Grp., lnc. v. United states, 93 Fed. Cl. 710,713 (2010). "Conclusory allegations of law and unwarranted irfeiences of fact do not suffice to support a claim." Bradlev v. Chiron Corp., 136 F.3d 1317,1322 (Fed. Cir. 1998); see also McZeal v. Sprint Nextel Corp.' 501 F.3d 1354' 1363 n.9 (Fed. Cir. 2007) (Dyk, J., concurring in part, dissenting in part) (quoting C. Wright and A. Miller, Federal Practice and Procedure S 1286 (3d ed.2004)). "A plaintiffs factual allegations must 'raise a right to relief above the speculative level' and cross 'the line from conceivable to plausible."'Three S Consulting v. United States, 104 Fed. Cl. 510' 523 (2012) (quoting Bell Atl. Corp. v. Twomblv, 550 U.S. at 555), affd, 562 F. App'x 964 (Fed. Cir.;, ien;q denieO (Fed. Cir. 2014). As stated in Ashcroft v. lqbal, "[a] pleading,that offers 'labels and conclusions' or 'a formulaic recitation of the elements of a cause of action will not do.' 550 u.s. at 555. Nor does a complaint suffice if it tenders'naked assertion[s]' devoid of 'further factual enhancement."' Ashcroft v. lqbal, 556 U S. at 678 (quoting Egll Atl. Corp. v. Twomblv, 550 U.S. at 555). The Tucker Act grants jurisdiction to this court as follows: The United States Court of Federal Claims shall have jurisdiction to render judgment upon any claim against the United States founded either upon the Constitution, or any Act of Congress or any regulation of an executive department, or upon any express or implied contract with the United States, or for liquidated or unliquidated damages in cases not sounding in tort. 28 U.S.C. S 1a91(aX1). As interpreted by the United States Supreme Court, the Tucker Act waives sovereign immunity to allow jurisdiction over claims against the United States (1) founded on an express or implied contract with the United States, (2) seeking a refund from a prior payment made to the government, or (3) based on federal constitutional, statutory, or regulatory law mandating compensation by the federal government for damages sustained. See United States v. Navaio Nation, 556 U.S. 287, 289-90 (2009); United States v. Mitchell, 463 U.S. 206,216 (1983); see also Greenlee Cntv., Ariz. v. United States,487 F.3d 871,875 (Fed. Cir.), reh'q and reh'q en bancdenied (Fed. Cir. 2007), cert. denied, 552 U.S. 11a2 QO08); Palmer v. United States, 168 F.3d 1310, 1314 (Fed. Cir. 1999). "Not every claim invoking the Constitution, a federal statute, or a regulation is cognizable under the Tucker Act. The claim must be one for money damages against the United States . . . ." United States v. Mitchell, 463 U.S. at 216; see also United States v. White Mountain Apache Tribe, 537 U.S. 465,472 (2003); Smith v. United States, 709 F.3d 1114, 1116 (Fed. Cir.), cert. denied, 134 S. Ct. 259 (2013); RadioShack Corp. v. United States,566 F.3d 1358, 1360 (Fed. Cir.2009); Rick's Mushroom Serv.. lnc. v. United States, 521 F.3d at 1343 ("[P]laintiff must . . . identify a substantive source of law that creates the right to recovery of money damages against the United States."); Golden v. United States, 118 Fed. Cl.764,768 (2014). In Ontario Power Generation, lnc. v. United States, the United States Court of Appeals for the Federal Circuit identified three types of monetary claims for which jurisdiction is lodged in the United States Court of Federal Claims. The court wrote: The underlying monetary claims are of three types. . . . First, claims alleging the existence of a contract between the plaintiff and the government fall within the Tucker Act's waiver. Second, the Tucker Act's waiver encompasses claims where "the plaintiff has paid money over to the Government, directly or in effect, and seeks return of all or part of that sum." Eastoort S.S. lCorp. v. United States, 178 Ct. Cl. 599,605-06'1 372F.2d [1002,] 1007-08 [(1967)] (describing illegal exaction claims as claims "in which 'the Government has the citizen's money in its pocket"' (quoting Clappv. United States,127 Ct. Cl.505, 117 F. Supp.576,580 (1954)... . Third, the Court of Federal Claims has jurisdiction over those claims where "money has not been paid but the plaintiff asserts that he is nevertheless entitled to a payment from the treasury." Eastport S.S., 372 F.2d at 1007 Claims in this third category, where no payment has been made to the government, either directly or in effect, require that the "particular provision of law relied upon grants the claimant, expressly or by implication, a right to be paid a certain sum." ld.; see also lUnited States v. lTestan, 424 U.S. 1392,1 401-02 [1976] ("Where the United States is the defendant and the plaintiff is not suing for money improperly exacted or retained, the basis of the federal claim-whether it be the Constitution, a statute, or a regulation- does not create a cause of action for money damages unless, as the Court of Claims has stated, that basis 'in itself . . can fairly be interpreted as mandating compensation by the Federal Government for the damage sustained."' (quoting Eastport S.S., 372 F.2d at 1009)). This category is commonly refened to as claims brought under a "money-mandating" statute. Ontario PowerGeneration, Inc. v. United States,369 F.3d 1298, 1301 (Fed. Cir.2004); see alsoTwp. of Saddle Brookv. United States, 104 Fed. Cl. 101, 106(2012)' To prove that a statute or regulation is money-mandating, a plaintiff must demonstrate that an independent source of substantive law relied upon "'can fairly be interpreted as mandating compensation by the Federal Government."' United States v. Navaio Nation, 556 U.S. at 290 (quoting United States v. Testan, 424 U 5. 392' 400 (1976)); see also United States v. White Mountain Apache Tribe, 537 U.S. at 472; United States v. Mitchell,463 U.S. at 217; Blueport Co., LLC v. United States' 533 F.3d 1374' 1383 (Fed. Cir. 2008), cert. denied, 555 U.S. 1153 (2009). The source of law granting monetary relief must be distinct from the Tucker Act itself. See United States v. Navaio Nation, 556 U.S. at 290 (The Tucker Act does not create "substantive rights; [it is simply a1 iurisdictional provision[] that operate[s] to waive sovereign immunity for claims premised on other sources of law (e.9., statutes or contracts)."). "'lf the statute is not money-mandating, the court of Federal claims lacks jurisdiction, and the dismissal should be for lack of subject matter lurisdiction."' Jan's Helicopter serv., lnc. v. Fed. AviationAdmin.,52S F.3d 1299, 1308 (Fed. Cir.2008) (quoting Greenlee.Cjtlv'-Ariz. v. United States, 487 F .3d at 876); Fisher v. United States , 402 F .3d 1167 , 1173 (Fed. Cir. 2005) (The absence of a money-mandating source is "fatal to the court's jurisdiction under the TuckerAct."); Peoples v. United States, 87 Fed. Cl. 553' 565-66 (2009). In his complaint, plaintiff asserted violations of his constitutional right to due orocess under the Fifth Amendment to the United States Constitution. Plaintiff alleges that this court has jurisdiction to hear his claims pursuant to the Tucker Act, 28 U.S.C. s 1491(a). Defendant argues, however, that "the Tucker Act does not, by itself, create a right to money damages against the united states," and that "the substantive right to money damages must extend from the constitutional provision, statute, contract, or regulation giving rise to the claim." Defendant contends that plaintiff "has failed to plead a iubstantive source of law that creates a right to recover money damages from the Government," because it is well established that the Due Process clause of the Fifth Amendment does not mandate the payment of money, and, thus, does not provide a cause of action under the Tucker Act. 7 Regarding plaintiff's claim for due process under the Fifth Amendment, the Federal Circuit has held that this court does not possess jurisdiction to consider claims arising under the Due Process Clauses of the Fifth and Fourteenth Amendments. See Smith v. United States, 709 F.3d 1114, 1116 (Fed. Cir.) ("The law is well settled that the Due Process clauses of both the Fifth and Fourteenth Amendments do not mandate the payment of money and thus do not provide a cause of action under the Tucker Act." (citing LeBlancv. United States,50 F.3d 1025, 1028 (Fed. Cir. 1995))), cert. denied, 134S. Ct. 259 (20'13); In re United States,463 F.3d 1328, 1335 n.5 (Fed. Cir.) ("[B]ecausethe Due Process Clause is not money-mandating, it may not provide the basis for jurisdiction under the Tucker Act."), reh'q and reh'q en banc denied (Fed. Cir. 2006) cert. denied sub nom. Scholl v. United States, 552 U.S. 9a0 (2007);Acadia Tech., Inc. & Global Win Tech., Ltd. v. United States,458 F.3d 1327,1334 (Fed. Cir. 2006); Crockerv. United States, 125 F.3d 1475, 1476 (Fed. Cir. 1997) (citing LeBlancv. United States,50 F.3d at 1028) (no jurisdiction over a due process violation underthe Fifth and Fourteenth Amendments); Collins v. United States, 67 F.3d 284,288 (Fed. Cir.) ("[T]he due process clause does not obligate the government to pay money damages.'), rc"h-g denied (Fed. Cir. 1995); Mullenberq v. United States,857 F.2d770,773 (Fed. Cir. 1988) (finding that the Due Process clauses "do not trigger Tucker Act jurisdiction in the courts"); Murrav v. United States, 817 F.2d 1580, 1583 (Fed. Cir. 1987) (noting that the Fifth Amendment Due Process clause does not include language mandating the payment of money damages); Harperv. United States, 104 Fed. C\|.287,291 n.5 (2012); Hampel v. United.States' 97 Fed. Cl. 235,238,aff'd,429 F. App'x 995 (Fed. Ctt.2011), cert. denied, 132 S. Ct. 1105 (2012); McCullouqh v. United States,76 Fed. Cl. 1,4 (2006), appeal dismissed,236 F. App'x 615 (Fed. Cir.), reh'q denied (Fed. Cir.), cert. denied, 552 U S. 1050 (2007) ("[N]either the Fifth Amendment Due Process Clause nor the Privileges and lmmunities Clause provides a basis for jurisdiction in this court because the Fifth Amendment is not a source that mandates the payment of money to plaintiff."). Due process claims "must be heard in District court." Kam-Almaz v. united states, 96 Fed. ct. 84, 89 (2011) (citing Acadia Tech.. lnc. & Global win Tech., Ltd. v. united States, 458 F.3d at 1334), alf'd,682 F.3d 1364 (Fed. Cir.2012); see also Hampel v. United States, 97 Fed. Cl. at 238. Therefore, to the extent that plaintiff is attempting to allege Due Process violations, no such cause of action can be brought in this court. In addition to monetary damages, plaintiff asks this court to declare that, [T]he actions of the Defendant in exercising authority under 28 U.S.C. 3231 io prohibit all beneficial uses of the "personal rights" constitute a taking of private property for public use in violation of Petitioner's rights under the Fifth Amendment to the United States Constitution of the Republic. . . [and] that the foregoing actions of Defendant under 28 U S'C. 3231 constitute a deprivation of Petitioner's property without due process of law in violation of Petitioner's rights under the Fifth Amendment to the United States Constitution. . . Plaintiffls allegations in no way can be considered a taking in violation of the Fifth Amendment to the United States Constitution. The Takings Clause of the Fifth Amendment to the United States Constitution provides in pertinent part: "nor shall private property be taken for public use without just compensation." U.S. Const. amend. V. The purpose of this Fifth Amendment provision is to prevent the government from "'forcing some people alone to bear public burdens which, in all fairness and justice, should be borne by the public as a whole."' Palazzolo v. Rhode lsland, 533 U.S. 606, 618 (2001) (quoting Armstronq v. United States, 364 U.S.40,49 (1960)); see also Penn Central Transp. Co. v. Citv of New York, 438 U.S. 104,123-24, reh'q denied, 439 U.S. 883 (1978); Linqle v. Chevron U.S.A. lnc., 544 U.S. 528, 536 (2005); E. Enters. v. Apfel, 524 U.S. 498,522 (1998); Rose Acre Farm. Inc. v. United States, 559 F.3d 1260,1266 (Fed. Cir.), reh'q en bancdenied (Fed. Cir.2009), cert. denied, 130 S. Ct. 1501 (2010); Janowskvv. United States, 133 F.3d 888,892 (Fed. Cir. 1998); Resource Invs., lnc. v. United States, 85 Fed. C'.447,469-70 (2009); Pumpellv v. Green Bav & Miss. Canal Co., 80 U.S. (13 Wall.) 166, 179 (1871\ (citing to principles which establish that "private property may be taken for public uses when public necessity or utility requires" and that there is a "clear principle of natural equity that the individual whose property is thus sacrificed must be indemnified."). A plaintiff must show that the government took a private property interest for public use without just compensation. See Adams v. United States, 391 F.3d 1212, 1218 (Fed. Cir. 2004), cert. denied, 546 U.S. 81 1 (2005); Arbelaez v. United States, 94 Fed. Cl. 753, 762(2010):Gahaqanv.UnitedStates,72Fed. Cl. 157, 162(2006). "Theissueofwhether a taking has occurred is a question of law based on factual underpinnings." Huntleiqh USA Corp v. United States, 525 F.3d 1370, 1377-78 (Fed. Cir.), cert. denied, 555 U.S. 1045 (2008) (citations omitted). The Federal Circuit has established a two-part test to determine whether governmental actions amount to taking of private property under the Fifth Amendment. See Klamath lrr. Dist. v. United States,635 F.3d 505,511 (Fed. Cir. 2011); Am. Pelaqic Fishinq Co. v. United States,379 F.3d 1363, 1372 (Fed. Cir.) (citing M & J Coal Co. v. United States , 47 F .3d 1148, 1153-54 (Fed. Cir. 1995)), reh'q en banc denied, (2004). A court first determines whether a plaintiff possesses a cognizable property interest in the subject of the alleged taking. Then, the court must determine whether the government action is a "'compensable taking of that property interest."' Huntleiqh USA Corp v. United States, 525 F.3d at'1377 (quoting Am. Pelaqic Fishinq Co.. L.P. v. United States, 379 F.3d a\| 1372). A plaintiff alleging a taking must have a legally cognizable property interest, such as the right of possession, use or disposal of the property. See Loretto v. Teleprompter Manhattan CATV Corp., 458 U.S. 419,435 (1982) (citing United States v. Gen. Motors Corp., 323 U.S. 373 (19a5)); CRV Enters., lnc. v. United States,626 F.3d 1241,1249 (Fed. Cir. 2010), cert. denied, 131 S. Ct.2459 (201 1); KarukTribe of Cal. v. Ammon, 209 F.3d 1366, 1374-75 (Fed. Cir.), reh'o denied and en banc suqqestion denied (Fed. Cir. 2OO0), cert. denied, 532 U.S. 941 (2OO1). "\|t is axiomatic that only persons with a valid property interest at the time of the taking are entitled to compensation." Am. Pelaoic Fishinq Co. v. United States, 379 F.3d al 1372 (quoting Wvatt v. United States , 27'l F .3d 1090, 1096 (Fed. Cir. 2001), cert. denied, 353 U.S. 1077 (2002) and citing Cavin v. United States, 956 F.2d 1131,1134 (Fed. Cir. 1992)). Therefore, "[i]f the claimant fails to demonstrate the existence of a legally cognizable property interest, the courts [sic] task is at an end." Am. Pelaqic Fishinq Co. v. United States, 379 F.3d at 1372 (citing Maritrans lnc. v. United States , 342 F.3d 1344, 1352 (Fed. Cir. 2003) and M & J Coal, 47 F.3d at 1 154). The court does not address the second step "without first identifying a cognizable property interest." Air Peqasus of D.C., lnc. v. United States, 424 F.3d 1206, 1213 (Fed. Cir.) (citing Am. Pelaqic Fishinq Co. v. United States, 379 F.3d at 1381 and Conti v. United States,291 F.3d 1334, 1340 (Fed. Cn.2002), cert. denied,537 U.S. 1112 (2003)), reh'o denied and reh'o en banc denied (Fed. Cir. 2005). Only if there is to be a next step, "after having identified a valid property interest, the court must determine whether the governmental action at issue amounted to a compensable taking of that property interest." Huntleiqh USA Corp. v. United States, 525 F.3d at 1378 (quoting Am. Pelaqic Fishinq Co. v. United States, 379 F.3d at 1372). To the extent plaintiff alleges that a violation of due process is a taking, plaintiff has not alleged a property interest subject to a constitutional taking claim nor has plaintiff alleged any cognizable taking. Moreover, the Federal Circuit has explained that the Court of Federal Claims "does not have general equity jurisdiction to grant injunctive relief." Shemonskv v. United States, 21 5 F.3d 1340 (Fed. Cir. 1999) (upholding decision of the Court of Federal Claims that the court's jurisdiction did not extend to injunctive or declaratory relief); see also Bank of Guam v. United States, 578 F.3d 1318, 1331 (Fed. Cir.) reh'q and reh'q en banc denied (Fed. Cir. 2009), cert. denied, 561 U.S. 1006 (2010); Martinez v. United States, 333 F'3d 1295, 1303 (Fed. Cir.2003); Choate v. United States,60 F.3d 840 (Fed. Cir. 1995) (holding that "[t]he Tucker Act does not provide a means by which the Court of Federal Claims may grant injunctive or declaratory relief where the suit does not involve a pre- award protestorthe application of section 7428 of lhe Internal Revenue Code"). As stated by the Federal Circuit: The Court of Federal Claims has never been granted general authority to issue declaratory judgments, and to hold that the Court of Federal Claims may issue a declaratory judgment in this case, unrelated to any money claim pending before it, would effectively override Congress's decision not to make the Declaratory Judgment Act applicable to the Court of Federal Claims. Nat'l AirTraffic Controllers Ass'n v. United States, 160 F.3d 714,716-17 (Fed. Cir.1998); see also United States v. Tohono O'Odham Nation,563 U.S.307 (2011) (The United States Court of Federal Claims "has no general power to provide equitable relief against the Government or its officers."). Moreover, in an action brought under 28 U.S.C. "as an incident of and S 1491(a), this court can only provide declaratory or injunctive relief collateral to" a judgment for money damages. 28 U.S.C. S 1a92(a)(2); see also Tavlor v' United states, 1 1 3 Fed. cl. 17 1, 173 (2013) (holding that the Tucker Act does not provide independent jurisdiction over claims for injunctive relief in contractual dispute cases). For example, in a case brought under the Tucker Act that did not involve a pre-award protest, once the Court of Federal Claims determined that it did not have jurisdiction to hear a plaintiffs claims for money damages, it necessarily followed that the court did not have authority to hear that plaintiffs claims for injunctive relief. See Kanemoto v. Reno, 41 10 F.3d 641, 645 (Fed. Cir. 1994); see also Tavlor v. United States, 113 Fed. Cl. at 173. Accordingly, plaintiff s demands for declaratory relief are dismissed for lack of jurisdiction. Plaintiff also contends in his complaint that the New Jersey District Court did not have subject matter or personal jurisdiction over him. To the extent that plaintiff is alleging that the New Jersey Diskict Court erred in exercising jurisdiction in the criminal action brought against him, such allegations also do not establish jurisdiction in this court. The Court of Federal Claims does not have authority to review decisions issued by judges of the United States District Courts. See Vereda, LTDA. v. United States,271 F.3d 1367, 1375 (Fed. Cir.2001); Joshua v. United States, 17 F.3d 378,379 (Fed Cir. 1994); see also Cooper v. United States, 104 Fed. Cl. 306, 312 (2012) (holding that "this court does not have jurisdiction over [plaintiffs] claims because the court may review neither criminal matters, northe decisions of district courts.") (internal citations omitted); Mendes v. United States, 88 Fed. Cl. 759,762, appeal dismissed ,375 F. App'x 4 (Fed. Cir' 2009); Huffed v. united states, 87 Fed. Cl. 696, 702 (2009) (holding that the united states court of feOerat Claims lacked jurisdiction over claims arising from the violation of a criminal statute); Matthews v. United states,72 Fed. cl. 274,282 (finding that the court lacked jurisdiction -lr/cCullouqhto consider plaintiffs criminal claims), recons. denied, 73 Fed. Cl. 524 (2006); v. United States, 76 Fed. Cl. at 4 (finding that the court lacked jurisdiction to consider platntiffb criminal claims). Therefore, this court lacks jurisdiction over any of plaintiff s claims. when filing his complaint, plaintiff also filed a motion to proceed .in forma pauperis. According to hislomplaint, plaintiff is an inmate at the Monmouth County Correctional Institutiori in Freehold, New Jersey. In order to provide access to this court to those who cannot pay the filing fees mandated in this court by RCFC 77.1(c) (2015), the statute at 2g U.S.C.-S 1g1S(2012) permits a court to allow plaintiffs to file a complaint without payment oifees or security, under specific circumstances. The standard in 28 U.S.C. pay such fees or give security b iglSt"ltr) for jn forma pauperis eligibility is "unable to iherefor.'; Determination of what constitutes "unable to pay" or unable to "give security paUperis is left to therefor," and therefore, whether to allow a plaintiff to proceed in forma the discretion of the presiding judge, based on the information submitted by the plaintiff or plaintiffs. See, eg, Rowiand v. Cal. Men's Colonv, 506 U.S. 194' 217-18 (1993); Fuentes v. United States, 100 Fed. Cl. 85, 92 (2011) when the person submitting a request to proceed i1 forma oauperig is a prisoner, 2g U.S.C. S 191 5iaX2) requires tnit ttre prisoner submit, along with the affidavit required by subsection (aX1), a certified copy of: [T]hetrustfundaccountstatement(orinstitutiona\|equiva\|ent)forthe brisone, for the 6_month period immediately preceding the filing of the complaint or notice of appeal, obtained from the appropriate official of each prison at which the prisoner is or was confined 28 U.S.C. S 191 5(aX2); see also Matthews v. United states , 72 Fed. cl. at 277. In the affidavit required under 28 U.S.C. S 1 915(aX1 ), a prisoner must further "state the nature of the action, defense or appeal and affiant's belief that the person is entitled to redress." 11 28 U.S.C. S 1915(aXl). In the above-captioned case, plaintiff submitted the required information and certification with his request to proceed in forma oauperis. Although Mr. El's income level may qualifo him for in forma oauoeris status, as discussed above, his complaint is being dismissed for lack of jurisdiction. CONCLUSION For all the reasons discussed above, plaintiffs complaint is DlsilllssED. The Clerk of the Court shall enter JUDGMENT consistent with this Order. IT IS SO ORDERED. lLrral* /MARIAN BLANK HORN 12	{ "pile_set_name": "FreeLaw" }
Structural determinants of factor IX(a) binding in nitrophorin 2, a lipocalin inhibitor of the intrinsic coagulation pathway. Nitrophorin 2 (NP2) is a salivary lipocalin from Rhodnius prolixus that binds with coagulation factors IX (fIX) and IXa (fIXa). Binding of NP2 with fIXa results in potent inhibition of the intrinsic factor Xase complex. A panel of site-directed surface mutants of NP2 was generated to locate determinants of high affinity fIX(a) binding. The locations of the mutations were based on comparisons with the related, but less potent, inhibitor nitrophorin 3 (NP3). Three point mutants (K21A, K92A, and V94A) were found that clearly reduced the inhibitory potency as measured by the activity of a reconstituted factor Xase system. Binding of NP2 with fIXa and fIX as measured by surface plasmon resonance and isothermal titration calorimetry was reduced in a similar manner. Of the three mutants, two (K92A and V94A) were located on the loop connecting beta-strands E and F of the lipocalin beta-barrel. The largest changes were seen with the K92A mutation, which lies at the apex of the loop, with a smaller effect being seen with mutation of Val(94). Combination of four E-F loop mutations (K92A, A93K, V94A, E97A) in a single mutant reduced the inhibitory potency and binding to levels similar to those seen with NP3 without affecting heme or histamine binding.	{ "pile_set_name": "PubMed Abstracts" }
Gait Patterns in Children With Cancer and Vincristine Neuropathy. Children treated with vincristine often develop chemotherapy-induced peripheral neuropathy (CIPN), but effects of CIPN on gait have not been reported. Gait variables of 52 children/adolescents treated for non-central nervous system cancers with CIPN were compared with an age- and sex-matched control group. Gait data were collected via GaitRite walkway before and after completing a 6-minute walk test (6MWT). Ankle range-of-motion (ROM) measures, balance, and strength tests were also completed. Participants with CIPN had decreased velocity and step length. Ankle ROM and balance explained variability in step length. Both groups increased self-selected velocity after the 6MWT, but participants with cancer walked with slower velocity, shorter step length, and decreased cadence. Strength, neuropathy, and self-selected velocity measured before the 6MWT explained variability in 6MWT scores. Ankle ROM and balance are important factors when treating step length deficits, whereas strength is also an important consideration for walking capacity.	{ "pile_set_name": "PubMed Abstracts" }
Soft tissue reaction to de novo plaque formation on implants and teeth. An experimental study in the dog. The aim of the present investigation was to assess the effect of de novo plaque formation on the gingiva and masticatory mucosa around teeth and implants. The study was performed in 5 beagle dogs which at the initiation of the experiment were 15 months old. During a preparatory period, the mandibular right premolars were extracted, 3 fixtures installed, abutment connection performed and a 4-month period of plaque control completed. A clinical examination was performed and biopsies of the second mandibular premolar (P2) and the contralateral implant site (2P) were sampled. The dogs were allowed to form plaque during a period of 3 weeks. The clinical examination was repeated and biopsies harvested from the 2 remaining implants and the contralateral tooth sites. The tissue samples were prepared for histometric and morphometric analysis. Both the masticatory mucosa at implants and the gingiva responded to de novo plaque formation with the development of an inflammatory lesion. The size as well as the composition of the lesions in the 2 tissues had many features in common. It was concluded that the mucosa around implants and the gingiva around teeth had a similar potential to respond to early plaque formation.	{ "pile_set_name": "PubMed Abstracts" }
"Motorola and Apple have been talking to Cingular about it using the iTunes phone," according to one of the publication's sources, who asked not to be named. Meanwhile, RBC Capital analyst Mark Sue said in a recent research note that Apple and Cingular were working out final details on revenue sharing pertinent to the arrangement. Representatives from both Apple and Cingular declined to comment on the report, but a Motorola spokeswoman said the company is on schedule to ship an iTunes phone in the third quarter. Apple and Motorola have been working to bringing Apple's popular iTunes service to mobile phones but Motorola has delayed unveiling its iTunes device so far amid speculation about a lack of interest from operators. According to a report in French newspaper Le Figaro, Apple and Motorola may announce the device, dubbed iPhone, as early as next Tuesday.	{ "pile_set_name": "Pile-CC" }
1. Introduction {#sec1-materials-12-03635} =============== Laser cutting is a process in which a (continuous or pulsed) laser beam melts, simultaneously melts and evaporates, or melts and/or burns away the material in the cut (the kerf). It is a thermal cutting process that uses a large amount of energy that is focused on a small area \[[@B1-materials-12-03635]\]. Along with the laser beam, a stream of gas is emitted coaxially; the gas might be reactive to the material being cut (oxygen, air) or it might be an inert gas (nitrogen, argon). The presence of a reactive gas in the kerf produces an exothermic reaction, which effectively increases the power of the laser, allowing it to cut material of considerable thickness \[[@B2-materials-12-03635],[@B3-materials-12-03635]\]. An inert gas is used to remove molten material and it also acts as a cooling and protective agent for the optical system of the laser cutter \[[@B3-materials-12-03635],[@B4-materials-12-03635]\]. Laser technologies can be used to section most engineering materials, e.g. unalloyed steel \[[@B1-materials-12-03635],[@B5-materials-12-03635]\], stainless steel \[[@B4-materials-12-03635],[@B6-materials-12-03635]\], aluminum alloys \[[@B7-materials-12-03635]\], nickel alloys \[[@B8-materials-12-03635]\], carbon fiber reinforced polymer composites \[[@B9-materials-12-03635]\], and also natural materials. The laser cutting of metallic materials is widely used in the automotive, chemical, marine, and aviation industries \[[@B10-materials-12-03635]\]. The strengths of this material separation technology include a high cutting speed (many times faster than the speeds used during machining), satisfactory dimensional accuracy, and low roughness of the cut surface, as well as a high level of automation and flexibility \[[@B1-materials-12-03635],[@B11-materials-12-03635],[@B12-materials-12-03635]\]. During laser cutting, the coherent laser beam might generate imperfections on the cut faces and edges. Imperfections, according to the EN ISO 17658: 2015 standard, are irregularities or deviations from the specified shape or location of cut. The laser cutting process directly causes them (no adverse phenomena resulting from external stresses or deformations are taken into account). The quality of the surface and the course of the cutting process depend on many factors with different levels of control. The most important of them include laser power, laser type and operation mode, position of laser beam focus relative to the surface of the workpiece, cutting speed, and assist gas type and pressure \[[@B10-materials-12-03635]\]. The quality of laser-cut edges and surfaces is evaluated quantitatively in accordance with EN ISO 9013:2017. In industrial practice, surface roughness, width of the heat affected zone, material phase changes, residual stresses in the vicinity of the kerf, and the formation of dross on the lower edges of the kerf define the quality of laser-cut surfaces \[[@B1-materials-12-03635],[@B9-materials-12-03635],[@B11-materials-12-03635],[@B13-materials-12-03635]\]. Surface roughness in laser cutting is the result of the overlapping effects of the thermal and hydrodynamic processes that are characteristic of laser processing and vibrations of the workpiece caused by the high-pressure gas jet \[[@B3-materials-12-03635]\]. The surface of the workpiece can be divided into two areas of different shape and quality. These zones are separated from each other by the so-called boundary layer separation (BLS). One zone is the laser beam entrance area and the other is the laser beam exit area; the latter has a much higher surface roughness \[[@B11-materials-12-03635],[@B14-materials-12-03635]\]. A characteristic feature of the laser cutting process is the formation of striations on the cut surface. The striations pattern forms as a consequence of hydrodynamic flow of molten material, laser power fluctuation, gas flow fluctuation, and laser head oscillation \[[@B15-materials-12-03635],[@B16-materials-12-03635]\]. During laser cutting, the heat that is delivered to the material and the high temperature gradient create a thin film of material with variable properties, called the heat affected zone, which is characterized by a small thickness, but locally, especially at the edges and corners, has high hardness. The differences in hardness may be associated with a non-uniform distribution of energy at beam cross-section and differences in heat dissipation rates \[[@B17-materials-12-03635],[@B18-materials-12-03635]\]. The high temperature in the cutting zone gives rise to thermal stresses. Heating of the material followed by rapid cooling with the assist gas leads to material shrinkage and phase changes. The dynamics of phase changes at different cooling rates lead to the formation of tensile residual stresses. The value and distribution of thermal stresses depend on the thermal conductivity of the material being cut \[[@B11-materials-12-03635],[@B13-materials-12-03635]\]. Non-alloy steel characteristically forms a thin surface layer of iron oxide phases FeO and Fe~3~O~4~, which change the color of the surface to bluish \[[@B19-materials-12-03635],[@B20-materials-12-03635],[@B21-materials-12-03635]\]. The resultant particles are about 50 µm in size and they most often have the form of iron spheres that are coated with iron oxide \[[@B3-materials-12-03635]\]. The presence of various imperfections on the surfaces and edges of laser-cut parts and the presence of drops of solidified material at their bottom edges calls for finishing treatment. Scale and outflow can be removed with hand files, wire brushes, deburring machines, and belt sanders. Mechanical grinding \[[@B22-materials-12-03635]\] and smoothing with diamond tools \[[@B23-materials-12-03635]\] can improve the geometrical structure of a laser-cut surface. Mechanical grinding allows for reducing surface roughness and the thickness of the hardened layer and introduces into the workpiece a state of tensile residual stress, which might lead to cracking of the surface. The resultant defects are loci of stress concentration and reduce the bearing capacity of the processed surface \[[@B22-materials-12-03635]\]. Smoothing with diamond tools allows for reducing surface roughness by several times. At the same time, it causes structural and phase changes in the material. As a result of smoothing, residual austenite is transformed into martensite. The surface layer is mechanically hardened. The use of a diamond tool is associated with the occurrence of sliding friction between the tool and the surface of the workpiece, which leads to an increase in temperature in the machining zone. Hardness in the edge zone is higher than that after laser cutting; discontinuities and microcracks also appear \[[@B23-materials-12-03635]\]. The condition of the surface and edges of a laser-cut semi-finished product does not allow or makes its use difficult in subsequent production stages. It is also difficult to apply paint coatings to it because of flaking. Centrifugal shot peening changes the properties of the surface layer of workpieces pre-machined by turning, milling, or grinding. Centrifugal shot peening of a pre-milled surface leads to the formation of a dimpled surface with elongated depressions that can retain additional lubricant \[[@B24-materials-12-03635]\]. The use of optimal centrifugal shot peening conditions during finish machining of turned C45 and 40H steel shafts, allows for obtaining a several-fold reduction in Ra. The positive effects of using shot peening for post-processing of machined parts encourage the use of centrifugal shot peening to modify the surface layer of laser-cut parts. Shot peening as a process used to finish machine parts allows for obtaining a surface that is characterized by a low roughness \[[@B25-materials-12-03635]\] and a large bearing area (material ratio). This is due to the more "streamlined" shape of the micro asperities that form during peening \[[@B25-materials-12-03635],[@B26-materials-12-03635]\]. Indentations imparted to the shot-peened surface serve as "reservoirs" for lubricants, allowing the formation of a protective layer that increases the resistance of these surfaces to abrasive wear \[[@B27-materials-12-03635]\]. The surface texture formed during processing affects the energy state and the adhesive properties of the surface \[[@B28-materials-12-03635],[@B29-materials-12-03635]\]. One positive effect of shot peening, apart from the fact that it improves the properties of the surface layer, is the rounding of the workpiece edges \[[@B30-materials-12-03635]\]. Centrifugal shot peening can also be combined in a single operation with grinding while using hybrid tools that enable the processing of difficult-to-cut materials, such as nickel and titanium alloys \[[@B31-materials-12-03635]\]. During shot peening, the concentration of crystal structure defects changes, as confirmed in tests that were carried out by annihilation techniques \[[@B32-materials-12-03635]\], as a result of which compressive stresses form in the surface layer of the workpiece \[[@B33-materials-12-03635]\]. Changes in the state of residual stress and the increase in the microhardness of the surface layer enhance the resistance of the material to abrasive and fatigue wear \[[@B34-materials-12-03635],[@B35-materials-12-03635]\]. A review of the literature shows that the laser cut parts require finishing steps. The beneficial effect of shot peening on the condition of the surface of machined and ground parts is well-known. The aim of the present study was to assess the impact of centrifugal shot peening on the surface layer of C45 steel parts that were cut with laser. 2. Materials and Methods {#sec2-materials-12-03635} ======================== The tests were performed while using specimens of non-alloy C45 steel (in accordance with EN ISO 683-2:2018). This steel grade is used in the machine industry for the manufacture of medium-load machine and equipment parts, such as spindles, shafts, axles, and unhardened gears. [Table 1](#materials-12-03635-t001){ref-type="table"} shows the chemical composition and strength properties of the steel tested (as per the technical specification sheet). Rectangular specimens, 5 mm × 8 mm × 100 mm in size, were cut in a LASER Amada 3000 W laser cutter from Amada America Inc. (Düsseldorf, Germany), while using oxygen as the assist gas and standard parameters: cutting speed v = 1150 mm/min., power 2.15 kW, frequency f~Hz~ = 1280 Hz, assist gas pressure 0.06 MPa, and laser beam focus position +13 mm. Laser-cut specimens were finish-machined using centrifugal shot peening, which consisted in impacting the surface of a workpiece with shot propelled from openings radially arranged in a rotating peening head. After the head is set into rotational motion, the shot particles hit the surface of the workpiece under the influence of centrifugal force. Upon brief contact with the shot, the surface of the workpiece undergoes plastic deformation. Peening reduces the surface roughness and hardens the surface layer of the processed material. After centrifugal shot peening, the resulting machining marks on the surface are organised in the longitudinal and transverse direction, which allows for obtaining a surface with less roughness. It can be used to process axially symmetrical parts, flat surfaces, and surfaces with complex shapes. Moreover centrifugal shot peening allows for the treatment of elements with low rigidity. Centrifugal shot peening tests were performed on an FV-580a (MOC Mechanicy Pruszków, Pruszków, Poland) vertical machining center while using a special peening head with an outer diameter of 70 mm with a blasting unit containing twelve (z~k~ =12) symmetrically arranged shot balls with a diameter d~k~ = 6.3 mm ([Figure 1](#materials-12-03635-f001){ref-type="fig"}). During shot peening, specimen (1) was clamped in a holder (2) so that the peened surface was parallel to the spindle axis. The holder was secured in the jaws of a vice (3) that was mounted on the table of the machine tool, which moved at speed v~f~ during peening. The body of special head has radial holes, in which the balls move. In the guiding part the balls can move freely along the parting axis. On the body of the head a ring with radially arranged holes in which the balls are located is mounted. On the circumference of the ring the diameter of the holes is smaller than the diameter of the balls, which prevents the balls from falling out. The peening head (4) rotated with speed n and simultaneously moved in the cross-feed direction f~p~. When the head was set in rotational motion, shot particles were blasted from it under the influence of centrifugal force and they impacted the surface of the workpiece. Upon impact, the particles bounced off the peened surface and were retracted into the seats in the head at distance g, referred to as "head infeed to the workpiece". Centrifugal shot peening was performed while using Mobile Cut cooling lubricant (Mobile, Germany) at a constant infeed g = 0.5 mm and a constant cross-feed f~p~ = 0.08 mm; the number of passes was i = 1. The parameters that were varied in the experiments included \- tangential speed of shot peening head: v~g~ = 528--1143 m/min. \- feed rate: v~f~ = 1368--10488 mm/min. The measurements of surface roughness and three-dimensional (3D) topography were performed while using a Hommel-Etamic T8000RC 120--140 device (Jenoptik, Villingen-Schwenningen, Germany). According to EN ISO 9013:2017, measurements of surface roughness should be made at a distance equal to 1/3 of the thickness of the workpiece, from the upper cut edge, when the thickness of the workpiece is above 2 mm. Following the guidelines of the EN ISO 9013:2017 standard, roughness was measured at a distance of 1/3 of workpiece thickness from the upper cutting edge, in the so-called "laser entrance zone" and at a distance of 1/3 of workpiece thickness from the bottom cut edge, in the so-called "laser exit zone". We decided to determine the roughness parameters in this lower area because the laser-cut parts had two characteristic zones with different roughness. [Figure 2](#materials-12-03635-f002){ref-type="fig"} schematically shows the place of roughness measurements. The same measurement scheme was used to measure the roughness of shot-peened specimens. Following the guidelines for roughness measurements outlined in EN ISO 4288:1997, the sampling length was set at lr = 2.5 mm. The material ratio Rmr was c = 2 µm below the highest peak of the roughness profile. The structure of the material was assessed while using a Nikon Eclipse MA 100 metallographic microscope (Nikon, London, UK). Microhardness measurements were made by the Vickers method in accordance with EN ISO 6507-1:2018 using transverse metallographic specimen surfaces that were prepared in a standard way. A LM Leco 700 at microhardness tester (Leco, San Jose, MI, USA) was used at an indenter load of 50 g (0.05 HV). Microhardness measurements were carried out according to the diagram shown in [Figure 3](#materials-12-03635-f003){ref-type="fig"}. A constant step from the surface after laser cutting or laser cutting and centrifugal shot peening was determined during the measurement. Measurements at a given depth were repeated 10 times. The maximum microhardness, which is just below the machined surface, was determined based on the micro hardness distribution. The results of the microhardness measurements served as a basis for calculating the degree of strain hardening of the surface layer e from the Equation (1):$$e = 100 \times \frac{{HV}_{\max} - {HV}_{0}}{{HV}_{0}},~\left( \% \right)$$ where: ${HV}_{\max}$---maximum microhardness of the surface layer after centrifugal shot peening,${HV}_{0}$---microhardness before shot peening. X-ray diffraction tests were performed with a Empyrean diffractometer (Panalytical, Malvern, UK). The experiments were carried out in the symmetrical Bragg-Brentano geometry, in which a divergent beam is used at room temperature and atmospheric pressure. Rectangular 5 × 8 × 15 mm^3^ specimens, which were cleaned in acetone prior to the experiments, were tested. A 3 kW copper anode lamp was used as the x-ray source. Anode radiation was filtered through nickel, which absorbs K~β~ radiation. Data were recorded in the angle range 2θ = 5--110° while using the step scan method. The scan step was 0.05° and the point exposure time was 3--5 s. Residual stresses in the surface layer of laser-cut-and laser-cut and shot-peened specimens were tested while using the mechanical method. They were calculated on the basis of measurements of specimen deformations that occurred when the sequential layers of the material in which these stresses resided were removed. 3. Results {#sec3-materials-12-03635} ========== 3.1. Surface Roughness {#sec3dot1-materials-12-03635} ---------------------- [Figure 4](#materials-12-03635-f004){ref-type="fig"}a shows the surface topography of a laser-cut specimen. The surface formed by cutting is characterized by large differences in height (there are numerous peaks and valleys). In the area where the laser beam entered the material (A), the surface is covered with fine, evenly spaced straight drag lines. The surface roughness parameters for this area are as follows: Ra = 2.49 ± 0.12 µm and Rz = 15.76 ± 1.33 µm. In the laser beam exit zone (B), the drag lines are curvilinear; they deviate from standard slope (the intended path of the beam). This indicates that the material "slips away" from the kerf. The roughness parameters in the exit zone are Ra = 5.04 ± 0.35 µm and Rz = 31.29 ± 2.27 µm. The values of the surface roughness parameters in the exit zone are about twice as high as those in the entrance zone. The same relationship was observed for the parameters of the Abbott--Firestone curve, which were Rpk = 2.43 ± 0.3µm, Rk = 8.14 ± 0.38 µm, and Rvk = 3.72 ± 0.93 µm in the entrance zone and Rpk = 4.98 ± 0.58 µm, Rk = 16.54 ± 1.17 µm, and Rvk = 7.31 ± 0.56 µm in the exit zone. An increase in the viscosity of the workpiece at the bottom cut edge causes the large differences in the quality of the surface. This enhances the thickness of the liquid metal layer, which, as a result, sticks to the surface being cut. This promotes the formation of large irregularities. During centrifugal shot peening, the shot balls impact the surface of the laser-cut workpiece, leading to the deformation of its geometric structure ([Figure 4](#materials-12-03635-f004){ref-type="fig"}b). As a consequence, the height of the drag lines is reduced and micro asperity summits are flattened. The difference between the height of the surface roughness profiles of the beam entrance zone (A) and the beam exit zone (B) is also reduced. [Figure 5](#materials-12-03635-f005){ref-type="fig"} shows the effect of tangential speed of the tool v~g~ and feed rate v~f~ on the roughness parameter Ra. As the rotational speed n, and thus tangential speed v~g~ increase, surface roughness decreases. This is due to the more intense smoothing of micro asperities on laser-cut surfaces as a result of increasing impact energy. The maximum decrease in Ra relative to the value recorded after laser cutting was four-fold. The changes in surface roughness that were obtained in the present study are more prominent when compared to those reported for vibratory-and-rotational shot peening of C45 steel specimens, for which a 30% reduction in surface roughness parameters \[[@B35-materials-12-03635]\] was observed, and for centrifugal shot peening of 41Cr4 steel specimens, where a three-fold reduction in Ra was obtained \[[@B26-materials-12-03635]\]. On the other hand, an increase in feed rate results in a decrease in the number of impacts per unit of exposure area, which leads to an increase in surface roughness. The nature of the changes in the surface roughness parameter Rz as a function of the analyzed technological parameters of centrifugal shot peening is similar to that observed for Ra. An analysis of the changes in the parameter Rz of the roughness profile as a function of feed rate showed that the maximum reduction in Rz after centrifugal shot peening was about three-fold for the beam entrance zone and about four-fold for the exit zone ([Figure 6](#materials-12-03635-f006){ref-type="fig"}). Cooperation between two surfaces is described by the parameters that are associated with the Abbott--Firestone curve (bearing curve). The bearing curve for laser-cut parts is a degressive-progressive curve ([Figure 7](#materials-12-03635-f007){ref-type="fig"}a). It has a large angle of inclination. The surface described by this curve has "sharp" micro-asperity summits and a small bearing area. The Abbott--Firestone curve for parts processed by centrifugal shot peening ([Figure 7](#materials-12-03635-f007){ref-type="fig"}b) is only slightly inclined, which might point to a high abrasion resistance of the tested surfaces. It is a curve with a "thick center" and it can therefore be classified as a progressive curve. The effect of tangential speed v~g~ on the parameters of the bearing curve was similar to that observed for Ra and Rz ([Figure 8](#materials-12-03635-f008){ref-type="fig"}). An increase in speed v~g~ caused more intense smoothing of micro asperities on the laser cut surface, which translated into a decrease in the values of Rpk, Rk, and Rvk parameters. Core roughness depth Rk, depending on the tangential speed of the peening head, decreased by two to seven times after shot peening. This means that centrifugal shot peening improves the so-called bearing capacity of the surface, or, to put it differently, that, after a grinding-in period, a substantial part of the surface will be in contact with the surface of the mating element. The Rpk parameter, which characterizes the behavior of a surface during grinding-in, decreased by 3 to 8.1 times relative to the value after laser cutting. The depth of the valleys (parameter Rvk) was reduced by centrifugal shot peening up to two times, depending on the tangential speed. This means that surfaces processed in this way may have lower oil retention capacity. The increase in the feed rate v~f~ ([Figure 9](#materials-12-03635-f009){ref-type="fig"}) translated into a smaller proportion of elastic-plastic strains, which caused an increase in the parameters of the bearing curve. Feed rate v~f~ had less effect on the Abbott--Firestone curve parameters than tangential speed v~g~. At feed rate v~f~ = 1368 mm/min "(least)", Rpk dropped up to six-fold when compared to the value that was obtained for laser-cut specimens. On the other hand, when a tangential speed v~g~ = 1143 m/min "(largest)" was used, an over eight-fold decrease in Rpk was observed. In the case of the roughness parameter Rk, the influence of tangential speed v~g~ was also more pronounced than the effect of feed rate v~f~. An up to five-fold reduction in Rk was obtained when compared to the value that was recorded for the laser cut specimens. After shot peening, parameter Rvk was reduced more than two-fold, a result that is similar to that obtained for tangential speed v~g~. The material ratio was increased for all of the investigated peening parameters when compared to the laser-cut surface ([Figure 10](#materials-12-03635-f010){ref-type="fig"}). The material ratio of the roughness profile of the laser-cut parts was Rmr~(2)~ = 6.08% ± 0.57% for the beam entrance zone and Rmr~(2)~ = 5.42% ± 0.42% for the beam exit zone. This means that the laser-cut surface had low abrasion resistance. Centrifugal shot peening increased the material ratio by 3.33 to over 14 times for the beam entrance zone and up to almost 12 times for the beam exit zone. This suggests that the process also increased the abrasion resistance of the surface. During centrifugal shot peening, there is a wide angle between the normal velocity vector and the peened surface. This promotes strong friction between the peened surface and the shot, which causes intense abrasion of the micro-asperity summits that previously formed on the hard material, decreasing the height of micro asperities and leading to the formation of micro-cavities on the peened surface. The favorable changes in the stereometric properties of the peened surface are likely to make the surface resistant to abrasive wear \[[@B35-materials-12-03635]\]. The "smoothing" of the surface as a result of deformation leads to an evening out of zones with varying surface roughness, which is likely to have a positive effect on the use of a given part in a next production stage (e.g. application of paint coatings). 3.2. Microstructure and Microhardness {#sec3dot2-materials-12-03635} ------------------------------------- Before the laser cutting process, the C45 steel specimens had a ferritic--pearlitic structure. As a result of strong heating with the laser beam followed by cooling down, the material around the cut edge became hardened. A heat affected zone was created ([Figure 11](#materials-12-03635-f011){ref-type="fig"}a), which differed in the structure and properties from the parent material. In the heat affected zone, martensite needles ([Figure 11](#materials-12-03635-f011){ref-type="fig"}b) are visible next to the edge of the cut, with ferrite, and then martensite located further away from the edge. The laser cutting process lasted too short for ferrite grains to become enriched with carbon, which resulted in the formation of low-carbon martensite. Perlite, on the other hand, transformed into high carbon martensite. The cyclic bombarding of the material with shot particles modifies its structure ([Figure 11](#materials-12-03635-f011){ref-type="fig"}c). The deformation of the material caused by centrifugal shot peening leads to an increase in dislocation density. The gliding dislocations are stopped when they encounter obstacles, such as other dislocations, grain boundaries, or cementite separation. The increase in the dislocation density has a hardening effect on the material. The resulting structure can be classified as fine-needle martensite. In laser-cut specimens, the zone of changes in microhardness is over 0.3 mm wide ([Figure 12](#materials-12-03635-f012){ref-type="fig"}). The microhardness of the surface layer increased more than twice in the area of the cut edge (approx. 560 HV0.05), and then slowly decreased to reach the core microhardness value (approx. 200 HV0.05). The literature only offers reports on the use of centrifugal shot peening for processing parts with hardnesses of 240 HV and 360 HV \[[@B26-materials-12-03635]\]. However, the present study shows that the laser-cut parts can also be treated by centrifugal shot peening. Though the hardness and thickness of the hardened layer only increase slightly during centrifugal shot peening ([Figure 12](#materials-12-03635-f012){ref-type="fig"}), the increase is likely to translate into increased resistance to abrasive wear \[[@B35-materials-12-03635]\]. The degree of strain hardening of the surface layer for laser-cut-and shot-peened C45 steel depend on the technological parameters of shot peening ([Figure 13](#materials-12-03635-f013){ref-type="fig"}). An increase in the tangential speed v~g~ leads to an increase in impact energy and it elevates the number of impacts per unit of area, which translates into a greater proportion of plastic-elastic deformations in the workpiece and thus increases the degree of strain hardening of the surface layer. The thickness of the hardened layer g~h~, in relation to the core, was max. 60 µm, as verified by a statistical test of means. The largest changes in microhardness occurred right next to the treated surface, where the most deformed grains were found ([Figure 11](#materials-12-03635-f011){ref-type="fig"}c). The degree of strain hardening increased as a function of tangential speed v~g~ from about 6% to 15% ([Figure 13](#materials-12-03635-f013){ref-type="fig"}a), with the dynamics of increase the degree of strain hardening declining in the velocity range v~g~ = 989--1143 m/min. A rise in feed rate increases the spacing between the indentations that are imparted by shot, which results in uneven deformation of the treated surface. The changes in the thickness of the hardened layer after centrifugal shot peening (for a variable feed rate v~f~) ranged from 15 µm to 56 µm. At low feed rates, the number of impacts per unit area was higher, which increased the thickness of the hardened layer. The increase in microhardness for v~f~ = 1368 mm/min relative to the value that was recorded after laser cutting was 16.4% ([Figure 13](#materials-12-03635-f013){ref-type="fig"}b). 3.3. XRD Tests {#sec3dot3-materials-12-03635} -------------- The presence of oxygen as the assist gas during cutting promotes an exothermic reaction, which leads to the formation of combustion products on the cut surface. This is confirmed by the qualitative change in the phase composition of the surface layer after laser cutting ([Figure 14](#materials-12-03635-f014){ref-type="fig"}). The XRD tests that we carried out allowed for us to identify the following phases on the surface of C45 steel: Fe~3~O~4~, Fe~2~O~3~, Fe~0.9~O ([Figure 14](#materials-12-03635-f014){ref-type="fig"}). [Table 2](#materials-12-03635-t002){ref-type="table"} shows the lattice parameters and angles for the compounds formed on the surface of metal as a result of laser cutting. XRD tests of a laser-cut-and shot-peened specimen demonstrated the presence of two compounds on the treated surface. The diffractogram ([Figure 15](#materials-12-03635-f015){ref-type="fig"}) shows two main dominant peaks, which correspond to ferrite and martensite in the diffraction database ([Table 3](#materials-12-03635-t003){ref-type="table"}). Shot balls, when impacting the surface of the workpiece, "blast off" and "shear off" oxide phases, removing this way the thin oxide film that had formed on the surface of the workpiece. The removal of combustion products from the treated surface reduces or eliminates the peeling of coatings applied to it. 3.4. Residual Stress {#sec3dot4-materials-12-03635} -------------------- Laser cutting is accompanied by the re-solidification of molten material, which takes place in unstable conditions. This promotes the occurrence of tensile residual stresses that reside in the specimens at a depth of about 0.05 mm from the surface of the workpiece ([Figure 16](#materials-12-03635-f016){ref-type="fig"}). Tensile stresses also originate from the structural changes that take place during the cutting process. In the process of centrifugal shot peening, compressive residual stresses are produced in the surface layer ([Figure 16](#materials-12-03635-f016){ref-type="fig"}), which improve the functional properties of the material, e.g. increase its resistance to fatigue wear \[[@B34-materials-12-03635]\]. Grinding, which has been used so far to finish laser cut parts, leads to the formation of tensile residual stresses in the surface layer \[[@B22-materials-12-03635]\]. An increase in the tangential speed of the tool increased the depth of compressive residual stresses ([Figure 16](#materials-12-03635-f016){ref-type="fig"}a), and also, though to a lesser extent, contributed to an increase in the value of these stresses. The absolute maximum value of compressive residual stress was found at a depth of 0.05--0.1 mm. On the other hand, as the feed rate increased, the absolute maximum value of compressive stresses produced in the peened part decreased. The depth of compressive residual stresses did not change significantly ([Figure 16](#materials-12-03635-f016){ref-type="fig"}b). The lowest residual stress was obtained at v~f~ = 10 488 mm/min and v~g~ = 835 m/min within the range of the technological parameters of the experiment. 4. Conclusions {#sec4-materials-12-03635} ============== The following conclusions can be drawn based on the centrifugal shot peening tests of the laser-cut C45 steel elements. 1. The use of centrifugal shot peening for finish machining of laser-cut C45 steel parts allowed for obtaining a four-fold reduction in the surface roughness parameters Ra and Rz. Centrifugal shot peening diminished the differences in Ra and Rz between the beam entrance zone and the beam exit zone. 2. An increase in the tangential speed of the tool v~g~ resulted in a decrease in roughness parameters, while an increase in feed rate v~f~ had the opposite effect, with the changes being more pronounced for the variable v~g~. 3. Centrifugal shot peening allowed for changing the nature of the bearing curve from a degressive--progressive curve to a degressive curve. A significant decline in Rpk, Rk, and Rvk parameters was observed as compared to the values that were obtained after laser cutting. 4. Centrifugal shot peening resulted in an up to 14-fold increase in the material ratio of the roughness profile as compared to the value obtained after laser cutting. 5. Peened workpieces had an up to 16% higher microhardness (v~g~ = 835 m/min, v~f~ = 1365 mm/min) and a 15 μm to 58 μm deep hardened layer. 6. As the surface of a workpiece was impacted by shot during centrifugal shot peening, oxide phases, which are combustion products, were blasted off and sheared off the surface, which caused the removal of Fe~3~O~4~, Fe~2~O~3~, and Fe~0.9~ oxides. 7. In the surface layer of the specimens, compressive residual stresses were formed during centrifugal shot peening, whose absolute maximum value varied from 450 MPa to 740 MPa. The stresses resided at a depth of 0.25--0.40 mm, depending on the technological parameters of the peening process. 8. As the tangential speed of the peening head v~g~ grew to 835 m/min, an increase was observed in the absolute value of compressive residual stresses and the depth of their accumulation; a further increase in tangential speed only increased the depth of the occurrence of compressive residual stresses. An increase in feed rate v~f~ caused a decrease in the absolute value of compressive residual stresses, but it did not significantly affect the depth of accumulation of residual stresses. 9. The results of the centrifugal shot peening experiments demonstrate that this cold working method can be successfully used with CNC machine tools. The CNC machine tools are equipped with control systems that can be used to guide the tool along a designated path, which allows for processing parts with complex shapes. Conceptualization, A.S. and K.Z.; Methodology, A.S.; formal analysis, K.Z.; investigation, A.S.; data curation, A.S.; writing---original draft preparation, A.S.; writing---review and editing, A.S. and K.Z.; visualization, A.S.; supervision, K.Z. The project/research was financed in the framework of the project Lublin University of Technology-Regional Excellence Initiative, funded by the Polish Ministry of Science and Higher Education (contract no. 030/RID/2018/19). The authors declare no conflict of interest. ![Scheme of centrifugal shot peening process on a vertical machining center (1---sample, 2---holder, 3---vice, 4---shot peening head): (a) shot peening kinematics; (b) principle of centrifugal shot peening (g---infeed, Rmax---maximum head radius).](materials-12-03635-g001){#materials-12-03635-f001} ![Surface view after laser cutting with marked roughness measurements point.](materials-12-03635-g002){#materials-12-03635-f002} ![Schema of microhardness measurements.](materials-12-03635-g003){#materials-12-03635-f003} ![Surface topography C45 steel after: (a) laser cutting; (b) laser cutting and centrifugal shot peening (v~g~ = 1143 m/min, v~f~ = 3648 mm/min).](materials-12-03635-g004){#materials-12-03635-f004} ![Effect of tangential speed v~g~ (v~f~ = 3648 mm/min) and feed rate v~f~ (v~g~ = 835 m/min) on the roughness parameter Ra specimens after laser cutting and centrifugal shot peening: (a) laser entrance zone; (b) laser exit zone.](materials-12-03635-g005){#materials-12-03635-f005} ![Effect of tangential speed v~g~ (v~f~ = 3648 mm/min) and feed rate v~f~ (v~g~ = 835 m/min) on the roughness parameter Rz specimens after laser cutting and centrifugal shot peening: (a) laser entrance zone; (b) laser exit zone.](materials-12-03635-g006){#materials-12-03635-f006} ![Curve of the material bearing for the "exit zone" after: (a) laser cutting; (b) laser cutting and centrifugal shot peening (v~g~ = 1143 m/min, v~f~ = 3648 mm/min).](materials-12-03635-g007){#materials-12-03635-f007} ![Effect of tangential speed v~g~ on the parameters Abbott-Firestone curve specimens after laser cutting and centrifugal shot peening (v~f~ = 3648 mm/min).](materials-12-03635-g008){#materials-12-03635-f008} ![Effect of feed rate v~f~ on the parameters Abbott-Firestone curve specimens after laser cutting and centrifugal shot peening (v~g~ = 835 m/min).](materials-12-03635-g009){#materials-12-03635-f009} ![Effect of tangential speed v~g~ (v~f~ = 3648 mm/min) and feed rate v~f~ (v~g~ = 835 m/min) on the material ration specimens after laser cutting and centrifugal shot peening: (a) laser entrance zone; (b) laser exit zone.](materials-12-03635-g010){#materials-12-03635-f010} ![Microstructure of C45 steel: (a) HAZ and transition zone; (b) HAZ after laser cutting; (c) HAZ after laser cutting and centrifugal shot peening (v~g~ = 1143 m/min, v~f~ = 3648 mm/min).](materials-12-03635-g011){#materials-12-03635-f011} ![Microhardness distribution of C45 steel element surface layer after laser cutting and laser cutting and centrifugal shot peening (v~g~ = 1143 m/min, v~f~ = 3648 mm/min).](materials-12-03635-g012){#materials-12-03635-f012} ![Degree of strain hardening e and hardening layer thickness g~h~ specimens after laser cutting and centrifugal shot peening as a function of: (a) tangential speed v~g~ (v~f~ = 3648 mm/min); (b) feed rate v~f~ (v~g~ = 835 m/min).](materials-12-03635-g013){#materials-12-03635-f013} ![Diffraction pattern with matching theoretical reflection obtained after analyzing the surface of C45 steel after laser cutting.](materials-12-03635-g014){#materials-12-03635-f014} ![Diffraction pattern with the matching theoretical reflection obtained after analyzing surface C45 steel after laser cutting and centrifugal shot peening (v~g~ = 835 m/min, v~f~ = 3648 mm/min).](materials-12-03635-g015){#materials-12-03635-f015} ![Distribution of residual stress in the function distance from sample surface after laser cutting and centrifugal shot peening at the varible: (a) tangential speed v~g~ (v~f~ = 5928 mm/min); (b) feed rate v~f~ (v~g~ = 835 m/min).](materials-12-03635-g016){#materials-12-03635-f016} materials-12-03635-t001_Table 1 ###### Chemical composition and selected properties of C45 steel. Chemical Composition, (%) --------------------------- ---------------- ------ ------- ------ ------ ------ ------- ------ 0.48 0.74 0.36 0.011 0.01 0.09 0.02 0.002 rest Yield point R~e~ = 430 MPa Tensile strength R~m~ = 740 MPa Hardness 250 HB materials-12-03635-t002_Table 2 ###### Data of identified compounds based on the diffraction pattern obtained after analysis surface of C45 after laser cutting. ------------------------------------------------------------------------------------ Number in the Base\ Chemical Formula Lattice Parameters (nm) Angles (°) ICDD PDF--4+ --------------------- ------------------ ------------------------- ----------------- 04-009-8436 Fe~3~O~4~ a = b = c = 0.8403 α = β = γ = 90° 04-006-9058 Fe~2~O~3~ a = b = 0.5350\ α = β = 90°\ c = 1.3720 γ = 120° 04-001-9267 Fe~0.9~O a = b = c = 0.4298 α = β = γ = 90° ------------------------------------------------------------------------------------ materials-12-03635-t003_Table 3 ###### Data identified phases based on the diffraction patter obtained after analysis surface of C45 after laser cutting and centrifugal shot peening. ------------------------------------------------------------------------------------ Number in the Base\ Chemical Formula Lattice Parameters (nm) Angles (°) ICDD PDF--4+ --------------------- ------------------ ------------------------- ----------------- 04-002-1061 Fe--α a = b = c = 0.2862 α = β = γ = 90° 00-044-1289 C~0.14~Fe~1.86~ a = b = 0.2846\ α = β = γ = 90° c = 0.3053 ------------------------------------------------------------------------------------	{ "pile_set_name": "PubMed Central" }
Immigration Minister Sylvi Listhaug said the migrant crisis is going to have violent consequences We will use your email address only for sending you newsletters. Please see our Privacy Notice for details of your data protection rights. Sign up fornow and never miss the top politics stories again. The centre-right Government is proposing tighter controls in order to avoid what the immigration minister described as "violent consequences" to the country's welfare system. The measures, which would need parliamentary approval, would make it difficult for refugees to bring family members to Norway and raise the requirements for permanent resident permits. The move comes after more than 30,000 asylum-seekers sought shelter in the country, which has a population of 5.1million people, this year. Immigration Minister Sylvi Listhaug, of the right-wing Progress Party, said the country’s influx of migrants was one of the highest numbers in Europe in relation to its population size.	{ "pile_set_name": "OpenWebText2" }
The only way I know how to add downloaded source code files is by loading them up in text editor somehow, and then copying and pasting into Palm desktop. This is WAY to difficult and too much hassle. Can't learn this way. There are too many files I need to sync, and takes too much time. Syncs are too frequent and run down my battery. Got a solution? Well there is no limit to how big they can be so you dont have to worry about breaking them up, futhermore you don't have to copy and paste into Palm Desktop, just select the file name and then choose auto-install to the user of your choice. OR you could stop complaining and being rude and just either take it like a man or use PDE. <blockquote id="quote"><font size="1" face="Verdana, Arial, Helvetica" id="quote">quote:<hr height="1" noshade id="quote"> Well there is no limit to how big they can be <hr height="1" noshade id="quote"></blockquote id="quote"></font id="quote"> Actually, there is a limit. According to the PocketC documentation: The uncompressed size of a doc file must be less than 64K. Hey, I'm just a happy lucky Palm guy. It seems to me like you may be answering a different question Nicholas. I've never used MakeDoc so I don't have experience with it. There's not enough messsage for me to reply. PDE does not solve the problem I have, which I would happily discuss with any folks who are interested. If you are using the desktop version of PocketC and have POSE running in the background, all the programs you compile get automatically uploaded to POSE. This makes desktop development with PocketC quite easy. Another approach is to use MakeDoc to convert your source textfiles into palm doc files. You can then drag and drop the resulting .prc file on to the emulator and do a compile. Since palm doc files can be quite large, this saves you the trouble of having to split the file up into smaller (4k) memo chunks. Alternatively, you can upload the doc file to POSE with a custom perl script. POSE has a scripting interface which allows you automate a number emulator tasks (including file installs). See the POSE documentation for more details. Below is a perl script I use to upload the PToolboxLib to POSE every time I recompile it. Also, after a hotsync you can retrieve your doc files by looking in your "Program Files/Palm/<user_name>/Backup" directory for them. All being said, a better solution to this problem would be for Jeremy to update the desktop compiler to be capable of reading and writing palm doc files directly. If you like this idea, I'd suggest putting in a request to Jeremy for it. The more responses he gets the greater the chances it will see the light of day. <blockquote id="quote"><font size="1" face="Verdana, Arial, Helvetica" id="quote">quote:<hr height="1" noshade id="quote"><i>Originally posted by jstadolnik</i> <hr height="1" noshade id="quote"></blockquote id="quote"></font id="quote"> >Another approach is to use MakeDoc to convert your source textfiles into palm doc files. You can then drag and drop the resulting .prc file on to the emulator and do a compile. Since palm doc files can be quite large, this saves you the trouble of having to split the file up into smaller (4k) memo chunks. So then this lets you, say if you wrote 7K of source, you don't have to break it in two? Now, if you have a file that #include's several other files or other libraries and a .h file, it is just a fair lot of steps in your way, as I mentioned. I've never used MakeDoc. If I ever tried it, probably it might have seemed to be one of those complicated contraptions from soundforge or whatever that make these Open Source hard to understand projects. I guess because I could already get files to read on the Palm, like with iSilo. If it takes several files and compresses them into one to pull apart on the Palm, that would be apropos. Just like, I never read PocketC will read anything other than some text files starting with // in Memopad. > Alternatively, you can upload the doc file to POSE with a custom perl script. POSE has a scripting interface which allows you automate a number emulator tasks (including file installs). See the POSE documentation for more details. Below is a perl script I use to upload the PToolboxLib to POSE every time I recompile it. I haven't learned Perl yet. Books on it seemed over my head. But Ok, so if I maybe take that code, cut it and save in Notepad, add an extension that Perl will recognize, then it will upload whatever file/path is in quotes - if I run the file? <blockquote id="quote"><font size="1" face="Verdana, Arial, Helvetica" id="quote">quote:<hr height="1" noshade id="quote"> So then this lets you, say if you wrote 7K of source, you don't have to break it in two? <hr height="1" noshade id="quote"></blockquote id="quote"></font id="quote"> Yes. <blockquote id="quote"><font size="1" face="Verdana, Arial, Helvetica" id="quote">quote:<hr height="1" noshade id="quote"> I've never used MakeDoc. If I ever tried it, probably it might have seemed to be one of those complicated contraptions from soundforge or whatever that make these Open Source hard to understand projects. I guess because I could already get files to read on the Palm, like with iSilo. <hr height="1" noshade id="quote"></blockquote id="quote"></font id="quote">	{ "pile_set_name": "Pile-CC" }
(Bloomberg) -- Hewlett Packard Enterprise Co. is losing business from Microsoft Corp., one of the world’s largest users of servers, the latest sign of trouble for the pioneering computer maker as it struggles with the rise of cloud services, people familiar with the matter said. Hewlett Packard Enterprise Chief Executive Officer Meg Whitman said last week her company saw "significantly lower demand" for servers from a tier-1 service provider, but without identifying the customer. Tier-1 service providers are typically major cloud and telecom companies. The softer demand came from Microsoft, the people said, as the software giant pushes for lower prices from hardware providers to help it efficiently expand its public cloud service and keep up with rivals Amazon.com Inc. and Alphabet Inc.’s Google. Spokeswomen for Microsoft and HPE declined to comment. Late last year, the Redmond, Washington-based company unveiled a new in-house cloud server design that it will require hardware vendors to follow. This forces HPE and rival Dell Technologies Inc. to compete against lower-cost generic, commodity manufacturers. Already, Microsoft has been using less-expensive gear for its data centers and the new design is set to be fully implemented later this year. "We will continue to meet customer demand by expanding data center capacity while driving efficiencies through new technologies," Microsoft Chief Financial Officer Amy Hood said on a call with analysts in January when it announced earnings. Microsoft’s Azure public cloud business reported a 93 percent revenue surge in the final quarter of 2016 as more businesses opted for the flexibility and ease of accessing computing power and storage over a network instead of building their own data centers. HPE has been a leading seller of servers that go in these corporate data centers. But the shift to the public cloud means businesses don’t need to buy their own servers anymore. Selling to the big cloud providers is harder, either because they demand more volume discounts, or increasingly they design their own cheaper servers. "Within Tier 1 we had a much lower demand from a single large customer," Whitman said on the call, noting that these types of deals aren’t as profitable as other parts of the server business. "The Tier 1 business is very competitive, and we’ll see what happens there." HPE last week cut its adjusted profit forecast for the current fiscal year and reported sales that missed analysts’ projections for the third consecutive quarter.	{ "pile_set_name": "OpenWebText2" }
Dennis Maddux Cleveland City Manager Janice Casteel The City of Cleveland has replaced its police chief after his first official day on the job. Cleveland City Manager Janice Casteel said in a release this afternoon that she will serve as head of the police department effective immediately. It was not immediately clear what will become of former chief Dennis Maddux. Sunday was Maddux's first official day on the job, though he's been acting chief since January. On Saturday, Maddux was found in a car with the wife of one of his officers before that officer was arrested, according to an affidavit. Casteel is appointing Capt. Mark Gibson to assist her with day-to-day operations at the Cleveland Police Department, she said in the release. Casteel will act as Cleveland's fourth police chief in just more than a year. She said she intends to ask the city council to form a committee to review the organizational structure and policies of the police department. Maddux replaced David Bishop, who took over in December 2013 but went on medical leave in October. Gibson acted as interim chief until Maddux was named to the position. The chief shuffle began with the abrupt retirement of longtime chief Wes Snyder at the end of 2013. Snyder retired after video footage surfaced showing him trysting with a woman not his wife at a warehouse storage unit in Cleveland. This is a developing story. Stay with the Times Free Press for more details.	{ "pile_set_name": "OpenWebText2" }
Land Transportation Office (Philippines) The Land Transportation Office ( or LTO) is an agency of the Philippine government under the Department of Transportation and is responsible for all land transportation in the Philippines. Functions of the Land Transportation Office include the inspection and registration of motor vehicles, issuance of license and permits, enforcement of land transportation rules and regulations, adjudication of traffic cases, and the collection of revenues for the government of the Philippines. Its primary mission is to rationalize the land transportation services and facilities and to effectively implement the various transportation laws, rules, and regulations. It believes that it is the responsibility of those involved in the public service to be more vigilant in their part in the over-all development scheme of national leadership. Hence, the promotion of safety and comfort in land travel is one of LTO's continuing commitments. It aims to be a frontline government agency that showcases fast and efficient public service for a progressive land transport sector. History There were several predecessors before the formation of the present Land Transportation Office. Automobile Section In order to regulate and license of operators for motor vehicles in the Philippines, Act No. 2159 was enacted in 1912 under the American colonial Insular Government. This was the first formal law on land transportation in the country. It created the Automobile Section under the Administrative Division of the Bureau of Public Works. In 1926, Act No. 3045 compiled and incorporated all laws governing motor vehicles. The Automobile Section was upgraded to the Automobile Division but still under the supervision of the Bureau of Public Works. Act No. 3992 (Revised Motor Vehicle Law) was enacted in 1933, amending Act No. 3045. The Automobile Division was renamed Division of Motor Vehicles. Motor Vehicles Office In 1945, the Department of Public Works and Highways issued Department Order No. 4 for the reorganization of the Division. It took effect after the Philippines were liberated from the Japanese during World War II. Executive Order No. 94 was promulgated in 1947 reorganizing the different executive departments, bureaus, and offices. Under Section 82 of E.O. 94, the Division of Motor Vehicles was upgraded into the Motor Vehicles Office (MVO) with the category of the Bureau. However, the Motor Vehicle Office was abolished in 1964 by Republic Act No. 4136 or the Land Transportation and Traffic Code. The Land Transportation and Traffic Code was an act that compiled all the laws relative to transportation and traffic rules, to create a land transportation commission and for other purposes. This act was eventually replaced by the Land Transportation Commission. The Land Transportation Commission was tasked with the registration and operation of motor vehicles and the licensing of conductors and drivers. In order for the commission to effectively carry out its duty, regional offices were established in various parts of the country. Additionally, the powers, functions, and duties previously conferred on the Chief of the Motor Vehicles are now performed by the Land Transportation Commissioner. Land Transportation Commission During the Marcos dictatorship, Executive Order No. 546 was promulgated in 1979, creating the Ministry of Transportation and Communications (MOTC). The Land Transportation Commission was renamed into Bureau of Land Transportation and was absorbed into that ministry. The creation of the Board of Transportation and the Bureau of Land Transportation was nullified in 1985 by Executive Order 1011. The E.O. established the Land Transportation Commission, which was tasked to perform functions such as registering motor vehicles, licensing of drivers and conductors, franchising of public utility vehicles and enforcing land transportation rules and regulations. Establishment of the Land Transportation Office The Land Transportation Commission was abolished in 1987, and two offices were created, namely the Land Transportation Office (LTO) and the Land Transportation Franchising and Regulatory Board (LTFRB). The LTO took over the functions of the former BLT while the LTFRB took over the functions of the BOT. The MOTC was likewise renamed as the Department of Transportation and Communications (DOTC). Functions and Mandate License and Permit Issuance The LTO is in charge of the issuance, renewal, and regulation of driver's licenses. It can issue licenses to both citizens and foreigners provided that they meet the requirements for those licenses. The LTO provides the non-professional driver's license, which allows holders to operate vehicles under the restriction codes 1,2, and 4. It also provides the professional driver's license, which allows the bearer to operate vehicles under all 8 restriction codes. The LTO also issues student permits (SP), which are a primary requirement for both the non-professional and professional driver's licenses. All applicants are also required to pass both a written and practical examination to be granted their license. The application process for any license or permit may be done at any LTO Licensing Center and District/Extension Office. Motor Vehicle Registration The LTO is responsible for the registration of motor vehicles and renewals. It can register brand new vehicles, including locally manufactured vehicles, imported vehicles, light electric vehicles, low speed vehicles, three wheeled vehicles, and tax exempt vehicles. Each category has an obligatory set of requirements and procedures to follow to be fully registered. Renewals are also obligated to submit the necessary requirements and to follow procedures to be fully renewed. The LTO also oversees miscellaneous transactions, specifically transactions requiring change of certificate of registration, and transactions that do not require the change of certificate of registration. Law Enforcement and Adjudication of Cases One of the main functions of the LTO is to strictly implement and enforce the laws regarding land transportation. Necessary requirements and procedures are to be followed in settlements of admitted cases and contested cases of motor vehicles, plates, and driver's licenses. The LTO also follows a strict procedure in settlements of impounded violations. It also allows private and for hire motor vehicles to apply for duplicate plates. Furthermore, the LTO is in charge of ensuring that public land transportation services abide by the fares set by the LTFRB. It has sanctions for the overcharging and undercharging of fares, and for non-issuance of fare tickets. Manufacturers, Assemblers, Importers, and Dealers Reporting (MAIDR) The LTO grants accreditation to certain manufacturers, assemblers, importers, and dealers who wish to transact business with the LTO. A firm, person, or corporation must file an application for accreditation with Assistant Secretary of the Land Transportation Office containing certain requirements and qualifications in order to transact business with the LTO relative to Motor Vehicles or its components. It also issues the Certificate of Stock Reported (CSR) which certifies that a motor vehicle or its component has already been reported by its manufacturer, assembler, or importer to MAIDRS. The LTO also requires sales reports that inform the LTO that the reported stock is already sold to the end-user. Sales reports include regular sales transactions and stock transfers. It also reports the issuance of a single CSR, of Motor Vehicles (MVs) formed out of combining components (new, used, or previously registered) MV/MC that are undocumented in a procedure called special reporting. The LTO also maintains the processes that facilitate requests for the approval of the MAID office with LTO related transactions. Such transactions include the issuance of a conduction sticker, a public bidding of a motor vehicle, and the stamping of chassis identification number. Organizational structure In 1987, under section 11 of Executive Order No. 125, the Regional Offices of the Land Transportation Commission were abolished and their functions were transferred to the Regional Offices for Land Transportation. The newly renamed Department of Transportation and Communication was placed under the authority of the Secretary of Transportation and Communication. As with the Land Transportation Commission before it, the Land Transportation Office is an office under the Department of Transportation and is headed by the Assistant Secretary for Land Transportation, who is appointed by the President upon the recommendation of the Secretary. The current LTO Board is listed below: Branches To service the entirety of the country, the LTO has established many branches in the different regions of the Philippines: Region I - Aguila Road, Brgy. Sevilla, San Fernando City, La Union Region II - San Gabriel, Tuguegarao, Cagayan Region III - Government Center, Brgy. Maimpis, City of San Fernando, Pampanga Region IVA - J. C. Abadilla Memorial Bldg., Old City Hall Compound, B. Morada Ave., Lipa City Batangas Region IVB - MIMAROPA – LTO Compd., East Avenue, QC. Region V - Regional Govt. Center Site, Rawis, Legaspi City Region VI - Tabuc-Suba, Jaro, Iloilo City Region VII - Natalio Bacalso Avenue, Cebu City Region VIII – Old Army Road, Tacloban City Region IX - Veterans Ave., Zamboanga City/Balangasan St., Pagadian Region X - MVIS Compound, Zone 7, Bulua, Cagayan de Oro City Region XI - Quimpo Blvd., Davao City Region XII - ARMM Compound, Cotabato City/No. 79 G. Del Pilar St., Koronadal City, South Cotabato National Capital Region (NCR) – #20 G. Araneta Avenue, Brgy. Sto Domingo, Q.C. Cordillera Administrative Region (CAR) - Engineer’s Hill, Baguio City/2nd Flr., Post Office Loop, Session Road, Baguio City CARAGA - J. Rosales Avenue, Butuan City For more detailed information, please visit the LTO Directory. Projects The Land Transportation Office is in charge of various projects which aims to improve its functions and mandates. Land Transportation Office Infrastructure and Information System (LTO-IIS) Project In 2012, the Department of Transportation and Communication and the Land Transportation Office introduced a project entitled the Land Transportation Office Infrastructure and Information System (LTO-IIS) which aims to utilize information and communication technology to enhance the functional efficiency of the LTO in the delivery of its front line services to the public and the performance of its mandate. LTO-IIS involves building an IT network for the LTO which can host and support its Front and Back Office Applications in order to computerize and automate its processes and services, develop its database information system, and provide interconnection between and among LTO offices nationwide. In short, the LTO-IIS serves as an efficient means to process motor vehicle registrations, renewal and application of drivers’ licenses and permits, back-end transactions, apprehensions, and such ancillary transactions or processes. This new LTO-IT system aimed to make accessing relevant vehicle information easier for authorities especially for tracing stolen vehicles while addressing issues including the involvement of third-party providers with key data such as vehicle registration. The IT system overhaul was estimated to cost P8.2-billion. 5-year Validity of Drivers’ Licenses August 29, 2017 marked the beginning of the five-year validity of drivers' licenses cards issued by the LTO. Applications were accepted beginning October 2016 for driver's licenses as well as renewals with five-year validity, however, issues with the printers' contracts stalled the card printing for almost one year resulting in a backlog of 3.6 million driver's licenses which only began to be processed September 2017. Republic Act (RA) No. 10930 effectively amended Section 23 of Republic Act No. 4136, otherwise known as the "Land Transportation (LTO) and Traffic Code" which was signed on August 2, 2017 by President Rodrigo Duterte to extend the validity of the driver's license from three years to five years. According to the law, "Except for student permits, all drivers' licenses shall be valid for five years reckoned from the birthdate of the licensee, unless sooner revoked or suspended." This amendment also added that holders of nonprofessional and professional driver's licenses who do not commit violations in RA 4136 or any other traffic laws during the five-year period "shall be entitled to a renewal of such license of 10 years, subject to the restrictions as may be imposed by the LTO." Besides the amendments to the law, new improvements to the quality and security of the license cards were introduced. The old cards with three-year validity were made from PVC, thermally printed, and had limited security features. The new plastic license cards are laser engraved and made of polycarbonate material which are more durable. The new cards also includes several new security features. The Transportation Secretary, Arthur Tugade, and LTO chief Assistant Secretary, Edgar Galvante, led the official rollout. Controversies Fixers The Land Transportation Office and their various constituent branches and offices are notorious for their corrupt employees who engage in the malpractice of hiring and colluding with ‘fixers’ to illicitly garner more income. Fixers litter the various branches of the LTO, offering faster and/or easier transaction and procurement of official LTO paraphernalia at a more expensive price than the standard fee. These fixers and their employers pocket the extra payment in exchange for placing a client higher up in the queuing system, falsifying official government documents, fabricating driving test and written test results, and many other illegal practices. Section 11 of the Republic Act No. 9485 or the “Anti-Red Tape Act of 2007” lists fixing and/or collusion with fixers as a grave offense, with the penalty being dismissal and perpetual disqualification from public service. The fixers themselves can receive a penalty of imprisonment not exceeding six years or a fine not less than Twenty Thousand Pesos (P20,000.00) but not more than Two Hundred Thousand Pesos (P200,000.00) or both fine and imprisonment at the discretion of the court. References External links Land Transportation Office Philippines website Category:Department of Transportation (Philippines)	{ "pile_set_name": "Wikipedia (en)" }
Q: Can Pentax and Canon mounts on Tamron lenses be swapped? I have two Tamron lenses. Lens A has a mount ring for Canon, and Lens B has a mount ring for Pentax. Lens A is broken. Can I swap the mount rings and use lens B for my Canon camera? A: If your lenses are older Tamron Adaptall style lenses, then yes, you can remove the Canon FD ring and replace it with the Pentax K-mount ring and use the lens with pentax cameras.	{ "pile_set_name": "StackExchange" }
import bpy bpy.context.scene.render.fps = 30 bpy.context.scene.render.fps_base = 1	{ "pile_set_name": "Github" }
United States Court of Appeals FOR THE EIGHTH CIRCUIT ___________ No. 02-2663 ___________ Tyrone A. Davis, * * Plaintiff - Appellant, * * Appeal from the United v. * States District Court for * the District of Nebraska. * Dr. Stephen G. Lemar, Tracey M. * (UNPUBLISHED) Curry, and Otha Serrell, * * Defendants - Appellees. * ___________ Submitted: January 14, 2003 Filed: January 27, 2003 ___________ Before WOLLMAN and MURPHY, Circuit Judges, and GRITZNER,1 District Judge. ___________ PER CURIAM. Tyrone A. Davis suffered an injury to his knee when another inmate accidentally fell onto him in the exercise yard of the Nebraska State Penitentiary, and he underwent knee surgery some four months later. Davis brought a civil rights action under 42 U.S.C. § 1983 against appellees, Stephen G. Lemar, Tracey Curry, 1 The Honorable James E. Gritzner, United States District Judge for the Southern District of Iowa, sitting by designation. and Otha Serrell, for violating his Eighth Amendment right to be free from cruel and unusual punishment. Davis alleged appellees were deliberately indifferent to his medical needs by failing to diagnose his injury earlier and by delaying treatment of it. The district court2 granted summary judgment to appellees after thoroughly discussing the record and concluding that Davis had not made a showing of deliberate indifference or that his Eighth Amendment rights had been violated. After hearing oral argument and reviewing the record, we affirm on the basis of the district court’s opinion. 8th Cir. R. 47B. A true copy. Attest: CLERK, U.S. COURT OF APPEALS, EIGHTH CIRCUIT. 2 The Honorable Richard G. Kopf, Chief Judge, United States District Court for the District of Nebraska. -2-	{ "pile_set_name": "FreeLaw" }
Hornady Z-Max was developed in response to the upcoming Zombie apocalypsewell not really. Hornady Z-Max puts a Hollywood spin on a old favorite Hornady V-Max. The Z-Max projectiles are actually identical to the ever popular V-Max projectiles, with one major difference; the iconic red polymer tip has been traded in for a Zombie green tip. Depending on the particular projectile Z-max has both flat base and boat tail offerings which are caliber specific and provide you with incredible in-flight stability for long range shooting. .17 Caliber .172" Diameter 20 Grain Z-Max Flat Base Ballistic Coefficient 0.185 Sectional Density 0.097 500 Count Per Box Note: These are bullets which are intended to be used in conjunction with reloading. This is NOT loaded ammunition.	{ "pile_set_name": "Pile-CC" }
Chelsea Walton Chelsea Walton (born July 11, 1983) is a mathematician whose research interests include noncommutative algebra, noncommutative algebraic geometry, symmetry in quantum mechanics, Hopf algebras, and quantum groups. She is an associate professor at the University of Illinois at Urbana–Champaign and a Sloan Research Fellow. Education and career Walton is African-American, originally from Detroit, and was educated in the Detroit public schools. As a child she made a letter frequency table from her children's dictionary, and as a high school student, seeking a way to "do logic puzzles all day and get paid for this", she was already planning a career as a mathematics professor. She graduated from Michigan State University in 2005, and completed her Ph.D. at the University of Michigan in 2011. Her dissertation, On Degenerations and Deformations of Sklyanin Algebras, was jointly supervised by and Karen E. Smith, and based in part on her work as a visiting student at the University of Manchester, where Stafford had moved. Walton did postdoctoral research at the University of Washington and the Mathematical Sciences Research Institute, and became a C. L. E. Moore instructor at the Massachusetts Institute of Technology from 2012 to 2015. She came to Temple University as Selma Lee Bloch Brown Assistant Professor of Mathematics in 2015. She moved to the University of Illinois in 2018. Recognition Walton was named a Sloan Fellow in 2017, becoming the fourth African-American to win a Sloan Fellowship in mathematics. In 2018 she won the Andre Lichnerowicz Prize in Poisson geometry, the first woman to be awarded this prize. The award citation noted her research on Sklyanin algebras in Poisson geometry, on the actions of Hopf algebras, and on the universal enveloping algebra of the Witt algebra. References Further reading External links Home page Category:1983 births Category:Living people Category:21st-century American mathematicians Category:American women mathematicians Category:African-American mathematicians Category:Michigan State University alumni Category:University of Michigan alumni Category:Massachusetts Institute of Technology faculty Category:Temple University faculty Category:University of Illinois at Urbana–Champaign faculty Category:Sloan Research Fellows Category:21st-century women mathematicians	{ "pile_set_name": "Wikipedia (en)" }
Knows Sirius Black didn't betray the Potters. Lets him get life sentence in Azkaban anyway. 792 shares	{ "pile_set_name": "OpenWebText2" }
// Copyright 2012-present Oliver Eilhard. All rights reserved. // Use of this source code is governed by a MIT-license. // See http://olivere.mit-license.org/license.txt for details. package elastic import ( "encoding/json" "testing" ) func TestInnerHitEmpty(t testing.T) { hit := NewInnerHit() src, err := hit.Source() if err != nil { t.Fatal(err) } data, err := json.Marshal(src) if err != nil { t.Fatalf("marshaling to JSON failed: %v", err) } got := string(data) expected := `{}` if got != expected { t.Errorf("expected\n%s\n,got:\n%s", expected, got) } } func TestInnerHitWithName(t testing.T) { hit := NewInnerHit().Name("comments") src, err := hit.Source() if err != nil { t.Fatal(err) } data, err := json.Marshal(src) if err != nil { t.Fatalf("marshaling to JSON failed: %v", err) } got := string(data) expected := `{"name":"comments"}` if got != expected { t.Errorf("expected\n%s\n,got:\n%s", expected, got) } }	{ "pile_set_name": "Github" }
All relevant data are within the paper and its Supporting Information files. Introduction {#sec004} ============ A population of pluripotent adult somatic stem cells, well known as "neoblasts", has been extensively characterized and documented for free-living platyhelminthes (flatworms) \[[@pntd.0005418.ref001]--[@pntd.0005418.ref005]\]. Neoblasts represent the only proliferative cell population, responsible for cell renewal during homeostasis, development and regeneration \[[@pntd.0005418.ref006]--[@pntd.0005418.ref011]\]. Like in its free-living relatives, neoblast-like stem cells and similar cell renewal mechanisms also exist in the other two main groups of flatworms, trematodes (flukes) and cestodes (tapeworms), both living a parasitic life \[[@pntd.0005418.ref003], [@pntd.0005418.ref012]--[@pntd.0005418.ref015]\]. In cestodes, a population of undifferentiated cells, called "germinative cells", is the only source for cell proliferation. These germinative cells are totipotent and are thought to drive growth and development throughout the life cycle of cestodes \[[@pntd.0005418.ref012], [@pntd.0005418.ref015]--[@pntd.0005418.ref016]\]. The larval stage of the cestode Echinococcus multilocularis is the causative agent of alveolar echinococcosis (AE), one of the most lethal human helminthiasis \[[@pntd.0005418.ref017]\]. An infection is initiated when the intermediate host (rodents, humans) ingests infective eggs produced by adult tapeworms. The oncosphere hatches from the egg and then develops in the liver into cyst-like metacestode vesicles, which grow like tumors and infiltrate host tissue, forming new vesicles and even metastasizes. The metacestode vesicles bud giving rise to brood capsules, which in turn generate protoscoleces by asexual multiplication. Protoscoleces can either mature into adult tapeworms if ingested by the definitive host (canids) or develop into metacestode vesicles when distributed in the intermediate host. This unique proliferation potential of E. multilocularis metacestode larvae is believed to be based upon the germinative cells, which are totipotent and have the ability for extensive self-renewal \[[@pntd.0005418.ref015]\]. It has been well documented that the maintenance of pluripotency and self-renewal capacity of stem cells requires a continuous input from cell-extrinsic signals \[[@pntd.0005418.ref018]--[@pntd.0005418.ref019]\]. The extrinsic factors initiate various intrinsic signaling cascades which in turn maintain stem cells and regulate their functions. Signaling axes including LIF/gp130/STAT3, BMPs/BMPRs/Smads, Wnt/Frizzled/β-catenin, PI3K/AKT, and FGF/FGFR have been extensively evidenced to participate in controlling the survival, self-renewal, and differentiation of stem cells \[[@pntd.0005418.ref019]--[@pntd.0005418.ref020]\]. Increasing evidence has shown that the epidermal growth factor receptor (EGFR)-dependent signaling pathways also play important roles in the maintenance and regulation of stem cells \[[@pntd.0005418.ref021]--[@pntd.0005418.ref025]\]. The metacestode larvae of E. multilocularis grow and proliferate in close contact with the intermediate host's tissues, mainly within the liver. The microenvironment for metacestode development involves a number of host-derived hormones and cytokines, such as insulin, bone morphogenetic protein (BMP), fibroblast growth factor (FGF) and epidermal growth factor (EGF) \[[@pntd.0005418.ref026]\]. These host-derived factors are thought to bind to parasite receptors and in turn influence the parasite's growth and development through exhibiting their impacts on the relevant evolutionarily conserved signaling systems within the parasite \[[@pntd.0005418.ref026]--[@pntd.0005418.ref027]\]. A recent study has evidenced that host insulin activates E. multilocularis PI3K/AKT signaling pathways and stimulates germinative cell proliferation and larval development \[[@pntd.0005418.ref028]\]. In addition, in vitro cultivation of E. multilocularis larvae and primary cells requires continuous presence of host cells as the feeder cells (like stem cell cultivation) or host cell-conditioned medium which contains host-derived growth factors \[[@pntd.0005418.ref029]\]. Together, lines of evidence offer compelling clues that the conserved signaling pathways in E. multilocularis could respond to host factors and may regulate germinative cells which are fundamental for the larval growth and development of the parasite \[[@pntd.0005418.ref030]\]. It has been shown that the Ras/Raf/MEK/ERK signaling in E. multilocularis is activated in response to a host-derived EGF signal, which is most probably mediated by the parasite's EGFR-like kinase \[[@pntd.0005418.ref031]--[@pntd.0005418.ref033]\]. Using in vitro cultivation systems we show here that E. multilocularis EGFR/ERK signaling pathway is activated upon addition of human EGF and promotes germinative cell proliferation during the parasite's larval growth and development. Methods {#sec005} ======= Ethics statement {#sec006} ---------------- All animal experiments were conducted in strict accordance with China regulations on the protection of experimental animals (Regulations for the Administration of Affairs Concerning Experimental Animals, version from July-18-2013) and specifically approved by the Institutional Animal Care and Use Committee of Xiamen University (Permit Number: 2013--0053). Parasite in vitro culture, growth and development assay, and treatment {#sec007} ---------------------------------------------------------------------- The parasite isolate used in this study was obtained from Hulunbeier Pasture of Inner Mongolia of China \[[@pntd.0005418.ref034]\] and maintained by in vivo propagation of the parasite material in mice (supplied by Xiamen University Laboratory Animals Center, XMULAC). In vitro cultivation of metacestode vesicles was performed using host cell conditioned medium according to a previously established protocol \[[@pntd.0005418.ref035]\] unless otherwise indicated. For the growth assay, vesicles (diameter \< 1mm) were manually picked up and cultured in 24-well cell culture plates supplemented with different media as indicated in the text. 100 ng/mL recombinant human EGF (PeproTech, Rocky Hill, NJ) was used for all experiments unless otherwise indicated. Parasite growth was determined by the measurement of vesicle's diameter under inverted microscope weekly. Each group contains at least 3 replicates and more than 150 vesicles in total for each group were analyzed. Two-three independent experiments were performed. For the treatment of inhibitors, the EGFR inhibitors CI-1033 and BIBW2992 or the MEK inhibitor U0126 (Selleck Chemicals, Houston, TX) was added into the culture medium at a final concentration as indicated. All experiments were performed with exchange of the medium containing the same ingredients every three days. Protoscoleces were collected from parasite material and in vitro cultured in conditioned medium. The vesicle formation process, in which protoscoleces dilate and vacuolate, were examined after 18 days of culture. 40 mM of hydroxyurea was used to treat metacestode vesicles as described before \[[@pntd.0005418.ref015]\]. BrdU incorporation assay, EdU labeling and EdU-BrdU dual labeling {#sec008} ----------------------------------------------------------------- Vesicles were incubated with BrdU for two days and chromosomal DNA was isolated for BrdU incorporation assay with the colorimetric BrdU ELISA kit (Roche, Mannheim, Germany) as described before \[[@pntd.0005418.ref028]\]. Vesicles were incubated with 50 μM of EdU for 4 hours and whole-mount prepared according to Cheng et al. \[[@pntd.0005418.ref036]\]. Click-iT-EdU Alexa Fluor 555 Imaging Kit (Life Technologies, Shanghai, China) was used for detection of EdU. For the EdU-BrdU dual labeling, vesicles were incubated with 10 μM EdU for 4 hours, washed and then cultured with no labeling for 44 hours. 10 μM BrdU was next used for continuous labeling for another 24 hours. Vesicles were fixed at the end of the labeling period and whole-mount prepared (see also [S2C Fig](#pntd.0005418.s002){ref-type="supplementary-material"}). After a 45-minute 2N HCl treatment, immunofluorescence was performed for BrdU detection using the anti-BrdU antibody (clone MoBU, Life Technologies) followed by EdU detection. DNA was counterstained with 4', 6-diamidino-2-phenylindole (DAPI) (Sigma, St. Louis, MO) for all labeling experiments. For the inhibitor experiments during the recovery from hydroxyurea treatment, vesicles were allowed to recover in the conditioned medium supplemented with EGF. 10 μM CI-1033 or 20 μM U0126 was then added into the medium immediately after the initial EdU pulse. Germinative cell proliferation was analyzed by EdU-BrdU dual labeling after 4 days of recovery. For the quantification of EdU^+^ or BrdU^+^ cells, at least 12 random microscopic fields from 4--6 vesicles were captured and the positive cells were manually counted. 3--5 labeling experiments were performed and analyzed for each control and treatment group. mRNA expression analysis of E. multilocularis EGFR members {#sec009} ------------------------------------------------------------ Total RNA was extracted from in vitro-cultivated protoscoleces or metacestode vesicles, treated with RNase-free DNase and reverse transcribed into cDNA. cDNAs were processed for RT-PCR analysis using the primers: EmER-qF2 (5'-GCG AAT GTA AGC ATT TCA AGT CA-3') and EmER-qR2 (5'-TTC ACA AAG TAG CAG AAA GCA CAT-3') for Emer; 617300-qF (5'-GCC GCA TCT ATG GAC ACGC-3') and 617300-qR (5'-AGT CAT CTT GTG GGA GGA ATCG-3') for Emuj_000617300; 969600-qF (5'-CTC TGG GGT GTC TGC TGT CC-3') and 969600-qR (5'-TCC CAC AGA GTC ACA CCG TAGG-3') for Emuj_000969600. Expression of EmER in Xenopus oocytes {#sec010} --------------------------------------- Expression of the parasite EGF receptor EmER in Xenopus oocytes was performed according to \[[@pntd.0005418.ref037]\]. Briefly, the full-length coding sequence of EmER was cloned into the Xenopus oocyte expression vector pXT7-flag (a gift from Dr. Li Guang in Xiamen University). Linearized plasmids were then used as the templates for capped mRNA (cRNA) synthesis using the T7 mMessage mMachine Kit (Ambion). Oocytes were obtained from Xenopus laevis (supported by Stem Cell Bank, Chinese Academy of Sciences) and then microinjected with EmER cRNA or water (noninjected control). Membrane proteins were extracted from 30 oocytes after 48 h of injection and immunoprecipitated by the anti-flag monoclonal antibody (Sigma) and analyzed by western blot. For EGF and CI-1033 treatment, oocytes that had been expressing EmER for 48 h were incubated with 10 μM of CI-1033 or DMSO for 4 h followed by 20 minutes of EGF stimulation. Membrane extracts were immunoprecipitated and analyzed by western blot using the anti-flag or anti-phospho-tyrosine (CST, Beverly, MA) monoclonal antibodies. Induction of GVBD (germinal vesicle breakdown) in EmER-expressing Xenopus oocytes was performed according to \[[@pntd.0005418.ref037]\]. Oocytes that had been expressing EmER for 48 h were pretreated with 10 μM of CI-1033 or DMSO for 4 h and then incubated with EGF. GVBD was monitored after 16 h of EGF incubation. As a positive control for GVBD, oocytes were stimulated with progesterone (PG), the natural inducer. 20--30 oocytes were used for each group and three independent experiments were performed. Western blot and immunofluorescence {#sec011} ----------------------------------- Lysates of in vitro-cultivated protoscoleces or metacestode vesicles were produced, separated on 12% acrylamide gels and transferred to PVDF membranes. Detection of E. multilocularis ERK and phosphorylated ERK was performed according to Spiliotis et al. \[[@pntd.0005418.ref032]\] using the polyclonal rabbit anti-ERK1/2 (Stressgen, Victoria, Canada) and anti-ERK1/2 \[pTpY^185/187^\] (Life Technologies) antibodies, respectively. The anti-rabbit IgG antibody conjugated with horseradish peroxidase (Theromo Scientific, Shanghai, China) was used as a secondary antibody. For all western blot experiments, detection of E. multilocularis actin was performed using the polyclonal anti-β-actin antibody (CST) as loading controls. Immunofluorescence was performed using the whole-mount prepared metacestode vesicles as described before \[[@pntd.0005418.ref036]\]. For Histone H3 detection, the anti-phospho-Histone H3 antibody (Ser10, 1:200) (CST) was used. For all immunofluorescence experiments, an Alexa 488-conjugated second antibody (Life Technologies) was used and DNA was counterstained with DAPI. Data analysis and statistics {#sec012} ---------------------------- Data of three or more experimental repeats are shown as mean ± SD as indicated in the respective figure legend unless otherwise indicated. The mean values of the data from the experimental groups were compared by performing two-tailed Student's t-test and the P values were indicated as those: \*P \< 0.05, \\P \< 0.01, and \\\*P \< 0.001. Results {#sec013} ======= EGF stimulates E. multilocularis larval growth and development {#sec014} ---------------------------------------------------------------- To examine the impacts of EGF on the larval growth of E. multilocularis, metacestode vesicles were incubated in host cell-conditioned medium supplemented with recombinant human EGF. We found that 10 ng/mL or higher concentrations of EGF can greatly promote vesicle's growth ([S1 Fig](#pntd.0005418.s001){ref-type="supplementary-material"}). We then used 100 ng/mL of EGF which showed the most significant effect on parasite's growth in vitro for further studies. The results show that addition of EGF can stimulate the growth of metacestode vesicles ([Fig 1A](#pntd.0005418.g001){ref-type="fig"}). Similar results were observed after EGF was added into the Dulbecco's modified eagle medium (DMEM) containing 10% serum ([Fig 1B](#pntd.0005418.g001){ref-type="fig"}). We also found that EGF greatly promoted the vesicle formation process, in which the protoscoleces dilated and vacuolated ([Fig 1C](#pntd.0005418.g001){ref-type="fig"}). These results illustrate that the larval growth and development of in vitro-cultivated E. multilocularis larvae could be stimulated by exogenously added EGF, which is probably mediated by an EGFR-dependent signaling in the parasite. ![EGF stimulates E. multilocularis larval growth and development.\ (A-B) Metacestode vesicles were cultivated in conditioned medium (C-medium) (A) or serum-containing DMEM (B) supplemented with 100 ng/mL recombinant human EGF or not. Vesicle growth is shown as the increase of vesicle diameter as compared to day 0. Comparison between the EGF group and the control group at the same timepoint was performed using two-tailed Student's t-test. (C) Vesicle formation from protoscolex in conditioned medium (control) with addition of EGF. Control was set to 1 and results were normalized against the control (right). Representative images of the formation process are shown (left). Bar = 50 μm. (D) Survival of the vesicles in serum-free DMEM supplemented with EGF. Data in (A-D) are shown as mean ± SD of at least three replicates, representative of 2--3 independent experiments.\*P \< 0.05, \\P \< 0.01, and \\\*P \< 0.001.](pntd.0005418.g001){#pntd.0005418.g001} Considering that either serum or conditioned medium contains complex ingredients, we then incubated vesicles in the serum-free DMEM only supplemented with EGF. Addition of EGF could not obviously stimulate the growth of vesicles, however, it remarkably promoted their survival ([Fig 1D](#pntd.0005418.g001){ref-type="fig"}). Although metacestode vesicles could not survive for long in this situation, the method excludes the influence of other host factors in serum and suggests that the parasite is responsive to host EGF stimulation. Further experiments in this study were all performed using host cell-conditioned medium unless otherwise indicated. EGF promotes the proliferation of E. multilocularis germinative cells {#sec015} ----------------------------------------------------------------------- Given that the germinative cells, a population of stem cell-like cells, drive larval growth and development of E. multilocularis, we then investigated the impacts of EGF on the germinative cells. Vesicles were incubated with 5-bromo-2'-deoxyuridine (BrdU), an analogue of thymidine used for studying cell proliferation by detecting its incorporation into the newly synthesized DNA of replicating cells. The result shows that addition of EGF greatly stimulated the BrdU incorporation in the vesicles ([Fig 2A](#pntd.0005418.g002){ref-type="fig"}). Since germinative cells are the only cells capable of proliferation in metacestode vesicles, this result suggests that EGF promotes germinative cell proliferation. ![EGF promotes E. multilocularis germinative cell proliferation.\ (A) Vesicles were incubated with BrdU for two days and chromosomal DNA was isolated for BrdU incorporation assay. Control was set to 1 and results were normalized against the control. Data are shown as mean ± SD of triplicates. \\ P \< 0.01. (B) Vesicles were in vitro cultivated under normal conditions and germinative cell proliferation was visualized by EdU-BrdU dual labeling. Insert shows the magnified view. Arrows indicate EdU^+^BrdU^+^ cells. Bar = 20 μm. (C) Vesicles were pretreated with 40 mM of hydroxyurea for three days and allowed for recovery in conditioned medium (control) supplemented with EGF. Germinative cell proliferation was analyzed after 4 days of removal of hydroxyurea and representative images are shown. Quantification of EdU^+^ and EdU^+^BrdU^+^ cells is shown in the right panel. Data are shown as mean ± SD of 3 separate labeling experiments. \* P \< 0.05; \\ P \< 0.01. Bar = 20 μm.](pntd.0005418.g002){#pntd.0005418.g002} A dual labeling method through sequential pulses of 5-ethylnyl-2'-deoxyuridine (EdU) and BrdU, which has been utilized to verify the self-renewal capacity of adult somatic stem cells in the human blood fluke Schistosoma mansoni \[[@pntd.0005418.ref014]\], was further applied to determine the effects of EGF on germinative cells. EdU is a newly found analogue of thymidine \[[@pntd.0005418.ref038]\], which has been shown to be incorporated by proliferating cells of E. multilocularis \[[@pntd.0005418.ref015]\]. Under normal in vitro cultivation conditions, we found that most of the dividing germinative cells in the vesicles could incorporate EdU before a chase period of 44 h, which was further used for EdU-BrdU dual labeling experiments ([S2A Fig](#pntd.0005418.s002){ref-type="supplementary-material"}). The results show that about 48% of cells that initially incorporate EdU are BrdU^+^ 3 days after an initial EdU pulse (749 EdU^+^BrdU^+^ / 1554 EdU^+^ nuclei, 4 independent labeling experiments) ([Fig 2B](#pntd.0005418.g002){ref-type="fig"}), indicating that these germinative cells divide and produce proliferation-competent daughter cells that initially incorporate EdU can incorporate BrdU during the second replication. It has been shown that depletion of the germinative cells in E. multilocularis vesicles could be achieved through hydroxyurea treatment for longer periods of time (e.g. seven days) and that the germinative cells undergo clonal expansion like stem cells after removal of hydroxyurea \[[@pntd.0005418.ref015]\]. We also performed similar experiments and found that the EdU^+^BrdU^+^ cells are highly presented in the clonally growing germinative cells ([S3 Fig](#pntd.0005418.s003){ref-type="supplementary-material"}), suggesting that the EdU^+^BrdU^+^ cells are extensively proliferating germinative cells and a part of them might be undergoing self-renewing divisions. To further investigate the effect of EGF on germinative cells, we treated vesicles with hydroxyurea to eliminate most germinative cells \[[@pntd.0005418.ref015]\]. After removal of hydroxyurea, germinative cell proliferation/self-renewal was allowed for recovery in the medium with addition of EGF or not for 4 days, and the EdU-BrdU sequential pulses began on the second day of the recovery (see also [S2B and S2C Fig](#pntd.0005418.s002){ref-type="supplementary-material"}). At the end of the dual labeling period, the results show that addition of EGF induced significantly more numbers of both EdU^+^ and EdU^+^BrdU^+^ cells in the vesicles compared to the controls ([Fig 2C](#pntd.0005418.g002){ref-type="fig"}). We found that the proportion of EdU^+^BrdU^+^ cells with respect to the number of EdU^+^ cells was also greatly increased (25.6% and 50.5% for the control and EGF-treated groups, respectively, statistical significance P = 0.00101), suggesting increased continuous proliferation, and possibly promoted self-renewal of the germinative cells upon EGF stimulation. Together, these results support the findings that addition of EGF can promote the proliferation of germinative cells ([Fig 2A](#pntd.0005418.g002){ref-type="fig"}), which subsequently drives the larval growth and development of E. multilocularis. These results also suggest that an EGFR-dependent signaling in the parasite may be involved in regulating germinative cell proliferation upon EGF stimulation. EGFR/ERK signaling contributes to germinative cell proliferation {#sec016} ---------------------------------------------------------------- E. multilocularis possesses an EGFR-like kinase (EmER) which is suggested to interact with host EGF \[[@pntd.0005418.ref031], [@pntd.0005418.ref033]\]. Besides EmER, we also found two additional E. multilocularis EGF receptor members (Emuj_000617300 and Emuj_000969600) by analyzing E. multilocularis genome sequence (<http://www.genedb.org/Homepage/Emultilocularis>) and fully cloned and sequenced the respective genes. Comparisons of the putative protein sequences reveal that these EGF receptor members exhibit significant homologies to human EGFR and the EGFR homologue of the closely related schistosome S. mansoni, especially in the tyrosine kinase domains ([S4A--S4D Fig](#pntd.0005418.s004){ref-type="supplementary-material"}). The results of mRNA expression analysis show that these EGF receptor homologues are constitutively present in E. multilocularis metacestode vesicles and protoscoleces ([S4E Fig](#pntd.0005418.s004){ref-type="supplementary-material"}). To investigate whether the E. multilocularis EGF receptor(s) respond to EGF stimulation or not, we utilized the Xenopus oocyte expression system, which is a powerful tool for receptor tyrosine kinase research and has been successfully used for studying the EGF receptor (SER) in S. mansoni \[[@pntd.0005418.ref037]\]. The results show that the parasite EGFR EmER could be efficiently expressed in Xenopus oocytes with a molecular weight approximately 200 kDa ([Fig 3A](#pntd.0005418.g003){ref-type="fig"}), and that addition of EGF resulted in the activation of EmER by detection of phosphorylated tyrosine ([Fig 3B](#pntd.0005418.g003){ref-type="fig"}). It has previously been shown that host EGF could induce germinal-vesicle breakdown (GVBD) in S. mansoni SER-expressing oocytes \[[@pntd.0005418.ref037]\]. Similar results were also observed in the EmER-expressing oocytes ([Fig 3C](#pntd.0005418.g003){ref-type="fig"}), suggesting that addition of exogenous EGF could activate the parasite EGFR in the oocytes and induce GVBD. Using the oocyte system, we also found that CI-1033 (canertinib), an irreversible inhibitor for human EGF receptors \[[@pntd.0005418.ref039]\], could effectively inhibit the EGF-induced activation of EmER and GVBD ([Fig 3B and 3C](#pntd.0005418.g003){ref-type="fig"}). ![EGF activates E. multilocularis EGF receptor EmER in Xenopus oocyte expression system.\ (A) Membrane extracts were prepared from Xenopus oocytes expressing EmER or not (NI, noninjected) and immunoprecipitated and analyzed by western blot using the anti-flag antibody. The results of two independent injection experiments are presented (I-1 and I-2). The bands exhibited a molecular mass larger than expected (178 kDa), which could be attributable to glycosylation. (B) Oocytes expressing EmER were incubated with CI-1033 or DMSO for 4 h followed by 20 minutes of EGF stimulation. Membrane extracts were immunoprecipitated by the anti-flag antibody and analyzed by western blot using the anti-flag or anti-phospho-tyrosine antibodies. (C) Induction of GVBD (germinal vesicle breakdown) in EmER-expressing Xenopus oocytes. Oocytes that had been expressing EmER for 48 h were pretreated with CI-1033 or DMSO for 4 h and then incubated with EGF. GVBD was monitored after 16 h of EGF incubation and the mean percentages of oocytes exhibiting GVBD for three separate experiments are shown. Noninjected oocytes were used as controls. DMSO (final concentration 0.25%) was found no effects on GVBD. PG: progesterone; "/": not tested.](pntd.0005418.g003){#pntd.0005418.g003} We wondered if the impaired EGFR activation would impact E. multilocularis germinative cell behaviors. To this end, we treated vesicles with CI-1033. The results show that CI-1033 significantly reduced the number of EdU^+^ cells in the vesicles ([Fig 4A](#pntd.0005418.g004){ref-type="fig"}). Another EGFR inhibitor, BIBW2992 (afatinib) \[[@pntd.0005418.ref040]\], also exhibited a similar effect to CI-1033 on the germinative cells ([S5A Fig](#pntd.0005418.s005){ref-type="supplementary-material"}). ![Inhibition of E. multilocularis EGFR impairs germinative cell proliferation.\ (A) Metacestode vesicles were treated with 10 μM CI-1033 and the representative images for day 0, 3 and 6 are shown on the left (red: EdU; blue: DAPI). Quantifications of the EdU^+^ germinative cells in the vesicles treated with 10 μM CI-1033 for indicated time (middle) and 5--10 μM CI-1033 or DMSO control (0) for 6 days (right) are shown. Values represent the mean ± SD of 5 separate labeling experiments. \* P \< 0.05; \\\* P \< 0.001. (B) Effects of CI-1033 on germinative cell proliferation during the recovery from hydroxyurea treatment. Vesicles were allowed for recovery in conditioned medium supplemented with EGF, and CI-1033 was added into the medium to a final concentration of 10 μM immediately after the initial EdU pulse. Germinative cell proliferation was analyzed by EdU-BrdU dual labeling after 4 days of recovery. Images show rare EdU^+^ and EdU^+^BrdU^+^ cells following CI-1033 treatment (see the text). (C) Representative images of the accumulations of EdU^+^ germinative cells in some cell aggregates (indicated by the dashed-line boxes) in the vesicles treated with DMSO (control) or 10 μM CI-1033 for 6 days (red: EdU; blue: DAPI). (D) Effects of CI-1033 on the larval growth and development. Vesicles or protoscoleces were cultivated in DMSO-containing conditioned medium (control) supplemented with the ingredients as indicated. Vesicle growth (left) and vesicle formation from protoscoleces (right) were analyzed after 28 days and 18 days of cultivation, respectively. Data are shown as mean ± SD of triplicates, representative of 3 independent experiments. \\ P \< 0.01 and \\\* P \< 0.001. Bar = 20 μm in (A), (B) and (C).](pntd.0005418.g004){#pntd.0005418.g004} In the EdU-BrdU dual labeling experiments, vesicles were allowed to recover from the hydroxyurea treatment with addition of EGF, and CI-1033 was administrated to the vesicles immediately after the initial EdU pulse. At the end of the labeling period, only 1.6‰ of total cells were EdU^+^ (11 EdU^+^ / 6737 DAPI nuclei), and the EdU^+^BrdU^+^ cells were hardly detected ([Fig 4B](#pntd.0005418.g004){ref-type="fig"}). There is a strong accumulation of EdU^+^ cells in numerous aggregates in some developing vesicles, which indicates that the active proliferation and extensive self-renewal of germinative cells may drive the development of brood capsule and protoscolex in the vesicles \[[@pntd.0005418.ref015]\]. Our studies show that CI-1033 can also abolish the accumulation of EdU^+^ cells in these cell aggregates ([Fig 4C](#pntd.0005418.g004){ref-type="fig"}). Further investigations showed that CI-1033 and BIBW2992 can significantly inhibit the larval growth and development upon EGF stimulation ([Fig 4D](#pntd.0005418.g004){ref-type="fig"} and [S5B Fig](#pntd.0005418.s005){ref-type="supplementary-material"}). These results suggest that an EGFR-dependent signaling in the parasite is required for the promoted germinative cell proliferation and larval growth, and that the signaling could probably be impaired by the EGFR inhibitors initially designed against human EGF receptors. In E. multilocularis, structural and functional homologues to mammalian MAP kinase cascade molecules, such as RAF, MEK and ERK, have been identified and characterized \[[@pntd.0005418.ref027]\]. Previous study has shown that host EGF could induce E. multilocularis ERK activation \[[@pntd.0005418.ref032]\]. Our data show that the basal level of ERK phosphorylation in the vesicles was down-regulated following CI-1033 treatment in a time-dependent manner ([Fig 5A](#pntd.0005418.g005){ref-type="fig"}). We also found that CI-1033 can inhibit the phosphorylation of ERK induced by EGF ([Fig 5B](#pntd.0005418.g005){ref-type="fig"}). These results suggest that the EGFR inhibitors could impair the activations of parasite's EGFR and ERK. ![EGFR/ERK signaling contributes to E. multilocularis germinative cell proliferation.\ (A) CI-1033 impairs E. multilocularis ERK phosphorylation. Vesicles were treated with 10 μM CI-1033 for indicated time. (B) CI-1033 inhibits EGF-stimulated E. multilocularis ERK phosphorylation. Vesicles were treated with 10 μM CI-1033 (+) or DMSO (-) for 6 h, followed by stimulation with EGF (+) or not (-) for 20 minutes. (C) U0126 impairs E. multilocularis ERK phosphorylation. Vesicles were treated with indicated concentrations of U0126 for 6 h. (D) U0126 inhibits EGF-stimulated E. multilocularis ERK phosphorylation. Vesicles were treated with 20 μM U0126 (+) or DMSO (-) for 6 h, followed by stimulation with EGF (+) or not (-) for 20 minutes. (E) U0126 reduces the number of EdU^+^ germinative cells. Vesicles were treated with 20 μM U0126 for 6 days. Representative images are shown on the left and the quantification is shown on the right. Values represent the mean ± SD of 4 separate labeling experiments. \\\* P \< 0.001. Bar = 20 μm. (F) Effects of U0126 on germinative cell proliferation during the recovery from hydroxyurea treatment. Vesicles were allowed for recovery in conditioned medium supplemented with EGF and U0126 was added into the medium to a final concentration of 20 μM immediately after the initial EdU pulse. Germinative cell proliferation was analyzed after 4 days of recovery. Images show rare EdU^+^ and EdU^+^BrdU^+^ cells following U0126 treatment (see the text). Bar = 20 μm. (G) Effects of U0126 on the larval growth. Vesicles were incubated with 10 μM U0126 and parasite growth was analyzed after 28 days of cultivation. Data are shown as mean ± SD of triplicates. \\ P \< 0.01.](pntd.0005418.g005){#pntd.0005418.g005} We then treated vesicles with U0126, a MEK inhibitor which effectively suppressed both of the basal and the addition of EGF-induced ERK phosphorylations in the parasite ([Fig 5C and 5D](#pntd.0005418.g005){ref-type="fig"}). Along with the inhibition of MEK/ERK activity, a remarkable decrease in the number of EdU^+^ germinative cells was observed in the vesicles ([Fig 5E](#pntd.0005418.g005){ref-type="fig"}). EdU-BrdU dual labeling experiments also indicate that U0126 has a comparable inhibition effect to CI-1033 on the EGF-promoted germinative cell proliferation (2.9‰ of the total cells were EdU^+^ and the EdU^+^BrdU^+^ cells were hardly detected) ([Fig 5F](#pntd.0005418.g005){ref-type="fig"}). Our further investigation shows that U0126 can significantly inhibit the EGF-stimulated vesicle growth ([Fig 5G](#pntd.0005418.g005){ref-type="fig"}). Taken together, these results suggest that the EGF-mediated EGFR/MEK/ERK signaling contributes to germinative cell proliferation during E. multilocularis larval growth and development. Discussion {#sec017} ========== Throughout the complex life cycle of E. multilocularis, the parasite always keeps a population of stem cell-like cells, the germinative cells, which are considered as one of the fundamental underpinnings of its growth and development in the host \[[@pntd.0005418.ref015]\]. Stem cell maintenance and functions are strictly controlled by signals from the local tissue microenvironments known as "niches", which have been widely characterized and elucidated for mammals and invertebrate model animals \[[@pntd.0005418.ref018]\]. Proliferation and differentiation of neoblasts, the stem cells in the free-living flatworm planarian, are also regulated by signals from the surrounding cells \[[@pntd.0005418.ref011]\]. However, the situation is somewhat different for the host liver tissue-dwelling metacestode larvae of E. multilocularis. Due to the intimate parasite-host contact, the parasite is believed to be able to sense signals derived not only from its own tissue but also from the host-derived hormones and cytokines, as these signaling receptors and downstream signaling cascades are evolutionarily conserved between the mammalian hosts and E. multilocularis \[[@pntd.0005418.ref027]\]. Thus it is tempting to suggest that E. multilocularis germinative cells should be regulated by the host-derived factors \[[@pntd.0005418.ref030]\]. In the present study, we show that germinative cell proliferation of the in vitro-cultivated E. multilocularis larvae is promoted upon the addition of human EGF, which in turn drives vesicle growth and vesicle formation from protoscolex (Figs [1](#pntd.0005418.g001){ref-type="fig"} and [2](#pntd.0005418.g002){ref-type="fig"}). These results suggest that the germinative cells are regulated by the signaling pathways within the parasite that could sense the host-derived EGF signal. A physiologically relevant concentration of EGF (1 ng/mL) showed a modest effect on the growth of in vitro-cultivated vesicles, while 10 ng/mL or higher concentrations of EGF greatly stimulated the growth ([S1 Fig](#pntd.0005418.s001){ref-type="supplementary-material"}). We then used 100 ng/mL of EGF, which exhibited the most significant effect on parasite's growth, for further in vitro studies. This concentration of EGF is considerable and widely used in human cancer cell research, and is relevant to those used in the in vitro studies of E. multilocularis and S. mansoni \[[@pntd.0005418.ref032], [@pntd.0005418.ref037]\]. However, it could exceed the physiological concentrations in liver. Considering that the larval development of E. multilocularis causes host liver tissue destruction and regeneration while EGF is continually made available to the liver and strongly produced during regeneration processes \[[@pntd.0005418.ref041]--[@pntd.0005418.ref042]\], it will be interesting to investigate the effects of host EGF observed in this study on the parasite in future using in vivo infection models. The promoted proliferation of germinative cells upon EGF stimulation was supported by our EdU-BrdU dual labeling experiments ([Fig 2B and 2C](#pntd.0005418.g002){ref-type="fig"} and [S3 Fig](#pntd.0005418.s003){ref-type="supplementary-material"}), which also suggest that the cell-cycle time for most of the actively proliferating germinative cells is less than three days ([S2 Fig](#pntd.0005418.s002){ref-type="supplementary-material"} and [Fig 2B](#pntd.0005418.g002){ref-type="fig"}), similar to that for S. mansoni adult somatic stem cells \[[@pntd.0005418.ref014]\]. It has been suggested that there are subpopulations of the germinative cells capable of maintaining their pluripotency and self-renewing like stem cells \[[@pntd.0005418.ref015]\]. Although our data suggest that the increased number of EdU^+^BrdU^+^ cells (as well as the increased ratio of EdU^+^BrdU^+^ cells to EdU^+^ cells) may also result from the promoted self-renewal of these stem-like cell populations, due to the limitations of the dual labeling experiments we still could not distinguish the self-renewing cells from the transit amplifying cells. Specific molecular markers of the stem-like cell populations that work independently of proliferation would be needed for further explorations to clarify the contribution of EGF to germinative cell self-renewal in E. multilocularis. Addition of host EGF promotes germinative cell proliferation, which then promotes in vitro-cultivated protoscoleces to form metacestode vesicles ([Fig 1C](#pntd.0005418.g001){ref-type="fig"}). The formation of vesicle from protoscolex occurs in vivo following the rupture of parasite cysts and distribution of protoscoleces, and is thought to contribute to prolonged parasite survival in the intermediate host \[[@pntd.0005418.ref019], [@pntd.0005418.ref028]\], which would result in a poor prognosis after surgery-induced rupture of parasite cysts in human echinococcosis, at least in cystic echinococcosis (CE). Interestingly, we found that addition of host EGF may not only promote this formation process but also initiate it by triggering activation of the germinative cells from a quiescent state in the developed protoscoleces ([S6 Fig](#pntd.0005418.s006){ref-type="supplementary-material"}). Koziol et al. \[[@pntd.0005418.ref015]\] indicated that there is a large population of germinative cells capable of proliferation in the developed protoscolex, but they remain in a quiescent state or with slow cell-cycle kinetics for as long as the protoscolex remains resting within the metacestode. These quiescent germinative cells were activated when the protoscoleces were activated by artificially mimicking the ingestion by the definitive host or when the protoscoleces were in vitro cultured in serum-containing DMEM. Thus it is tempting to suggest that host factors activate the quiescent germinative cells to re-enter the cell-cycle for proliferation and self-renewal, which may further stimulate the protoscoleces to mature into adults within the definitive host's intestine or to form metacestode vesicles in the intermediate host's liver and other tissues. It is still unclear how protoscoleces alternate between developmental fates: the adult or metacestode vesicle. In any case, this unique development potential for protoscoleces is based on the germinative cells, which may response to different host-derived signals from different host tissue microenvironments. Our data suggest that host factors may play a vital role in host-parasite interaction via mediating the relative signaling pathways in the parasite to regulate germinative cell functions. It has been suggested that host EGF could activate the highly conserved Ras/Raf/MEK/ERK signaling cascade in E. multilocularis, which is probably mediated by the parasite's EGF-receptor-like kinase \[[@pntd.0005418.ref031]--[@pntd.0005418.ref033]\]. Besides EmER, the first EGFR homologue identified in E. multilocularis \[[@pntd.0005418.ref031]\], two additional members of the EGFR family could be identified, which display significant homologies with human EGFR and S. mansoni EGFR in the functional domains ([S4A--S4D Fig](#pntd.0005418.s004){ref-type="supplementary-material"}). Like EmER, these two EGFR homologues are continually expressed in E. multilocularis metacestode vesicles and protoscoleces ([S4E Fig](#pntd.0005418.s004){ref-type="supplementary-material"}). It will be interesting to clarify their roles as the EGF receptor kinase in the parasite's development within the host in the future work. In this study, using the Xenopus oocyte expression system we show that one of the EGFR homologues EmER can be activated by host EGF ([Fig 3](#pntd.0005418.g003){ref-type="fig"}). We also show here that inhibition of the MEK/ERK signaling activation by either the EGFR inhibitors CI-1033 and BIBW2992 or the MEK inhibitor U0126 significantly impaired E. multilocularis germinative cell proliferation, larval growth and development (Figs [4](#pntd.0005418.g004){ref-type="fig"} and [5](#pntd.0005418.g005){ref-type="fig"} and [S5 Fig](#pntd.0005418.s005){ref-type="supplementary-material"}). In mammals, the MEK/ERK pathway plays a critical role in regulating stem cells. For example, it is required for maintenance of stemness and self-renewal of mouse neural stem/precursor cells \[[@pntd.0005418.ref043]--[@pntd.0005418.ref044]\]. The role of MEK/ERK pathway in mammalian embryonic stem cells (ESCs) is much more complex. The MEK/ERK signaling plays a functional role in promoting differentiation of mouse ESCs, while it promotes self-renewal of human ESCs (reviewed in \[[@pntd.0005418.ref020]\] and \[[@pntd.0005418.ref045]\]). In invertebrates, much of what is known about the role of the MEK/ERK signaling in regulation of stem cells derives from the studies of Drosophila. It has been extensively documented that the EGFR-dependent activation of the MEK/ERK signaling pathway is essential for promoting the maintenance and self-renewal of various types of adult somatic stem cells in Drosophila \[[@pntd.0005418.ref023]--[@pntd.0005418.ref024], [@pntd.0005418.ref046]\]. Our data also show that EGF-promoted germinative cell proliferation and larval growth rely on the activation of the parasite's EGFR/ERK signaling. However, it still remains unclear that the contribution of EGFR/ERK signaling to the promoted proliferation is attributed to the direct response in the germinative cells or to the indirect response to a second signal produced by their surrounding differentiated cells upon EGF stimulation, or to both. Further analysis of the EGFR activation in proliferating germinative cells would be needed to clarify this issue. While downstream of EGFR lies the PI3K/AKT, MEK/ERK and STAT3 pathways, our findings define here that the MEK/ERK pathway contributes to the role of EGFR signaling in regulating E. multilocularis germinative cell proliferation. Considering that the PI3K/AKT pathway in E. multilocularis was recently suggested to be involved in the host insulin-stimulated germinative cell proliferation \[[@pntd.0005418.ref028]\], we also treated metacestode vesicles with the PI3K inhibitor LY294002, which was shown to effectively inhibit E. multilocularis PI3K activity \[[@pntd.0005418.ref028]\]. We found that LY294002 did not exhibit as obvious an inhibitory effect as U0126 on the proliferation of germinative cells but slightly decreased the number of EdU^+^ cells in metacestode vesicles. Future work would be required for evaluating the contribution of EGFR/AKT/PI3K signaling to the regulation of E. multilocularis germinative cells. Increasing evidence has shown that the inhibitors originally designed against the human kinases can effectively inhibit the activity of related kinases in E. multilocularis \[[@pntd.0005418.ref028], [@pntd.0005418.ref047]--[@pntd.0005418.ref049]\]. Based on the evolutionary conservation among the kinases of vertebrates and invertebrates (including invertebrate parasites), it has been widely considered that small molecules that target human kinases are promising drug candidates for treating human helminthiasis, including echinococcosis \[[@pntd.0005418.ref050]--[@pntd.0005418.ref051]\]. Considering our observations that both basal ERK phosphorylation and ERK phosphorylation induced by administered EGF were effectively suppressed by either CI-1033 or U0126 ([Fig 5A--5D](#pntd.0005418.g005){ref-type="fig"}), it is therefore conceivable that the kinase inhibitors used in this study could impair EGFR/ERK signaling in E. multilocularis. Although these inhibitors were used within the range concentrations required for these compounds to specifically inhibit their respective targets in humans, it is possible that they may also have cellular targets other than EGFR/ERK signaling in E. multilocularis. Our findings suggest that exogenous EGF-activated EGFR/ERK signaling in the parasite was inhibited by CI-1033. Given that the long-term in vitro maintenance of E. multilocularis larvae and primary cells requires continuous presence of host cell-derived growth factors \[[@pntd.0005418.ref029]\], it is reasonable to assume that EGFR inhibitors could impair the activation of parasite's EGFR upon host EGF stimulation, which might be the main reason for the diminished germinative cell proliferation and the impaired larval growth and development ([Fig 4](#pntd.0005418.g004){ref-type="fig"} and [S5 Fig](#pntd.0005418.s005){ref-type="supplementary-material"}). However, our data could not exclude the possibility that EGFR inhibitors may also impair the parasite's EGFR activation mediated by its own EGF molecules. It has been recently shown that neoblast clonal expansion in the free-living flatworm planarian is regulated by its own EGF-mediated EGFR signaling \[[@pntd.0005418.ref052]\]. Since the parasite also possesses a putative EGF homologue \[[@pntd.0005418.ref053]\], this endogenous EGFR ligand-mediated signaling might also be involved in regulating E. multilocularis germinative cells. Improved in vitro cultivation systems and methods that could avoid/reduce the impacts of host factors will be helpful to investigate the roles of this endogenous signaling in germinative cell regulation. Although stem cell-like germinative cells has been widely described in tapeworms and their roles in the parasite's development within the host are thought to be of fundamental importance, there are still long standing gaps in our knowledge of mechanisms controlling the behavior of these cells. This study defines an essential role for the EGF-mediated EGFR/ERK signaling in promoting germinative cell proliferation in E. multilocularis. It makes an effort to unravel the mechanisms of regulation of tapeworm germinative cells in response to host-derived growth factors, and helps in understanding the delicate developmental strategies of these parasites within the host. Targeting the signaling pathways involved in regulating germinative cells may provide a novel therapeutic strategy against echinococcosis and other human cestodiasis. Supporting information {#sec018} ====================== ###### Effects of EGF on E. multilocularis larval growth. Metacestode vesicles were cultivated in conditioned medium (Control) supplemented with 1--100 ng/mL recombinant human EGF for 49 days. Vesicle growth is shown as the increase of vesicle diameter as compared to day 0 for each group. Data are shown as mean ± SD of triplicates, representative of two independent experiments. (TIF) ###### Click here for additional data file. ###### EdU-BrdU dual labeling experiments. $A$ EdU labeling and phospho-Histone H3 (Ser10, p-H3) immunofluorescence. Metacestode vesicles were administrated to a 4-h EdU pulse, and after 44 hours of pulse about 88% (365/413) of p-H3^+^ mitotic cells are EdU^+^. The chase period of 44 h was then used for EdU-BrdU dual labeling experiments. Note the low percentage of p-H3^+^ cells, which is consistent with a previous report \[[@pntd.0005418.ref015]\]. (B) Analysis of proliferating germinative cells by EdU labeling in hydroxyurea (HU) treatment experiments. Metacestode vesicles were treated with 40 mM of hydroxyurea for three days and then allowed for recovery in conditioned medium. Representative images are shown as: no treatment control, hydroxyurea treatment, and 4 days of recovery after removal of hydroxyurea (red: EdU; blue: DAPI). Bar = 40 μm. (C) Timeline for hydroxyurea treatment and EdU-BrdU dual labeling. Metacestode vesicles were pretreated with 40 mM of hydroxyurea for three days. EGF was immediately added into the conditioned medium after removal of hydroxyurea. Sequential pulses of EdU and BrdU began at 96 h after removal of hydroxyurea. Dual labeling under normal culture conditions (related to [Fig 2B](#pntd.0005418.g002){ref-type="fig"}) was carried out without hydroxyurea treatment, which is: EdU label for 4 hours, no label for 44 hours, and BrdU label for 24 hours. (TIF) ###### Click here for additional data file. ###### EdU^+^BrdU^+^ cells are highly presented in the clonally proliferating germinative cells. Metacestode vesicles were treated with 40 mM hydroxyurea (HU) for seven days and then transferred to HU-free medium. Samples were administrated to EdU-BrdU dual labeling at the day 3 after HU removal. Dashed line boxes in (A) indicate patches of EdU^+^ cells clonally growing. Bar = 100 μm. The magnified views are shown as in (B). Bar = 20 μm. (TIF) ###### Click here for additional data file. ###### Analysis of amino acid sequence and mRNA expression of the EGF receptor members of E. multilocularis. (A)-(C) Amino acid sequence analysis of the receptor-L-domain 1 (A), receptor-L-domain 2 (B) and Pkinase \_Tyr domain (C) of human (Hs), S. mansoni (Sm) and E. multilocularis (Em) EGF receptors. Domains are predicted using the online software (<http://scansite3.mit.edu/>). Positions at which all of the residues are conserved are shaded in black. (D) Similarities of E. multilocularis EGF receptor members to human EGFR. Similarity values to the L-C-L domain (two receptor L domains separated by a cysteine-rich furin-like region) and the kinase domain are indicated below as % identical residues (not bracketed) and % similar residues (bracketed). Further indicated are the similarities of overall protein sequences. (E) RT-PCR analysis of mRNA expression of E. multilocularis EGF receptor members in protoscoleces (lane 1--3) and metacestode vesicles (lane 4--6). Lane 1 and 4: EmER. Lane 2 and 5: Em_000617300. Lane 3 and 6: Em_000969600. M indicates the DNA marker. (TIF) ###### Click here for additional data file. ###### BIBW2992 impairs E. multilocularis germinative cell proliferation, larval growth and development. $A$ Representative images of EdU^+^ germinative cells in the metacestode vesicles following treatment of 5 μM BIBW2992 or DMSO control for 3 days (red: EdU; blue: DAPI). Bar = 20 μm. (B) Effects of BIBW2992 on the larval growth and development. Vesicles or protoscoleces were cultivated in the DMSO-containing conditioned medium (control) supplemented with the ingredients as indicated. Vesicle growth (left) and vesicle formation from protoscoleces (right) were analyzed after 28 days and 18 days of cultivation, respectively. Data are shown as mean ± SD of triplicates, representative of 2--3 independent experiments. \\\* P \< 0.001. (TIF) ###### Click here for additional data file. ###### EGF stimulates the quiescent germinative cells in the developed protoscoleces. Protoscoleces freshly isolated from the metacestode material were in vitro maintained in PBS supplemented with EGF or not for 12h followed by a 4-hour pulse of EdU. Few EdU^+^ cells presented in the developed protoscoleces (arrows), however, the number of EdU^+^ cells dramatically increased after EGF stimulation. The arrow head indicates a developing protoscolex which possesses plenty of EdU^+^ cells. Bar = 100 μm. (TIF) ###### Click here for additional data file. The authors would like to thank Prof. Tang Chongti for providing the parasite material, Dr. Li Guang and Prof. Zuo Zhenghong for their assistance in Xenopus oocyte microinjection. [^1]: The authors have declared that no competing interests exist. [^2]: Conceptualization: ZC YW.Formal analysis: ZC FL XL.Funding acquisition: YW ZC FL HT.Investigation: ZC FL XL MD JW XG HT ZH YL XC.Methodology: ZC FL XL XG HT ZH YL.Project administration: YW ZC.Resources: ZC FL XL MD JW XG HT ZH YL XC.Supervision: YW.Validation: YW.Writing -- original draft: ZC FL YW.	{ "pile_set_name": "PubMed Central" }
In the computer world, there is a constant battle occurring between bad actors that want to attack computing platforms and good actors who try to prevent the same. Unfortunately, the complexity of such computer attacks in constantly increasing, so technology needs to be employed that understands the complexity of these attacks and is capable of addressing the same. Additionally, the use of Artificial Intelligence (AI) and Machine Learning (ML) has revolutionized the manner in which large quantities of content may be processed so that information may be extracted that is not readily discernible to a human user. Accordingly and though the use of AI/ML, the good actors may gain the upper hand in this never ending battle.	{ "pile_set_name": "USPTO Backgrounds" }
Q: Minimum Gower's distance for two data frames I am looking for an implementation that determines the minimum value of Gower's distance for all records in one (say, test) data frame to any record in a second (say, training) data frame. The result is a vector with one element for each row in test. The data are categorical with unordered categorical attributes, and can be generated, for example, like this: set.seed(20130926L) DIMS <- 12 CATS <- 2 create.data <- function(SPARSITY) { sparse.data <- rbinom(CATS ** DIMS, 1, SPARSITY) sparse.array <- array(sparse.data, dim=rep(CATS, DIMS)) sparse.table <- as.table(sparse.array) sparse.df <- as.data.frame(sparse.table) sparse.df <- subset(sparse.df, Freq > 0, select=-Freq) sparse.df } data.train <- create.data(0.001) data.test <- create.data(0.01) head(data.train, 3) ## Var1 Var2 Var3 Var4 Var5 Var6 Var7 Var8 Var9 Var10 Var11 Var12 ## 745 A A A B A B B B A B A A ## 1156 B B A A A A A B A A B A ## 1574 B A B A A B A A A B B A summary(data.test) ## Var1 Var2 Var3 Var4 Var5 Var6 Var7 Var8 Var9 Var10 ## A:24 A:31 A:23 A:20 A:30 A:27 A:22 A:20 A:26 A:23 ## B:24 B:17 B:25 B:28 B:18 B:21 B:26 B:28 B:22 B:25 ## Var11 Var12 ## A:24 A:22 ## B:24 B:26 How do I find, for all rows in data.test, the row in data.training where Gower's distance is minimal (or at least the distance to that particular row)? The code below works, but needs too much memory already for 20 attributes or for more than 2 categories: nrow(data.test) ## [1] 48 library(StatMatch, quietly=T, warn.conflicts=F) apply(gower.dist(data.train, data.test), 2, min) ## [1] 0.3333 0.4167 0.2500 0.5000 0.3333 0.4167 0.2500 0.3333 0.2500 0.4167 ## [11] 0.5000 0.3333 0.3333 0.3333 0.4167 0.4167 0.2500 0.4167 0.1667 0.3333 ## [21] 0.4167 0.3333 0.4167 0.5000 0.3333 0.5000 0.5000 0.4167 0.3333 0.3333 ## [31] 0.2500 0.4167 0.5000 0.4167 0.3333 0.5000 0.3333 0.4167 0.3333 0.3333 ## [41] 0.5000 0.5833 0.5000 0.2500 0.3333 0.4167 0.3333 0.5000 The function cluster::daisy() also returns a matrix of distances. Similar: How to calculate Euclidean distance (and save only summaries) for large data frames. There, it is suggested to call the distance function several times for subsets of data.train. I can do that, but the computation time is still prohibitive. After all, the definition of Gower's distance permits a more efficient algorithm, perhaps a recursive divide-and-conquer approach that operates attribute by attribute and calls itself on subsets. Recall that Gower's distance is a (weighted) sum of attribute-wise distances, which is defined for categorical attributes: 0 if equal, 1 otherwise for ordered attributes: 0 if equal, proportional to rank distance otherwise for continuous attributes (not needed here): proportional to ratio of distance and range of the attribute The following is a simple demonstration where Gower's distance between (A, A) and all combinations of A and B is computed. Rows that differ on one attribute have a distance of 0.5, the row that differs on both attribute gets the maximal distance of 1.0: (ex.train <- expand.grid(Var1=LETTERS[1:2], Var2=LETTERS[1:2])) ## Var1 Var2 ## 1 A A ## 2 B A ## 3 A B ## 4 B B ex.test <- ex.train[1, ] gower.dist(ex.train, ex.test) ## [,1] ## [1,] 0.0 ## [2,] 0.5 ## [3,] 0.5 ## [4,] 1.0 If both train.data and test.data are analyzed column-wise, a possible implementation might look like this: For all value levels v of the first column choose subset of test.data where first column has value v choose subset of train.data where first column has value v call procedure recursively to obtain an upper bound for the minimum choose subset of train.data where first column has value <> v call procedure recursively using the previously obtained upper bound for early cut-off Is there really no implementation around, or perhaps a paper that describes such an algorithm? A: I'm not familiar with Gower's distance, but from what you describe, it appears that, for unordered categorical attributes, Gower's distance is equivalent to the Hamming distance divided by the length of the vector. In other words, the Gower distance between vectors x and y is simply mean(x!=y). In this situation, you can save a significant amount of computation time by avoiding computing the entire distance matrix, and instead using colSums. Here is an example, with three levels and 10000 training rows: > set.seed(123) > train.rows<-10000 > test.rows<-100 > cols<-20 > levels<-c("a","b","c") > train.set<-sample(levels,train.rowscols,T) > dim(train.set)<-c(train.rows,cols) > test.set<-sample(levels,test.rowscols,T) > dim(test.set)<-c(test.rows,cols) > system.time(gdist<-apply(gower.dist(train.set,test.set),2,min)) user system elapsed 13.396 0.324 13.745 > system.time(hdist<-apply(test.set,1,function(x) min(colSums(x!=t(train.set))/cols))) user system elapsed 0.492 0.008 0.504 > identical(hdist,gdist) [1] TRUE If the data is not discrete and unordered, then the formula for Gower's distance is different, but I suspect that there is a similar way to compute this more efficiently without computing the entire distance matrix via gower.dist. Update: this can be made more efficient by using @Frank's suggestion, and generating t(train.set) upfront rather than within the function: require(microbenchmark) ttrain.set<-t(train.set) microbenchmark( a=apply(test.set,1,function(x) min(colSums(x!=t(train.set))/cols)), b=apply(test.set,1,function(x) min(colSums(x!=ttrain.set)/cols))) ## Unit: milliseconds ## expr min lq median uq max neval ## a 523.3781 533.2950 589.0048 620.4411 725.0183 100 ## b 367.5428 371.6004 396.7590 408.9804 496.4001 100	{ "pile_set_name": "StackExchange" }
The head of the Calgary Airport Authority says more changes will be coming to the new international terminal over the next two years. Bob Sartor, CEO of the Calgary Airport Authority, addressed some of the complaints about the new terminal at the authority's annual general meeting Thursday. The new facility opened last fall, but there were complaints from travellers about a lack of seating at the gates, the distance travellers have to go to make their connections, and the fact there are no water fountains. Sartor says seating has been added. Some water stations have been installed, but the issue will take some time to fix. "What we're doing is adding not only water fountains but water stations where you can fill a bottle," Sartor says. "Some of them take time because there's more plumbing to it than I thought," he says. Sartor also says a number of moving sidewalks are being added to shorten the walking time from one end of the building to the other. "We will not be finished completely until 2019 because the connectivity program which brings all of the incremental moving sidewalks from the domestic part of the terminal all the way to D and E won't be finished until then," he says. "We'll open it up in sections but the final section will be in 2019." Sartor says work is going on this year to extend a new baggage handling system into all areas of the old terminal so it connects better with the new terminal.	{ "pile_set_name": "OpenWebText2" }
Rolf Aurness Rolf Aurness was born on February 18, 1952 in Santa Monica, California. He won the 1970 World Surfing Championships held at Johanna in Victoria, Australia, beating Midget Farrelly in the finals. Surfing career When he was nine Aurness suffered a skull fracture after falling from a tree. His father, reported to be an enthusiastic surfer, used surfing to help his son recover. He implemented a strict training regime of dawn sessions at beaches, long distance swimming and weekend beach trips, including the Hollister Ranch. Several times a year they visited Hawaii, renting accommodation on Mākaha beach. Personal life Aurness is the son of Gunsmoke actor James Arness and nephew of Mission Impossible actor Peter Graves. In the decade following his World Surfing Championship win Aurness fell out of surfing as his wife, mother and sister all died. His wife died in 1978 from cancer, his mother Virginia (née Chapman) died in 1976,and his sister Jenny Lee Aurness committed suicide on May 12, 1975. His half-brother Craig founded the stock photography agency Westlight and also was a photographer for National Geographic. His father, well known Western and Gunsmoke television show actor James Arness, died on June 3, 2011. See also References External links The Ranch www.surfline.com. Greg Heller, November 2000 Carroll: Swimming with Marshal Dillon Orange County Register. December 28, 2010 Category:Living people Category:1952 births Category:American surfers	{ "pile_set_name": "Wikipedia (en)" }
Federal investigators close off access to the offices of the New England Compounding Center in Framingham, Massachusetts, October 16, 2012. Criminal investigators from the FDA were on site at the center on Tuesday in the first public indication that the government was preparing a criminal case against the company linked to a deadly outbreak of meningitis. (Photo: Barry Chin / Boston Globe)More Americans than died in the attacks on the World Trade Center die every year from contaminated medicine, food, air and water. Yet the GOP argues we cannot afford the promulgation and enforcement of the regulations that would save these lives. I doubt Lilian Cary, 67, would be voting for Mitt Romney or any Republican this year. Actually, Lilian won’t be voting for anyone next week, because she died of meningitis, weeks after being injected with steroids for back pain. She is one of 25 people so far to have died from contaminated vials of epidural steroids. Her husband was also given those injections at the same clinic and could be facing a similar fate. As of late October, 338 people have been stricken by infection. In all, over 14,000 people have been injected with these contaminated steroids. There will undoubtedly be more illnesses and deaths before this mass tragedy is over. The tainted vials were made by a pharmacy, New England Compounding Center (NECC). A deadly black fungus, Exserohilum, found in the vials was also found throughout the facility. “The entire pharmacy was an incubator of bacteria and fungus. The pharmacy knew this through monitoring results, and chose to do nothing,” said former FDA officer Sarah Sellers who left the agency in 2008 after unsuccessfully pushing it to increase regulation of compounding pharmacies. Employees knew about the unsterile conditions and violated established protocols, but the pharmacy chose to do nothing about it. Inspectors found multiple breaks in sterile techniques, including that employees had turned off the air conditioning at night to save the company money, undermining the conditions needed to prevent growth of contaminating organisms. Every drug the company made is now suspect. Drug manufacturing companies such as NECC, prepare medications for clinics and doctors largely outside federal control. The FDA says that the law does not give it adequate authority to establish oversight, leaving regulation largely to the states. In my state, there is exactly zero oversight of this kind of product. In my speciality, anesthesiology, it isn’t just contaminated drugs; there’s also a shortage of critical drugs. For the last several years, anesthesiologists have had to resort to suboptimal drug selection because some of our most important drugs have simply been unavailable. It seems that the free market doesn’t compel drug companies to make drugs if they don’t want to, or don’t find it profitable, or they can create artificial shortages to drive up prices. Welcome to a glimpse of Mitt Romney’s and the Republican’s Party’s Holy Grail for economic revival – a free market on steroids, and a war on regulation. Virtually all government regulatory functions are under attack. Drug manufacturing, food safety, environmental protections, health care, banking and financial regulations, work place safety, and non-discrimination protection have all been placed on the altar of the free market under the cloak of “freedom, job creation” or the discipline and joy of “personal responsibility.” Mitt Romney types can say that NECC will probably go out of business and in that respect it affirms that the free market works. But in this case – and so many others – the road to free market vindication is littered with dead bodies, lost jobs, lost life savings, and widespread suffering. Lilian Cary and the other NECC victims paid the ultimate price for poor regulations. I suppose in a sick way you could rationalize that even this is a manifestation of how well the free market works, like the “free market” of the Serengeti – the weak and the unlucky fall victim to predators in the jungle of corporate negligence and profiteering. And who could dispute that weak regulation has helped NECC create jobs – although at the morgue. While the Republican’s wage an overt war on regulation, they also wage a covert war by demanding budget cuts to virtually all regulatory agencies, further neutering their ability to function. With the GOP demand for budget austerity, first on their list of things to cut is “discretionary spending,” which, among other things, means spending on enforcing regulations. For example, even though President Obama signed the Food Safety Modernization Act into law – giving the FDA wider power to prevent food-borne illness outbreaks – the number of Americans who became sick or died because of contaminated food increased 44% over the last two years, according to a new report from the US Public Interest Research Group. But the FDA remains unable to implement the reforms because of Republican refusal to fund them. Typical of the Republican thought process is this statement last year from Rep. Jack Kingston (R-GA): “The food supply in America is very safe because the private sector self-polices, because they have the highest motivation. They don’t want to be sued; they don’t want to go broke. They want their customers to be healthy and happy.” This is how wildly successful this self policing strategy is: 76 million illnesses occur annually from contaminated food, with 128,000 requiring hospitalization and 3,000 people dying – every year – as many as died in the terrorist attacks at the World Trade Center (remember contaminated spinach, peanut butter, hamburger, and cantaloupes?). We have spent trillions of dollars on the “War on Terror,” the wars in Iraq and Afghanistan, homeland security, and Mitt Romney wants the sky to be the limit on even more defense spending, yet spending a few million to prevent contaminated food – which kills far more people – is something that the entire Republican Party just can’t tolerate. Every five years, the Clean Air Act requires the EPA to revise national air quality standards to reflect current science. Current science indicates that existing standards are not strict enough and that standards 25% stricter would save about 36,000 lives – over ten times the number of deaths on 9/11 – every year (1). You have a far greater likelihood of dying from air pollution than you do from a terrorist attack. But Mitt Romney and the Republicans will not let up on the idea that clean air and clean water are killing jobs and the big polluters can’t afford it. Voting for them this fall means you will be voting to gut the Clean Air Act and the EPA’s budget, and that of the FDA, the SEC, and virtually every federal regulatory agency whose job it is to protect our citizens. For the Republicans, deregulation has such mythical power they would even have us be fooled by Wall Street into letting them police themselves again. In the last few years, millions of Americans have lost their financial nest egg, the equity in their home, their job, health or even their lives because of the greed and excesses of business. And the Republicans would have you believe that somehow you are better off for it. In the immortal words mangled by George W. Bush, “Fool us once, shame on them, fool us twice, shame on us.” Lilian Cary was fooled into thinking her government was looking out for her. It wasn’t. And if Romney and the Republicans fool Americans into voting for their war on regulation, we will have become a ship of fools indeed. References 1. Report by the American Lung Association, Earthjustice, and Clean Air Task Force -How the EPA can save lives by cleaning up fine particle pollution. 2011.	{ "pile_set_name": "OpenWebText2" }
Prediction of CYP-mediated drug interactions in vivo using in vitro data. Over the past 15 years, a concerted effort has been undertaken by the pharmaceutical industry to reduce the attrition of clinical candidates resulting from undesirable ADME characteristics. Increasing regulatory and competitive pressures demand that pharmaceutical products brought to the market possess pristine safety and drug co-administration profiles for most therapeutic areas. The high-profile withdrawal of drugs such as mibefradil from the market because of unfavorable drug-drug interaction profiles has focused efforts on screening for cytochrome P450 (CYP)-mediated drug interactions early in the discovery paradigm and on predicting the impact of inhibition on the in vivo situation. This paper discusses current practices used to screen for CYP-mediated drug-drug interactions in vitro (inhibition and induction) and how these data are being used to predict whether a clinically relevant drug-drug interaction is likely to occur in vivo.	{ "pile_set_name": "PubMed Abstracts" }
<?php /* Copyright (c) 1998-2009 ILIAS open source, Extended GPL, see docs/LICENSE / /* * Saves (mostly asynchronously) user properties of accordions * * @author Alex Killing <alex.killing@gmx.de> * @version $Id$ * @ingroup ServicesAccordion * @ilCtrl_Calls ilAccordionPropertiesStorage: / class ilAccordionPropertiesStorage { /* * @var ilObjUser / protected $user; /* * @var ilCtrl / protected $ctrl; /* * @var ilDB / protected $db; /* * Constructor / public function __construct() { global $DIC; $this->user = $DIC->user(); $this->ctrl = $DIC->ctrl(); $this->db = $DIC->database(); } public $properties = array( "opened" => array("storage" => "session") ); /* * execute command / public function &executeCommand() { $ilUser = $this->user; $ilCtrl = $this->ctrl; $cmd = $ilCtrl->getCmd(); // $next_class = $this->ctrl->getNextClass($this); $this->$cmd(); } /* * Show Filter / public function setOpenedTab() { $ilUser = $this->user; if ($_GET["user_id"] == $ilUser->getId()) { switch ($_GET["act"]) { case "add": $cur = $this->getProperty( $_GET["accordion_id"], (int) $_GET["user_id"], "opened" ); $cur_arr = explode(";", $cur); if (!in_array((int) $_GET["tab_nr"], $cur_arr)) { $cur_arr[] = (int) $_GET["tab_nr"]; } $this->storeProperty( $_GET["accordion_id"], (int) $_GET["user_id"], "opened", implode($cur_arr, ";") ); break; case "rem": $cur = $this->getProperty( $_GET["accordion_id"], (int) $_GET["user_id"], "opened" ); $cur_arr = explode(";", $cur); if (($key = array_search((int) $_GET["tab_nr"], $cur_arr)) !== false) { unset($cur_arr[$key]); } $this->storeProperty( $_GET["accordion_id"], (int) $_GET["user_id"], "opened", implode($cur_arr, ";") ); break; case "clear": $this->storeProperty( $_GET["accordion_id"], (int) $_GET["user_id"], "opened", "" ); break; case "set": default: $this->storeProperty( $_GET["accordion_id"], (int) $_GET["user_id"], "opened", $_GET["tab_nr"] ); break; } } } /* * Store property in session or db / public function storeProperty( $a_table_id, $a_user_id, $a_property, $a_value ) { $ilDB = $this->db; switch ($this->properties[$a_property]["storage"]) { case "session": $_SESSION["accordion"][$a_table_id][$a_user_id][$a_property] = $a_value; break; case "db": / $ilDB->replace("table_properties", array( "table_id" => array("text", $a_table_id), "user_id" => array("integer", $a_user_id), "property" => array("text", $a_property)), array( "value" => array("text", $a_value) )); / } } /* * Get property in session or db / public function getProperty($a_table_id, $a_user_id, $a_property) { $ilDB = $this->db; switch ($this->properties[$a_property]["storage"]) { case "session": $r = $_SESSION["accordion"][$a_table_id][$a_user_id][$a_property]; //echo "<br><br><br><br><br><br><br><br>get-".$r; return $r; break; case "db": / $set = $ilDB->query("SELECT value FROM table_properties ". " WHERE table_id = ".$ilDB->quote($a_table_id, "text"). " AND user_id = ".$ilDB->quote($a_user_id, "integer"). " AND property = ".$ilDB->quote($a_property, "text") ); $rec = $ilDB->fetchAssoc($set); return $rec["value"]; break; */ } } }	{ "pile_set_name": "Github" }
A dual-sensitive mesoporous silica nanoparticle based drug carrier for cancer synergetic therapy. A multifunctional envelope-type mesoporous silica nanoparticle (MSN) was delicately designed for subcellular co-delivery of drug and therapeutic peptide to tumor cells. Firstly, a kind of cell apoptosis peptide (KLAKLAK)2 (KLA) was anchored on surface of MSN via disulfide bond to obtain MSN-SS-KLA. Subsequently, anticancer drug doxorubicin hydrochloride (DOX) was loaded into the pores of MSN-SS-KLA. Then, the drug loaded MSN-SS-KLA (DOX@MSN-SS-KLA) was further coated with bovine serum albumin (BSA) to obtain a biological media stable MSN based drug delivery system (DDS), DOX@MSN-SS-KLA/BSA, for cancer synergetic therapy. The results show that stability of the DOX@MSN-SS-KLA/BSA is much better than that of DOX@MSN-SS-KLA and it could keep well dispersed in serum for more than 24 h. After accumulating at tumor site by EPR effect, the DOX@MSN-SS-KLA/BSA could be effectively phagocytosed by HeLa cells and release apoptotic peptide KLA as well as DOX simultaneously responding to reductive stimulus inside the cells. In vitro cell experiment results show that the DOX@MSN-SS-KLA/BSA complex exhibits much better inhibition on HeLa cells compared with pure DOX, indicating that co-delivery of KLA and DOX is expected to achieve synergetic therapy of cancer.	{ "pile_set_name": "PubMed Abstracts" }
Q: Merging a vector layer with a Raster layer I have a Tiff file that has an aerial view and also a shapefile which has contours of the same area. How can I put the contours in the tiff file so that you can view the TIFF and see them both in one picture. I am using QGIS. A: Bring in a shapefile of your contours and make sure they have an elevation attribute with them. Go to Raster > Conversion > Rasterize and select your shapefile, the elevation attribute, and then your existing Tiff file you want to put the contours on. Press run and you will get a tiff with the contours on top of the layer. The following link is very helpful to accomplish this : How to control line width when merging vector files into raster? . Thank you @Pooneil	{ "pile_set_name": "StackExchange" }
Description Both cozy and chic, these faux animal fur blankets and plush friends are great for everyday outings, and make a wonderful welcome gift for a new baby. Plush: 14" wide x 20" high x 5" deep Stroller Blanket: 30 ...	{ "pile_set_name": "Pile-CC" }
Yeah, and a big thanks to the people that didn't make an OST for that game. I had to record the whole song via an extension cable running from my PC to my line-in on my stereo. It took an hour to set up, and then another half hour to get right.	{ "pile_set_name": "Pile-CC" }
Maintaining standards of aesthetic practice in trainees subject to NHS restrictions. The Specialist Advisory Committee (SAC) in plastic surgery within the United Kingdom (UK) recommends a modular training programme to include aesthetic surgery. The intercollegiate board examinations test candidates on all aspects of aesthetic practice yet there is no formal, national aesthetic training in the UK. Closure of National Health Service (NHS) private patient facilities has reduced training opportunity [Nicolle FV. Sir Harold Gillies Memorial Lecture; Aesthetic plastic surgery and the future plastic surgeon. Br J Plast Surg 1998;51:419-24.] Calmanisation [Hospital doctors: training for the future. The Report of the Working Group on Specialist Medical Training (The Calman Report). London: HMSO; 1993.], the European Working Time Directive (EWTD) [; Phillips H, Fleet Z, Bowman K. The European Working time Directive-interim report and guidance from The Royal College of Surgeons of England working party chaired by Mr Hugh Phillips; 2003 []; Chesser S, Bowman K, Phillips H. The European Working Time Directive and the training of surgeons. BMJ Careers Focus 2002;s69-7.], and more importantly the implementation of "local" aesthetic guidelines have placed further pressures on training. Reductions of NHS case mix will ultimately lead to a reduction in trainee experience. With increasing regulatory pressure from the Commission for Healthcare Improvement, standards of aesthetic practice can only be maintained by increasing private/independent sector involvement. At present a disparity exists between the demand and provision of aesthetic surgery training in the UK. Aesthetic surgery forms part of the training curriculum for plastic surgery and as such remains a training issue. A review of aesthetic surgery training is needed in the UK through consultation with trainers and trainee representatives.	{ "pile_set_name": "PubMed Abstracts" }
File this under I had no idea this existed. During the early 20th century residents of Fort Bragg, California chose to dispose of their waste by hurling it off the cliffs above a beach. No object was too toxic or too large as household appliances, automobiles, and all matter of trash were tossed into the crashing waves below, eventually earning it the name The Dumps. In 1967 the North Coast Water Quality Board closed the area completely and initiated a series of cleanups to slowly reverse decades of pollution and environmental damage. But there was one thing too costly (or perhaps impossible) to tackle: the millions of tiny glass shards churning in the surf. Over time the unrelenting ocean waves have, in a sense, cleansed the beach, turning the sand into a sparkling, multicolored bed of smooth glass stones now known as Glass Beach. The beach is now an unofficial tourist attraction and the California State Park System has gone so far as purchasing the property and incorporating it into surrounding MacKerricher State Park. (images courtesy digggs, matthew high, meganpru, lee rentz and linked to sources. via kuriositas) Do stories and artists like this matter to you? Become a Colossal Member and support independent arts publishing. Join a community of like-minded readers who are passionate about contemporary art, help support our interview series, gain access to partner discounts, and much more. Join now!	{ "pile_set_name": "OpenWebText2" }
Q: UIControl Subclass - Events called twice I'm currently working on a custom UIControl Subclass. To track the touches I use the following Method: - (BOOL)beginTrackingWithTouch:(UITouch )touch withEvent:(UIEvent )event { NSLog(@"Start"); CGPoint location = [touch locationInView:self]; if ([self touchIsInside:location] == YES) { //Touch Down [self sendActionsForControlEvents:UIControlEventTouchDown]; return YES; } else { return NO; } } This works as expected and @"Start" is loged exactely once. The next step is that I add a Target and a Selector with UIControlEventTouchDown. [markItem addTarget:self action:@selector(action:) forControlEvents:UIControlEventTouchUpInside]; This works also and the action: method is called. But that's my problem. The action is called twice. What am I doing wrong? I just use [self sendActionsForControlEvents:UIControlEventTouchDown]; once and the target action is called twice. What's wrong with my code? Sandro Meier A: The first call to the action method happens automatically by the event dispatcher once you've called: [markItem addTarget:self action:@selector(action:) forControlEvents:UIControlEventTouchUpInside]; to register the handler. So when you then call: //Touch Down [self sendActionsForControlEvents:UIControlEventTouchDown]; you are generating the second call to your handler (and any others that are hooked up). So it seems like you don't need both the action handler and the beginTracking - use one or the other. Update: Given your comment and further thought: since you are a subclass of UIControl, I think you probably don't want to be registering for event handlers for yourself. Instead you should exclusively use: - (BOOL)beginTrackingWithTouch:(UITouch )touch withEvent:(UIEvent )event; - (BOOL)continueTrackingWithTouch:(UITouch )touch withEvent:(UIEvent )event; - (void)endTrackingWithTouch:(UITouch )touch withEvent:(UIEvent )event; - (void)cancelTrackingWithEvent:(UIEvent *)event; // event may be nil if cancelled for non-event reasons, e.g. removed from window Also the tracking instance variable. So I think you should not be posting events or listening to events. Further, is it actually possible to get a beginTrackingWithTouch event if it's not in your view? Doesn't seem like it would be. So I don't think you need the testing to see if it's in your view. So I think it might be worth stepping back and thinking about what you are trying to do and re-reading UIControl documentation. Specifically: Subclassing Notes You may want to extend a UIControl subclass for either of two reasons: To observe or modify the dispatch of action messages to targets for particular events To do this, override sendAction:to:forEvent:, evaluate the passed-in selector, target object, or UIControlEvents bit mask, and proceed as required. To provide custom tracking behavior (for example, to change the highlight appearance) To do this, override one or all of the following methods: beginTrackingWithTouch:withEvent:, continueTrackingWithTouch:withEvent:, endTrackingWithTouch:withEvent:. The first part is for having your UIControl subclass do non-standard handling of target action processing for clients or users of your control (that doesn't sound like what you are trying to do, though you didn't really give a high-level description). The second part sounds more like what you are wanting to do - custom tracking within your UIControl subclass.	{ "pile_set_name": "StackExchange" }
15 2 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3	{ "pile_set_name": "Github" }
Bioruns private limited. OMORFEE, an Indo – German venture of Bioruns Private Limited, is an organic personal care brand well known for its purity and efficacy. It offers a luxurious collection of personal care products made from organic botanicals. The rich scents and therapeutic formulations of the products help to heal, restore and renew the skin, body and soul. FOLLOW US ON FACEBOOK KEEP IN TOUCH Franchising Roots is a venture of FR Business Services PVT. LTD.The information contained herein is of generic nature only and Franchising Roots is not by means of this website or any part thereof rendering professional advice or services. With respect to use with the website, kindly visit www.franchisingroots.com/terms-of-service/	{ "pile_set_name": "Pile-CC" }
Q: How would you do multiple operations on a java 8 stream? You have a list of a pojo you want to convert to a JSONObject for instance. You have a list of pojo. But in order to convert to a JSONObject you need to use the JSONObject put method. JSONObject personJson = new JSONObject(); for(Person person : personList){ personJson.put("firstName", person.firstName); personJson.put("lastName", person.lastname); ... } If it were just one operation I wanted to do, then I could do personList.stream.map(personJson.put("firstName", person.firstName)); A: JSONArray array=new JSONArray(); personList.stream().forEach(element -> { JSONObject personJson = new JSONObject(); personJson.put("firstName", element.firstName); personJson.put("lastName", element.lastname); array.add(personJson); }); A: The solutions mentioned here are working but they are mutating objects outside the streams context, which should be avoided. So instead of using .forEach() which is a terminating operation returning void. The correct approach would be to use .map() which, like the name says, maps one value to another. In your case the first operation you want to do is to map a Person to a JSONObject. The second operation is a reducer function where you want to reduce all JSONObjects to one JSONArray object. public JSONArray mapListToJsonArray(List<Person> persons) { List<JSONObject> jsonObjects = persons .stream() .map(person -> { JSONObject json = new JSONObject(); json.put("firstName", person.getFirstName()); json.put("lastName", person.getLastName()); return json; }) .collect(Collectors.toList()); return new JSONArray(jsonObjects); } Unfortunately the json.org implementation of JSONArray does not have a way to easily merge two arrays. So I instead of reducing to a JSONArray, I first collected all JSONObjects as a List and created a JSONArray from that. The solution looks even nicer if you replace the lambda expression with a method reference. public JSONArray mapListToJsonArray(List<Person> persons) { List<JSONObject> jsonObjects = persons .stream() .map(this::mapPersonToJsonObject) .collect(Collectors.toList()); return new JSONArray(jsonObjects); } public JSONObject mapPersonToJsonObject(Person person) { JSONObject json = new JSONObject(); json.put("firstName", person.getFirstName()); json.put("lastName", person.getLastName()); return json; } A: personList.forEach(person -> { personJson.put("firstName",person.firstName); personJson.put("lastName", person.lastName); });	{ "pile_set_name": "StackExchange" }
Q: Initialization Pattern? Say I have class A: public class A { private B _b; public class A(B b) { Assert.That(b != null); _b = b; } } And object b needs some complex initialization, like: b.Prop1 = ... b.Prop2 = ... b.Prop3 = ... int answerToSomeComplexFormula = PerformComplexFormula(); b.Prop4 = answerToSomeCopmlexFormula etc... I do not want to perform this initialization in the constructor. Is there a name for some pattern that describes returning an object that has complex initialization? Something like: public class BInitializer { public B Create() { B b = new B(); // set up properties return b; } } BInitializer initializer = new BInitializer(); B b = initializer.Create(); A a = new A(b) Thanks! A: Your solution with BInitializer is very good and it is called Factory Method design pattern. Here you can find some common creational design patterns: https://sourcemaking.com/design_patterns/creational_patterns	{ "pile_set_name": "StackExchange" }
Q: How should I get rid of TFS Branches properly? I am wondering how I should properly get rid of branches that no longer have any purpose. Right now even if i delete them and commit they are still listed as branches in the properties windows for a particular branching root (directory). If I select merge I don't get an option to merge to the deleted branch which obviously is as expected but therefore I am puzzled about the branch still showing up in the properties window. Any explanation on this behavior would be greatly appreciated. A: I had a situation where a branch had been deleted and there was no purpose for it to stick around. I couldn't get tf destroy to work until I found out the deletion number of the directory like Damien mentioned. I couldn't get that ID with tf properties since there was no local copy and no server copy based on the error messages. I was able to get the full TFS path by using: tf dir $/MyPathTo/TheParent/Directory /deleted Then I found the postfixed ";Ident" to the directory and could issue: tf destroy $/MyPathTo/TheParent/Directory/TheDirectoryToGetRidOff;Ident Damien your answer helped me out - thanks. I thought I'd include the syntax that got me through it to go along with your post. A: The deleted branch is only marked as deleted at a moment in time and it's possible to still get to the code if you sync to a changeset or time before that delete operation. If you do not want to have the file in the branch in the database at all you can use the tf destroy command line: http://msdn.microsoft.com/en-us/library/bb386005.aspx As your branch is already deleted you'll need to use tf dir /deleted to find the deletion number of that branch in order to destroy the files.	{ "pile_set_name": "StackExchange" }
Q: How to fix Permission denied (publickey,password) in gitlab Ci/CD? I have below simple script in .gitlab-cl.yml file: build_deploy_stage: stage: build environment: Staging only: - master script: - mkdir -p ~/.ssh - echo "$PRIVATE_KEY" >> ~/.ssh/id_dsa - cat ~/.ssh/id_dsa - chmod 600 ~/.ssh/id_dsa - echo -e "Host *\n\tStrictHostKeyChecking no\n\n" > ~/.ssh/config - cat ~/.ssh/config - scp myfile.js user@example.com:~ But I get this error when job is run, executing the last line (scp command): Warning: Permanently added 'example.com' (ECDSA) to the list of known hosts. Permission denied, please try again. Permission denied, please try again. Permission denied (publickey,password). I spent whole day but could not fix it. I verified that $PRIVATE_KEY exists. I generated key pair while logged into example.com copying the generated private to PRIVATE_KEY variable on gitlab. How to fix this problem? Note that it is dsa key. A: Check your permission for ~/.ssh (700) and all the files in them (600) Your config file, for instance, might have default permissions that are too large. If you can, activate a debug session in the sshd of the remote server: you will see if an dsa key is accepted (for recent version of sshd, that might be restricted). rsa would be better. As seen here, OpenSSH 7.0 and higher no longer accept DSA keys by default. The OP ace confirms in the comments: I fixed the problem when I regenerated tsa key pairs instead of dsa keys	{ "pile_set_name": "StackExchange" }
Q: Framework7 dynamic AJAX loaded pages not working I am building a framework7 app and since lots of editing has been done to one page, the AJAX loaded in pages have stopped working. I have a feeling this is to do with the structure as I ran into this problem once before, but I need someone with more experience or a better brain to help me solve this. The problem is with the links at the end of the code that link to the chat.html page. They just do not work. Thanks A: After trial and error I found that enabling inline pages fixed my problem even though the pages I am loading in are not inline. var mainView = myApp.addView('.view-main', { domCache: true //enable inline pages });	{ "pile_set_name": "StackExchange" }
This invention relates generally to the storage and subsequent recovery of compressed air energy and more specifically to the retrofit of simple cycle gas turbine engines for compressed air energy storage application. It will be appreciated that there exists a significant daily variation in the total amount of power required from a baseload power plant. However, it would be most costly and wasteful to provide for the peak demands of short duration with the same expensive, although efficient, baseload power plant machinery which is justified for baseload power generation. In the past, power utilities have addressed the problem of providing additional non-baseload peaking power in an overall economical manner through the use of simple cycle gas turbines (SCGTs). Simple cycle gas turbines (SCGTs) are state-of-the-art turbomachinery designed for peaking duty operation. Their main elements are an air compressor, a turbine and a combustor. To meet consumer demand, SCGTs are coupled to electric generators to provide additional power to baseload power plants during peak loads. Most recently, power utilities have addressed the aforementioned problem of supplying peaking power in an overall cost effective manner through the use of Compressed Air Energy Storage (CAES) systems. CAES plants store off-peak energy from relatively inexpensive energy sources such as coal and nuclear baseload plants by compressing air into storage devices such as underground caverns or reservoirs. Underground storage can be developed in hard rock, bedded salt, salt dome or aquifer media. Following off-peak storage, the air is withdrawn from storage, heated, combined with fuel and expanded through expanders, i.e., turbines, to provide needed peaking or intermediate power. Since inexpensive off-peak energy is used to compress the air, the need for premium fuels, such as natural gas and imported oil, is reduced by as much as about two-thirds compared with conventional gas turbines. In such circumstances, utilization of non-peak energy for the subsequent production of peak energy is clearly desirable, especially since non-peak energy can often be obtained for far less than the cost of obtaining peak energy. Compressors and turbines in CAES plants are each connected to a synchronous electrical machine such as a generator/motor device through respective clutches, permitting operation either solely of the compressors or solely of the turbines during appropriate selected time periods. During off-peak periods (i.e., nights and weekends), the compressor train is driven through its clutch by the generator/motor. In this scheme, the generator/motor functions as a motor, drawing power from a power grid. The compressed air is then cooled and delivered to underground storage. During peak/intermediate periods, with the turbine clutch engaged, air is withdrawn from storage and provided to a combustor. The combustor combines the pre-heated compressed air with a fuel, such as No. 2 fuel oil, and expands the mixture of fuel and compressed air in a turbine, which provides power by driving the generator/motor. In this scheme, the generator/motor functions as a generator, providing power to a power grid. To improve the CAES heat rate, waste heat from a low pressure turbine exhaust is used to pre-heat high pressure turbine inlet air in a recuperator. For a more complete discussion of CAES systems, see Nakhamkin, M. et al. "Compressed Air Energy Storage: Plant Integration, Turbomachinery Development", ASME International Gas Turbine Symposium and Exhibition, Beijing, Peoples' Republic of China, 1985 and Nakhamkin, M. et al. "Compressed Air Energy Storage (CAES): Overview, Performance and Cost Data for 25 MW to 220 MW Plants", Joint Power Generation Conference, Toronto, Canada 1984, both incorporated herein by reference. Unfortunately, SCGTs require the use of significantly more fuel than CAES systems in order to provide a like quantity of peaking power. Additional considerations such as that of fuel quality required for proper operation and efficient use of previously stored energy also tend to favor CAES systems. However, numerous SCGT peaking power plants are presently in existence and do provide a means for supplying peaking power, albeit at considerable expense. Due to the considerable expense in constructing new CAES plants it is most desirable to convert existing simple cycle gas turbine plants to CAES plants. Unfortunately, direct use of SCGTs in the compression and expansion trains of CAES systems such as those described in the cross-referenced applications presents difficulty due to SCGT maximum cycle pressures which are generally too low for typical underground storage depths and corresponding compressed air storage pressures required in typical CAES air storage and operation. An additional difficulty encountered in direct use of SCGTs in CAES systems relates to pressure losses generally occurring in underground air storage devices.	{ "pile_set_name": "USPTO Backgrounds" }
Q: Display solution/file path in the Visual Studio IDE I frequently work with multiple instances of Visual Studio, often working on different branches of the same solution. Visual C++ 6.0 used to display the full path of the current source file in its title bar, but Visual Studio 2005 doesn't appear to do this. This makes it slightly more awkward than it should be to work out which branch of the solution I'm currently looking at (the quickest way I know of is to hover over a tab so you get the source file's path as a tooltip). Is there a way to get the full solution or file path into the title bar, or at least somewhere that's always visible, so I can quickly tell which branch is loaded into each instance? A: This is a extension available in the online gallery specifically tailored for this job. Checkout Labs > Visual Studio Extension: Customize Visual Studio Window Title. A: There is not a native way to do it, but you can achieve it with a macro. The details are described here in full: How To Show Full File Path (or Anything Else) in VS 2005 Title Bar You just have to add a little Visual Basic macro to the EvironmentEvents macro section and restart Visual Studio. Note: The path will not show up when you first load Visual Studio, but it will whenever you change which file you are viewing. There is probably a way to fix this, but it doesn't seem like a big deal. A: Check out the latest release of VSCommands 2010 Lite. It introduced a feature called Friendly Solution Name where you can set it to display the solution file path (or any part of it) in Visual Studio's main window title. More details: http://vscommands.com/releasenotes/3.6.8.0 and http://vscommands.com/releasenotes/3.6.9.0	{ "pile_set_name": "StackExchange" }
Leo Cantor Leo Cantor (February 28, 1919 – June 4, 1995) was an American football defensive back. He played for the New York Giants in 1942 and for the Chicago Bears in 1945. References Category:1919 births Category:1995 deaths Category:American football defensive backs Category:UCLA Bruins football players Category:New York Giants players Category:Chicago Bears players	{ "pile_set_name": "Wikipedia (en)" }
Excelsior College - Engineering Ranking 2015 in the USAExcelsior College is a very large not-for-profit college with some focus on engineering programs and located in Albany, New York. The school was opened in 1971 and is presently offering associate's and bachelor's degrees in 4 engineering programs. According to recent analysis, Excelsior College area is safe: the college is reported to have an excellent rating for on-campus secutiy. Excelsior College engineering program ranks #365 (out of 1871; top 20%) in the United States and #30 in New York. Major competing engineering colleges for this school are Massachusetts Institute of Technology in Cambridge and Stevens Institute of Technology in Hoboken. See the details about all twelve competing engineering colleges as well as list of offered engineering majors and minors below. Engineering programs ratings 2014-2015: Overall rating: Curriculum rating: Teaching rating: Safety rating: Overall school rankings 2015: Ranked #30 Engineering School in New YorkRanked #89 Engineering School in The NortheastRanked #365 Engineering School in USA	{ "pile_set_name": "Pile-CC" }
Героїзація Бандери ніяк не вплине на відносини з Україною - посол Польщі 23 лютого 2017, 18:42 Вы также можете прочесть этот материал на русском языке Пекло: Питання Бандери - це минуле, важливіше майбутнє (Фото: Фото: polskieradio.p) Україні та Польщі необхідно думати про майбутнє, а питання Степана Бандери - це минуле, заявив посол Польщі в Україні Ян Пєкло Таку позицію дипломат висловив у відповідь на запитання журналіста з проханням прокоментувати нещодавню заяву лідера правлячої партії Польщі Право і Справедливість (Піс) Ярослава Качинського про те, що "Україна не ввійде в Європу з Бандерою", передає Liga.net. "Це історія. Історія важлива для поляків, і для українців. Але є і майбутнє, і майбутнє важливіше. Нам потрібно думати про майбутнє. Качиньський також думає про майбутнє... Багато хто, напевно, вже забули, як він під час Революції Гідності був на Майдані і кричав "Слава Україні!". Це яскравий приклад, що він проукраїнський, а не навпаки", - відповів Пекло. Польський дипломат підкреслив, що питання героїзації Степана Бандери в Україні ніяк не вплине на відносини двох держав. Глава Інституту національної пам'яті Володимир В'ятрович прокоментував різкі випади польського лідера. На його думку, Качиньський намагається нав'язати українцям власне викривлене уявлення про історію свого народу.	{ "pile_set_name": "OpenWebText2" }
Q: Usage of "call" when arranging a phone call Imagine I'm one the phone with somebody and want to postpone the call. I know i can say I'll call you tomorrow or Call me tomorrow But what if I don't want to specify who is calling who? Can I use "call" too? Like Let's call tomorrow What would be the common term to express something like this? A: You can say "Let's have a call tomorrow" or "Let's call each other tomorrow". Note that "have a call" has a slight sense of this being a business call. If you want to be less formal, you might use "let's talk tomorrow" instead. A: Why would you EVER want to not specify who is making the call? If you don't specify, then in all likelihood, you are going to wait for them to call and they are going to wait for you to call, meaning the call will never happen. I suggest, instead, ask this; We should talk about this tomorrow, what would be a good time for me to call?	{ "pile_set_name": "StackExchange" }
--- abstract: 'We study strongly correlated phases of a pseudo-spin-1/2 Bose gas in an artificial gauge field using the exact diagonalization method. The atoms are confined in two dimensions and interact via a two-body contact potential. In Abelian gauge fields, pseudo-spin singlets are favored by pseudo-spin independent interactions. We find a series of incompressible phases at fillings $\nu=2k/3$. By comparison with the non-Abelian spin singlet (NASS) states, constructed as zero-energy eigenstates of a $(k+1)$-body contact interaction, we classify the non-trivial topology of the states. An additional spin-orbit coupling is shown to switch between NASS-like states and spin-polarized phases from the Read-Rezayi series.' author: - 'T. Graß$^1$, B. Juliá-Díaz$^{1}$, N. Barberán$^2$, and M. Lewenstein$^{1,3}$' bibliography: - 'lett.bib' title: 'Non-Abelian spin singlet states of two-component Bose gases in artificial gauge fields' --- Topological phases of matter attract a great deal of attention since the late 1980s, when the fractional quantum Hall (FQH) effect of electrons in two dimensions (2D) subjected to a strong perpendicular magnetic field was recognized as a paradigm of such phases [@wen-niu]. It was then pointed out that amongst these states there might be some which support non-Abelian excitations, where the exchange of two quasiparticles is described by some unitary matrix, as it shifts between different degenerate ground states [@moore-read]. Recently, this property has triggered the interest in topological phases, as quantum gates operating with non-Abelian anyons might pave the way to fault-tolerant quantum computers [@nayak]. For this purpose, the possibility of controlling or even designing quantum many-body states seems to be advantageous if not inevitable. A very important tool to do so are cold atoms which can, in various ways, be manipulated through interactions with light [@mlbook]. Despite being electroneutral, the atoms may even couple to external gauge potentials, which, instead of being true electromagnetic vector potentials, have to be synthesized. Artificial magnetic fields have been achieved experimentally by rotating the atomic cloud [@schweikhard] (see also [@cooper-aip] for a review), though instabilities at high rotation frequencies have so far prevented from reaching the highly correlated regime. A promising alternative pursues the generation of gauge fields by a laser dressing of the atoms [@dalibard; @brunoPRA; @brunoNJP]. This method has already been proven experimentally [@lin; @spielmanPRL]. It is readily generalized to non-Abelian gauge fields [@fleischhauer], and has recently allowed for synthesizing a spin-orbit coupling [@spielman-sobec]. The strongly correlated states occurring in such systems have been numerically investigated for the case of spinless or spin-polarized bosons [@cooper-wilkin-gunn], in which the incompressible phases are well described by the Read-Rezayi (RR) series of parafermionic states [@read-rezayi], including the Laughlin wave function [@laughlin], and the Moore-Read (MR) wave function [@moore-read]. Although also bosons with a pseudo-spin-1/2 degree of freedom have already been condensed [@ketterle-2comp; @wieman-2comp], and their behavior under rotation has been studied both theoretically [@mueller-ho-vortices; @reiman-1vortex; @ueda-vortices] and experimentally [@cornell-vortices; @cornell-vortex-lattice], few attention has yet been paid to the strongly correlated regime in such systems. For the electronic FQH effect, however, a variety of states have been proposed for systems with spin, which despite the strong magnetic field might be in a singlet phase due to the small gyromagnetic ratio of electrons in many solids. Within a conformal field theory (CFT) framework, a generalization of the RR series to the so-called non-Abelian spin singlet (NASS) series [@ardonne-schoutens; @nass-nucl; @estienne-bernevig] has been formulated. Here the term “non-Abelian” refers to the nature of the excitations. For bosons, this series of topological states occurs at fillings $\nu=2k/3$, and includes the Halperin state [@halperin] with Abelian excitations at $k=1$. Like the RR states, the NASS states are constructed as the zero-energy eigenstates of a repulsive $(k+1)$-body contact interaction. In this paper we show, using exact diagonalization, that also two-body contact (2BC) interaction leads to the formation of incompressible states precisely at the NASS filling factors in a 2D Bose gas exposed to an Abelian gauge field. For $k=1$, this is trivially the Halperin state, but also for $k=2$ significant overlap with the corresponding NASS state and a similar spectral structure indicate that the system can be brought into the NASS phase within this realistic setup. This finding is in close analogy to the spin-polarized case with RR-like states [@cooper-wilkin-gunn] at filling $\nu=k/2$. However, as it is experimentally difficult to get into the regime of low filling, the two-component system benefits from supporting analogs of the RR states at higher filling. Furthermore, its possibility of tuning independently inter-spin and intra-spin interactions provides additional control. For pseudo-spin-independent repulsive 2BC interactions, a mechanism is discussed which favors pseudo-spin singlet states. We show that, counterintuitively, states with zero interspin interaction energy are obtained by increasing the interspin interaction strength. Finally, effects stemming from a spin-orbit (SO) coupling can be integrated in the framework of two-component gases by generalizing to a non-Abelian gauge field. We investigate this scenario, and find a transition from the single-component RR states [@trombettoni; @trombettoniPRA; @palmer-pachos; @komineas-cooper] to the NASS states at SO coupling strengths corresponding to a Landau level (LL) degeneracy. We consider a two-component Bose gas in 2D, which on the single-particle level is described by the Hamiltonian $H_{\mathrm{sp}}=(\vec{p}-\vec{A})^2/(2m)$. The vector potential $\vec{A}$ is given by $$\begin{aligned} \label{A} \vec{A} = B (0,x,0) \mathbb{1} + q (\sigma_x,\sigma_y,0).\end{aligned}$$ The first term, acting equally on both pseudo-spin components, describes a magnetic field of strength $B$ in the Landau gauge. The second term, with $\sigma_i$ being the Pauli matrices, accounts for a Rashba SO coupling controlled by the coupling strength $q$, which we set to zero in the first part of this work. A detailed description how to implement this Hamiltonian for electroneutral atoms can be found in the supplementary material of Ref. [@trombettoni]. As the boundary conditions we choose a periodic square of length $a$. This eliminates edge effects from our study, and, moreover, generates a non-trivial toroidal geometry, on which topological phases can be classified by the number of degenerate ground states [@wen-niu]. Solving the single-particle Hamiltonian yields, for $q=0$, the usual LL structure [@yoshioka-halperin-lee], where every state ${\left\|n,j,s\right\rangle}$ within a fully degenerate Landau level $n$ is characterized by a momentum quantum number $j$ and an additional two-fold degenerate spin quantum number $s=\uparrow,\downarrow$. Since an energy gap $2B$ separates the different LLs, we restrict ourselves to the lowest LL, $n=0$. Fixing the number of fluxes per unit cell, $N_{\Phi}$, restricts $j$ to values running from 1 to $N_{\Phi}$, and the many-body Hamiltonian is, up to a constant amounting for the kinetic energy, given by $$\begin{aligned} \label{H} {\ensuremath{\Hat{\mathrm{H}}}} = \sum_{\{j,s\}} V_{\{j,s\}} {\ensuremath{\Hat{\mathrm{a}}}}_{j_1s_1}^{\dagger} {\ensuremath{\Hat{\mathrm{a}}}}_{j_2s_2}^{\dagger} {\ensuremath{\Hat{\mathrm{a}}}}_{j_3s_3}{\ensuremath{\Hat{\mathrm{a}}}}_{j_4s_4},\end{aligned}$$ where ${\ensuremath{\Hat{\mathrm{a}}}}_{js}$ annihilates a particle in state ${\left\|0,j,s\right\rangle}$, and $\{j,s\}$ denotes the set of quantum numbers $j_1,\dots,j_4$ and $s_1,\dots,s_4$. The interaction conserves total momentum $J=\sum_i j_i \ \mathrm{mod} \ N_{\Phi}$, and the spin of the particles, so the coefficients read $$\begin{aligned} \label{V} V_{\{j,s\}} &=& \delta'_{j_1+j_2,j_3+j_4} \delta_{s_1,s_3} \delta_{s_2,s_4} \times \\ \nonumber && {\left\langlej_1,s_1\right\|}\otimes{\left\langlej_2,s_2\right\|} g_{s_1s_2} \delta(z_1-z_2) {\left\|j_3,s_3\right\rangle}\otimes{\left\|j_4,s_4\right\rangle},\end{aligned}$$ with $g_{s_1s_2}$ the spin-dependent contact interaction strength of two particles, and $\delta'$ a Kronecker delta modulo $N_{\Phi}$. The Hamiltonian is diagonalized in blocks with a fixed particle number $N$, defining the filling factor $\nu=N/N_{\Phi}$, fixed Haldane momentum $\vec{K}$ [@haldane], and a fixed spin polarization $S=N_{\uparrow}-N_{\downarrow}$, where $N_{\uparrow}$ ($N_{\downarrow}$) denotes the number of spin-up (spin-down) particles. First, we show that for spin-independent interactions, i.e. $g_{s_1s_2}=g$, the singlet state is energetically favored. This configuration is a natural choice, as many experiments [@ketterle-2comp; @wieman-2comp] work with the ${\left\|F=1, m_F=0,1\right\rangle}$ states of $^{23}$Na, or the ${\left\|F=1,m_F=-1\right\rangle}$ and ${\left\|F=2,m_F=1\right\rangle}$ states of $^{87}$Rb with almost equal s-wave scattering lengths within and between the components. We start by contrasting the singlet to the fully polarized situation, $S=N$, in which we recover the results from Ref. [@cooper-wilkin-gunn], with incompressible phases at $\nu=k/2$, forming the RR series. The fully polarized zero-energy state with highest filling has $\nu=1/2$, and is the Laughlin state. In the opposite limit of a fully unpolarized system, zero-energy states occur up to $\nu=2/3$, where the 221-Halperin state is the exact unique ground state of the 2BC interaction. The analytic expression of this state on a disc reads $$\begin{aligned} \Psi_{\mathrm{H}} = \prod_{i<j}^{N_{\uparrow}} (z_{i \uparrow}-z_{j \uparrow})^2 \prod_{k<l}^{N_{\downarrow}} (z_{k \downarrow}-z_{l \downarrow})^2 \prod_{i,k} (z_{i \uparrow} - z_{k \downarrow}),\end{aligned}$$ where we omit the irrelevant exponential term $\exp\left(-\sum \|z_i\|^2/4\right)$. This wave function vanishes whenever two particles are at the same position, and it is symmetric under exchange of two spin-up or two spin-down particles. However, it is antisymmetric under exchange of particles with different spin, and thus cannot be used for describing spinless bosons. For this case, also the terms in the last product had to be squared, which would result in a less dense Laughlin-like wave function. Generalizing this observation, we state that any spatial wave function which solves the fully polarized problem must also be a solution for $\|S\|<N$, while the opposite is not true. Thus, for a spin-independent interaction, it strictly follows for the ground state energies $E(S_i)$ with $\|S_1\|<\|S_2\|$ that $E(S_1) \leq E(S_2)$. Our numerical data, partially shown in the left side of Fig. \[filling\], supports this finding. In fact, in all cases we have investigated, the ground state of the $S=2$ subspace occurs in the spectrum of $S=0$ as a low-lying excitation, except for $N=6$ and $N_{\Phi}=8$, where we have a degeneracy, $E(S=0)=E(S=2)$. In most cases, thus, the ground state is a singlet, which is understood by noting that interactions of a $\uparrow\downarrow$-pair can be fully avoided by a single zero of the form $z_{i \uparrow} - z_{k \downarrow}$ in the wave function, while at least two zeros must be spent if a pair of particles with equal spin is to be anticorrelated in a similar way. ![\[filling\] (Color online) Left: Ground state energies, not showing possible degeneracies, at $N_{\Phi}=6$ for different $N$ and $S$ subspaces with $g_{s_1s_2}=g$. Even (odd) values of $S$ correspond to even (odd) $N$. Right: $\Delta$ vs. $\nu$ for $N_{\Phi}=6$ and $g_{s_1s_2}=g$.](fig1.eps) Feshbach resonances allow to tune the spin-dependency of interactions (cf. [@mlbook]). Increasing $g_{\uparrow\downarrow}$ while leaving $g_{\uparrow\uparrow}=g_{\downarrow\downarrow}=g$ should favor spin polarizations with less $\uparrow \downarrow$-pairs. However, this does not necessarily drive the system into the fully polarized configuration: As shown in Fig. \[eud\] for $N=6$ at $\nu=1$, the energy of even the fully unpolarized system saturates for large $g_{\uparrow\downarrow}$. In this limit, the real ground state has $S=2$ and $E=0.449$ (in units of $gB$), close to the $S=0$ and $S=4$ ground states with $E=0.466$ and $E=0.475$. All of them are well separated from the fully-polarized ground state with $E=0.581$. In contrast to this, the energy of spin-unpolarized systems will never saturate at larger filling, as is shown in Fig. \[eud\] for $\nu=6/5$. This different behavior is understood by noting that for $\nu=1$ the number of available zeros in the wave function is sufficient to completely suppress interactions between pairs of different spins, $\langle E_{\uparrow\downarrow} \rangle \rightarrow 0$, while it is not for larger $\nu$. Jumps in the curve of $\langle E_{\uparrow\downarrow} \rangle$ as a function of $g_{\uparrow\downarrow}$ show that the states with $\langle E_{\uparrow\downarrow} \rangle = 0$ are reached by several abrupt re-organizations of the ground states. We note that once the state with $\langle E_{\uparrow\downarrow}\rangle=0$ is obtained, this state must be an eigenstate for arbitrary $g_{\uparrow\downarrow}$. Thus, by abruptly switching off the interspin interaction, one could produce binary mixtures of highly entangled, non-interacting systems. ![\[eud\] (Color online) Interaction energy for $N=6$ and $S=0,2$ as a function of $g_{\uparrow\downarrow}$. Left: $N_{\Phi}=6$. Right: $N_{\Phi}=5$.](fig2.eps){width="9cm"} Coming back to spin-independent interactions, we ask now the question whether, in analogy to the RR states for spin polarized systems, incompressible states can be found amongst the spin-unpolarized ground states. Since incompressibility is connected to a discontinuity in the chemical potential, we define the particle-hole excitation gap as $\Delta(N) = N [ E(N+1)/(N+1) + E(N-1)/(N-1) - 2 E(N)/N ]$, which is plotted for $N_{\Phi}=6$ in Fig. \[filling\] (right). Upwards peaks correspond to downward cusps in the energy as a function of $\nu$, and signal incompressible phases. They occur at fillings $\nu=2k/3$ with $k\in \mathbb{N}$. For these fillings, a series of singlet states being exact zero-energy eigenstates of a $(k+1)$-body contact interaction is known [@nass-nucl]. For $k=1$, this is the 221-Halperin state, which has Abelian excitations, while the states for $k>1$ are predicted to have non-Abelian excitations, which established the name NASS series. ![(Color online) Energy spectra of singlet states for $N=12$ particles at filling $\nu=4/3$. Black crosses (with the left energy scale) show the results for an SU(2)-symmetric 2BC interaction, while red triangles (with the right energy scale) are the NASS spectrum for any three-body interaction with $g_{s_1s_2s_3}>0$. We do not show the three-fold center-of-mass degeneracy, but visualize further degeneracies by slight shifts in $\vec{K}$. \[2spectra\]](fig3.eps) We first construct these states for $k=2$ and 3 by diagonalizing the three- and four-body contact interaction for up to $N=12$ particles: As predicted by CFT, these states are characterized by a $(k+1)(k+2)/2$-fold topological degeneracy on the torus. A subtlety occurs at $k=3$, where the CFT construction [@nass-nucl], being restricted to $N=2kp$ with $p \in \mathbb{N}$, is not able to predict zero-energy ground states for all $N$ possible at $\nu=2$. In fact, we find zero-energy states also for $N=8$ and 10, but, in contrast to the states with $N=6$ and 12 which have the predicted ten-fold degeneracy, they are non-degenerate, and thus belong to a different topological phase. The overlaps of these states with the ground states of Eq. (\[H\]), given in Table \[ov\], turn out to be significant: For $k=2$, the states are exactly equal for $N=4$, and even for a relatively large-sized system with $N=12$ the overlaps are still around 0.8. Such a decrease of the overlap with large $N$ also occurs in the case of the RR states in single-component systems with 2BC interaction [@cooper-aip], suggesting the use of ground state degeneracies to characterize the topological phase in the thermodynamic limit [@wen-niu]. Topological degeneracies are usually lifted in finite systems, as also happens here. In the case of $\nu=4/3$ we may consider the fact that the six lowest eigenstates of ${\ensuremath{\Hat{\mathrm{H}}}}$ agree in their Haldane momenta $\vec{K}$ with the ones of the sixfold degenerate three-body eigenstates as a signal for belonging to the same topological phase. Both spectra are shown for $N=12$ in Fig. \[2spectra\]. We note that both the degeneracy splitting $\Delta_{\mathrm{GS}}$ and the gap to the excited band $\Delta_{\mathrm{EX}}$ behave non-monotonic with $N$. The situation is different for $\nu=2$, where the candidates for the degenerate ground state manifold mix up with excited states. Our results therefore suggests that only the $\nu=4/3$-NASS phase can be produced with 2BC interaction. For the experimental feasibility of this state to be high, it should also be robust with respect to deviations from the SU(2)-symmetric interaction. In the case of a $(k+1)$-body contact interaction at $\nu=2k/3$, the numerical values of each interaction parameter do not influence the ground states. Though this is different for 2BC interaction at $\nu=4/3$, the overlaps with the NASS states remain high within a wide range of $g_{\uparrow\downarrow}/g$, and have a maximum in the SU(2)-symmetric configuration. For $N=8$, all overlaps are above 0.8 if $g_{\uparrow\downarrow}/g \in [0.8,1.6].$ -------------- ----------- ------------------------------------------------------- ------------------- -- -------------- ----------- ------------------------------------------------------- ------------------- $N,N_{\Phi}$ $\vec{K}$ $\|{\left< \mathrm{2b} \vphantom{\mathrm{3b}} \right\| $E_{2\mathrm{b}}$ $N,N_{\Phi}$ $\vec{K}$ $\|{\left< \mathrm{2b} \vphantom{\mathrm{4b}} \right\| $E_{2\mathrm{b}}$ \left. \mathrm{3b} \vphantom{\mathrm{2b}} \right>}\|$ \left. \mathrm{4b} \vphantom{\mathrm{2b}} \right>}\|$ 4,3 (0,0) 1 0.435 12,6 (0,0) 0.334 2.604 1 0.520 0.828 2.677 8,6 (0,0) 0.906 0.901 (0,2) 0.775 2.708 0.920 0.908 (0,4) 0.775 2.708 12,9 (0,0) 0.729 1.289 (2,0) 0.775 2.708 0.885 1.333 (2,2) 0.646 2.685 (2,4) 0.646 2.285 (4,0) 0.775 2.708 (4,2) 0.646 2.685 (4,4) 0.646 2.685 -------------- ----------- ------------------------------------------------------- ------------------- -- -------------- ----------- ------------------------------------------------------- ------------------- : Overlaps between zero-energy eigenstates of $(k+1)$-body interaction with corresponding eigenstates of the 2BC interaction at energy $E_{2\mathrm{b}}$ (in units of $gB$) and Haldane momentum $\vec{K}$ (in units of $2\pi \hbar/a$). All states at filling $\nu=4/3$ have an additional three-fold center-of-mass degeneracy. \[ov\] Finally, we consider an additional SO coupling with $q>0$ in Eq. (\[A\]). This coupling mixes internal and external degrees of freedom on the single-particle level, such that the LLs become superpositions of the form ${\left\|n,j,\pm\right\rangle} \equiv \alpha_{n}^{\pm} {\left\|n-1,j,\uparrow\right\rangle} + \beta_{n}^{\pm} {\left\|n,j,\downarrow\right\rangle}$ where $\alpha_{n}^{\pm}$ and $\beta_{n}^{\pm}$ are functions of $q$ [@trombettoni]. For general $q$, this lifts the previous spin-degeneracy, as the single-particle energies now read $E_{n}^{\pm} = 2Bn +2q^2 \pm \sqrt{B^2+8Bq^2n}$. As has been pointed out in Refs. [@trombettoni; @palmer-pachos], the system then behaves basically like a single-component system, and the incompressible states are described by the RR series projected into the lowest LL of the SO coupled system. However, for $q^2/B = 2n+1$, a degeneracy between $E_{n}^-$ and $E_{n+1}^-$ occurs as the single-particle ground state switches from ${\left\|n,j,-\right\rangle}$ to ${\left\|n+1,j,-\right\rangle}$. The Hamiltonian from Eq. (\[H\]) can then be adapted to the system with SO coupling by identifying the $s$ quantum number with the two-fold degeneracy between $n$ and $n+1$, and by re-defining $V_{\{js\}}$ accordingly. Note that in this case $S$ does not correspond to the spin polarization, but quantifies the population imbalance between ${\left\|n,j,-\right\rangle}$ and ${\left\|n+1,j,-\right\rangle}$. As observed in Ref. [@palmer-pachos] within a lowest LL approximation, the projected RR states do not describe the system close to these degeneracy points. For $q^2/B = 3$, the system becomes gapless at both the Laughlin and the MR filling $\nu=1/2$ and 1. However, we find a clear gap in the spectrum at $\nu=2/3$ with $N=6$, and a small gap at $\nu=4/3$ with $N=8$. Projecting this $\nu=2/3$ state into the lowest LL of the system without SO coupling, and comparing with the 221-Halperin state, we find an overlap of only 0.19. It should be noted that, in contrast to the previous case of a pure spin degeneracy, interactions now can induce flips within the two-fold degenerate manifold, and therefore $S$ is not conserved. For a complete description of the state, we use generalizations of the Halperin state, obtained by diagonalizing the 2BC interaction at $\nu=2/3$ in an Abelian field ($q=0$) within different spin subspaces. Then the highest overlap of 0.60 is obtained with the state having $S=-6$, which corresponds to all particles being in ${\left\|0,j,-\right\rangle}$. This is also the most populated LL, showing that, despite the degeneracy on the single-particle level, interactions favor the LL with lower $n$. The total fidelity for $-6 \leq S \leq 6$ is 0.82. For $\nu=4/3$ with $N=8$, but now comparing the non-degenerate ground state of the SO coupled system with the ground states of a three-body contact interaction at $q=0$ for different $S$, we still obtain a total fidelity of 0.52, summing from $-8 \leq S \leq 8$. We have investigated two-component Bose gases in artificial gauge fields. The possibility of tuning the interaction constants allows to control and manipulate the strongly correlated nature of the eigenstates. Our main finding is the formation of incompressible phases described by the NASS series within the experimentally feasible setup of particles interacting via two-body contact interaction. The internal degree of freedom permits also to realize systems with spin-orbit coupling. This work has been supported by EU (NAMEQUAM, AQUTE), ERC (QUAGATUA), Spanish MINCIN (FIS2008-00784, FIS2010-16185, FIS2008-01661, 2009 SGR1289), Alexander von Humboldt Stiftung, and AAII-Hubbard. B. J.-D. is supported by the Ramón y Cajal program. M. L. acknowledges Hamburg Theory Award. Note added: After submission of this article, a preprint by S. Furukawa and M. Ueda appeared presenting additional numerical evidences for the NASS phase [@furukawa-ueda].	{ "pile_set_name": "ArXiv" }
Downtown Three Rivers Commercial Historic District The Downtown Three Rivers Commercial Historic District is a commercial historic district located along North Main Street, between Michigan and Portage Avenues, in Three Rivers, Michigan. It was listed on the National Register of Historic Places in 1982. History The original section of Three Rivers, including what is now the downtown and some surrounding residential areas, was platted in 1836 by John H. Bowman. This location immediately began to serve as the commercial center of the settlement, with modest wooden buildings constructed along what is now Main Street as early as the 1830s. A number of mills were constructed in the area in the 1830s and 1840s, and in 1851 the Lockport Hydraulic Company was formed to dam the river and construct a power canal. This was followed in 1853 by the construction of a railroad line through the village. As a result, Three Rivers boomed in the latter half of the 19th century, with a wave of new construction starting in the 1850s. The oldest extant buildings in the district are the Kelsey block at 39-43 N. Main (built in 1854), the Whitesell and McMurtrie block at 109 Portage (built in 1859), and the 1861 block at 18 N. Main. Further construction filled the downtown in the 1860s, 70s, and 80s. These buildings were constructed almost exclusively by local investors, primarily merchants. Seven of the commercial blocks, however, were built by farmer Isaac Null over the period of 1863-1886. In 1884, Null also built his house, located within the district at 105 W. Michigan. The downtown area remains one of the most intact nineteenth and early twentieth-century downtowns in southwest Michigan Description The Downtown Three Rivers Commercial Historic District contains all the structures remaining from Three Rivers' late 19th century commercial center. The district contains 48 structures, including one residence, the former public library, and 46 one- to three-story commercial buildings. The commercial buildings are predominantly Commercial Italianate structures, and are set side by side and directly on the sidewalk. The visually unified and harmonious Late Victorian streetscape gives the district its character. Significant buildings include: Prutzman Block (22-24 North Main), a lavishly detailed two story, seven bay Italianate structure. Three blocks (34, 36 and 38 North Main) with mansard roofs constructed in the 1870s. 52-56 North Main, a broad commercial block constructed in 1890. Riviera Theatre (50 N. Main), a classical theatre with a projecting Art Moderne marquee, constructed in the 1920s in the shell of an 1860s block. Three Rivers Savings and Loan "south building" (101 S. Main), a single story buff brick Neo-Classical building trimmed with terra cotta, constructed in 1916. Public Library (107 N. Main), a 2-1/2 story cross-gable building constructed of rubble stone with shingled gables. It was built in 1904, and designed by A.W. Rush & Co. of Grand Rapids. Gallery References Category:National Register of Historic Places in St. Joseph County, Michigan Category:Italianate architecture in Michigan Category:Historic districts on the National Register of Historic Places in Michigan	{ "pile_set_name": "Wikipedia (en)" }
/* * Copyright (C) 2013 Christoph Leitner <c.leitner@student.uibk.ac.at> * * This program is free software: you can redistribute it and/or modify * it under the terms of the GNU General Public License as published by * the Free Software Foundation, either version 3 of the License, or * (at your option) any later version. * * This program is distributed in the hope that it will be useful, * but WITHOUT ANY WARRANTY; without even the implied warranty of * MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the * GNU General Public License for more details. * * You should have received a copy of the GNU General Public License * along with this program. If not, see <http://www.gnu.org/licenses/>. / #include "ruc_connection.h" #include "stubborn_sender.h" #include "uc_connection.h" #define DEBUG 1 #define dout DEBUG && std::cout using namespace gr::ieee802_15_4; ruc_connection::ruc_connection(rime_stack block, uint16_t channel, pmt::pmt_t inport, pmt::pmt_t outport, const uint8_t rime_add_mine[2]) : rime_connection(block, channel, inport, outport, rime_add_mine), d_stubborn_sender(block, this, d_mac_outport), d_send_seqno(0), d_recv_seqno(0) { d_stubborn_sender.start(); } std::array<uint8_t, 256> ruc_connection::make_msgbuf(uint16_t channel, bool ack, int seqno, const uint8_t src[2], const uint8_t dest[2]) { std::array<uint8_t,256> buf; buf[0] = channel & 0xff; buf[1] = (channel >> 8) & 0xff; if(ack){ buf[2] = (1 << 7) & 0xff; //ack packet } else { buf[2] = 0; //data packet } buf[2] \|= seqno << (7 - seqno_bits); //set seq.no. buf[2] \|= dest[0] >> (1 + seqno_bits); buf[3] = (dest[0] << (7 - seqno_bits)) & 0xff; buf[3] \|= dest[1] >> (1 + seqno_bits); buf[4] = (dest[1] << (7 - seqno_bits)) & 0xff; buf[4] \|= src[0] >> (1 + seqno_bits); buf[5] = (src[0] << (7 - seqno_bits)) & 0xff; buf[5] \|= src[1] >> (1 + seqno_bits); buf[6] = (src[1] << (7 - seqno_bits)) & 0xff; return buf; } void ruc_connection::pack(pmt::pmt_t msg) { assert(d_send_seqno < (1 << seqno_bits)); if(pmt::is_eof_object(msg)){ d_block->message_port_pub(d_mac_outport, pmt::PMT_EOF); d_block->detail().get()->set_done(true); return; } std::string tmp = rime_connection::msg_to_string(msg); uint8_t dest[2]; if(!uc_connection::rime_add_from_string(tmp, dest)){ std::cerr << "Warning: invalid target RIME-Address for runicast on channel "; std::cerr << static_cast<unsigned>(d_channel); std::cerr << ". Message will not be sent." << std::endl; return; } std::array<uint8_t, 256> buf = ruc_connection::make_msgbuf(d_channel, false, d_send_seqno, d_rime_add_mine, dest); size_t data_len = tmp.length(); assert(data_len); assert(data_len < 256 - header_length); std::memcpy(buf.data() + header_length, tmp.data(), data_len); pmt::pmt_t rime_msg = pmt::make_blob(buf.data(), data_len + header_length); pmt::pmt_t dict = pmt::make_dict(); dict = pmt::dict_add(dict, pmt::mp("seqno"), pmt::from_long(d_send_seqno)); d_stubborn_sender.enqueue(pmt::cons(dict, rime_msg)); d_send_seqno = (d_send_seqno + 1)%(1 << seqno_bits); } void ruc_connection::unpack(pmt::pmt_t msg) { uint8_t buf[256]; uint8_t target_rime_zero, target_rime_one; uint8_t sender_rime_zero, sender_rime_one; bool is_ack = false; uint8_t packet_seqno; size_t data_len = pmt::blob_length(msg); std::memcpy(buf, pmt::blob_data(msg), data_len); target_rime_zero = buf[2] << (1 + seqno_bits); target_rime_zero \|= buf[3] >> (7 - seqno_bits); target_rime_one = buf[3] << (1 + seqno_bits); target_rime_one \|= buf[4] >> (7 - seqno_bits); sender_rime_zero = buf[4] << (1 + seqno_bits); sender_rime_zero \|= buf[5] >> (7 - seqno_bits); sender_rime_one = buf[5] << (1 + seqno_bits); sender_rime_one \|= buf[6] >> (7 - seqno_bits); dout << "[" << static_cast<int>(d_rime_add_mine[0]) << "."; dout << static_cast<int>(d_rime_add_mine[1]) << "]: "; //this block is not the destination of the message if(target_rime_zero != d_rime_add_mine[0] \|\| target_rime_one != d_rime_add_mine[1]){ dout << "received packet with wrong receiver, discarding"; dout << "(" << static_cast<int>(target_rime_zero) << "."; dout << static_cast<int>(target_rime_one) << ")" << std::endl; return; } if((buf[2] & 0x80) > 0){ is_ack = true; buf[2] &= 0x7f; //reset ack-flag } packet_seqno = buf[2] >> (7 - seqno_bits); if(is_ack){ if(packet_seqno != recv_seqno()){ //ignore duplicate packets dout << "received duplicate ack "; dout << static_cast<int>(packet_seqno); dout << " (should be: "; dout << recv_seqno() << ")"<< std::endl; return; } dout << "received ack for seqno "; dout << static_cast<int>(packet_seqno) << std::endl; d_stubborn_sender.stop(); inc_recv_seqno(); dout << "expected next seqno: " << d_recv_seqno << std::endl; } else { //output message pmt::pmt_t rime_payload = pmt::make_blob(buf + header_length, data_len - header_length); d_block->message_port_pub(d_outport, pmt::cons(pmt::PMT_NIL, rime_payload)); //send ack uint8_t dest[] = {sender_rime_zero, sender_rime_one}; std::array<uint8_t, 256> buf = make_msgbuf(d_channel, true, packet_seqno, d_rime_add_mine, dest); dout << "sent ack message for seqno " << static_cast<int>(packet_seqno); dout << " to "; dout << static_cast<int>(dest[0]) << "."; dout << static_cast<int>(dest[1]) << std::endl; pmt::pmt_t ack_msg = pmt::make_blob(buf.data(), header_length); d_block->message_port_pub(d_mac_outport, pmt::cons(pmt::PMT_NIL, ack_msg)); } } void ruc_connection::inc_recv_seqno() { gr::thread::scoped_lock lock(d_mutex); d_recv_seqno = (d_recv_seqno + 1)%(1 << seqno_bits); } int ruc_connection::recv_seqno() { gr::thread::scoped_lock lock(d_mutex); return d_recv_seqno; }	{ "pile_set_name": "Github" }
Terminal ligature of inferior thyroid artery branches during total thyroidectomy for multinodular goiter is associated with higher postoperative calcium and PTH levels. To evaluate the impact of truncal versus terminal branch ligature of the inferior thyroid artery (ITA) on postoperative calcium and PTH plasma levels in patients undergoing total thyroidectomy for multinodular goiter. A prospective randomized study was performed comparing a group of patients that underwent either truncal ligature of the ITA (group 1) or terminal ligature of ITA branches (group 2). A series of 126 consecutive patients with non-toxic euthyroid multinodular goiter underwent total thyroidectomy. Truncal ligature of the ITA was performed in 63 patients (group 1) and terminal branch ITA ligature in 63 patients (group 2). Postoperative ionized serum calcium (mmol/L) at 24 hours was significantly lower in group 1 than in group 2 patients (1.22 ± 0.06 vs. 1.25 ± 0.05, P<0.05) and at 48 hours (1.20 ± 0.05 vs. 1.23 ± 0.05, P<0.05). Mean postoperative PTH levels (pg/mL) at 4 hours after thyroidectomy were significantly lower in group 1 than in group 2 patients (22.32 ± 11.64 vs. 25.82 ± 12.87, P=0.044). Mean hospital stay (hours) was higher in group 1 than in group 2 patients (87.47 ± 41.04 vs. 70.34 ± 24.82, P<0.05). This study shows that terminal ligature of ITA branches during total thyroidectomy for multinodular goiter is associated with higher mean postoperative calcium and PTH levels, and shorter hospital stay. However, no significant difference in terms of permanent hypoparathyroidism was observed between the two groups.	{ "pile_set_name": "PubMed Abstracts" }
"Temos uma terra a qual nunca vamos renunciar", disse o presidente A Argentina relançou nesta sexta-feira (3) sua campanha pela soberania das Malvinas (Falklands, para os britânicos). “Temos uma terra a qual nunca vamos renunciar. Os britânicos invadiram as Malvinas e, nesta primeira vez que me refiro ao tema como presidente, quero enfatizar que nossa convicção em lutar por recuperar este território continua sendo contundente”, afirmou o presidente Alberto Fernández, em discurso realizado no Porto de Buenos Aires. No mesmo evento, o chanceler Felipe Solá assegurou que a campanha internacional pela soberania argentina voltou a ser “prioridade” da política externa argentina. “Queremos uma solução pacífica e definitiva para a disputa”, disse. (Com agências internacionais) (Estadão Conteúdo)	{ "pile_set_name": "OpenWebText2" }
There’s no strong evidence that if you warm up, you will reduce your risk of injury, but there is evidence that warming up can improve your performance. Five to 10 minutes of aerobic exercise will raise your muscle temperature and your heart rate so that more oxygen reaches your muscles. Warming up also provides an opportunity to incorporate beneficial exercises such as strengthening and control work. Whether or not you warm up will probably depend on how long and how hard you plan to work out. If you have 20 minutes for a run, you could just put your shoes on and go. If you are about to take part in a race, such as a Parkrun-type event, you might want to warm up for 15 or 20 minutes. Elite athletes will warm up for 45 minutes to an hour before they go into competition, and may do specific exercises such as “strides” – race-pace speeds to prepare the body. You don’t need to do that if you’re going for a gentle jog around the park. If you’re really unfit, you could warm up with a walk-run. As far as stretching is concerned, it also doesn’t prevent injury, and there is a school of thought that if you do lots of sustained static stretching – holding a position – it can dampen down some of the force your muscles can produce. Most athletes will do more “dynamic mobility” work, where you take your joints through their range of motion, rather than holding a stretch. Toby Smith is technical lead physiotherapist at the English Institute of Sport. He was talking to Emine Saner	{ "pile_set_name": "OpenWebText2" }
<!DOCTYPE html PUBLIC "-//W3C//DTD HTML 4.01 Transitional//EN" "http://www.w3.org/TR/html4/loose.dtd"> <html> <head> <meta http-equiv="Content-Type" content="text/html; charset=UTF-8"> <script src="/benchmark/js/jquery.min.js"></script> <script type="text/javascript" src="/benchmark/js/js.cookie.js"></script> <title>BenchmarkTest02202</title> </head> <body> <form action="/benchmark/pathtraver-02/BenchmarkTest02202" method="POST" id="FormBenchmarkTest02202"> <p>Please make your car selection, and edit the value to be sent [value]:</p> <select name="BenchmarkTest02202" multiple> <option value="FileName" selected>Audi. [FileName]</option> <option value="Ford">Ford. [Ford]</option> <option value="Opel">Opel. [Opel]</option> <option value="Subaru">Subaru. [Subaru]</option> </select><br/><br/> <input type="submit" value="Submit"> </form> </body> </html>	{ "pile_set_name": "Github" }
Q: Can i get the generic type of an object? this is my method: GetListItemsPainted<T>(List<T> list) and i don't know what type is that list of, how can i create new list that would have the passed list type? something like this: List<list.GetType()> newList = new List<list.GetType()>(); how can i cast my list to the real type so i would have all his properties etc.? thanks A: You can create a list using T: List<T> newList = new List<T>(); If you must get the type, you can use typeof. This is similar to what you asked for, but has other uses, you don't need it to make generics work: Type myType = typeof(T); A: You do not have to create a new List, you already have one. If you require a specific type, then constrain your generic type parameters with a where constraint If you intend to react to a wide variety of arbitrary types, which is a bad design decision in my opinion, then you will need to use a conditional with .Cast<T>() something like: Type myListType = list.GetType().GetGenericArguments()[0]; // or Type myListType = typeof(T); as stated by Kobi if(myListType == typeof(SomeArbitraryType)) { var typedList = list.Cast<SomeArbitraryType>(); // do something interesting with your new typed list. } But again, I would consider using a constraint.	{ "pile_set_name": "StackExchange" }
Q: Parallellizable Algorithms to traverse a 2D matrix being aware of both col/row-wise neighborhood I have a fairly large NN integer matrix Matrix2D (assume sufficient memory), 1, within each row/column, I need to record the col/row index of an element if it's value is different than it's right/lower neighbour. 2, I want to find an optimal algorithms that is parallellizable, ideally by OMP. So, finally I will have some data structures like, std::vector<std::vector<int>> RowWiseDiscontinuity(N);// N= #of rows std::vector<std::vector<int>> ColWiseDiscontinuity(N);// N= #of cols where inner std::vector<int> records the row/col indices. I put my serial version here but find it difficult to be OMP parallelized... Someone could provide some idea how to implement the traversing over this 2D matrix with omp? code snippet, std::vector<std::vector<int>> RowWiseDiscontinuity(N);// N= #of rows std::vector<std::vector<int>> ColWiseDiscontinuity(N);// N= #of cols std::vector<int> TempX1; std::vector<int> TempX2; for (int y=0; y<N; ++y) { TempX1.clear(); for (int x =0; x<N; ++x) { int value = Matrix2D(x,y); TempX1.push_back(value); } auto iter1 = TempX1.begin(); auto iter2 = TempX2.begin(); if (y>0) for (int x =0; x<N; ++x) { if (iter1 !=(iter1+1)) { RowWiseDiscontinuity[y].push_back(x); //Critical for OMP } ++iter1; ++iter2; if (iter1 != *iter2) { ColWiseDiscontinuity[x].push_back(y); //Critical for OMP } } TempX2.swap(TempX1); // proceed to next row, remember previous } A: I would create another array that holds the nearest neighbor both column and row-wise. This would have to be done as a first pass, obviously. I recommend creating a 2d array of pairs (pair) that holds the indices you want. Instead of two vectors, I would do a vector of pairs. Pairs are parallellizable and easily sorted. vector<vector<pair<int, int>>> elements(N);	{ "pile_set_name": "StackExchange" }
591 F.2d 101 U. S.v.Dow*** No. 78-5457 United States Court of Appeals, Fifth Circuit 2/23/79 1 S.D.Tex. AFFIRMED * Summary Calendar case; Rule 18, 5 Cir.; see Isbell Enterprises, Inc. v. Citizens Casualty Co. of New York et al., 5 Cir., 1970, 431 F.2d 409 ** Local Rule 21 case; see NLRB v. Amalgamated Clothing Workers of America, 5 Cir., 1970, 430 F.2d 966	{ "pile_set_name": "FreeLaw" }
The role of dietary fat in adipose tissue metabolism. Energy intake and expenditure tend on average to remain adjusted to each other in order to maintain a stable body weight, which is only likely to be sustained if the fuel mix oxidised is equivalent to the nutrient content of the diet. Whereas protein and carbohydrate degradation and oxidation are closely adjusted to their intakes, fat balance regulation is less precise and that fat is more likely to be stored than oxidised. It has been demonstrated that dietary fatty acids have an influence not only on the fatty acid composition of membrane phospholipids, thus modulating several metabolic processes that take place in the adipocyte, but also on the composition and the quantity of different fatty acids in adipose tissue. Moreover, dietary fatty acids also modulate eicosanoid presence, which have hormone-like activities in lipid metabolism regulation in adipose tissue. Until recently, the adipocyte has been considered to be no more than a passive tissue for storage of excess energy. However, there is now compelling evidence that adipocytes have a role as endocrine secretory cells. Some of the adipokines produced by adipose tissue, such as leptin and adiponectin, act on adipose tissue in an autocrine/paracrine manner to regulate adipocyte metabolism. Furthermore, dietary fatty acids may influence the expression of adipokines. The nutrients are among the most influential of the environmental factors that determine the way adipose tissue genes are expressed by functioning as regulators of gene transcription. Therefore, not only dietary fat amount but also dietary fat composition influence adipose tissue metabolism.	{ "pile_set_name": "PubMed Abstracts" }
Impairment of fronto-striatal and parietal cerebral networks correlates with attention deficit hyperactivity disorder (ADHD) psychopathology in adults - a functional magnetic resonance imaging (fMRI) study. Attention deficit hyperactivity disorder (ADHD) is a common, genetically transmitted common childhood-onset disorder with a high rate of persistence in adulthood. Although many studies have shown anatomical and functional abnormalities in children and adolescents, studies with adult patients are rare. Nineteen adults with ADHD (11 ADHD, combined type; 8 ADHD, partially remitted) and 17 controls were included in this functional magnetic resonance imaging (fMRI) study. Brain activation was investigated with a continuous performance test (CPT). Impaired activation of a fronto-striatal and a parietal attentional network was observed during the NoGo condition in ADHD subjects. Correlations of reduced activity of the caudate nuclei, the anterior cingulate cortex, and parietal cortical structures, as well as increased activity in the insular cortex, with inattention and impulsivity symptom scores were found. The activation patterns were similar to those known from children and adolescents with ADHD. In conclusion we found not only a widespread dysfunction of brain regions that are involved in cognitive processing in adults with ADHD compared with controls, but also correlations between symptom severity and dysfunction of neuronal systems across adult subjects with a history of ADHD in childhood but whose symptoms did (persistent ADHD) and did not (not persistent ADHD) qualify for a full diagnosis of ADHD in adulthood.	{ "pile_set_name": "PubMed Abstracts" }
[Identification of the mode of stab-and-hack wound infliction to the chest based on wound features]. The objective of this study was to obtain morphological characteristics of stab-and-hack wounds inflicted by falling down on the knife blade. A formula is derived based on the results of discrimination analysis that can be used in combination with wound morphological features to differentiate between chest stabbing modes.	{ "pile_set_name": "PubMed Abstracts" }
Blueberry muffin baby Blueberry muffin baby is the characteristic distributed purpura occurring as a result of extramedullary hematopoiesis found in infants. The purpura are often generalized, but occur more often on the trunk, head, and neck. The name is from the superficial similarity to a blueberry muffin. Causes The condition was originally considered characteristic of rubella, but is now considered to be potentially associated with many other conditions, such as cytomegalovirus, metastatic neuroblastoma, and Congenital Leukemia. Diagnosis See also TORCH complex List of cutaneous conditions References External links Category:Vascular-related cutaneous conditions	{ "pile_set_name": "Wikipedia (en)" }
It made Mr Becker a sport and culture attache to the EU in April 2018. Mr Becker’s defence has been lodged in the High Court. His lawyers maintain he cannot be made subject to any legal process unless CAR President Faustin-Archange Touadera specifically lifts his immunity at the request of the British government. His legal team said: “This means he cannot be subject to legal process in the courts of any country for so long as he remains a recognised diplomatic agent.” The former tennis champion said the proceedings were “unjustified and unjust” and being declared bankrupt “inflicted a whole heap of damage on me”. He said he was asserting diplomatic immunity to “bring this farce to an end” and stop “the gravy train for the suits”. He added: “I am immensely proud of my appointment [by] the Central African Republic… sport is incredibly important in Africa and is fast becoming a universal language.” An Indian and a Kazakh businessman, as well as a former adviser to the late Libyan leader Muammar Gaddafi, have all at various times attempted to avoid legal action in Europe by citing diplomatic immunity from the CAR. Central African Republic: Rich in diamonds, gold, oil and uranium, but has one of the world’s poorest populations Unstable since independence from France in 1960; renewed crisis since rebels seized power in 2013 27% of the population, 1.2m people, have fled their homes; 2m need food aid, 45% of the population Government only controls the capital; 14,000 UN peacekeepers in the country Life expectancy is 48 (men) and 51 (women) Mr Becker was declared bankrupt after Arbuthnot Latham claimed he owed them a large sum for more than two years. At the time, he said: “This order relates to one disputed loan which I was due to repay in full in one month’s time.” But the registrar said Mr Becker gave the impression of “a man with his head in the sand”. He has appointed Ben Emmerson QC, who has previously represented Julian Assange and Marina Litvinenko, the widow of Alexander Litvinenko.	{ "pile_set_name": "Pile-CC" }
Hotel Splendid Venice All the Magic of Venice Just a few steps from some of the world’s greatest sights, the Splendid Venice is a charming luxury hotel, as elegant as Venice itself. In the sixteenth century, it was an inn; today, it is a residence of rare refinement overlooking the canals and the Mercerie, the age-old streets, or calli, that link Piazza San Marco and the Rialto bridge. The Splendid Venice is within walking distance of the city’s most impressive destinations Hotel Splendid Venice stands in the heart of Venice, in a quiet yet central area near the city’s main landmarks and many hidden gems.This sublime setting, immersed in the peace of the calli and canals, offers guests a rare privilege: the frisson of discovering all the real beauty of Venice, beyond the clichés and away from thronging crowds.	{ "pile_set_name": "Pile-CC" }
Formal Ontology in Information Systems N. Guarino (Ed.) !OS Press, 1998 Pattern as an Ontological Category Ingvar Johansson Department of Philosophy and Philosophy of Science Umea University S-901 87 Umea, Sweden lngvar.J ohansson@philos.umu.se Abstract. The paper argues that causal systems and spatial patterns are species of the same genus, namely pattern, and that a clear view of spatial patterns throws light on some aspects of the ontological nature of causal systems. In particular, it is argued that all patterns (and systems) depend on a fiat delimitation of something which in itself is a unity without borders. Pattern realism is true. Keywords: Foundations, Kinds of entity, Top-level ontological taxonomies, Patterns. Most top-level ontologies in the history of philosophy reckon in some kind of primary substance (be it things, events, or something else), properties, and relations, and they have some place for space and time. Some other categories like causality and quantity are also almost universal in this sense. Very seldom, however, has pattern got a place high up in any ontological hierarchy. The aim of this paper is to show that patterns really deserve such a place. Of course, patterns are always patterns of something. Therefore, pattern as a category cannot be logically basic, but it is important nonetheless. In some modem ontologies, systems are given a prominent place. In particular, this is true of Mario Bunge's Treatise of Basic Philosophy. The fourth volume of this treatise is given the subtitle Ontology II: A World of Systems, and is wholly dedicated to systems. It opens with the following two sentences: "Every science studies systems of some kind, whether natural (physical, chemical, biological, or social) or artificial (technical). Moreover most sciences study nothing but systems"([ I] p. l ). I agree whole-heartedly. However, in my view, systems make up some special species of the genus pattern. And that fact and some other things are not made clear by Bunge. I will here try to develop further some ideas about patterns and systems which I have put forward earlier (see [2) chapters 4, 6, 7, 14, and 15). Whatever the practical use of ontologies is, and will be, now when information technology revolutionises the world, one of their theoretical uses is to create a bird's eye view from which one can correct various forms of myopia. I will try to show that one gets a better ontological understanding of some aspects of systems when they are seen as species of the category of pattern. The simplest kind of patterns are the spatial patterns, and the ontological structure of such patterns is the topic for the first four sections of this paper. Then there are short sections on spatiotemporal patterns and causal systems. Whether or not my analysis is of direct relevance to the ontology of information systems, I am not able to tell. I look upon the world of research as an extensive division of labour which encompasses not only the sciences and technology but philosophy as well. Philosophy can interact with both science and technology, and I hope that this paper will do so, but the interaction is not spelled out in the paper in itself. 1 The ontological point of departure Every pattern has some kind of components. Patterns, as I have said, are always patterns of something. Therefore, I need some ontological categories before I can start my discussion of patterns. Firstly, I will kind of unity wbi and structure of ** aggregate of p Secondly, Thirdly, properties do of will regard, for regard density, small parts of * Fourthly, s* both between * certain distance instances. Pro will be used as a. Fifthly, I grounded rel . exists thanks to than-', 'has a than-', 'is hea not regard sp * (e.g. 'is to property-groun . '-R-'. The ontolo epistemological things a and b between these round and b is 1 between a and * property-grou 'S -', betw account). The deriv existentially . ( e.g. round an independently o being of differe The exis • S-' is quite co things both in abstract spaces also can cons space. For some realist opinion)' Such relations by statements relation ' has round) and b ( . true by facts w . the form 'aSb' in if f I. Johansson I Pattern as an Ontological Category 87 Firstly, I will rely on a rather common-sensical conception of space. Space is a special kind of unity which contains things, persons, states of affairs, etc. Problems about the limits and structure of space are disregarded. The important thing is that space is not regarded as an aggregate of part-spaces. Secondly, as primary substances I will rely on ordinary material macro things. Thirdly, material things will be assumed to have properties inhering in them. 1 Such properties do of course inherit mind-independence from the material things they inhere in. I will regard, for instance, shape, size, and mass as surely being such properties. I will also regard density, i.e. mass per unit volume, as a real property which can inhere in arbitrary small parts of things. When needed, 'P' will be used as a variable for such properties. Fourthly, since things and property instances exist in space, there are spatial relations both between things and property instances. A property instance can lie beside and be at a certain distance from another property instance. Also, it can lie between two other property instances. Property instances can stand in all kind of spatial relations. When needed, '-S-' will be used as a variable for such relations. Fifthly, I want to introduce a special class of relations which I will call propertygrounded relations (see [2] chapter 8). A property-grounded relation is a relation which exists thanks to some properties which inhere in the things related. Examples are: 'is larger than -', ' has a size between that of and -', ' has another colour hue than -', ' is brighter than -', ' is heavier than -', 'has a higher density than -', ' is similar to -', and so on. I do not regard spatial location as a property, which means that I do not regard spatial relations (e.g. 'is to the left of-', 'is five metres from-', 'is situated between and -') as property-grounded relations . As a variable for binary property-grounded relations, I will use '-R-'. The ontological difference between relations of type '-R-' and '-S-' is reflected in an epistemological difference. If one has knowledge of all the properties which inhere in two things a and h ( = the relata), one can derive all the property-grounded relations that exist between these relata, but no spatial relation is derivable. For instance, if one knows that a is round and h is square, one can derive that a and b have different shapes, but the distance between a and b is not derivable. Nor, of course, are spatial relations derivable from property-grounded relations. In every statement of the form 'R -', the spatial relations, i.e. 'S -', between the relata have been abstncted away (if they have ever been taken into account). The derivation of property-grounded relations is possible because such relations are existentially dependent upon some properties of the relata. Each ?-instance in each relata (e.g. round and square) can exist independently of the other, and therefore it can also exist independently of the grounded relation (being of different shape). But the grounded relation being of different shape cannot possibly exist if there are no property instances of shape. The existence of an ontological and an epistemological difference between '-R-' and ' S-' is quite consistent with the fact that one certainly can construct location functions for things both in relation to real space and in relation to abstract property spaces. In my view, all abstract spaces are conceptual constructs but real space is not. The confusing thing is that we also can construct a conceptual abstract "spatial space" whose metric mirrors relations in real space. For some of my later claims about patterns, it is important to note that (in my immanent realist opinion) a property-grounded relation exists only in the property instances related. Such relations have what might be called scattered instantiation. States of affairs described by statements of the form 'aRb' exist only in a and b. My last example of property-grounded relation 'has a shape different from-' exists nowhere else than in the things a (which is round) and b (which is a square) together; i.e. 'a has another shape than b', if true, is made true by facts wholly contained in a and h. Spatial relations behave differently. Statements of the form 'aSb' describe states of affairs which contain not only a and b, but also some part of space. The statement 'a is five metres from b', if true, is made true by a, b, and the spatial distance between them. The fact that a is five metres from bis instantiated in a spatial unity, 1 I am an immanent realist, but I think that this paper could equally well be written from the point of view of a Platonist or a "trope thinker". 88 I. Johansson I Pattern as an Ontological Category whereas the fact that a has another shape than b is instantiated in a spatial scatter. 2 Spatial patterns The paradigm examples of patterns are colour patterns. Let us for instance think of the simple chequered pattern of the chessboard. In common sense, this pattern of black and white squares is thought of as inhering in a thing, the board. Such a pattern would of course not exist if there were no shapes. Colours are not enough. In order for a shape to exist it must have a border, and the borders of the squares of this pattern are constituted by the different colours of the neighbouring squares; the outermost squares have of course one side constituted by the encompassing colour. If the colours did not differ, there would be no small squares. On the other hand, if there were no shapes nor would there be different colours. A colour pattern contains not only the property of colour, necessarily it also contains the property of shape. Its components are colours-with-shapes. Generalised, the last two statements supply me with my first proposal for one of seven "spatial pattern truths": 1. A spatial property pattern contains not only the property in question, necessarily it also contains shapes; the components of a 5patial property pattern are properties-withshapes. In the sixty-four squares (i.e. colours-with-shapes) of the chessboard, there are a lot of property-grounded relations instantiated. If we name the squares, as in chess, from al to g8, then some of these relations can be described by sentences like the following: 'al has the same shape as bl'. 'b8 is darker than c2', 'f2 has the same size as d3', and so on. These relations exist as scattered in the relata (=colours-with-shapes) they relate. I find it fairly obvious that no "sum" of property-grounded relations can be identical with the corresponding pattern. Such a "sum" is either a set or an aggregate of property-grounded relations. By 'aggregate' I mean "a collection of items not held together by bonds, and /which/ therefore lacks integrity or unity" ([ 1] pp.3-4 ). If the "sum" is a set it is a non-spatiotemporal entity, but the pattern is spatiotemporal, so a set cannot possibly be a pattern. Certainly, an aggregate is a spatiotemporal entity, but since by definition aggregates contain no inner unity, such an aggregate cannot be a pattern either. A colour pattern is a unity in space, and that unity cannot be identical with a number of relations that have scattered instantiation. From these remarks the second of my claims about spatial patterns follows: 2. A spatial property pattern contains a lot of property-grounded relations, but it is not identical with a set or an aggregate of such relations. My third claim is similar to the second one, but it is not equally obvious. It is about spatial relations like 'al is to the left of a2', 'a2 is between al and a3', etc. It says: 3. A spatial property pattern contains a lot of spatial relations, but it is not identical with a set or an aggregate of such relations. To repeat, a set (=non-spatiotemporal entity) of relations between colours-with-shapes cannot be identical with a colour pattern (=spatioternporal entity). But what about, for instance, the aggregate of all the spatial relations between all the sixty-four squares of the chessboard pattern? If, in the aggregate in question, we have all relations like 'c4 is in space close to b4, c3, c5, d4 ' with their relata (and the property-grounded relations they constitute), do we not then have the chessboard pattern as well? In my opinion, we have all the parts needed for the pattern, but we do not have the unity necessary for the pattern itself. Not even this aggregate has the kind of inner unity which patterns have. An aggregate does by definition not take the Gestalt character of a pattern into account. Spatial patterns are spatially inclusive unities. The last-mentioned feature comes out more clearly in the following thought experiment. Let us assume that the sixty-four squares of the chessboard are spread out in such a way that each square is one centimetre away from all the neighbouring squares. This rearrangement of the components of the old pattern changes a lot of spatial relations, at the same time as it le happens: New ~ space which no1 Spatial patterns• scattered. They , regarded as ~ chessboard beC<1 properties of sel hole property. n 4. A spatial pr~ I According t properties and' relations, and (l for a spatial pã claim below, t:11 normally result i size of all the i.m sizes are not cw affords me with' I 5. The type-idel 3 The miñ There is a põ proposed, namel have written, tJI today we kno• Therefore, the ~ patterns, note~ Of course! also defend the j 6. Mind-indep~ patterns. 1 I will not d that I take it fo• mind-indepeñ strength are tlm scalar quantitid patterns, althouj In order tG realistically, õ hand there is th! the other hand j densities are ñ differs from thd construct. It has realistically as ~ has in each spat A specific, colour hue, or I instance is extd reaches the end 2 There are of coli f the simple . and white course not :xist it must he different e one side be no small colours. A :ontains the te last two s": :essarily it mies-withare a lot of n al to g8, 'al has the , on. These nd it fairly Tesponding :lations. By h/ therefore >oral entity, n aggregate .ty, such an i that unity From these ut it is not It is about it identical .vi th-shapes about, for 1ares of the is in space ations they we have all lttern itself. :regate does patterns are ~g thought read out in uares. This 1ons, at the I. Johansson I Pattern as an Ontological Category 89 same time as it leaves all the old property-grounded relations unchanged. But something more happens: New components come into existence. The new pattern necessarily makes all the space which now exists between the sixty-four squares into one or several components.2 Spatial patterns cannot, like for instance property-grounded relations and aggregates, exist as scattered. They can only exist in a closed area. If a patch outside a chessboard shall be regarded as part of the chessboard pattern, then the space between the patch and the chessboard becomes part of the pattern, too. In topological terms, a pattern has to have the properties of self-connectedness and 0-connectivity, i.e. the one-piece property and the nohole property. The following statement is, I think, true: 4. A spatial property pattern contains a specific kind of spatial unity. According to the four "spatial pattern-truths" stated above, a spatial pattern contains (i) properties and properties-with-shapes, (ii) property-grounded relations, (iii) spatial relations, and (iv) a specific kind of spatial unity. All these parts are necessary conditions for a spatial pattern to exist. (Together with a further requirement, formulated in the seventh claim below, they constitute a sufficient condition.) A change in any of the parts will normally result in a change of the pattern. A special kind of change occurs when the absolute size of all the instances of properties-with-shapes are changed in such a way that their relative sizes are not changed. Then we have the same pattern as before, only smaller in size. This affords me with yet another claim: 5. The type-identity of a spatial property pattern is size independent. 3 The mind-independence of spatial patterns There is a possible overall objection to all the five "spatial pattern-truths" that I have proposed, namely that they have to be placed within some kind of subject-object distinction. I have written, the objection continues, as if colours are mind-independent properties. But today we know that perceived and sensed colours exist only in the eye of the beholder. Therefore, the objection ends, I have shown nothing whatsoever about mind-independent patterns, not even their existence. Of course I think that this objection fails and I will, at the same time as I explain why, also defend the following representation thesis: 6. Mind-independent spatial patterns can be represented by mind-dependent spatial patterns. I will not discuss realism in general, only realism for property patterns. This means that I take it for granted that some of the properties which physics talks about do exist in a mind-independent world. Density (=mass density), charge density, and electromagnetic field strength are three such properties which can constitute property patterns. The first two are scalar quantities whereas the third is a vector quantity. There are scalar as well as vector patterns, although I will reason only about scalar quantities. In order to realise that densities-with-shapes and density patterns can be understood realistically, one has to grasp a distinction between real and nominal density. On the one hand there is the real density of each compact and homogeneous part of a body with mass, on the other hand there is the (nominal) mean value density. In homogeneous things the two densities are numerically identical; in heterogeneous things the mean value density always differs from the real density of at least some parts. Mean value density is merely a conceptual construct. It has to be conceived nominalistically. Real density, however, can be conceived as realistically as mass can. As a moving body has in each instant a velocity, a body with mass has in each spatial point a density. A specific colour hue instance is extended in space until it "meets" an instance of another colour hue, or reaches the end of the thing it inheres in. Similarly, a specific real density instance is extended in space until it "meets" an instance of another specific real density, or reaches the end of the thing it inheres in. All real densities that are not infinitesimally small, 2 There are of course new spatial relations, too; but that it is not the important point. 90 I. Johansson I Pattern as an Ontological Category have such borders and are always instantiated as densities-with-shapes. This means that they can be components in spatial property patterns. Between two ( or more) densities-withshapes, there are always property-grounded relations as well as spatial relations. As soon as the densities-with-shapes come together in such a way that they inherit a spatial unity from that part of the space in which they exist, a spatial density pattern comes into existence. An example of a density pattern could be a sheet of wood "divided" into sixty-four squares, thirty-two with "white" density and thirty-two with "black" density. This mindindependent chequered density pattern would not be perceptually apprehensible, but it would be just as real as the mind-dependent chequered colour pattern. Obviously, the visual chessboard pattern can be used to represent the density pattern just described. There are in the natural world (mass) density patterns, charge density patterns, electromagnetic field strength patterns, and other kinds of objective spatial property patterns. Most of them seem to lend themselves to representation by means of subjective colour patterns. In passing I want to remark that all physical properties cannot constitute property patterns. Mass, in contradistinction to mass density, cannot. But that is a problem I have pursued elsewhere (see [2] chapter 4, and [3]). 1 4 The conventionality of spatial patterns There are in the world not only on the one hand mind-independent phenomena and on the other hand mind-dependent phenomena, there are fusions of mind-independent and minddependent phenomena as well. A special kind of such fusions occur when, by a man-made decision, some mind-independent entity is divided into parts. The ownership of a bit of land is such a fusion. Although the land in itself is mind-independent, the borders are man-made and in that sense conventional. An estate is afiat object with a fiat boundary. (The concept of fiat objects and boundaries is taken from some of B. Smith's papers; see e.g [5].) Conventions can exist in relation to both mind-dependent and mind-independent phenomena. Phenomenal colours, to take an example, are subjective, but the propertygrounded relations they ground are objective. (Objective grounded relations can be grounded in subjective phenomena.) The property-grounded relations of similarity and dissimilarity among phenomenal colour hues can constitute a scale, the spectrum. And, trivially, the spectrum can be conventionally divided into a number of colour hues. Likewise, the density scale can be conventionally divided into, for example, very low density, low density, mean density, high density, and very high density. Continuous scales contain no natural borders. Therefore, for pragmatic reasons, consciously, half-consciously or unconsciously we create fiat boundaries. Like continuous scales, mind-independent spatial property patterns have no natural borders. Since zero density can be part of density patterns, the whole universe may be said to make up one spatial density pattern. All objective spatial property patterns are merely parts of the corresponding pattern of the universe. A specific density instance, on the other hand, has a natural border. It ends where another density is instantiated. But, obviously, a density pattern can not end in this way. The density on the other side of the presumed border can very well be made part of the pattern. The borders of ordinary spatial mind-independent patterns cannot but be conventional. Or, in other words: Spatial mind-independent patterns (which are smaller than the whole universe) are fiat objects. This conventionality highlights the spatial unity spoken of in claim number four. In fact, it is the unity of space itself which creates the unity of each spatial pattern. I have taken a socalled container conception of space for granted (see [2] chapter 10), and in such a space pure spatial parts are necessarily fiat parts. Such a space is neither constituted by relations between things or events in space, nor by its parts since the parts have no identity of their own. The parts exist merely as parts of space. The conventional border of a pattern does not create the unity within the border. It merely "carves out" a part of space which already has unity, and that unity "unites" all the included properties-with-shapes and all their relations into a spatial property pattern. 'In my terminology in [2] and [3], exclusive properties can constitute patterns but inclusive properties cannot; mass is an inclusive property. The ( sixth) 1 dependent spati• creating in the ' Mind-de they can be co clear, our perce patterns. There What our perce has to do in re In thought instance that choose the table the room in w * on. The border The claims' Spatial patterns properties-with Spatial unificati A spatial patte extends beyond existence. Non and think of te Consider changing colo chessboard p chessboard pa are seen as rep A lot of o non-changing let the shapes vary with time. squares const last thought e spatiotemporal the clockwork.* The comp where the co time changing (Real spa* impossible to argued th~t if~--geometry ma The seven following co but some, I ho I. A spatiotd. changing pro * • shapes. . Comment: S square which spatial and te 1at they :s-with- ;oon as 'J from :ty-four : mind- : would visual atterns, atterns. colour roperty I have on the I mindn-made of land n-made :oncept Jendent opertyounded nilarity lly, the density ,, mean ,orders. ~ create natural said to Jarts of nd, has density der can Jendent patterns In fact, :n a so- .ce pure elations of their oes not 1dy has elations :s I. Johansson I Paltern as an Ontological Category 91 The (sixth) claim that mind-independent spatial patterns can be represented by minddependent spatial patterns, relies on the fact that the space of visual perception is unitycreating in the same way as objective space is. Mind-dependent spatial patterns, like the chessboard pattern, are conventional, too. But they can be conventional in a, so to speak, natural way. As Gestalt psychology once made clear, our perceptual system construes unities. And, obviously, this applies to perceptions of patterns. Therefore, visual patterns can (falsely') appear as naturally given "brute facts". What our perceptual system does in relation to mind-dependent patterns, our cognitive system has w do in relation to mind-independent patterns. In thought we are able to transcend perceptually given pattern borders. Assume for instance that the chessboard I have talked about is lying on a table. In thought we can then choose the table as the border, and if we do, we will get another colour pattern. If we choose the room in which the table is placed as the border, we will get still another pattern. And so on. The border of the chessboard pattern must be regarded as conventional. Except for the universe as a whole. it is true that: 7. Spatial patterns are necessarily fiat ohjects. The claims number seven and number four should be seen in the light of each other. Spatial patterns need both a delimitation and a unification. A delimitation of instances of properties-with-shapes without any unification would give us an aggregate, not a pattern. Spatial unification without delimitation would give us the universe only. S Spatiotemporal patterns A spatial pattern can exist as a whole. but this is not possible for a temporal pattern which extends beyond "the now". The past no longer exists, and the future has not yet come into existence. Nonetheless we can represent time by means of a spatial line, and we can imagine and think of temporal patterns in analogy with spatial patterns. Consider again the chessboard, but now imagine that the squares, as time goes by, are changing colours, and that they change in such a way that at every moment there is a chessboard pattern. In time, this spatial pattern gives rise to a temporal pattern. This chessboard pattern with its colour changes is a spatiotemporal colour pattern. If the colours are seen as representing different densities. then we have a spatiotemporal density pattern. A lot of other but similar patterns can jusst as easily be thought of. Above, we had (a) non-changing shapes but changing colours, but we can also (b) keep the colours constant and let the shapes of the pattern vary with time. We can (c) let both the colours and the shapes vary with time. Still another opportunity is (d) to keep both the shapes and the colours of the squares constant, but let them move around with time. We then get a movement pattern. The last thought experiment becomes easier if one thinks of each square as a thing. Such spatiotemporal patterns make up the non-causal part of mechanisms. Think for instance of the clockwork. The complexity of spatiotemporal patterns can be even greater. Try to think of a case (e) where the components of the pattern are moving and changing their shapes, while at the same time changing some of their other properties too. (Real spatiotemporal patterns have of course causes, but that fact does not make it impossible to describe the patterns independently of their causes. I have in another paper, [4], argued that if the causes of mechanisms are abstracted away, then one can study mechanism geometry in a way similar to classical purely spatial geometry.) The seven "spatial pattern truths" I have put forward can be transformed into the following corresponding "spatiotemporal pattern truths". Some of them need a comment, but some, I hope, need not. I. A spatiotemporal property pattern contains properties-with-shapes that are either changing properties, or changini shapes, or moving; this gives rise to spatiotemporal shapes. Comment: Spatiotemporal shapes can be four-dimensional. A two-dimensional chessboard square which continuously changes shape into a circle, gives rise to a three-dimensional spatial and temporal shape: a cube which continuously changes shape into a sphere, gives rise 92 I. Johansson I Pattern as an Ontological Category to a four-dimensional spatiotemporal shape. 2. A spatiotemporal property pattern contains a lot of property-grounded relations, but it is not identical with a set or an aggregate of such relations; the instantiations of these relations can he hoth .1patially and temporally scattered. 3. A spatiotemporal property pattern contains a lot of both spatial and temporal relations, hut it is not identical with a set or an aggregate of such relations. 4. A spatiotemporal property pattern contains a specific kind of spatial unity and a specific kind of temporal unity. Comment: The spatial unity is the same as earlier described. Time, however, has not the same kind of unity, since it is only "the now" which exists. Since this paper is not a paper in the philosophy of time, I will rest content with two remarks. (i) Temporal unity is not a really existing unity: in one sense it is an imagined and fiat unity, but in another sense it is not an imagined unity since the imagined events have had existence. (ii) When we represent time as a spatial line, our representation of time inherits the kind of unity which space has. This means, among other things, that, as spatial patterns are necessarily "one-piece and no-hole" areas or volumes, (spatio)temporal patterns cannot possibly have temporal holes. If, to take an example, a square during t 1 tot, changes shape into a circle, next, from t, to t1 remains a circle, and from t1 to t4 changes -into an ellipse, then the non-change be-tween t2 and t, becomes part of the temporal pattern, too. If this non-change is not regarded as part of the pattern, then we would have one pattern between 11 and t2, and another pattern between t1 and t4 . The first pattern can not be connected with the last pattern into one single pattern without bringing in also what happens during 12 to t 1• Therefore, I regard time as unitycreating in about the same way as container space is unity-creating. 5. The t_vpe-identity of' a spatiotelll/}()rlll property pattern is independent of spatiotemporal si::e. Comment: Both specific kinds of changes and specific kinds of movements can take place in a longer or a shorter temporal interval: both of them have a change intensity. Just as a purely spatial pattern can be spatially diminished without losing its qualitative identity, a spatiotemporal pattern can be spatiotcmporally diminished without losing its qualitative identity. 6. Mind-independent spatiotemporal patterns can he represented by mind-dependent spatiotemporal patterns. 7. Spatiotemporal patterns are necessarily .fiat ohjects. Comment: This claim is a corollary of what was said about spatiotemporal unity in the comment to the fourth claim, and what was earlier said about fiat objects. Spatiotemporally based unities have no natural boundaries, except, of course, the universe and its whole history: if that totality has boundaries. Taking the peculiarity of time's existence into account, one might say that spatiotemporal patterns arc more fiat than purely spatial patterns because they are more dependent upon subjective representation. 6 Causal systems In order for a pattern to exist, a unity must be created. In spatial patterns this unity arises thanks to the unity of space and a fiat delimitation of one part of space; in spatiotemporal patterns the unity arises thanks to a spatiotemporal unity and a fiat delimitation. What distinguishes causal patterns (=systems) from spatial and spatiotemporal patterns is primarily the way their unity is created. Let us imagine an old wire-based telephone system in a little village with fifteen cottages. The components of this causal system are fifteen telephone apparatuses, a telephone exchange, and the telephone-wires. All the components are things with properties. Scattered in these components there are a lot of property-grounded relations, and between them there are a lot of spatiotemporal relations. Metaphorically, on top of this there are a lot of causal relations. Howe create the neces1 my arguments. J The unity, causal interactici telephone systell actual causal inti the wired systell used the compa between the con My cond causal interactio causality and a themselves, the, respects I am a f A telepho cannot be used f system. Nonethc is not part of the: phone and thro1 functioning. In f to affect each ot kinds of causal i The kindl chosen from all fiat we decide to before, conventi systems, just lik1 When, nc order to be brief 1. A causal syst spatiotemporal 2. A causal syst with a set or an 3. A causal syst not identical wi 4. A causal sysi 5. The type-ide1 6. Causal syste1 7. Causal syste1 Once agai be seen in the lit A delimitation o a system. Caus large systems. 7 Degrees ol I have distingui 4 In fact, I think the causal and function pattern, intentionali unity-making factc A group of people, different in charact1 i but it Fthese and a ie sarne r in the l really not an time as s. This D-hole" to take nains a and t, of the 1,1een t3 pattern unitymt of ilace in . purely ttity, a liitative 1endent in the tporally whole ccount, t>ecause r arises :mporal 1. What rimarily fifteen lephone cattered m there 'causal I. Johansson I Pattern as an Ontological Category 93 relations. However, as I have shown before, neither a set nor an aggregate of relations can create the necessary unity; that now the relevant relations are causal makes no difference to my arguments. The unity of the imagined telephone system can neither consist in the unity of actual causal interaction nor in merely the spatiotemporal unity of the system. Firstly, the telephone system exists even when nobody is using it, i.e. it exists even when there is no actual causal interaction among the components. Secondly, there is a telephone system even if the wired system is replaced by a radio telephone system, and when such a system is not used the components exist merely as an aggregate. There are then no spatial connections between the components. My conclusion is that causal systems get their identity from some kind of possible causal interaction. Realism with regards to causal systems requires both a realist view of causality and a belief in de re possibility. Since these are big philosophical issues in themselves, they cannot be dealt with here. There is only space for confession: In both respects I am a believer. A telephone which is not connected by a wire to any of the other fifteen telephones cannot be used for making calls within the system, and cannot therefore be part of this causal system. Nonetheless, this telephone may causally effect the system in some other way which is not part of the functioning of the telephone system. For instance, someone may take this phone and throw it at a functioning phone, and so make the whole phone system nonfunctioning. In fact, most things on earth seem, in some far-fetched way or other, to be able to affect each other. Therefore, the unity of a causal system is always a unity of certain kinds of causal interactions. The kinds of possible causal interactions which constitute a causal system have to be chosen from all the possible ones. This means that every causal system is a fiat system. By fiat we decide to focus attention only on some kinds of interactions. However, as remarked before, conventionality can fuse with mind-independent phenomena. The "fiatness" of causal systems, just like the "fiatness" of spatial patterns, does not cancel mind-independence. When, now, 1 am once again listing my "seven pillars of pattern wisdom", I will, in order to be brief, do it with only one comment. 1. A causal system contains things with properties and corresponding spatial and spatiotemporal patterns. 2. A causal system contains a lot olproperty-grounded relations, but it is not identical with a set or an aggregate ol such relations. 3. A causal system contains a lot of both spatial, temporal, and causal relations, but it is not identical with a set or an aggregate of such relations. 4. A causal system contains a specific kind of causal unity. 5. The type-identity of a causal system is independent of its spatiotemporal size. 6. Causal systems can he represented by mind-dependent patterns. 7. Causal systems are fiat objects. Once again I would like to say that the claims number seven and number four should be seen in the light of each other. Causal systems need both a delimitation and a unification. A delimitation of causal components without any unification would give us an aggregate, not a system. Causal unification without delimitation would afford us with incomprehensibly large systems. 7 Degrees of self-sufficiency and degrees of fiatness I have distinguished three species of the category of pattern.4 Often, when there is talk of 4 In fact, I think there are four. If one believes, as I do, that intentional states cannot be identical with pure causal and functional data processing systems, then there is still another important species of the category of pattern, intentionality. As I hope is clear by now, what really differentiates the species of patterns is their unity-making factor. In my view, intentionality can be unity-making, just like space, time, and causality can. A group of people who mutually perceive each other, or talk with each other, make up a system which is different in character from a purely causal system. The unity of this system is created by nested intentionality. 94 I. Johansson I Pattern as an Ontological Category species and genera, as in botany taxonomy, the species come out as logically independent of each other. They exist, one may say, side by side. But this is not true for the species of pattern. There is a kind of order of existential dependence among the unity-making factors. In my view, causality requires spatiotemporality, and, of course, spatiotemporality requires spatiality; but not the other way round. Spatial unity is more existentially self-sufficient than spatiotemporal unity, which, in turn, is more self-sufficient than causal unity. This order is an order among the "fiatindependent" factor of a pattern unity. However, it may well be argued that this order implies a corresponding ordering of and degrees of fiatness. That is: spatial patterns are less fiat than spatiotemporal patterns which are less fiat than causal systems'. But all of them can, independently of their degree of fiatness, have a mind-independent part. Pattern realism is true. There are patterns and systems in the world. References [I] M. Bunge, Treatise of Basic Philosophy, vol. 4. Ontology II: A World of Systems, Reidel: Dordrecht 1979. [2] I. Johansson, Ontological Investigations, Routledge: London 1989. [3] I. Johansson. "Physical Addition", in R. Poli & P. Simons (eds.), Formal Ontology, Kluwer: Dordrccht 1996. [4] I. Johansson, "The Unnoticed Regional Ontology of Mechanisms", Axiomathes volume VIII (1997); special issue for the 10th anniversary of the Centro Studi per la Filosofia [5] B. Smith, "On Drawing Lines on a Map". in A.U. Frank, W. Kuhn and D.M. Mark (eds.), Spatial Information Theory. Proceedings of COS IT '95, Springer: Berlin 1995. See [2], chapter 15. 5 Also, I think that causal systems are less fiat than intentional systems, see note 4. Formal Ontology in I'! N. Guarino (Ed.) /OS Press, 1998 De Ge1 Abstract b of specifial! reusablespj type of situi a novels~ this pattern obligations; obligations.: appropriate, designers ii pattern are~ ' 1 Introduction * 1 When constructinj representing reqw as time consumili requirements relllll for this is that reqi for each new syst duplication of effo knowledge-intensr software engineen There are IIl8II and reused. One 1 invoking each od realized through d copied into anoth exchange of techni instead, the soluti facilitates its re-im Essential to al reusable modules. searched through I as suggested in [2 effectively suppor [16], overcome sc functional features	{ "pile_set_name": "PhilPapers" }
Health care auxiliaries in the detection and prevention of oral cancer. Oral cancer is one among the few human cancers with a vast potential for prevention. One of the operational strategies considered to translate preventive measures into practice in developing countries has been the use of community health workers and other health auxiliaries of the primary health care system to disseminate anti-tobacco health education messages and to provide mouth examinations in high-risk individuals during their routine home visits and community meetings. Studies conducted in India and Sri Lanka to address the role of the above approach indicate that it is feasible to train community health workers and other health auxiliaries in primary prevention and early detection of oral cancer and precancerous lesions. However, no evidence of the efficacy of such an approach in reducing the incidence and mortality from oral cancer is yet available. Sufficient evidence in terms of efficacy and cost effectiveness is needed to justifiably convince health administrators for the inclusion of non-communicable disease control in general and oral cancer screening in particular as part of the primary health care delivery by community health workers and other health auxiliaries especially when considering the burden of already existing work responsibilities. The need for studies in this direction is very obvious. However, the opportunities for 'case-finding' and health education should be utilised when encountering high-risk subjects both in primary medical and health care.	{ "pile_set_name": "PubMed Abstracts" }
This undated photo provided by the Texas Department of Criminal Justice shows Billy Jack Crutsinger. Texas Dept. of Criminal Justice A Texas death row inmate was executed Wednesday for fatally stabbing an 89-year-old woman and her daughter more than 16 years ago after entering their Fort Worth home under the pretense of doing some work for them. Billy Jack Crutsinger, 64, received a lethal injection Wednesday evening at the state penitentiary in Huntsville for the 2003 killings of Pearl Magouirk and her 71-year-old daughter Patricia Syren. Authorities said Crutsinger killed the two women and stole Syren's car and credit card. Crutsinger was arrested three days later at a bar in Galveston, more than 300 miles away. In a final statement that lasted four minutes, Crutsinger thanked three friends who witnessed the execution. Get Breaking News Delivered to Your Inbox "I'm at peace now and ready to go and be with Jesus and my family," Crutsinger said. Then as the lethal dose of pentobarbital began, he said he could feel it "in my left arm. It's kind of burning." Crutsinger then began coughing and breathing heavily and then made snoring noises at least 29 times before he stopped moving. At 6:40 p.m. CT — 13 minutes after the lethal dose started — Crutsinger was pronounced dead. No family members of Magouirk and Syren witnessed the execution. Crutsinger did not mention the two women during his final statement. "The defendant stabbed two elderly women to death in their own home. They had offered him a chance at honest work. The loss of mother and daughter Pearl Magouirk and Pat Syren is still felt deeply by their family and the Fort Worth community. Our sympathy and thoughts continue to be with them," Michele Hartmann, one of the prosecutors with the Tarrant County Criminal District Attorney's Office who convicted Crutsinger, said in a statement. Crutsinger was the 14th inmate put to death this year in the U.S. and the fifth in Texas, the nation's busiest capital punishment state. Ten more executions are scheduled in Texas this year. The U.S. Supreme Court declined a request by Crutsinger's attorney to stop the execution. Crutsinger's attorney had alleged his previous lawyer had a long history of incompetent work in death penalty cases. "The jury heard nothing from the defense that provided an explanation about the disease of alcoholism in relation to the offense conduct," including such things as "a history of domestic violence and abuse, and repeated losses of significant friends and relatives," Lydia Brandt, Crutsinger's current attorney, wrote in her one of her Supreme Court petitions. Brandt also argued lower courts had wrongly denied Crutsinger funding to investigate competency and mental health claims that were not sufficiently reviewed by prior attorneys. Lower appeals courts and the Texas Board of Pardons and Paroles had also declined to stop the execution. At trial, Hartmann told jurors Crutsinger's actions had nothing to do with alcohol but were the result of "evil." Friends and family described Magouirk, known as "R.D.," as an avid gardener. Syren volunteered as a receptionist at her church. Both women were retired and lived together. Crutsinger had been "spiraling downward much of his adult life," including three failed marriages and a propensity for violence when he drank, according to a report by a forensic psychologist hired by his trial attorneys. In the months before the slayings, Crutsinger became homeless and increasingly desperate after his wife kicked him out of their home and his mother, who had enabled his behavior, stopped helping him, according to the report. When Crutsinger realized Magouirk and Syren didn't have enough work to give him much financial relief, he flew into an alcoholic rage, the report said. "All of his anger at being left to fend for himself and of having his safety net taken from him was then brought to bear on the victims," according to the report. Magouirk was stabbed at least seven times while her daughter was stabbed at least nine times. Authorities said DNA evidence tied Crutsinger to the killings and he confessed to the crime. In an email, Brandt described Crutsinger's previous appellate lawyer, Richard Alley, as a "great word processor" who cut and pasted "worthless" legal arguments from other cases and who was removed from another death row client's case and suspended from practicing in federal court. Brandt alleged Alley performed similar shoddy work in at least six other death penalty cases. Four of those inmates have been executed. An attorney for former death row inmate Bobby Woods also alleged incompetent work by Alley before Woods was executed in 2009. In 2006, the Texas Court of Criminal Appeals removed Alley from its list of lawyers eligible to represent death row inmates in their appeals. Alley died in 2017. "I do the best I possibly can on all these cases," Alley told the Austin American-Statesman in a 2006 story that was part of a series that looked at the bad work by court-appointed attorneys in appeals of death penalty cases. The Texas Attorney General's Office called Crutsinger's allegations against Alley "speculative" because he had not identified any claim that Alley should have raised but did not. The attorney general's office also said Crutsinger's case has received an "extensive review" during his appeals process.	{ "pile_set_name": "OpenWebText2" }
Q: How should I proceed after hours of not finding balance in this design? Problem / Question: I designed a logo for a little project I just began contributing on, but there's a problem: The F was unbalanced, meaning there was no way I could use this logo on rectangular button or shape. Ultimately the balance makes it a poorly designed logo. I've been working for hours to balance this thing, but I simply cannot figure out how to create an "F" that is balanced with the "I" & "X" using the style shown here (this is horrid): Is there any way that I can achieve symmetric balance with the letter F in this style? How should I proceed after hours of not finding balance in this design? Result after building upon advice here: Note: I'm still not liking the balance / shape that I've achieved. I'll keep trying. Thanks to everyone who helped here, and feel free to chime in in the future with advice that might help others with this kind of problem. This is posted as an answer, but just to make sure everyone sees it: I think the lesson I learned from this may benefit others in the future: When it just isn't working, try re-thinking your design. Sometimes it's good to just play it simple. Although the simplest solutions can often be the hardest to reach: While this isn't related to the question directly, it's indirectly important: I kept working a design that didn't seem to want to be what I needed. In the end the answer was to revisit my objective and ask: Am I trying to create something because I like the style, or because it's the right style for the job at hand? The answer was obvious in this case. A: Can you use one of the front slash line from the x letter and work something out with that? It could give the logo a nice flow. My suggestion is a 5 minute work with bad proportions, but it might give you an idea of something.... A: I think the lesson I learned from this may benefit others in the future: When it just isn't working, try re-thinking your design. Sometimes it's good to just play it simple. Although the simplest solutions can often be the hardest to reach: While this isn't related to the question directly, it's indirectly important: I kept working a design that didn't seem to want to be what I needed. In the end the answer was to revisit my objective and ask: Am I trying to create something because I like the style, or because it's the right style for the job at hand? The answer was obvious in this case.	{ "pile_set_name": "StackExchange" }
All relevant data are within the paper and its Supporting Information files. Introduction {#sec001} ============ Acid hot springs provide unique environments for the evolution and establishment of microbial communities and their response to various biogeochemical and metabolic processes involving hydrogen (H~2~), sulfur (S), iron (Fe) and arsenic (As) \[[@pone.0146331.ref001]--[@pone.0146331.ref006]\]. Generally, all acid hot springs are classified into two different water types: acid sulfate-Cl water and sulfate water with low Cl \[[@pone.0146331.ref007], [@pone.0146331.ref008]\]. Acid sulfate-Cl hot springs are viewed to form by sulfide oxidation after saturated Na-Cl geothermal water is exposed to the Earth\'s surface, and thus have decreased pH values but retain high K, Na, F, Cl, Li, B and As concentrations as geothermal reservoir water \[[@pone.0146331.ref007]\]. The high As concentration of these environments has attracted attention of several groups studying As redox speciation \[[@pone.0146331.ref001], [@pone.0146331.ref009]--[@pone.0146331.ref011]\]. Results of previous studies illustrated that arsenite (As(III)) was predominant in the source water of acid sulfate-Cl hot springs, and was oxidized to arsenate (As(V)) once being discharged along the outlet by bacterial populations such as Hydrogenobacter, Hydrogenobaculum, Sulfurihydrogenibium, and Thiomonas \[[@pone.0146331.ref001], [@pone.0146331.ref009], [@pone.0146331.ref011], [@pone.0146331.ref012]\]. No As(III) oxidation in the source water of acid sulfate-Cl hot springs was observed because of sulfide inhibition by inactivating expressed As(III) oxidase enzyme (Aio) \[[@pone.0146331.ref004], [@pone.0146331.ref013]\]. Due to high concentrations of sulfide, Fe and As in those acid sulfate-Cl hot springs, elemental S and Fe depositions rich in As successively appeared along the outflow channel \[[@pone.0146331.ref009], [@pone.0146331.ref010], [@pone.0146331.ref014]\]. Various microbial populations such as Sulfolobus, Sulfobacillus, Metallosphaera, Sufurihydrogenibium, Hydrogenobaculum, Thiomonas and Acidicaldus had been found to respond to sulfide and Fe oxidations in those acid hot springs \[[@pone.0146331.ref001], [@pone.0146331.ref009], [@pone.0146331.ref014]--[@pone.0146331.ref016]\]. Comparatively, acid sulfate hot springs with low Cl have not studied systematically on As biogeochemistry. These geothermal features form by separation of the vapor phase rich in H~2~S from the reservoir and subsequent condensation and oxidation in shallow oxygen-rich groundwater or surface water \[[@pone.0146331.ref017]\]. Tengchong, located in southwestern of China, is a typical volcanic geothermal area and has abundant geothermal resources \[[@pone.0146331.ref018]\]. Zhenzhuquan in the Rehai geothermal field of Tengchong is a representative acid vapor-formed sulfate hot spring low in Cl, with 128.2 mg/L sulfate, 39.2 mg/L Cl and 71.1 μg/L As, as well as low concentrations of K, Na, Li and B \[[@pone.0146331.ref019]\]. This geochemistry is distinctly different from the previously studied acid sulfate-Cl hot springs (sulfate: 21.0--144.9 mg/L; Cl: 473.6--1907.0 mg/L; As: 1.8--5.3 mg/L) \[[@pone.0146331.ref001], [@pone.0146331.ref009]--[@pone.0146331.ref011]\]. The As concentrations in the sediments of Zhenzhuquan outflow channel occur at up to 16.44 g/kg, which substantially exceeded the terrestrial abundance of As (1.5--3 mg/kg) and could pose a potential environmental risk \[[@pone.0146331.ref020]\]. Previous studies indicated that different microbial communities inhabited geothermal environments with distinct geochemistry, including different media such as water and sediment at the same sites of hot springs \[[@pone.0146331.ref021], [@pone.0146331.ref022]\]. Though some microbial studies on As in mats or sediments along those acid sulfate-Cl hot springs outlet had been conducted by clone library and metagenome sequencing \[[@pone.0146331.ref001], [@pone.0146331.ref009], [@pone.0146331.ref010], [@pone.0146331.ref023]\], arsenic geochemistry and corresponding microbial communities in acid sulfate hot spring with low Cl have yet to be fully understood. Therefore, the objectives of this study were to: (1) investigate the As geochemistry and microbial community structures both in water and sediments along the outflow channel of a typical acid sulfate hot spring with low Cl; (2) evaluate the potential microbially-mediated As oxidation process; and (3) assess the environmental factors shaping the microbial community structures. Materials and Methods {#sec002} ===================== Site description {#sec003} ---------------- No specific permission was required for the described field studies because no animal or human subjects were involved in this research. The sampling locations are not privately owned or protected in any way. The field studies were not involved in endangered or protected species. As mentioned above, Zhenzhuquan (N24.9511°, E98.4361°) located in the Rehai geothermal field of Tengchong geothermal area in Yunnan, southwestern China, was selected for this study ([Fig 1A](#pone.0146331.g001){ref-type="fig"}). This spring is a heart-shaped acidic pool with a depth of 6--7 cm and a length of 4.36 m and is fed by numerous very small vigorous degassing vents ([Fig 1B](#pone.0146331.g001){ref-type="fig"}) \[[@pone.0146331.ref022]\]. It has a seasonally fluctuating temperature from 89.1 to 93.3°C and pH from 3.50 to 6.42, which results from the dilution of rainfall \[[@pone.0146331.ref017], [@pone.0146331.ref024]\]. Zhenzhuquan has a low discharge rate of 0.2 L/S and abundant reddish-brown sediments occur downstream from the source ([Fig 1C](#pone.0146331.g001){ref-type="fig"}). These sediments contain large amounts of As with concentrations up to 16.44 g/kg, even though As concentration is extremely low (48.16 μg/L) in the pool water \[[@pone.0146331.ref025]\]. Consequently, a transect of six sampling sites was established along Zhenzhuquan's outflow channel and each sampling site was assigned a name according to the relative distance from commenced discharge point (0 m), such as 3 m, 6 m and 9 m downstream and -2 m and -1 m in the pool ([Fig 1B](#pone.0146331.g001){ref-type="fig"}). Parallel water and sediment samples at each site were collected in August, 2014. ![A map showing the location of Tengchong in China (A), Zhenzhuquan located in Rehai, Tengchong geothermal area and sampling sites distribution (B), and magnified photograph from sediment at the -6 m downstream (C).](pone.0146331.g001){#pone.0146331.g001} Field measurements and sample collection {#sec004} ---------------------------------------- Water temperature, pH and dissolved oxygen (DO) were measured in the field at the site of water collection using a hand-held meter. Concentrations of sulfide, ammonium, ferrous iron (Fe(II)) and total iron (Fe~Tot~) were also determined in the field with a Hach spectrophotometer (DR850, Hach Corp., USA) according to the manufacturer's instructions. Water samples for laboratory measurements, e.g., anions, cations and dissolved organic carbon (DOC) were collected into 50 mL acid-washed polypropylene bottles and brown glass bottles respectively by filtration of spring water through 0.22 μm syringe polyethersulfone (PES) membrane filters (Pall Corp., NY, USA). Filtered biomass-containing membranes were placed into 15 mL sterile polypropylene tubes and immediately stored in dry ice. Water samples for cations and DOC were acidified with 1% v/v HNO~3~. Arsenic species separation was done on site, following the method reported by Le et al. \[[@pone.0146331.ref026]\]. Briefly, 10 mL of each water sample was passed through a silica-based strong anion-exchange cartridge (Supelco, USA) preconditioned with 50% methanol and deionized water before use. As(V) was retained in the cartridge and As(III) remained in the filtered solution. Subsequently, the cartridge was eluted with 10 mL 1 M HCl to release the bound As(V) to eluate samples. Sediment samples were collected in sterile 50 mL polypropylene tubes in duplicate by using sterile spoons and stored in ice. All samples for microbial community analysis (sediments and biomass-containing membranes) were stored in dry ice in the field and during transport, and then stored at -80°C in the laboratory until further analyses. Laboratory geochemical analysis {#sec005} ------------------------------- The cation and anion concentrations were measured by inductively coupled plasma-optical emission spectrometry (CAP6300, Thermo, USA) and ion chromatography (ICS1100, Dionex, USA), respectively. As~Tot~ and Fe~Tot~ in the sediments were extracted by 1:1 aqua regia digestion method in a water bath \[[@pone.0146331.ref027]\]. Pre-separated As(III) and As(V) from hot spring waters and extracted As~Tot~ from sediments were determined using liquid chromatography-hydride generation -atomic fluorescence spectrometry (LC-HG-AFS, Haiguang AFS-9780, Beijing) according to Jiang et al. \[[@pone.0146331.ref025]\]. Extracted Fe~Tot~ from sediments was determined by the 1,10-Phenanthroline-based assay: 10 mL extracted solutions was mixed with 5 mL acetate-sodium acetate buffer (pH = 4.6), 2.5 mL 1% hydroxylamine hydrochloride solution and 5 mL 0.1% 1,10-phenanthroline solution in 50 mL volumetric flask. The mixtures were made up to a volume of 50 mL with deionized water and allowed to stand for 10 min. The absorbance of each solution at 510 nm was measured with a spectrophotometer (UV1750, Shimadzu, Japan). Dissolved organic carton (DOC) of water samples and total organic carbon (TOC) of sediment samples were determined using a TOC analyzer (TOC-V~CPH~, Shimadzu, Japan) and a Macro elemental analyzer (Multi EA 4000, Analytik Jena, Germany), respectively. DNA extraction, amplification and sequencing {#sec006} -------------------------------------------- DNA was extracted from biomass-containing filters or from 0.5 g sediment samples using the FastDNA SPIN Kit for Soil (MP Biomedical, OH, USA). DNA concentrations were measured by Pico Green using a FLUOstar OPTIMA fluorescence plate reader (BMG LABTECH, Jena, Germany). The V4 region of 16S rRNA gene was amplified using the normal primer pair 515F (`5’-GTGCCAGCMGCCGCGGTAA-3’`) and 806R (`5’-GGACTACHVGGGTWTCTAAT-3’`) combined with Illumina adapter sequences, a pad and a linker of two bases, as well as barcodes on the reverse primers \[[@pone.0146331.ref028]\]. PCR amplification was carried out in a 25 μL reaction containing 2.5 μL 10× PCR buffer II (including dNTPs) (Invitrogen, Grand Island, NY), 0.4 μM of both forward and reverse primers, 10--15 ng DNA and 0.25 U high fidelity AccuPrime^™^ Taq DNA polymerase (Life Technologies) under the following program: initial denaturation at 94°C for 1 min, followed by 30 cycles of 94°C for 20 s, 53°C for 25 s, and 68°C for 45 s, and then a final extension at 68°C for 10 min. Reactions were performed in triplicate. Amplicons of each sample were combined and confirmed positive PCR products by agarose gel electrophoresis, and then quantified with PicoGreen. Finally, a total of 200 ng PCR product of each sample was pooled together and purified through QIAquick Gel Extraction Kit (Qiagen, Valencia, CA) and then was re-quantified with PicoGreen. Sample 16S rRNA clone libraries for sequencing were prepared according to the MiSeqTM Reagent Kit Preparation Guide (Illumina, San Diego, CA, USA) and the protocol described previously \[[@pone.0146331.ref029]\]. Briefly, sample denaturation was performed by mixing 10 μL of combined PCR products (2 nM) and 10 μL 0.2 M NaOH and incubated for 8 min at room temperature. Denatured DNA was diluted to 15 pM using HT1 buffer and mixed with a PhiX DNA library (final concentration 14.3%). A total of 600 μL sample mixture, together with customized sequencing primers for forward, reverse, and index reads, were loaded into the corresponding wells on the reagent cartridge of a 500-cycle v2 MiSeq kit. Sequencing was performed for 251, 12, and 251 cycles, respectively for forward, index, and reverse reads on a Illumina MiSeq system (Illumina, San Diego, CA). Sequence data preprocessing and statistical analysis {#sec007} ---------------------------------------------------- Raw sequences with perfect matches to barcodes were split to sample 16S rRNA clone libraries and were trimmed using Btrim with threshold of QC higher than 25 over 5 bp window size and the minimum length of 150 bp \[[@pone.0146331.ref030]\]. Forward and reverse reads with at least 50 bp overlap and lower than 5% mismatches were joined using Fast Length Adjustment of SHort reads (FLASH) \[[@pone.0146331.ref031]\]. After trimming of ambiguous bases (i.e. N), joined sequences with lengths between 247 and 258 bp were subjected to chimera removal by Uchime \[[@pone.0146331.ref032]\]. Operational taxonomic units (OTUs) clustering was through Uclust at 97% similarity level \[[@pone.0146331.ref033]\], and taxonomic assignment was through Ribosomal Database Project (RDP) classifier \[[@pone.0146331.ref034]\] with a minimal 50% confidence estimate. The above steps were performed through the Galaxy pipeline of Institute for Environmental Genomics in University of Oklahoma (<http://zhoulab5.rccc.ou.edu/>). Samples were rarefied at 12 955 sequences per sample. Singletons of generated OTU table were removed for downstream analyses. All statistical analysis in this study was performed based on genus-level OTUs at the 97% similarity level under the Vegan package in R (<http://www.r-project.org/>), unless otherwise stated. A variety of alpha diversity indices were calculated including Chao1, Shannon and Equitability. Hierarchical cluster tree using unweighted pair group method with arithmetic means (UPGMA), principal coordinates analysis (PCoA) and non-metric dimensional scaling (NMDS) ordination were built to depict the community composition structure based on the Bray-Curtis dissimilarity matrix of detected OTUs. The Envfit function in the package of Vegan was used to overlay the significant environmental variables on the NMDS ordination. Analyses of similarity (ANOSIM), non-parametric multivariate ANOVA (ADONIS), multi-response permutation procedure (MRPP) and Mantel were performed to test for significant differences in microbial community compositions between sample types (i.e., water vs. sediment) and different locations (i.e., pool vs. downstream). The DNA sequences were deposited to the Short Read Archive database at NCBI (Accession number: SRP056673). Results {#sec008} ======= Water and sediment geochemistry {#sec009} ------------------------------- Three water samples (-2 m, -1 m and 0 m) located in the pool of Zhenzhuquan showed similar physical and chemical conditions, with a slight fluctuation in temperature (84.3--91.3°C), pH (3.58--4.33), DO (0.23--0.33 mg/L), DOC (0.36--0.52 mg/L) and concentrations of various ions ([Table 1](#pone.0146331.t001){ref-type="table"} and [S1 Fig](#pone.0146331.s006){ref-type="supplementary-material"}), which suggested that Zhenzhuquan was derived by vigorous degassing from the same recharged source. The pool with a low DO average of 0.28 mg/L had correspondingly low ratios of Fe(III) and Fe~Tot~ (Fe(III)/Fe~Tot~: \~ 0.15), but contained high ratios of As(V) and As~Tot~ (As(V)/As~Tot~: \~ 0.81). Once the source water was discharged from the pool, most of physical-chemical parameters dramatically changed ([Fig 2](#pone.0146331.g002){ref-type="fig"}). Temperature ranged from 84.3°C to 49.3°C. The pH values showed a slight decline (pH = 3.89 at 0 m and pH = 3.69 at 12 m), which was consistent with acidic Succession Spring in Yellowstone National Park (YNP) \[[@pone.0146331.ref009]\]. The decrease in pH was primarily caused by evaporation, suggested by slight increase of F and Cl concentrations along the drainage ([S1 Fig](#pone.0146331.s006){ref-type="supplementary-material"}). Across sampling intervals, DO, sulfate and ratios of Fe(III) to Fe~Tot~ (Fe(III)/Fe~Tot~) significantly increased from 0.29 to 4.74 mg/L, 120.05 to 158.21 mg/L and 0.09 to 0.34, respectively. Dissolved Fe~Tot~ and As~Tot~ concentrations in water samples gradually declined from 0.64 to 0.45 mg/L and 66.83 to 44.83 μg/L respectively, coupled with significantly elevated concentrations of Fe~Tot~ (0.05--4.55 g/kg), As~Tot~ (10.53--16.44 g/kg) and TOC (1.37%-2.81%) in the downstream sediments. 10.1371/journal.pone.0146331.t001 ###### Geochemistry of water and sediment samples along Zhenzhuquan\'s outflow channel in Tengchong geothermal area. ![](pone.0146331.t001){#pone.0146331.t001g} Distance from discharge (m) Aqueous phase Solid phase ----------------------------- --------------- ------------- ------ ------ ------ ------ ------ -------- ------ ------ ------ ------ ------- ------- ------- ------ ------ ------- ------ -2 89.9 3.58 0.33 0.52 3.00 bdl 0.03 112.66 0.45 0.07 0.51 0.13 17.29 46.55 63.84 0.73 0.05 0.50 0.27 -1 91.3 4.33 0.23 0.36 3.30 bdl 0.03 114.37 0.40 0.11 0.51 0.22 9.27 52.99 62.26 0.85 0.07 0.36 0.09 0 84.3 3.89 0.29 0.52 3.10 bdl 0.05 120.05 0.58 0.06 0.64 0.09 9.20 57.63 66.83 0.86 0.09 0.28 0.57 3 76.2 3.64 0.30 1.00 3.00 0.24 0.02 129.20 0.47 0.06 0.53 0.10 8.10 49.67 57.77 0.86 4.55 16.44 1.37 6 56.8 3.69 2.38 0.60 3.40 bdl 0.03 150.95 0.36 0.10 0.45 0.21 7.33 38.53 45.86 0.84 1.20 10.53 2.72 9 49.3 3.69 4.74 0.47 3.40 0.38 0.01 158.21 0.31 0.16 0.47 0.34 7.06 33.78 40.83 0.83 1.29 11.64 2.81 bdl:below detection limit (1 μg/L) ![Distribution of selected geochemical data in water (A-E) and sediment (F) samples along the outflow channel of Zhenzhuquan.\ There were two more sampling sites (4.5 m and 7.5 m downstream) of DO. Error bars in D-F represented the standard deviations of duplicates.](pone.0146331.g002){#pone.0146331.g002} Alpha diversity of microbial communities {#sec010} ---------------------------------------- A total of 304 743 passing sequences were obtained from six-pair parallel water and sediment samples. After rarefaction at 12 955 sequences per sample, OTU clustering at 97% similarity level and removal of singletons, 155 050 sequences remained. A variety of taxa were present, with 123--834 observed and 172--1173 predicted OTUs (based on Chao1) and coverage values ranging from 47.04% to 83.08% ([S1 Table](#pone.0146331.s001){ref-type="supplementary-material"}). Along the outlet, microbial community richness, Shannon diversity and equitability from water samples significantly increased, whereas those indices of sediment samples had a decline with remarkable minimums at 3 m ([Fig 3](#pone.0146331.g003){ref-type="fig"}). Additionally, in the pool of Zhenzhuquan, richness, diversity and equitability in sediment samples were much higher than those in water samples. However, these indices were distinctly lower in sediment samples than those of water samples downstream. These alpha diversity differences between water and sediment samples can also be seen in their correlation with physical-chemical parameters: Shannon diversity and equitability of water samples were significantly correlated with DO, sulfate, Fe(III)/Fe~Tot~, Fe(II), dissolved As~Tot~, temperature and TOC, while these indices of sediment samples were significantly correlated with DOC, solid Fe~Tot~ and As~Tot~ in the sediments ([Table 2](#pone.0146331.t002){ref-type="table"}). ![Alpha diversity indices distribution of microbial community structures of water and sediment samples along the outflow channel of Zhenzhuquan.\ The solid circles and open circles represented sediment and water samples respectively.](pone.0146331.g003){#pone.0146331.g003} 10.1371/journal.pone.0146331.t002 ###### Correlation between alpha diversity indices at 97% similarity OTU level and environment factors. ![](pone.0146331.t002){#pone.0146331.t002g} Environment factors Water samples Sediment samples --------------------- --------------- ----------------------------------------------- ------------------------------------------------- ------------------------------------------------- --------------------------------------------- --------------------------------------------- ----------------------------------------------- Aqueous phase DO +0.492 +0.978[\\\](#t002fn003){ref-type="table-fn"} +0.991[\\\](#t002fn003){ref-type="table-fn"} -0.284 -0.201 -0.134 Sulfate +0.511 +0.911[\](#t002fn001){ref-type="table-fn"} +0.935[\\](#t002fn002){ref-type="table-fn"} -0.456 -0.445 -0.404 Fe(III)/Fe~Tot~ +0.742 +0.896[\](#t002fn001){ref-type="table-fn"} +0.829[\](#t002fn001){ref-type="table-fn"} +0.181 +0.127 +0.105 Fe(II) -0.756 -0.895[\](#t002fn001){ref-type="table-fn"} -0.829[\](#t002fn001){ref-type="table-fn"} -0.144 -0.045 +0.023 As~Tot~ -0.655 -0.952[\\](#t002fn002){ref-type="table-fn"} -0.938[\\](#t002fn002){ref-type="table-fn"} +0.233 +0.277 +0.287 T -0.469 -0.938[\\](#t002fn002){ref-type="table-fn"} -0.939[\\](#t002fn002){ref-type="table-fn"} +0.486 +0.460 +0.411 DOC -0.448 -0.221 -0.122 -0.717 -0.857[\](#t002fn001){ref-type="table-fn"} -0.896[\](#t002fn001){ref-type="table-fn"} Solid phase Fe~Tot~ -0.218 +0.031 +0.107 -0.679 -0.895[\](#t002fn001){ref-type="table-fn"} -0.964[\\](#t002fn002){ref-type="table-fn"} As~Tot~ +0.107 +0.480 +0.544 -0.694 -0.856[\](#t002fn001){ref-type="table-fn"} -0.897[\](#t002fn001){ref-type="table-fn"} TOC +0.461 +0.863[\](#t002fn001){ref-type="table-fn"} +0.896[\](#t002fn001){ref-type="table-fn"} -0.508 -0.504 -0.496 \p\<0.05, \\p\<0.01, \\\p\<0.001 Microbial community compositions {#sec011} -------------------------------- Microbial community compositions were distinctly different between water and sediment samples ([Fig 4](#pone.0146331.g004){ref-type="fig"}). In the pool, water sample 16S rRNA clone libraries were dominated by Crenarchaeota (48.90 to 93.92%) with the rest mainly composed of Actinobacteria (1.42--36.14%) and Proteobacteria (4.12--14.57%), while sediment sample 16S rRNA clone libraries at -2 m and -1 m were predominated by Proteobacteria (74.36--83.31%). Along the outlet from 0 m to 9 m, Crenarchaeota dominated in water samples gradually declined from 93.92% to 34.79%, which was coupled with slight increase of Acidobacteria (0.07--6.06%), Actinobacteria (1.42--2.80%), Armatimonadetes (0.01--5.13%), Bacteroidetes (0.07--8.18%), Firmicutes (0.22--4.36%) and Proteobacteria (4.12--23.10%). However, initial dominant Crenarchaeota in sediment samples was generally substituted by Chloroflexi along the outflow channel, ranging from 0.13% to 37.76%. There were some unclassified phyla in downstream samples 16S rRNA clone libraries, especially the sediment at 9 m ([Fig 4](#pone.0146331.g004){ref-type="fig"}). Generally, Acidobacteria, Armatimonadetes, Bacteroidetes and Chloroflexi positively correlated with DO, ammonia, sulfate, Fe(III)/Fe~Tot~, dissolved As~Tot~ and TOC, and negatively correlated with temperature, aqueous Fe(II), solid Fe~Tot~ and As~Tot~ ([S2 Table](#pone.0146331.s002){ref-type="supplementary-material"}). ![Microbial community structures distribution of water and sediment samples at phylum level.](pone.0146331.g004){#pone.0146331.g004} At the genus level, water sample 16S rRNA clone libraries in the pool were composed of the Crenarchaeota Sulfolobus (41.38--85.95%), the Actinobacteria Nocardia (1.17--34.90%) and the Proteobacteria Ralstonia (1.66--4.66%), Delftia (0.51--2.76%) and Acinetobacter (0.58--2.57%) ([Fig 5](#pone.0146331.g005){ref-type="fig"}, [S3 Table](#pone.0146331.s003){ref-type="supplementary-material"}). In contrast, sediment sample 16S rRNA clone libraries in the pool (-2 m and -1 m) dominated by Proteobacteria correspondingly were harbored by more Ralstonia (14.96--19.76%), Delftia (20.13--23.29%), Acinetobacter (4.10--6.99%), Undibacterium (7.84--9.29%) and Pseudomonas (2.89--2.92%), and less Sulfolobus (0.30--8.89%). Along the outflow channel, Sulfolobus in water samples gradually decreased from 85.95% to 24.94%, coupled with elevated Gp3 (0.02--4.22%) of Acidobacteria, Chthonomonas (0--5.11%) of Armatimonadetes, Alicyclobacillus (0.02--1.93%) of Firmicutes and Acidicaldus (0--10.48%) of Proteobacteria. However, in downstream sediment samples, 73.25--96.21% sequences did not belong to any known genus except for minor Gp3 (0.01--5.05%), Chthonomonas (0.03--10.77%) and Acidicaldus (0.01--4.86%). Most of unknown-genus sequences were from class Thermoprotei (86.48--97.18%) of Crenarchaeota at 0 m and 3 m and class Ktedonobacteria (31.73--37.34%) of Chloroflexi at 6 m and 9 m ([S2 Fig](#pone.0146331.s007){ref-type="supplementary-material"}). Moreover, at the class level, Thermoprotei was predominant in all water samples and sediment samples at 0 m and 3 m, but replaced by Ktedonobacteria in sediment samples at 6 m and 9 m ([S3 Fig](#pone.0146331.s008){ref-type="supplementary-material"}). Acidicaldus, Gp3, Acidisoma and Chthonomonas positively correlated with DO, ammonia, sulfate, Fe(III)/Fe~Tot~, dissolved As~Tot~ and TOC, and negatively correlated with temperature, aqueous Fe(II) and As~Tot~ ([S4 Table](#pone.0146331.s004){ref-type="supplementary-material"}). ![Microbial community structures distribution of water and sediment samples at genus level.\ The relative abundances of all genera in different samples were displayed in [S3 Table](#pone.0146331.s003){ref-type="supplementary-material"}.](pone.0146331.g005){#pone.0146331.g005} Microbial community structure statistics {#sec012} ---------------------------------------- Based on Bray-Curtis dissimilarity at the 97% similarity OTU level, an UPGMA cluster tree of the microbial community populations showed that sediment samples were divided into two groups (pool and downstream), and distinctly separated from the water samples ([Fig 6A](#pone.0146331.g006){ref-type="fig"}). A similar result was also revealed by PCoA analysis with explained 68.5% of the observed variation ([Fig 6B](#pone.0146331.g006){ref-type="fig"}). Four complimentary non-parametric multivariate statistical tests including ADONIS, ANOSIM, MRPP and Mantel further confirmed the significant differences of microbial communities between not only pool and downstream samples, but also water and sediment samples ([S5 Table](#pone.0146331.s005){ref-type="supplementary-material"}). Results of Envfit function indicated that six geochemical parameters were significantly correlated (P\<0.05) with microbial community structure along the outlet, including temperature, DO, sulfate, aqueous As~Tot~, solid TOC and As~Tot~ with R^2^ values of 0.63, 0.53, 0.62, 0.60, 0.65 and 0.57, respectively ([Fig 6C](#pone.0146331.g006){ref-type="fig"}). The similar directions of temperature and aqueous As~Tot~, and the opposite directions of DO, sulfate and TOC indicated correlations among these variables and did not necessarily suggest that all environmental factors were responsible driving forces of community structure. Significant environmental factors shaping microbial communities were selected according to general physiological niche of microbial populations. ![Microbial community distribution patterns at the 97% similarity OTU level.\ All analyses were performed based on Bray-Curtis dissimilarity of normalized OTU abundances of samples. The solid circles and open circles represented sediment and water samples respectively. A: The hierarchical cluster tree using unweighted pair group method with arithmetic means (UPGMA). B: Principal coordinates analysis (PCoA) scatter plot. The first two factors of PCoA1 and PCoA2 could explain 41.13% and 27.37% variations, respectively. C: Non-metric multidimensional scaling (NMDS) ordination plot. A biplot was overlaid on the ordination to identify environmental factors that were correlated with microbial community structure. The length of the line corresponds to the degree of the correlation. Only variables that had a significant correlation (P\< 0.05) are depicted.](pone.0146331.g006){#pone.0146331.g006} Discussion {#sec013} ========== Potential As, S and Fe oxidation processes {#sec014} ------------------------------------------ In the Zhenzhuquan pool, the remarkably high As(V)/As~Tot~ ratio (0.73--0.86) suggested that As(III) oxidation occurred at the discharge source, which were distinctly different from previous studies that showed As(III) was predominate in the source water of acid sulfate-Cl hot springs, such as Dragon Spring (As(V)/As~Tot~ = 0.05) \[[@pone.0146331.ref011]\], Beowulf Spring (As(V)/As~Tot~ = 0.04) \[[@pone.0146331.ref010]\] and Succession Spring (As(V)/As~Tot~ = 0.04) \[[@pone.0146331.ref009]\] in YNP and Champagne Pool in New Zealand (As(V)/As~Tot~ = 0.04) \[[@pone.0146331.ref001]\]. The As oxidation in the pool might be mediated microbially. Previous studies demonstrated that sulfide was a potent inhibitor of microbially-mediated As(III) oxidation in acid systems by inactivating expressed As(III) oxidase enzyme (Aio) \[[@pone.0146331.ref002], [@pone.0146331.ref004], [@pone.0146331.ref013]\]. Consequently, different from acid sulfate-Cl hot springs with sulfide concentrations of 2.02--12.6 mg/L in those previous studies, the distinctly low concentrations of sulfide (0.03--0.05 mg/L) in the Zhenzhuquan pool allowed for microbial As(III) oxidation. Our previous study on aioA genes had demonstrated the presence of several groups of As(III)-oxidizing microorganisms in the pool, including a few unidentified families of Aquificae and some postulated archaea \[[@pone.0146331.ref025]\]. And in this study, based on 16S rRNA sequences, some microbial populations in sample 16S rRNA clone libraries were also highly similar with bacteria Pseudomonas and Ralstonia which were found to be capable of As oxidation in geothermal and other environments \[[@pone.0146331.ref035], [@pone.0146331.ref036]\]. Except for degassing in part, the lack of sulfide (0.03--0.05 mg/L) in the pool was primarily due to microbial sulfide oxidation, suggested by high sulfate concentrations (112.66--120.05 mg/L) and the presence of abundant Sulfolobus (41.38--85.95% in water sample libraries), a putative thermoacidophilic surfur-oxidizing archaeon ([Table 1](#pone.0146331.t001){ref-type="table"} and [Fig 5](#pone.0146331.g005){ref-type="fig"}) \[[@pone.0146331.ref022], [@pone.0146331.ref037], [@pone.0146331.ref038]\]. Concurrent with sulfide oxidation, Fe oxidation happened in the outflow channel, indicated by the decline of Fe(II) concentrations and increase of Fe(III)/Fe~Tot~ ([Fig 2](#pone.0146331.g002){ref-type="fig"}). Previous studies displayed that Fe(II) oxidation in acid hot springs was mediated by microorganisms, such as Metallosphaera str. MK1, Sulfobacillus str. MK2, Sulfolobus str. MK3, Sulfolobales str. MK5 and Acidicaldus str. MK6 \[[@pone.0146331.ref009], [@pone.0146331.ref014]\]. In this study, some microbial populations belonging to Thermoprotei (such as Sulfolobus, and Metallosphaera) and Acidicaldus dominanated in downstream sample 16S rRNA clone libraries and were probably responsible for Fe(II) oxidation in the outflow channel ([Fig 2](#pone.0146331.g002){ref-type="fig"}) \[[@pone.0146331.ref014]--[@pone.0146331.ref016]\]. Due to lack of sulfide, yellow crystalline elemental S deposit presented in those acid sulfate-Cl hot springs did not appear downstream in this study \[[@pone.0146331.ref001], [@pone.0146331.ref009], [@pone.0146331.ref011]\]. Coupled Fe and As concentrations decline in water samples and increase in sediments suggested the co-deposition of As and Fe ([Fig 2](#pone.0146331.g002){ref-type="fig"}). However, extremely high As concentrations (up to 16.44 g/kg) and As/Fe mole ratios (2.70--6.72) in the sediments was significantly different from previous results from the acid sulfate-Cl hot springs (As/Fe mole ratios: 0.60 to 0.74) \[[@pone.0146331.ref009]--[@pone.0146331.ref011], [@pone.0146331.ref039]--[@pone.0146331.ref041]\]. Previous studies documented the clay minerals in geothermal areas, such as smectite and kaolinite, could host As concentrations up to 4 g/kg \[[@pone.0146331.ref008], [@pone.0146331.ref042]--[@pone.0146331.ref044]\]. Coincidentally, smectite and kaolinite were also detected in the downstream sediments of Zhenzhuquan by X-ray diffraction, which suggested that As might be also adsorbed on the clay mineral. Further investigation on mineralogy of the As-rich sediments downstream is warranted. Significant environmental factors shaping the microbial community structure {#sec015} --------------------------------------------------------------------------- Generally, water and sediment sample 16S rRNA clone libraries along the outflow channel of Zhenzhuquan were dominated by Thermoprotei (mostly comprised of the genus Sulfolobus) ([Fig 4](#pone.0146331.g004){ref-type="fig"} and [S2 Fig](#pone.0146331.s007){ref-type="supplementary-material"}), which was significantly different with sediments or waters from acid sulfate-Cl hot springs, mainly colonized by Hydrogenobacter, Hydrogenobaculum and Sulfurihydrogenibium of Aquificae \[[@pone.0146331.ref001], [@pone.0146331.ref009], [@pone.0146331.ref012]\]. This difference was probably derived by their distinct temperature and pH. Hydrogenobacter and Sulfurihydrogenibium favor circumneutral pH and Hydrogenobaculum has an optimal growth temperature of 60--70°C \[[@pone.0146331.ref045], [@pone.0146331.ref046]\], and microaerophilic acidophilic Sulfolobus prefer higher temperature of 65--85°C and more acidic pH of 2--3. These are all conditions similar to the Zhenzhuquan pool above at the 3 m site (DO: 0.23--0.33 mg/L; temperature: 76.2--91.3°C; pH: 3.58--4.33) ([Table 1](#pone.0146331.t001){ref-type="table"}) \[[@pone.0146331.ref022]\]. This result was also consistent with distribution of Sulfolobus in acidic solfataras and hot springs in YNP \[[@pone.0146331.ref047], [@pone.0146331.ref048]\]. As revealed by non-parametric multivariate statistical analysis, there were significant differences in microbial community structures between not only pool and downstream samples, but also water and sediment samples along the outflow channel ([S5 Table](#pone.0146331.s005){ref-type="supplementary-material"}). Temperature, DO and TOC significantly shaped microbial community structure of upstream and downstream samples ([Fig 6](#pone.0146331.g006){ref-type="fig"}). Responding to the relatively low DO (0.23--0.30 mg/L) and high temperature (76.2--91.3°C) in upstream ([Table 1](#pone.0146331.t001){ref-type="table"}), most microbial populations in 16S rRNA clone libraries were microaerophilic/anaerobic, thermophiles and hyperthermophiles, such as the dominant class Thermoprotei (48.90--97.18%, mainly comprised of Sulfolobus in water samples) \[[@pone.0146331.ref049]\], Nocardia (34.90%) in the water sample at -1 m \[[@pone.0146331.ref050]\], Fervidicoccus (4.46%) in sediment sample at 0 m \[[@pone.0146331.ref051]\], Delftia (20.13--23.29%) and Ralstonia (14.96--19.76%) in sediment samples at -1 m and -2 m \[[@pone.0146331.ref052]\] ([Fig 5](#pone.0146331.g005){ref-type="fig"} and [S2 Fig](#pone.0146331.s007){ref-type="supplementary-material"}). With a decline in temperature (56.8--49.3°C) and increased DO (2.38--4.74 mg/L) as well as TOC (2.72--2.81%), more aerobic heterotrophic mesophiles and thermophiles correspondingly inhabited downstream samples libraries, such as Ktedonobacteria (31.72--37.34% in sediment samples) \[[@pone.0146331.ref053]\], Acidobacteria Gp3 (0.91--5.05%) \[[@pone.0146331.ref054]\], Acidicaldus (1.98--10.48%) \[[@pone.0146331.ref055]\], Chthonomonas (2.12--10.16%) \[[@pone.0146331.ref056]\] and Sphingobacteria (4.94--10.40%) \[[@pone.0146331.ref057]\]. A significant positive correlation between sulfate and TOC (r = 0.989, p\<0.001) along the outlet ([Fig 2](#pone.0146331.g002){ref-type="fig"}) implied that sulfide oxidation might be coupled to carbon fixation by facultatively chemoautotrophic archaea, such as above Sulfolobus and unclassified genera within the Thermoprotei, leading to TOC accumulation in downstream sediments \[[@pone.0146331.ref023]\]. Previous studies demonstrated that microbial community diversity increased with temperature decline and TOC or DOC increase in geothermal environments \[[@pone.0146331.ref021], [@pone.0146331.ref022]\]. However, in this study, the remarkably high As concentrations (up to 16.4 g/kg) and abnormal low alpha diversity in the downstream sediments (much lower than those of water at the same site) suggested that solid As~Tot~ in the sediments probably played a significant role in shaping microbial community structure ([Fig 3](#pone.0146331.g003){ref-type="fig"}) \[[@pone.0146331.ref058], [@pone.0146331.ref059]\]. Otherwise, it should be noted that even at the cutoff of 0.07, most sequences from the sediments 16S rRNA clone libraries (72.41--95.91%) could still not be assigned to a known genus, which suggested that some novel species possibly inhabit the high As sediments downstream ([Fig 5](#pone.0146331.g005){ref-type="fig"} and [S2 Fig](#pone.0146331.s007){ref-type="supplementary-material"}). Conclusions {#sec016} =========== Arsenic oxidation mainly occurred in acid sulfate Zhenzhuquan pool with low chloride. Coupled with iron and sulfur oxidation along the outflow channel, arsenic was substantially accumulated in downstream sediments and appeared to significantly constrain their microbial community diversity. Temperature, total organic carbon and dissolved oxygen significantly shaped the different microbial communities between upstream and downstream samples of Zhenzhuquan. Some putative functional microbial populations were possibly related to arsenic oxidation (Aquificae and Pseudomonas), sulfur oxidation (Sulfolobus) and iron oxidation (Sulfolobus, Metallosphaera and Acidicaldus). A total of 72.41--95.91% unassigned-genus sequences in downstream high arsenic sediment 16S rRNA clone libraries probably implied the presence of some novel genera. Supporting Information {#sec017} ====================== ###### Distribution of alpha diversity indices at the 97% similarity OTU level by re-sampling 12 955 reads in each sample. (DOC) ###### Click here for additional data file. ###### Correlation between phylum at 97% similarity OTU level and environment factors. Only phylum significantly correlated with environment factors were displayed. (DOC) ###### Click here for additional data file. ###### The relative abundances of all genera in different samples. The unit is %. (DOC) ###### Click here for additional data file. ###### Correlation between genera at 97% similarity OTU level and environment factors. Only genera significantly correlated with environment factors were displayed. (DOC) ###### Click here for additional data file. ###### Significance tests of microbial community structures between different groups with four different statistical approaches. (DOC) ###### Click here for additional data file. ###### Normalized main cations and anions concentrations (divided by the sum) variation along the outflow channel of Zhenzhuquan. The values in parenthesis were averaged concentrations of ions with a unit of mg/L. (TIF) ###### Click here for additional data file. ###### Distribution of microbial community compositions from water sediment samples at the class level. The ratios which exceeded 0.5% were displayed in this figure. (TIF) ###### Click here for additional data file. ###### Change of main microbial community compositions at class level from water samples (A) and sediment samples (B) along the outflow channel of Zhenzhuquan. (TIF) ###### Click here for additional data file. We are grateful to the staff from the Yunnan Tengchong Volcano and Spa Tourist Attraction Development Corporation for their assistance. [^1]: Competing Interests:The authors have declared that no competing interests exist. [^2]: Conceived and designed the experiments: ZJ PL YXW. Performed the experiments: ZJ PL DJ. Analyzed the data: ZJ PL. Contributed reagents/materials/analysis tools: ZJ PL DJ XD RZ YHW. Wrote the paper: ZJ PL YXW.	{ "pile_set_name": "PubMed Central" }
Highlands and Islands Airports This is for the company operating airports in Scotland, for the operator for Hyderabad Airport, see Rajiv Gandhi International Airport Highlands and Islands Airports Limited (HIAL) is the company that owns and operates 11 airports in the Scottish Highlands, the Northern Isles and the Western Isles. The company is wholly owned by the Scottish Government, and is categorised as Executive Non Departmental Public Body (ENDPB) of the Scottish Government. The company is based at Inverness Airport. History Highlands and Islands Airports Limited was incorporated on 4 March 1986 by the Civil Aviation Authority. In 1995, ownership transferred from the CAA to the Secretary of State for Scotland, and to the Scottish Ministers upon devolution. The company was criticised for a PFI deal signed to build a new terminal at Inverness Airport, which meant that HIAL had to pay £3.50 to the PFI operator for every passenger flying from the airport. In 2006, the PFI deal was cancelled, costing the Scottish Executive £27.5m. Funding It receives subsidies from public funds under terms of the Civil Aviation Act 1982. For the year ending 31 March 2018, HIAL received £29.2 million of public money, of £20.6m was classed as revenue subsidy, and £8.6m was capital investment. Airports HIAL operates the following airports: Barra Airport Benbecula Airport Campbeltown Airport Dundee Airport Inverness Airport Islay Airport Kirkwall Airport Stornoway Airport Sumburgh Airport Tiree Airport Wick Airport References External links Official website Category:Companies established in 1986 Category:Aviation in Scotland Category:1986 establishments in Scotland Category:Companies based in Highland (council area)	{ "pile_set_name": "Wikipedia (en)" }
[Results of volar transposition of the ulnar nerve in cubital tunnel syndrome]. 66 volar transpositions of the ulnar nerve were performed in 63 patients with cubital tunnel syndrome. 53 patients were male, 10 female. The average age was 49.2 years ranging from 21-74 years. 64 patients had a neurological deficit, 2 merely suffered from pain. The nerve conduction velocity was delayed in all patients. 14 patients had had a cubital fracture in their history, 6 patients showed an arthrosis of the cubital joint. 5 patients suffered from a habitual luxation of the ulnar nerve. In 14 cases there had been recurrent distorsions of the cubital joint. 4 patients were diabetics, 2 alcoholics. In 31 cases no pathogenetic factor could be found. A deep intramuscular transposition with insertion of the intramuscular septa was performed. There were excellent and good results in 82% of all cases. 15% remained unchanged. In 3% a deterioration was observed. The shorter the preoperative history the better the prognosis. Complete recovery was less frequent in patient with severe neurological deficit than with slight. The results were better in younger patients than in elder. But long lasting severe atrophies in elder patients recovered completely in some cases. Diabetes mellitus and alcoholism could not be proved as contributing factors but seemed to cause poorer results.	{ "pile_set_name": "PubMed Abstracts" }
Q: php Invalid argument supplied for foreach(); Есть скрипт, который пингует все IP адреса в столбце "ip_address". Нужно чтобы после пинга записывались результат в столбец "status". Т.е. если компьютер находится онлайн, в столбец "status" писать значение 1, а если оффлайн - 0. Пример базы: Вот мой скрипт: <?php $link = mysqli_connect("192.168.10.56", "test", "test", "test"); if (!$link) { echo "Error: Unable to connect to MySQL." . PHP_EOL; echo "Debugging errno: " . mysqli_connect_errno() . PHP_EOL; echo "Debugging error: " . mysqli_connect_error() . PHP_EOL; exit; } echo "Работает: Подключение с базой данных имеется!" . PHP_EOL; echo "Информация о хосте: " . mysqli_get_host_info($link) . PHP_EOL; foreach($devices as $value){ exec("ping -c 1 " . $value->ip_address, $output, $result); if ($result == 0) { $insert_sql = mysqli_query($link, "UPDATE 'test' SET 'status'='1' WHERE ip_addres = '".$value->ip_address."'"); }else{ $insert_sql = mysqli_query($link, "UPDATE 'test' SET 'status'='0' WHERE ip_addres = '".$value->ip_address."'"); } } mysqli_close($link); ?> Меня удивляет то, что, когда я запускаю скрипт в самом сервере, он не показывает никаких ошибок, но и не пишет в столбец. Но когда запускаю его на виртуальном сервере, то вот что: A: Вообще-то правильно он все пишет. Invalid argument supplied for foreach() - он ссылается на строку foreach($devices as $value). Судя по коду, переменная $devices нигде не создается, вот он ее и не видит. Предположим, что $devices это массив с IP адресами. В таком случае Ваш код будет выглядить вот так: // берем из таблицы список ip адресов (тут можно и WHERE добавить) $result = mysqli_query($link, "SELECT ip_address FROM devices"); if ($result) { // если список есть $ok = $bad = Array(); // в этих массивах будем хранить списки адресов после проверки while ($row = mysql_fetch_assoc($result)) { // прокручиваемся через него по одному exec("ping -c 1 " . $row["ip_address"], $output, $result); // пингуем адрес if ($result == 0) { // в зависимости от результатов пинга заносим адрес в массив $ok[] = $row["ip_address"]; } else { $bad[] = $row["ip_address"]; } } // если есть OK адреса - обновлем все сразу одним запросом if (Count($ok)) { mysqli_query($link, "UPDATE test SET `status`=1 WHERE `ip_address` IN ('".implode("', '", $ok)."')"); } // если есть BAD адреса - обновлем все сразу одним запросом if (Count($bad)) { mysqli_query($link, "UPDATE test SET `status`=0 WHERE `ip_address` IN ('".implode("', '", $bad)."')"); } } А если Вас удивляет то, что ошибки показываются на одном сервере и не показываются на другом, так это вполне могут быть просто настройки. Отключено показывание сообщений об ошибках. Поставьте вот это error_reporting(E_ALL & ~E_NOTICE); ini_set('display_errors', '1'); в первых строчках кода. И тогда ошибки будут отображаться везде.	{ "pile_set_name": "StackExchange" }
INTRODUCTION ============ The ribonucleoprotein enzyme ribonuclease P (RNase P) throughout phylogeny removes 5′ leader sequences of precursor tRNAs (ptRNAs) to generate mature tRNA 5′ ends. The enzyme harbors a single RNA subunit (P RNA) of common ancestry, but only bacterial P RNAs are efficient catalysts in vitro in the absence of the single bacterial protein cofactor ([@B1],[@B2]). Very low RNA-alone activity of archaeal, eukaryal and organellar P RNAs ([@B3; @B4; @B5]) correlates with the presence of multiple protein subunits (usually 4 in Archaea and 9--10 in Eukarya). Bacterial P RNAs are composed of two independent folding domains, the catalytic (C-) domain and the specificity (S-) domain (6,7; [Figure 1](#F1){ref-type="fig"}), the latter playing a crucial role in substrate binding via its interaction with the T stem-loop region of ptRNAs (for review, see [@B8]). The single bacterial RNase P protein (P protein), dispensable in vitro but essential in vivo ([@B9],[@B10]), binds to the P2--J2/3--J3/4--P4--J18/2 region in the C-domain of P RNA ([@B11; @B12; @B13; @B14]). The protein increases substrate affinity through interaction with the 5′-leader ([@B14],[@B15]), which entails tighter binding of key metal ions involved in substrate positioning and catalysis ([@B16]). Figure 1.Secondary structure presentation according to Massire et al. ([@B20]) of M. thermoautotrophicus ΔH RNase P RNA (MM), E. coli P RNA (EE), and chimeras with the C-domain from M. thermoautotrophicus and the S-domain from E. coli (ME) and vice versa (EM). The 13 nucleotide exchanges and the single 1-nt insertion in P RNA of strain M. thermoautotrophicus strain Marburg (MM-Marburg in the tables) relative to M. thermoautotrophicus ΔH are indicated along the sequence in the secondary structure of MM RNA. The C-domain is depicted in green and the S-domain is depicted in blue. Grey nucleotides at the 5′- and 3′-ends indicate non-natural additions present in the T7 in vitro transcripts. Activity of archaeal P RNA in the absence of the archaeal RNase P protein cofactors Pop5, Rpp21, Rpp29 and Rpp30 was observed solely for one of the two types of archaeal P RNAs, type A, but not type M ([@B3]). Archaeal type A RNAs are structurally related to bacterial P RNAs, but lack the P13/14 stem and helix-P18 among several other deviations from the bacterial consensus (17; [Figure 1](#F1){ref-type="fig"}). Type A archaeal P RNA-alone activity was observed to be low and limited to conditions of very high ionic strength for optimal activity, such as 3 M NH~4~^+^ and 0.3 M Mg^2+^ in the case of P RNA from the euryarchaeon Methanothermobacter thermoautotrophicus ([@B3]). Addition of the P protein from Bacillus subtilis activated the archaeal P RNA to some extent at lower ionic strength ([@B3]). For bacterial type A RNA, three loop-helix interdomain contacts (L8--P4, L9--P1 and L18--P8) have been identified ([@B18; @B19; @B20; @B21]) and are thought to orient the C- and S-domains toward each other ([Figure 1](#F1){ref-type="fig"}). Only one of these contacts, L8--P4, can form in the M. thermoautotrophicus P RNA structure, since the P18 element is missing and the reported 5′- and 3′-ends of the RNA ([@B3]) only permit formation of a P1 helix too short to serve as a receptor site for L9 ([Figure 1](#F1){ref-type="fig"}). Previous attempts to increase the RNA-alone activity of archaeal or organellar P RNAs by insertion of structural elements or by in vitro selection have failed to identify variants with improved RNA-alone activity relative to the corresponding wild-type RNA ([@B5],[@B17],[@B22]). We have reinvestigated this issue in a more systematic manner by constructing a series of archaeal C-domain variants with confined alterations towards the bacterial P RNA consensus. The aim of this approach was to identify specific deficits of archaeal P RNAs that limit their RNA-alone activity and make them depend so much more on protein cofactors than their bacterial homologs. Combining the archaeal C-domain from M. thermoautotrophicus P RNA with the S-domain of Escherichia coli P RNA we were able to increase activity---up to three orders of magnitude---by simultaneously introducing a few minor alterations into the archaeal C-domain. All variants were screened at two Mg^2+^ concentrations, 100 mM in the RNA-alone reaction and 4.5 mM in the holoenzyme reaction. Observed activity gains may well include contributions from improved Mg^2+^ binding, which in turn could have affected folding, substrate binding, catalysis or binding of the RNase P protein. Thus, activity differences between tested P RNA variants might be mitigated to some extent at Mg^2+^ concentrations higher than those applied here (see Discussion section). We further provide evidence that the genuine 5′-and 3′-ends of P RNA from Methanothermobacter are longer than previously determined ([@B3]). This longer P1, which resulted in 100--200-fold higher RNA-alone activity, may exert its effect primarily through formation of the L9--P1 contact. Finally, analysis of ternary complex formation (P RNA, P protein, ptRNA) revealed that the archaeal S-domain has lost the capacity of bacterial S-domains to confer affine and productive interaction with the substrate, thereby masking the catalytic capacity of archaeal C-domains. MATERIAL AND METHODS ==================== Construction of plasmids for complementation and in vitro transcription ------------------------------------------------------------------------- Plasmids were constructed by standard cloning and PCR methods as described in ([@B5]). For specific details of plasmid construction and the oligonucleotides used, see [Supplementary Information](http://nar.oxfordjournals.org/cgi/content/full/gkn915/DC1). In vitro transcription and 5′ endlabelling of P RNAs and substrate ptRNA -------------------------------------------------------------------------- Run-off transcription with bacteriophage T7 RNA polymerase and 5′ and 3′ endlabelling with T4 polynucleotide kinase and T4 RNA ligase were performed essentially as described ([@B23],[@B24]). Templates used were pSBpt3′HH linearized with BamHI for the Thermus thermophilus ptRNA^Gly^ substrate ([@B23]), pDW98 linearized with BsaAI for E. coli P RNA ([@B24]) and pUC119_T7_M.th.\_rnpB linearized with BsaI for M. thermoautotrophicus wild-type P RNA ([@B3]). Mutants of the latter were transcribed from the pUC19 constructs described in [Supplementary Information](http://nar.oxfordjournals.org/cgi/content/full/gkn915/DC1); all pUC19 derivatives were linearized with EcoRI. Expression and purification of recombinant E. coli RNase P protein -------------------------------------------------------------------- E. coli RNase P protein (tagged with an N-terminal hexahistidyl peptide leader MRGSHHHHHHGS, encoded in plasmid pQE-30) was recombinantly expressed in E. coli JM109 and purifed by Ni-NTA affinity chromatography as described ([@B25]). RNase P processing assays ------------------------- The ptRNA^Gly^ substrate ([@B23]) was preincubated in assay buffer for 5 min at 55°C and 25 min at 37°C, P RNAs (separately) for 5 min at 55°C and 35 min at 37°C. For holoenzyme reactions, E. coli P protein was added to the P RNA 5 min before combining enzyme and substrate. Conditions of RNA-alone reactions were single turnover with trace amounts (\<1 nM) of 5′-^32^P-labelled substrate, 300 nM or 10 µM P RNA, 100 mM Mg(OAc)~2~, 100 mM NH~4~OAc, 50 mM MES and 2 mM EDTA, pH 6.0 and 37°C. Holoenzyme reactions were performed as multiple turnover reactions at concentrations of 10 nM P RNA, 50 nM P protein, 100 nM ptRNA^Gly^, and buffer KN (20 mM HEPES-KOH pH 7.4, 4.5 mM Mg\[OAc\]~2~, 150 mM NH~4~OAc, 2 mM spermidine, 0.05 mM spermine and 4 mM β-mercaptoethanol) at 37°C. Analysis of reactions by denaturing PAGE and evaluation of data was done as described ([@B23]); all quantifications were based on at least three independent experiments. Folding analysis by native PAGE ------------------------------- 3′-End-labelled P RNAs were preincubated at a concentration of 10 nM in buffer KN for 5 min at 55°C and then 50 min at 37°C. After addition of 1 volume of gel loading buffer (10% \[v/v\] glycerol, 4.5 mM MgCl~2~, xylene cyanol and bromophenol blue) at room temperature, the samples were immediately loaded onto non-denaturing polyacrylamide gels (11.25% \[v/v\] polyacrylamide, 66 mM HEPES, 33 mM Tris \[pH 7.4\], 0.1 mM EDTA, 100 mM NH~4~OAc and 4.5 mM MgCl~2~). Gels were run and the RNAs visualized as described ([@B25]). UV melting profiles ------------------- UV melting profiles were recorded at 260 nm on a CARY 100 Bio UV-Visible spectrophotometer, using the Cary Win UV software version 3.00. Bacterial strains ----------------- Total RNAs for RACE experiments were prepared from cells of M. thermoautotrophicus strain Marburg (DSM 2133, syn. M. marburgensis), whereas the Methanothermobacter RNase P RNA used throughout this study is that of M. thermoautotrophicus strain ΔH (DSM 1053). Determination of equilibrium dissociation constants --------------------------------------------------- Equilibrium dissociation constants (K~d~) of complex formation between RNase P holoenzymes and ptRNA substrate were determined by spin column assays essentially as described ([@B26],[@B27]) in a binding buffer containing 50 mM Tris-acetate, pH 7.1, 200 mM NH~4~OAc, 0.05% Nonidet P-40, and 15 mM Ca(OAc)~2~. Ca^2+^ was used instead of Mg^2+^ to prevent substrate cleavage during binding and the concentration of 15 mM was chosen to increase sensitivity of the assay, because the K~d~\'s measured at this Ca^2+^ concentration are substantially lower than those obtained at lower concentrations (e.g. 5 mM Ca^2+^). Before the assay, holoenzymes were reconstituted from P RNA variants and E. coli P protein by incubating 1 µM P RNA in binding buffer for 1 h at 37°C and then adding E. coli P protein to 5 µM, followed by another 15 min at 37°C. Holoenzymes were then diluted in binding buffer to desired concentrations, incubated for 5 min at 37°C, mixed with trace amounts (\<1 nM) of 5′-^32^P-labelled substrate (preincubated for 30 min at 37°C in binding buffer) and incubated at 37°C for 5 min. RESULTS ======= Analysis of E. coli and M. themoautotrophicus wild-type and chimeric P RNAs ------------------------------------------------------------------------------- Starting from M. thermoautotrophicus wild-type P RNA (henceforth called MM RNA) and E. coli wild-type P RNA (termed EE RNA; [Figure 1](#F1){ref-type="fig"}), we constructed P RNA chimeras by combining the S-domain of E. coli P RNA with the C-domain of M. thermoautotrophicus (termed ME RNA) or the E. coli C-domain with the M. thermoautotrophicus S-domain (termed EM RNA; [Figure 1](#F1){ref-type="fig"}). The chimeric P RNAs allowed us to address if the heterologous domains are able to cooperate and if there is a major asymmetry in the contribution of the archaeal C- and S-domains to the low RNA-alone activity of the archaeal P RNA. RNA-alone single turnover reactions (E\>\>S) at pH 6.0 in the presence of 300 nM P RNA, trace amounts (\<1 nM) ptRNA^Gly^ and 0.1 M each Mg^2+^ and NH~4~^+^ revealed only very weak cleavage by EM RNA (2 × 10^−4^ min^−1^) and no activity at all with ME and MM P RNA ([Table 1](#T1){ref-type="table"}). The activity of EM RNA increased linearly with concentration (tested from 300 nM to 10 µM, data not shown), indicating low substrate affinity. At 10 µM P RNA concentration, activities were detectable for all P RNA variants, with the chimeras EM and ME being 34--44-fold more active than the archaeal MM RNA ([Table 1](#T1){ref-type="table"}). The finding of activity at 10 µM but not at 0.3 µM for the MM and ME variants suggested that weak substrate affinity is also a major cause for the poor performance of these two P RNAs. The lower RNA-alone activity of MM versus EM and ME RNA ([Table 1](#T1){ref-type="table"}) showed that deficits of MM RNA are located in its S- as well as its C-domain. Table 1.ptRNA processing activity of E. coli/M. thermoautotrophicus P RNA domain swop chimeras in RNA-alone and holoenzyme assaysP RNA aloneHoloenzymeP RNAk~obs~ \[min^--1^ × 10^−4^\]P RNAk~obs~ \[min^--1^ × 10^−4^\]EE (0.3 μM)51 000 ± 4000EE (10 nM)82 000 ± 2000EM (0.3 μM)2 ± 1EMNot analyzedME (0.3 μM)Not detectableMENot analyzedMM (0.3 μM)Not detectableMMNot analyzedEM (10 μM)88 ± 19EM (10 nM)70 ± 10ME (10 μM)67 ± 17ME (10 nM)2 ± 1MM (10 μM)2 ± 0.1MM (10 nM)2 ± 1Ecat (10 μM)Not detectableEcat (10 nM)77 ± 6[^1] As in the RNA-alone reactions, RNase P holoenzymes reconstituted with the E. coli P protein and tested for multiple turnover under near-physiological salt conditions (buffer KN, 4.5 mM Mg^2+^, 10 nM P RNA, 50 nM P protein, 100 nM ptRNA^Gly^) yielded very low cleavage rates for MM RNA and the ME chimera ([Table 1](#T1){ref-type="table"}). The EM variant was 35-fold more active than MM and ME, though still a factor of \>1000 slower than EE RNA. Higher activity of EM RNA, harbouring the E. coli C-domain, was anticipated because the primary binding site of the bacterial P protein is in the C-domain ([@B11; @B12; @B13; @B14]). The fact that EM RNA activity equaled that of the isolated C-domain of E. coli P RNA (Ecat; [Table 1](#T1){ref-type="table"}) indicates that the archaeal S-domain made no functional contribution under the applied conditions of the holoenzyme assay. Not unexpected, alleles encoding MM, EM and ME RNAs were unable to rescue the mutant phenotype of a bacterial RNase P mutant strain (Table S3). ME RNA variants with specific structural alterations in the archaeal C-domain ----------------------------------------------------------------------------- With its substantial increase in RNA-alone activity compared to MM RNA ([Table 1](#T1){ref-type="table"}), we assumed ME RNA to be more sensitive toward activating changes in the archaeal C-domain. We thus generated an initial set of ME derivatives that were mutated towards the bacterial type A RNase P RNA consensus ([@B28]) by mutations ranging from single nucleotide exchanges to exchange of defined modules ([Figure 2](#F2){ref-type="fig"}). RNA-alone and holoenzyme assays were conducted as specified in [Table 1](#T1){ref-type="table"}, and activities of the ME variants illustrated in [Figure 2](#F2){ref-type="fig"} are summarized in [Table 2](#T2){ref-type="table"}. Table 2.Activity of ME variants in RNA-alone and holoenzyme assaysRNase P RNARNA-aloneHoloenzymek~obs~ \[min^--1^ × 10^−4^\]Changek~obs~ \[min^--1^ × 10^−4^\]ChangeME67 ± 17 12.0 ± 1.01ME-mP10.4 ± 0.1↓ 176×1.1 ± 0.1↓ 2×ME-mP29.7 ± 1.2↓ 7×1.1 ± 0.2↓ 2×ME-mP42.8 ± 1.8↓ 24×1.2 ± 0.1↓ 2×ME-mP541 ± 5↓ 2×1.2 ± 0.1↓ 2×ME-mP17Not detectable1.1 ± 0.1↓ 2×ME-mP1839 ± 6↓ 2×4.0 ± 0.3↑ 2×ME-mJ15/18250 ± 45↑ 4×6.5 ± 0.3↑ 3×ME-mJ2/37.7 ± 1.2↓ 9×52 ± 6↑ 26×ME-mJ15/18/P10.5 ± 0.1↓ 143×1.3 ± 0.2↓ 2×ME-mJ15/18/P229 ± 7.2↓ 2×3.6 ± 0.3↑ 2×ME-mJ15/18/P4160 ± 15↑ 2×3.2 ± 0.3↑ 2×ME-mJ15/18/P5100 ± 25 1×3.2 ± 0.4↑ 2×ME-mJ15/18/P171.6 ± 1.2↓ 42×1.3 ± 0.1↓ 2×ME-mJ15/18/P183250 ± 400↑ 49×12 ± 0.2↑ 6×ME-mJ15/18/2/333 ± 3.5↓ 2×320 ± 30↑ 160×ME-mJ2/3/P21350 ± 240↑ 20×900 ± 200↑ 450×ME-mJ2/3/P2-2480 ± 120↑ 7×480 ± 50↑ 240×ME-mJ15/18/2/3/P25400 ± 910↑ 81×930 ± 20↑ 465×ME-mJ15/18/2/3/P2/nP19880 ± 2520↑ 147×3000 ± 500↑1500×[^2] Figure 2.Secondary structure presentation of the C-domain from M. thermoautotrophicus according to Massire et al. ([@B20]) with mutations introduced during this work. Regions subjected to mutation are circled and highlighted in grey; sequences of the respective mutants (indicated by the prefix 'm', e.g. mP1) are given in the adjacent boxes with the altered nucleotides coloured in red. Throughout the paper, combinations of mutations are indicated by a slash inbetween, e.g. ME-mJ2/3/P2 refers to mutation mJ2/3 as well as mutation mP2 within one construct, with the ME in front indicating that the construct possesses an M. thermoautotrophicus C-domain and an E. coli S-domain. Grey nucleotides at the 5′- and 3′-ends indicate non-natural additions present in the T7 in vitro transcripts. Nucleotide numbering is that of M. thermoautotrophicus ΔH P RNA. RNA-alone activity ------------------ Among the initial set of eight variants, only a single variant, with a point mutation in the region that joins P15 and P18 in bacterial P RNA, had increased RNA-alone activity relative to ME RNA (ME-mJ15/18, 4-fold increase; [Table 2](#T2){ref-type="table"}). The other variants displayed RNA-alone cleavage rates decreased from 2-fold to below the level of detection in the case of ME-mP17. The latter alteration introduced the P6/16/17 elements of E. coli P RNA, based on the idea that this may help to position the L15 loop for interaction with the tRNA 3′-CCA end. The next most prominent impairments were seen for ME-mP1 (176-fold), which introduced the E. coli P1 helix intended to restore the L9--P1 interdomain contact, and for ME-mP4 (24-fold) with three Watson-Crick base pair identities in P4 changed towards the bacterial consensus ([Figure 2](#F2){ref-type="fig"}, [Table 2](#T2){ref-type="table"}). Likewise, insertion of E. coli P18 (ME-mP18) to restore the L18--P8 interdomain contact failed to improve activity, in line with a previous study ([@B17]). In a second round, the variant with increased activity, ME-mJ15/18, was combined with each of the other structural alterations. RNA-alone activities of the double mutants either equalled that of ME RNA (within a factor of two) or remained substantially lower (ME-mJ15/18/P1, ME-mJ15/18/P17), with one notable exception: incorporation of E. coli P18 (ME-mJ15/18/P18) significantly increased activity compared to ME and also ME-mJ15/18 ([Table 2](#T2){ref-type="table"}). If the individual changes (mJ15/18 and mP18) were independent and additive, one would have expected a roughly 2-fold increase in activity. However, the 49-fold increase for ME-mJ15/18/P18 versus ME indicates positive cooperativity between the two structural alterations. Activity with the bacterial P protein as cofactor ------------------------------------------------- When we assayed P RNA mutants for holoenzyme activity using the E. coli P protein as cofactor ([Table 2](#T2){ref-type="table"}, right columns), again only a single variant showed a marked increase in activity (26-fold) over ME RNA (ME-mJ2/3, with J2/3 reduced from 3 to 1 nt; [Figure 2](#F2){ref-type="fig"}). The fact that this small alteration caused a 9-fold activity decrease in the RNA-alone reaction, but 26-fold increase in the holoenzyme reaction suggested an effect on P protein binding, which is consistent with current knowledge about the P RNA binding interface of the bacterial P protein (Figure S1). All other individual changes had little effect (within 3-fold) on the holoenzyme reaction. Combination of the J15/18 mutation with one of the other structural alterations in the second round revealed two variants with substantially improved activity in the holoenzyme reaction: ME-mJ15/18/P18 (6-fold) and ME-mJ15/18/J2/3 (160-fold; [Table 2](#T2){ref-type="table"}). Changes in P2--P3 and the triple mutant ME-J15/18/2/3/P2 -------------------------------------------------------- The activity increase in the holoenzyme reaction owing to deletion of two nucleotides in J2/3 prompted us to combine changes towards the bacterial consensus in J2/3 and P2, aiming at a further improvement of E. coli P protein binding to the ME chimera. Helix-P2, which is part of the bacterial P protein binding site (see Introduction section and Figure S1), consists of 6 bp in MM RNA compared with 7 bp in bacterial P RNAs. We thus constructed P RNA variants that combine the J2/3 mutation with a P2 helix extended by 1 bp. In the first mutant an U--A bp was inserted between the fifth and the sixth base pair (counted from the 5′-end; [Figure 2](#F2){ref-type="fig"}) of the archaeal P2 helix. This increased RNA-alone activity 20-fold (ME-mJ2/3/P2), whereas these alterations individually impaired activity 9-fold (mJ2/3) and 7-fold (mP2; [Table 2](#T2){ref-type="table"}). In the holoenzyme reaction, the activity gain for variant ME-mJ2/3/P2 was 450-fold over ME RNA, compared with a 26-fold rate increase for variant ME-mJ2/3 and a 2-fold drop for variant ME-mP2. Thus, we observed positive cooperativity effects for the combined mJ2/3/P2 changes in the RNA-alone and holoenzyme assays, indicating that the P2--J2/3--P3 region not only contributes to bacterial P protein binding, but also to folding of the catalytic core. We further constructed variant ME-mJ2/3/P2-2 with the goal to restore the bacterial consensus with minimal structural alteration, solely deleting 1 nt in J2/3 and introducing a point mutation that creates an additional U--A bp at the end of P2. This variant improved RNA-alone activity of ME RNA 7-fold and raised substrate turnover 240-fold in the holoenzyme reaction ([Table 2](#T2){ref-type="table"}). The 2--3-fold weaker performance of variant ME-mJ2/3/P2-2 relative to ME-mJ2/3/P2 suggests that a terminal U--A bp at the P2/P3 junction has a slightly destabilizing effect compared with a closing C--G bp. We then combined the mJ15/18 point mutation with the mJ2/3/P2 alteration to generate variant ME-J15/18/2/3/P2. This further increased activity roughly 4-fold relative to ME-mJ2/3/P2 in the RNA-alone reaction, but showed little additional effect in the holoenzyme reaction ([Table 2](#T2){ref-type="table"}), for unknown reasons. Length of P1 and the L9--P1 interdomain contact ----------------------------------------------- In some bacterial P RNAs, a long-range tertiary interaction between loop L9 docking onto helix P1 serves as an interdomain strut of major importance for the global RNA fold ([@B20],[@B29]). The mature 5′- and 3′-ends of MM RNA, mapped in a previous study ([@B3]), would be too short to allow formation of such an L9--P1 contact. However, some archaeal P RNAs seem to have an extended P1 helix compatible with formation of the L9--P1 contact, and even the MM RNA gene sequence encodes the potential for an extended P1 helix that, if part of the P RNA transcript, may enable formation of the P1--L9 interaction. ([@B17]). We thus constructed an MM RNA variant with P1 extended by 7 bp based on the sequence of the P RNA (rnpB) gene of M. thermautotrophicus strain ΔH. This increased the RNA-alone activity of MM RNA 200-fold ([Table 3](#T3){ref-type="table"}). Introduction of this P1 extension into variant ME-mJ15/18/2/3/P2 to generate ME-mJ15/18/2/3/P2/nP1 further increased activity to 147-fold (RNA-alone) and 1500-fold (holoenzyme) over ME RNA ([Table 2](#T2){ref-type="table"}). Table 3.Activity of MM variants in RNA-alone and holoenzyme assaysRNase P RNARNA-aloneHoloenzymek~obs~ \[min^--1^ × 10^−4^\]Changek~obs~ \[min^−1^ × 10^−4^\]ChangeMM1.7 ± 0.1 12 ± 1 1MM-nP1340 ± 60↑ 200×1.3 ± 0.1↓ 2×MM-mJ15/18/P182 ± 1 11.2 ± 0.1↓ 2×MM-mJ2/3/P2230 ± 50↑ 135×7 ± 1↑ 4×MM-mJ2/3/P2-2100 ± 20↑ 59×6.7 ± 0.8↑ 4×MM-mJ15/18/2/3/P2270 ± 60↑ 159×80 ± 10↑ 40×MM-mJ15/18/2/ 3/P2/nP1740 ± 60↑ 435×75 ± 4↑ 38×MM-Marburg3.3 ± 0.6Not detectableMM-Marburg-nP1310 ± 202.1 ± 0.9MM-Marburg- mapped ends240 ± 201.0 ± 0.2[^3] The stimulating effect of extension nP1 on MM RNA-alone activity prompted us to reinvestigate the 5′- and 3′-boundaries of MM RNA by RACE. This was done with the closely related M. thermoautotrophicus strain Marburg instead of the original source strain M. thermautotrophicus ΔH, because we had access to freshly grown cells of the former only. Owing to the switch of strains we performed several control experiments before conducting the RACE experiment: (i) sequencing of the gene encoding the MM-Marburg RNA revealed a few nucleotide changes relative to MM RNA of strain ΔH ([Figure 1](#F1){ref-type="fig"}), but in vitro transcribed MM-Marburg RNA with the short P1 helix as in MM RNA ([Figure 1](#F1){ref-type="fig"}) showed RNA-alone activity very similar to that of MM RNA ([Table 3](#T3){ref-type="table"}); (ii) introducing the same 7-bp extension as present in MM-nP1 into the MM-Marburg RNA, resulting in variant MM-Marburg-nP1 (Figure S2B), indeed revealed a similar activity increase (ca. 100-fold) as seen for MM-nP1 RNA ([Table 3](#T3){ref-type="table"}). Having confirmed that MM-Marburg RNA is an appropriate mimic of MM RNA, we carried out 5′- and 3′-RACE (Figure S2A), which unveiled that in vivo transcripts of MM-Marburg RNA, and by inference of MM RNA, carry 5′- and 3′-ends that permit formation of an extended P1 helix (Figure S2B) to provide a receptor site for loop L9. Construction and analysis of MM variants ---------------------------------------- Next we addressed the question if the activating structural changes nP1, mJ2/3, mP2 and mJ15/18, initially explored in the context of variant ME, would be beneficial in the natural context of MM RNA as well. Indeed, RNA-alone activity of MM RNA increased 435-fold by introducing these four minor changes ([Table 3](#T3){ref-type="table"}), while the activity gain was only 38-fold in the holoenzyme reaction ([Table 3](#T3){ref-type="table"}, columns on the right). Noteworthy, based on the comparison of cleavage rates (k~obs~) for P RNA MM-mJ15/18/2/3/P2/nP1 ([Table 3](#T3){ref-type="table"}) and its counterpart ME-mJ15/18/2/3/P2/nP1 ([Table 2](#T2){ref-type="table"}), the presence of the archaeal S-domain in the MM variant reduced activity 13- and 40-fold in the RNA-alone and holoenzyme reactions, respectively. Measurement of holoenzyme-substrate affinity -------------------------------------------- The relatively low catalytic performance of variant MM-mJ15/18/2/3/P2/nP1, particularly in the holoenzyme reaction, led us to analyze substrate affinities of P RNA variants reconstituted with the E. coli P protein. We initially tried direct measurements of affinity between P RNA and protein, but different approaches were unsuccessful in our hands. We thus switched to measuring ternary complex formation between holoenzyme and substrate ([Table 4](#T4){ref-type="table"}). As complex formation depended on P protein binding to P RNA under the applied low salt concentrations, the assay indirectly measured P RNA--protein affinities. Table 4.Affinity of reconstituted RNase P holoenzymes for the ptRNA^Gly^ substrateRNase P RNAK~d~ (nM)K~d~ changek~obs~ change (holoenzymes)MMn.d. 1MM-mJ2/3/P2n.d.↑ 4×MM-mJ15/18/2/3/P2n.d.↑ 40×MM-mJ15/18/2/3/P2/nP1n.d.↑ 38×MEn.d. 1ME-mJ2/3448 ± 30↑ 895×↑ 2×ME-mJ2/3/P247 ± 11↑ 94×↑ 450×ME-mJ15/18/2/348 ± 4↑ 96×↑ 160×ME-mJ15/18/2/3/P24.2 ± 1.2↑ 8×↑ 465×ME-mJ15/18/2/3/P2/nP15.0 ± 1.0↑ 10×↑ 1500×EE0.5 ± 0.1 1↑ 41 000×[^4] For the original ME chimera as well as MM RNA and all variants thereof ptRNA affinity was too low for K~d~ determination ([Table 4](#T4){ref-type="table"}). This changed with variant ME-mJ2/3, for which a K~d~ of 450 nM could be determined. Additional introduction of the J15/18 mutation increased affinity roughly tenfold. Similarly, the P2 mutation introduced into either the single mutant ME-mJ2/3 or the double mutant ME-mJ15/18/2/3 in either case led to a tenfold increase in affinity. The lowest K~d~ (4.2 nM) was obtained for the ME-mJ15/18/2/3/P2 RNA; additional incorporation of the nP1 extension did not further reduce K~d~ (5 nM). Thus, ptRNA affinities obtained with the best ME variants were 10-fold lower than that of the E. coli holoenzyme (EE RNA, K~d~ = 0.5 nM; [Table 4](#T4){ref-type="table"}). For the ME variants, increases in affinity (reductions in K~d~) nicely correlated with increases in bacterial holoenzyme activity, with the exception of the nP1 extension which enhanced activity ca. 3-fold when added to variant ME-mJ15/18/2/3/P2, but left substrate affinity essentially unaffected ([Table 4](#T4){ref-type="table"}). Low substrate affinity of the MM variants explains why activity gains in the bacterial holoenzyme assay were moderate (4--40-fold): since the archaeal S-domain is defective in substrate binding based on our K~d~ measurements and thus limits the rate of the cleavage reaction, beneficial effects of alterations mJ2/3, mJ15/18 and mP2 in the archaeal C-domain could not be coaxed into activities as high as in the presence of the E. coli S-domain. Folding analysis of P RNA variants by native PAGE ------------------------------------------------- Previous studies have shown that increased activity of P RNAs often goes along with compacted conformers becoming more prominent in native PAA gels ([@B5],[@B25],[@B29],[@B30]). Consistent with this observation, MM RNA showed the most diffuse migrational behaviour in native gels, pointing to a multitude of coexisting conformations ([Figure 3](#F3){ref-type="fig"}A). All other P RNAs analyzed, with a gain in activity compared to that of MM, displayed more distinct bands and some additional compaction, at least after the preincubation step to that allows RNA folding. Replacement of either domain of MM with the counterpart of EE RNA (variants ME and EM) resulted in stabilization of distinct folds, although EM RNA suffered from substantial aggregation as inferred from RNA retained in the gel pocket ([Figure 3](#F3){ref-type="fig"}A). Extension of the P1 helix (MM-nP1) obviously supports tertiary folding ([Figure 3](#F3){ref-type="fig"}B), as distinct conformers appeared relative to MM RNA. Interestingly, RNAs MM-nP1 and MM-M-nP1 differ in that one major conformer appears for the latter compared with two conformers in the case of RNA MM-nP1 ([Figure 3](#F3){ref-type="fig"}B). This finding shows that even minor changes between two related RNAs may have profound effects on conformational equilibria. Gel mobility of variants ME-mJ15/18/2/3/P2/nP1 and MM-mJ15/18/2/3/P2/nP1 differed substantially, with the former resembling ME RNA in showing a single major band and the latter displaying at least two major conformers, similar to MM-nP1 RNA. However, the additional changes mJ15/18, mJ2/3 and mP2 further affected conformation relative to RNA MM-nP1, as inferred from weaker relative intensity of the upper conformer and faster migration of the lower conformer in the case of RNA MM-mJ15/18/2/3/P2/nP1. Figure 3.Native 11.25% PAGE analyses to compare the conformations of P RNA variants. (A) P RNAs from M. thermoautotrophicus (MM), E. coli (EE) and E. coli/M. thermoautotrophicus chimeras (EM and ME); (B) MM and MM-Marburg P RNA variants with P1 extension (MM-nP1, MM-M-nP1), as well as corresponding C-domain mutants of ME and MM RNA. Assay conditions were the same as in the holoenzyme activity assay (see legend to Table 1 and Material and methods section). Samples were loaded onto the gel either after storage for 55 min on ice (−) or after preincubation for 5 min at 55°C and 50 min at 37°C (+). For further details, see Materials and methods section. UV melting profiles of selected P RNA variants ---------------------------------------------- UV melting profiles were analyzed for P RNA variants ME, MM and MM-nP1, as well as the mJ15/18/2/3/P2/nP1 variants of ME and MM RNA. Conditions were 4.5 mM Mg^2+^ and 100 mM NH~4~^+^ ([Figure 4](#F4){ref-type="fig"}), resembling those of our kinetic holoenzyme assay. Variant MM (red curve) had the lowest T~m~ value (73.1°C), which increased to 74.4°C when helix P1 was extended to its natural length (MM-nP1, black curve). Thus, helix P1 has an overall stabilizing effect, in line with large activity losses of bacterial P RNA variants with disrupted P1 elements ([@B31]). In addition, there is only one major unfolding transition for MM-nP1, based on the first derivative of the melting profile (dA~260~/dT), whereas more unfolding of substructures occurred in MM RNA (shoulders between 60°C and 65°C and at approx. 77°C) aside from the major transition at 73.1°C. Variant ME (blue curve) showed a major unfolding transition around 76°C, but also substantial unfolding of substructures between 50°C and 70°C. In contrast, much less unfolding of substructures at lower temperatures was seen for variant ME-mJ15/18/2/3/P2/nP1 (gray curve), for which the P1 extension might be the major cause, in analogy to the profiles of MM versus MM-nP1 RNA. Figure 4.UV melting profiles of selected P RNA variants. The shown curves represent the mean of at least three measurements; assay conditions: 250 nM P RNA, 50 mM MES pH 6.0, 2 mM EDTA, 100 mM NH~4~OAc, 4.5 mM Mg(OAc)~2~; for UV absorbance (260 nm) melting profiles (left), the y-axis of every species was normalized to 0.0 − 1.0 (normalized Abs); normalized melting curve shapes were not affected by variation of P RNA concentration, indicating that intramolecular unfolding was measured; 'd normalized Abs/dT' (right panel): first derivatives of the melting curves on the left; the measuring program consisted of three steps: step 1 (preincubation): heating of the sample to 55°C at a rate of 10°C/min, incubation for 5 min at 55°C, cooling of the sample to 37°C at a rate of 10°C/min, incubation at 37°C for 35 min; step 2 (measuring of melting profile): heating of the sample from 37 to 90°C at a rate of 0.5°C/min with data recording every 0.1°C; stage 3: immediately after reaching 90°C in step 2, the sample was cooled to 25°C at a rate of 2°C/min and kept at 25°C for 5 min; the next round of measurement was again started with step 1. It is further instructive to compare variant MM-mJ15/18/2/3/P2/nP1 with MM-nP1. The former (green curve) has a similar T~m~ and derivative melting profile as RNA MM-nP1, but shows more unfolding of local structures below 65°C than MM-nP1. Thus, introducing the set of minor changes (mJ15/18/2/3/P2) towards the bacterial consensus into the concertedly folding MM-nP1 RNA uncoupled unfolding of some local structures from unstacking of the bulk of RNA structure. Notably, the derivative melting profiles of variants MM-mJ15/18/2/3/P2/nP1 and ME-mJ15/18/2/3/P2/nP1 were identical up to 65°C, but the major unfolding transition for the latter was broader and shifted to higher temperatures (T~m~ = 76.3°C versus 75.0°C). Therefore, replacement of the archaeal S-domain with the E. coli counterpart increased overall folding stability but reduced the extent of coordinate unstacking. Kinetic parameters of ME-mJ15/18/2/3/P2/nP1 and its MM counterpart ------------------------------------------------------------------ We determined the RNA-alone single turnover kinetic parameters for the best-performing variant ME-mJ15/18/2/3/P2/nP1 to further explore the catalytic capacity of the engineered archaeal C-domain. This analysis revealed a maximum rate (k~react~) of approx. 1.4 min^−1^ and a K~m(sto)~ of about 4 μM for the P RNA ME-mJ15/18/2/3/P2/nP1 ([Figure 5](#F5){ref-type="fig"}A). For comparison, k~react~ and K~m(sto)~ were determined as 10 min^−1^ and 0.24 μM, respectively, for E. coli wild-type P RNA under the same conditions ([@B32]). Thus, P RNA ME-mJ15/18/2/3/P2/nP1 differs from E. coli P RNA by only a 7-fold lower maximum cleavage rate and a 17-fold higher K~m(sto)~. In contrast, the rate of cleavage by variant MM-mJ15/18/2/3/P2/nP1 showed a linear dependence on P RNA concentration up to 10 μM ([Figure 5](#F5){ref-type="fig"}B), indicative of enzyme concentration being subsaturating and consistent with low substrate affinity in the presence of the archaeal S-domain. Figure 5.RNA-alone single turnover kinetics of P RNAs (A) ME-mJ15/18/2/3/P2/nP1 and (B) MM-mJ15/18/2/3/P2/nP1 as a function of the P RNA concentration. Processing assays were performed in the presence of \<1 nM 5′-endlabelled ptRNA, 100 mM Mg(OAc)~2~, 100 mM NH~4~OAc, 50 mM MES and 2 mM EDTA, at pH 6.0 and 37°C. The k~obs~ values are mean values from at least three independent experiments, with error bars indicating the standard error of the mean. For P RNA ME-m J15/18/2/3/P2/nP1, the single turnover kinetic parameters could be derived. K~m(sto)~ is the single turnover K~m~ (describes the enzyme concentration at which the half maximum rate under conditions of \[E\] \>\> \[S\] is achieved), and k~react~ is the single turnover V~max~. For further details, see Materials and methods section. We further tested P RNA ME-mJ15/18/2/3/P2/nP1 for function in B. subtilis and E. coli rnpB mutant strains, also under conditions of simultaneous overexpression of bacterial P proteins to compensate for reduced P RNA-P protein affinity. However, P RNA ME-mJ15/18/2/3/P2/nP1 was unable to rescue the mutant phenotypes (Table S3). DISCUSSION ========== We have shown here that a few mutations toward the bacterial P RNA consensus substantially activate the C-domain of archaeal P RNA from Methanothermobacter, in the absence and presence of the bacterial RNase P protein. All variants were screened at two Mg^2+^ concentrations, 100 mM in the RNA-alone reaction and 4.5 mM in the holoenzyme reaction. Activity measurements at the two Mg^2+^ concentrations provide snapshots, as the individual variants may have different Mg^2+^ optima. Such potential differences in Mg^2+^ requirements imply that the structural changes introduced into the archaeal C-domain may exert their effects on folding, substrate binding, catalysis or binding of the RNase P protein at least partly through changes in binding of crucial Mg^2+^ ions, which has to be considered when interpreting the data. In a recent study on the RNA-alone reaction catalyzed by E. coli RNase P RNA, Kirsebom and coworkers demonstrated that the Mg^2+^ requirements with respect to cleavage increased when substrate contacts to the C-domain (involving substrate functional groups near the cleavage site) as well as contacts to the S-domain (involving the T-stem/loop region of the substrate) are weakened ([@B33]). The improvement in substrate positioning seen for variant ME-mJ15/18/2/3/P2 in our probing experiments (see below) may thus well include contributions from improved Mg^2+^ binding to P RNA regions that mediate contacts to the substrate. Along these lines, we showed previously for E. coli and B. subtilis P RNAs with mutations in the substrate 3′-CCA binding region that in vitro processing defects in the holoenzyme reaction were exacerbated when Mg^2+^ concentrations were lowered from e.g. 4.5 mM to 2 mM ([@B34],[@B35]). Likewise, P RNAs with structural deletions ([@B36]) or substrates with ribose 2′-substitutions at strategic positions were reported to cause an increase in the Mg^2+^ requirement of P RNA-alone reactions ([@B37],[@B38]). On the basis of these findings it is reasonable to predict that low activity P RNA variants in the study presented here will tend to perform relatively better at higher Mg^2+^ concentrations compared to the variants with improved catalytic performance. A central finding of our study was that ribozyme activity of M. thermoautotrophicus P RNA is much higher (100 to 200-fold) than previously assumed owing to the presence of an extended P1 helix ([Table 3](#T3){ref-type="table"}, [Figure 2](#F2){ref-type="fig"}). This P1 extension also stabilizes the global fold as inferred from native PAGE analysis ([Figure 3](#F3){ref-type="fig"}) and UV melting profiles ([Figure 4](#F4){ref-type="fig"}). A likely explanation is that the extension creates a receptor site for loop L9. Since the L18--P8 interdomain contact is lacking, the archaeal RNA may strongly depend on the L9--P1 contact for interdomain orientation of its type A architecture. The presence of an extended P1 helix is also likely for other archaeal P RNAs (Figure S3). Interestingly, a common motif of archaeal P1 elements is an A:A or C:A mismatch at identical position in type A and type M archaeal P RNAs proposed to interact with the third G residue of a conserved 5′-GAGA L9 tetraloop ([@B17]). We have recently demonstrated that a special L9--P1 interaction, occurring in some bacterial P RNAs from thermophiles, represents a major determinant for thermostability of the P RNAs, for RNase P holoenzyme activity at physiological Mg^2+^-concentrations and for conformational compaction ([@B29]). Of note, some bacterial P RNAs harbouring this thermostable P1--P9 module also lack the L18--P8 interaction and have apparently compensated for its absence by strengthening the L9--P1 interdomain contact ([@B29],[@B32]). We recently reported that the P1--L9 interaction is degenerate in E. coli-like mesophilic type A RNase P RNAs, and thus unlikely forms at all, because mutations intended to disrupt this contact remained without effect on E. coli P RNA function in vitro as well as in vivo ([@B29]). This finding explains why variant ME-mP1, designed to restore the E. coli-like L9--P1 contact when its degeneracy was still unknown, failed to improve RNA-alone activity ([Table 2](#T2){ref-type="table"}). However, it remains unclear why the impairment of activity was this severe (176-fold). A pattern emerging from our reactivation study is that mutations within a single structural element were usually insufficient to improve RNA-alone activity. With the exception of the P1 extension, it took the cooperative effect of at least two alterations to be substantially beneficial for ribozyme activity, such as mP18 plus mJ15/18, mP2 plus mJ2/3 or mP2 plus mJ2/3 plus mJ15/18 ([Table 2](#T2){ref-type="table"}). Successful double mutants combined introduction (mP18) or alteration (mP2) of a helix and fine adjustments within adjacent joining regions likely involved in proper positioning of these structural elements. The stimulatory effects of combined changes mJ2/3/P2 on RNA-alone activity were, however, unexpected. Yet, since P2 and P3 are part of the catalytic core, length changes in J2/3 may affect coaxial stacking of P2 and P3 and thereby, combined with the P2 extension, induce subtle changes in folding of the entire core comprising the P1/P4/P5 and P2/P3 stacks as well as the respective joining segments. Surprisingly, in several cases activity increases in the RNA-alone and holoenzyme assays differed substantially. The combined changes in J2/3 and P2 activated RNA-alone activity of variant ME-mJ2/3/P2 20-fold (relative to ME RNA; [Table 2](#T2){ref-type="table"}) and that of variant MM-mJ2/3/P2 135-fold (relative to MM RNA; [Table 3](#T3){ref-type="table"}); however, in the holoenzyme reaction activity stimulation was 450-fold for variant ME-mJ2/3/P2, but only 4-fold for P RNA MM-mJ2/3/P2 ([Table 2](#T2){ref-type="table"} and [3](#T3){ref-type="table"}). A similar situation was seen for the mJ15/18/2/3/P2/nP1 mutation (RNA-alone reaction: 147-fold in the ME, 435-fold in the MM context; holoenzyme reaction: 1500-fold in the ME, 38-fold in the MM context). Such relatively low stimulatory effect on MM variants in the holoenzyme reaction could be attributed to the failure of the archaeal S-domain to confer ternary complex formation at the low metal ion and enzyme concentrations present in the holoenzyme reactions. Indeed, throughout low activities for variants carrying the archaeal instead of the E. coli S-domain ([Tables 2](#T2){ref-type="table"} and [3](#T3){ref-type="table"}, [Figure 5](#F5){ref-type="fig"}) were generally paralleled by very little formation of holoenzyme-substrate complexes ([Table 4](#T4){ref-type="table"}). The less severe phenotype of variants harboring the archaeal S-domain in the RNA-alone relative to the holoenzyme reaction can be attributed to the fact that RNA-alone assays were performed at very high P RNA concentrations (10 μM) and high Mg^2+^ (100 mM), which apparently mitigated the effect of low substrate affinity. These findings pinpoint the S-domain as the limiting part of the archaeal P RNA. In contrast, the archaeal C-domain with a few mutations not only displayed substantial catalytic proficiency, but also highly productive interaction with the bacterial P protein. It remains to be shown whether minor mutations will be able to partially cure the S-domain\'s defect, or if larger architectural alterations, for example, the incorporation of additional structural elements such as P13/P14, will be required to restore S-domain function in the absence of archaeal protein cofactors. The more pronounced activity gain of the mJ15/18/2/3/P2/nP1 mutation in the ME versus MM context (1500-fold versus 38-fold; [Table 2](#T2){ref-type="table"} and [3](#T3){ref-type="table"}) is of further interest in view of the fact that RNAs MM and ME had equal activities in the holoenzyme reaction (2 × 10^−4^ min^−1^; [Table 2](#T2){ref-type="table"} and [3](#T3){ref-type="table"}). This discrepancy can be explained by interdomain cooperativity between the E. coli S-domain and the engineered archaeal C-domain carrying the mJ15/18/2/3/P2/nP1 mutations. Indeed, our structure probing data indicate changes in the E. coli S-domain between variants ME and ME-mJ15/18/2/3/P2 (see below). The C to U exchange in J15/18 was the only single change that had a beneficial effect on the RNA-alone reaction by itself ([Table 2](#T2){ref-type="table"}). The molecular basis of this effect is not immediately clear, since the corresponding nucleotide in the crystal structure of the type A RNase P RNA from Thermotoga maritima is not resolved ([@B21]) and type B RNase P RNAs have a purine at this position ([@B28],[@B39]). In E. coli P RNA, a 2′-deoxy modification at this U residue and Rp-phosphorothioate as well as inosine modification at the conserved 5′-terminal G of J15/18, two nucleotides upstream of the U residue ([Figure 2](#F2){ref-type="fig"}), interfered with tRNA binding ([@B24],[@B40]). This G residue forms a trans Hoogsteen/sugar edge base pair with the 3′-terminal A residue in J5/15 in the crystal structure of type B RNase P RNA from B. stearothermophilus ([@B39]). These findings suggest the J15/18 region to be involved in a structurally and functionally important network of interactions. Comparative probing of P RNAs ME and ME-mJ15/18/2/3/P2 with RNase T1 and Pb^2+^ ions under native conditions unveiled distinct structural differences between the two RNA variants, surprisingly spanning the entire subregion P6, J6/16, P16, J16/15, P15, J15/18 and J5/15 ([Supplementary Figure S4](http://nar.oxfordjournals.org/cgi/content/full/gkn915/DC1)). J16/15 is the site of interaction with the tRNA 3′-CCA end. With respect to J15/18 itself, the 5′-terminal G residue was observed to be more accessible to RNase T1 cleavage in ME versus ME-mJ15/18/2/3/P2 RNA, whereas the site of mutation and the following C residue were more accessible to Pb^2+^-induced hydrolysis in ME-mJ15/18/2/3/P2 relative to ME RNA (Figure S4). Surprisingly, differences in accessibility were also seen in L11/12 and P11, although these regions are identical in both P RNA variants. This indicates that changes mJ15/18/2/3/P2 in the C-domain have long-range effects on S-domain structure. L11/12 and P11 are close to the T-loop region in the P RNA-tRNA complex model based on the T. maritima P RNA X-ray structure ([@B21]). In conclusion, changes mJ15/18/2/3/P2 seem to affect regions that interact with the substrate (J15/16) or which are close to the substrate binding interface (L11/12 and P11). The beneficial effects of these changes on P RNA activity thus seem to stem from improvements in substrate positioning. The structural probing data raise the question if the mutations introduced in P RNA ME-mJ15/18/2/3/P2 will have similar structural and functional consequences when placed into other archaeal C-domains. With respect to evolutionary implications of our approach, several findings are remarkable. First, only three minor changes, in J2/3, J15/18 and P2, were required to stimulate RNA-alone activity 80-fold in the ME and 160-fold in the MM context (465- and 40-fold, respectively, in the holoenzyme assay). Second, combining the changes in J2/3 and P2 improved activity in the absence as well as presence of the bacterial P protein. Thus, the latter exerts its function by making use of a structural element crucially important for RNA-alone activity. This may suggest an evolutionary scenario according to which the bacterial P protein was recruited to the catalytic RNA and adapted to it without eliciting significant co-evolution of the RNA subunit. In contrast, the strong dependence of archaeal/eukaryal P RNAs on their protein subunits for enzyme function indicates RNA/protein co-evolution, during which the archaeal P RNA subunit adapted to the protein components concomitant with substantial losses of RNA-alone activity. From a mirror-image perspective, it will be intriguing to explore if, for example, P RNA ME-mJ2/3/P2 may be impaired in its capacity to cooperate with the archaeal C-domain binding proteins Pop5 and Rpp30 ([@B41]). Along these lines, the substantial activation of the archaeal C-domain by surprisingly little changes suggests only minor differences in RNA conformation between bacterial and archaeal C-domains, in line with the structure probing results (see above). This may imply that the main function of the archaeal C-domain binding proteins Pop5 and Rpp30 for the C-domain itself is to merely fine-tune the conformation of the catalytic core. Finally, the maximum single turnover cleavage rate of variant ME-mJ15/18/2/3/P2/nP1 was only 7-fold lower and the K~m(sto)~ only 17-fold higher than the corresponding values measured for E. coli P RNA. It will be interesting to see if restoring the L9--P1 and/or L18--P8 interdomain struts in this ME chimera will further reduce these differences. In conclusion, the profound activation of the archaeal C-domain by minor changes demonstrates the evolutionary closeness of the archaeal and bacterial P RNAs, at least in terms of C-domain structure and function. SUPPLEMENTARY DATA ================== [Supplementary Data](http://nar.oxfordjournals.org/cgi/content/full/gkn915/DC1) are available at NAR Online. FUNDING ======= This work was supported by the Deutsche Forschungsgemeinschaft (HA 1672/14-1 and GK 1384). Funding for open access charge: DFG/GK 1384. Conflict of interest statement. None declared. Supplementary Material ====================== ###### \[Supplementary Data\] The gifts of pUC119_T7_M.th.\_rnpB (James W. Brown, North Carolina State University) and cells of M. thermoautotrophicus strain Marburg (Rolf Thauer, Max Planck Institute for Terrestrial Microbiology, Marburg) are gratefully acknowledged. [^1]: Assay conditions for the RNA-alone reaction were 50 mM MES pH 6.0 at 37°C, 2 mM EDTA, 100 mM NH~4~OAc, 100 mM Mg(OAc)~2~, trace amounts of ptRNA and 10 µM P RNA unless otherwise indicated. Holoenzyme activity assays were performed at 20 mM HEPES pH 7.4 at 37°C, 2 mM spermidine, 0.05 mM spermine, 4 mM β-mercaptoethanol, 150 mM NH~4~OAc, 4.5 mM Mg(OAc)~2~, 100 nM ptRNA, 10 nM P RNA and 50 nM recombinant E. coli P protein with N-terminal His tag. Errors are standard deviations; all values are based on at least three independent experiments. For further experimental details see Materials and methods section. Values for EE and Ecat RNA are taken from ref. ([@B5]). [^2]: For assay conditions and experimental details, see legend to Table 1 and Materials and methods section. [^3]: For assay conditions and experimental details, see legend to Table 1 and Materials and methods section. For structural details on P1 elements in RNAs MM-Marburg, MM-Marburg-nP1 and MM-Marburg-mapped ends, see Figure S2B. [^4]: Errors for K~d~ are standard errors of the curve fit; n.d., non-determinable because of affinities too low for K~d~ determination by this assay. K~d~ changes are given as n-fold increase in K~d~ relative to the E. coli holoenzyme (EE). Note that in our assay for all variants ptRNA binding to P RNA in the absence of protein did not exceed background values (ptRNA alone). Thus, the binding data for the ternary complex indirectly permit to assess P protein binding to P RNA; k~obs~ changes listed for comparison are taken from Tables 2 and 3.	{ "pile_set_name": "PubMed Central" }
Q: How to achieve the desired output in SQL Server 2008? tbl_Fuel #Temp1 CommodityID Commodity Fuel_Sector Fuel_Rate Fuel_Min Fuel_EffectiveDate ---------------------------------------------------------------------------------------------------- 1 General Cargo OTHERS 38.00 0.00 01/Apr/2019 1 General Cargo GULF+M.EAST 27.00 0.00 01/Apr/2019 1 General Cargo C/D 16.00 0.00 01/Apr/2019 2 Perishable - General OTHERS 7.00 0.00 15/Jun/2015 2 Perishable - General GULF+M.EAST 7.00 0.00 15/Jun/2015 2 Perishable - General C/D 7.00 0.00 15/Jun/2015 tbl_War #Temp2 CommodityID Commodity War_Sector War_Rate War_Min War_EffectiveDate -------------------------------------------------------------------------------------------- 1 General Cargo OTHERS 7.00 0.00 01/Apr/2019 1 General Cargo GULF+M.EAST 5.00 0.00 01/Apr/2019 1 General Cargo C\D 5.00 0.00 01/Apr/2019 1 General Cargo BGW 5.00 0.00 01/Apr/2019 3 Cut Flowers OTHERS 2.00 0.00 15/Jun/2015 3 Cut Flowers GULF+M.EAST 2.00 0.00 15/Jun/2015 tbl_XRay #Temp3 CommodityID Commodity XRay_Sector XRay_Rate XRay_Min XRay_EffectiveDate -------------------------------------------------------------------------------------------- 1 General Cargo ALL 2.00 225.00 16/Nov/2019 9 Pharma & Chemicals ALL 2.00 225.00 01/Jun/2011 Desired output: CommodityID Commodity \| Fuel_Sector \| Fuel_Rate \| Fuel_Min \| Fuel_EffectiveDate \|War_Sector \| War_Rate \| War_Min \| War_EffectiveDate \| XRay_Sector \| XRay_Rate \| XRay_Min \| XRay_EffectiveDate 1 General Cargo \| OTHERS \| 38.00 \| 0.00 \| 01/Apr/2019 \| OTHERS \| 7.00 \| 0.00 \| 01/Apr/2019 \| ALL \| 2.00 \| 225.00 \| 16/Nov/2019 1 General Cargo \| GULF+M.EAST \| 27.00 \| 0.00 \| 01/Apr/2019 \| GULF+M.EAST\| 5.00 \| 0.00 \| 01/Apr/2019 \| \| \| \| 1 General Cargo \| C/D \| 16.00 \| 0.00 \| 01/Apr/2019 \| C\D \| 5.00 \| 0.00 \| 01/Apr/2019 \| \| \| \| 1 General Cargo \| NULL \| NULL \| NULL \| NULL \| BGW \| 5.00 \| 0.00 \| 01/Apr/2019 \| \| \| \| 2 Perishable - General \| OTHERS \| 7.00 \| 0.00 \| 15/Jun/2015 \| \| \| \| \| \| \| \| 2 Perishable - General \| GULF+M.EAST \| 7.00 \| 0.00 \| 15/Jun/2015 \| \| \| \| \| \| \| \| 2 Perishable - General \| C/D \| 7.00 \| 0.00 \| 15/Jun/2015 \| \| \| \| \| \| \| \| 3 Cut Flowers \| \| \| \| \|OTHERS \| 2.00 \| 0.00 \| 15/Jun/2015 \| \| \| \| 3 Cut Flowers \| \| \| \| \|GULF+M.EAST \| 2.00 \| 0.00 \| 15/Jun/2015 \| \| \| \| 9 Pharma & Chemicals \| \| \| \| \| \| \| \| \| ALL \| 2.00 \| 225.00 \| 01/Jun/2011 After so many Condn.. I get the data in three different temp tables... War, Fuel, XRay... I want to show the data commodity wise, if available.... I tried this query: SELECT * INTO #Temp4 FROM (SELECT DISTINCT AirLine, CommodityId FROM #Temp1 UNION ALL SELECT DISTINCT AirLine, CommodityId FROM #Temp2 UNION ALL SELECT DISTINCT AirLine, CommodityId FROM #Temp3) a SELECT * FROM #Temp4 SELECT * FROM #Temp4 a1 LEFT OUTER JOIN #Temp1 a2 ON a1.AirLine = a2.AirLine AND a1.CommodityId = a2.CommodityId LEFT OUTER JOIN #Temp2 a3 ON a1.AirLine = a3.AirLine AND a1.CommodityId = a3.CommodityId LEFT OUTER JOIN #Temp3 a4 ON a1.AirLine = a4.AirLine AND a1.CommodityId = a4.CommodityId This query first DISTINCT data hold in another temp table, then I'm doing a few Left Joins with all temp tables, but it's not returning the correct output. A: You can use full join, but to avoid a Cartesian product for each commodityID, you need a sequence number: select coalesce(f.commodityID, w.commodityID, x.commodityID) as commodityID, coalesce(f.commodity, w.commodity, x.commodity) as commodity, f.fuel_sector, f.fule_rate, . . . -- other columns from "tbl_fuel" w.war_sector, . . . x.xray_sector, . . . from (select f., row_number() over (partition by commodityID order by commodityID) as seqnum from tbl_fuel f ) f full join (select w., row_number() over (partition by commodityID order by commodityID) as seqnum from tbl_war w ) w on w.commodityID = f.commodityID and w.seqnum = f.seqnum full join (select x.*, row_number() over (partition by commodityID order by commodityID) as seqnum from tbl_xray x ) x on x.commodityID = coalesce(f.commodityID, w.commodityID) and x.seqnum = coalesce(f.seqnum, w.seqnum);	{ "pile_set_name": "StackExchange" }
Diamond Platnumz is in trouble for a kiss By Sammy Awami BBC Africa, Dar es Salaam Published duration 17 April 2018 One of Africa's top musicians - Diamond Platnumz - has been questioned by Tanzanian police after posting a video clip of himself playfully kissing a woman on Instagram, which authorities say is indecent. It comes a few weeks after the award-winning "bongo flava" hip hop star threatened to leave the East African nation when his songs were included in list of more than 10 banned by the government for having sexually suggestive video images or lyrics. What law is he accused of breaking? Information Minister Harrison Mwakyembe told MPs the musician had fallen foul of the new Electronic and Postal Communications (Online Content) Regulations, introduced a few weeks ago. He said the authorities plan to file charges against Diamond Platnumz, who has since removed the offending Instagram post. Five things about Diamond Platnumz image copyright AFP One of East Africa's richest musicians Popularised "bongo flava", Tanzanian hip hop Sold second-hand clothes before he found fame Recently launched his own TV and radio station 4.5 million followers on Instagram If found guilty he could face a fine of at least five million Tanzanian shillings ($2,200; £1,500) or a prison sentence of a minimum of 12 months, or both. Mr Mwakyembe said the government was also monitoring other artists who had been engaging in "decadent behaviour", and warned they would be brought to justice irrespective of their popularity. What else do the regulations cover? Apart from censoring obscene content, the legislation requires bloggers to pay a hefty registration fee of more than $900. They must also fulfil a long list of other requirements, like submitting staff CVs, to qualify for a licence to operate. Many activists, online content providers and ordinary users have accused the government of using these regulations to suppress freedom of expression. But the government says they are to protect the nation's "culture". What's behind the crackdown? Since coming to power in 2015, President John Magufuli, nicknamed "The Bulldozer" , has taken a no-nonsense approach to running affairs - cracking down on corruption and government waste. image copyright Reuters image caption President John Magufuli is accused of having little tolerance for dissent His authoritarian nature has won him fans, but critics have also accused him of trying to silence dissenting voices. These online regulations follow the arrests of several people charged with "abusing" the president for criticising him on Facebook and WhatsApp.	{ "pile_set_name": "OpenWebText2" }
Tacky Glowing Valve Caps Look Cheap, Are Cheap Valve-cap LEDs may be tacky, but they'll also make you safer at night Here’s a case where tacky novelty can actually result in something that keeps you safe and also makes you look cool. The Flash Tire Wheel Valve Cap Lights pretty much sum up their function in the name: they are little LED lamps that replace your bike valve dust caps. The little battery-powered lights screw onto a Schrader valve (the fat kind also found on cars and motorbikes) and glow like tiny Lightsabers. For such a cheap item (just $3 per pair on Amazon), they’re actually pretty smart. Instead of switches, the lights have motion and light sensors so they only turn on when you’re moving and it’s dark. Once you get going, they’ll paint a virtual circle of light in the air. Be careful, though. Fellow gadget blogger and Wired.com alumnus John Brownlee put something similar onto his bike when he lived in Berlin, Germany. His lights were bigger and flashier, but the effects on the normally calm and bike-friendly population of Berlin were startling. Poor John was heckled and even had beer bottles thrown at him on one night ride. If you decide to risk it, strap on a helmet and grab a pair in red, green or blue.	{ "pile_set_name": "Pile-CC" }
Swatch TV ARTYOU 2011 PRESENTED BY SWATCH TRAILER Designed to bring together both established and emerging talented artists, Artyou is an annual exhibition event organized by Artsts growing urban art network. This year is Artyou's sixth edition and as part of a collaboration with Swatch plans are underway for 14 artists to both exhibit and show livepaintings in combination with a music/club-program at the city's renowned arts venue Ackermannshof Basel (Switzerland).	{ "pile_set_name": "Pile-CC" }
Q: C++ Template: typename and function to map to int I'm writing a C++ template that needs two params: typename T, and an arbitrary function that maps T to an unsigned int. How can I declare and use a template that can do that? I'd like to keep it simple, so that any dumb function can be used. UPDATE: Here is an example of what I'd like to do: template<typename T, function f> // f has signature: unsigned int f(T); class SortedContainer { ... } And, in this file: unsigned int weight(Package p) { return p.w; } SortedContainer<Package, &weight> sc; UPDATE upon writing code Based on the answers, I tried writing code, but it won't compile. Or rather, the template will compile, but not the test which invokes it. The template code looks like this: template<typename T, typename f> class C { ...f(T)... ... The invocation code looks like: struct S { int operator()(const int n) { return n; // Dummy test code } }; ...C<int, S>&... The error message is: error: no matching function for call to 'S::S(const int&)' note: candidates are: note: S::S() It seems like it's trying to use S's constructor for some reason, as opposed to using the operator() which I want it to do. The purpose of the f parameter is that the SortedContainer needs to be able to position T by an integer value. T is not necessarily an integer or even Comparable, so the caller, when instantiating a SortedContainer, needs to pass not only type T, but a function f to transform T to an integer. A: IMO, you don't require a separate template argument for Function F. template<typename T> // F not required! class SortedContainer { ... }; Choose a good name and use that function by overloading it for various cases. e.g. to_uint() Since you want to map (i.e. relate) a type to an unsigned int (uint), use following function in global scope: template<typename T> uint to_uint (const T& t) { return t.to_uint(); // Must have `uint to_uint() const` member, else error } // Overloads of `to_uint()` for PODs (if needed) template<typename T> // For all kinds of pointers uint to_uint (const T* const pT) { if(pT == nullptr) <error handling>; return to_uint(*pT); } Scenario: For Sorted_Container<X>, whenever to_uint(x) is invoked, then: If X is a class, then it must have uint to_uint() const method Else if X is a POD, then you may have to overload to_uint() for that type Else, the compiler will generate an error	{ "pile_set_name": "StackExchange" }
Take-away and delivered food is extremely popular around the globe, particularly in the UK and USA. Customers increasingly favour the convenience of being able to enjoy their favourite food either delivered out to them by car, motorbike, scooter or bicycle or collected by themselves to then be enjoyed in the comfort of their own homes, workplaces or offices. Whilst once primarily the domain of Pizza restaurants, the takeaway industry is now moving towards many types of cuisine such as Chinese, Indian, Burgers and hot sandwiches. Unlike other cuisines, the nature of burgers and sandwiches make them much harder to transport from the kitchen or restaurant's premises to the customers' homes. The multiple layers of food in a burger often come apart as the food is shaken. Furthermore, when subject to impact, traditional packaging solutions deform causing subsequent deformation and deterioration to the food. Upon arrival the bread may well be soggy or ‘steamed’, and the food can become so disturbed by the journey that it is no longer recognisable or appetising, and barely edible.	{ "pile_set_name": "USPTO Backgrounds" }
Computer Science Degree Getting your computer science degree is one of the best decisions that you can make to ensure a future of prosperity and job security. With a couple of years of study, you can learn the skills that companies all around the world are looking for in their employees. As technology finds its way into more and more parts of our world, those who know how to tame it and make it work to satisfy our needs will be the ones in demand in a job market that is becoming more competitive with every passing day. If you’re unsure about how important it is to understand the role of computers in our everyday lives, take a look around. Twenty years ago, the average house had one computer, if it had one at all. These days, many houses have at least one computer for every person in the house. There are more computers in the average household than there are cars – and most of the newer cars made today have built-in dashboard computers that connect the driver to traffic updates, weather forecasts, and satellite radio. Behind each of these computers is a team of men and women who built the system, programmed the software, and designed the user interface. These men and women all have one thing in common: a computer science degree. Computer science is often defined as the study of computing systems and computational processes. In reality, though, a computer science degree opens you up for a job focusing on any one of a number of specific career paths. Jobs from Information Management to Cyber security, from Database Administration to Computer Programming, are all based in skills and concepts that are mastered as part of the computer science degree program of study. With training in computer science, you become skilled in design methodology, concept testing, system analysis, and project implementation. What does this all mean? With a computer science degree, you will be able to look at a technical problem within a company, diagnose the cause of that problem, analyze the appropriate technical solution, and then build the product that provides the solution. If it sounds like an employee with an education in computer science is a master problem solver, it’s because he or she is. Although a computer science degree provides a good solid base of the concepts of computing, you are not limited to that broad scope. Many computer scientists specialize in fields that provide job security, excellent financial compensation, and a sense that they are a vital part of keeping their business data safe. Information Security personnel use their computer science skills to protect the confidential data of their employer. Employee information, customer records, financial data, and other forms of information are tempting for hackers to try to obtain. While some attempt to steal this data just to prove, to themselves or others, that they can, others will attempt to sell that data or exploit it at the harm of the company they steal it from. A company’s Information Security team prevents such theft, and in order to do that, a firm grasp of computer science is needed. Computer engineering is another popular field in which skills in computer science are required. Computer engineers are often tasked with designing and building the hardware that goes into all of our computers, smart-phones, automobile dashboards, and everything else that computers have found their way inside. This can range from designing the architecture of the latest, thinnest laptop to designing the circuit boards that go inside them. Computer engineers also work on the software side as well, designing the operating system that manipulates the hardware to perform the tasks that the user needs, whether it’s a serious spreadsheet calculation or shooting aliens from the sky in the latest video game. These are only a few of the many types of jobs that are available to you with a computer science degree. You owe it to yourself and to your family to check out the lucrative opportunities that await you after just a few short years of study. The world is not going to suddenly give up on computers. We rely on our gadgets and gizmos more and more every day. Technologically savvy men and women will continue to be in demand in today’s demanding job market. With a computer science degree, you can make sure that you are in demand as well.	{ "pile_set_name": "Pile-CC" }
Session Expired Your session has expired due to inactivity or because your browser does not accept cookies. If the time away from your search was more than 30 minutes, please start your search again. If you do not have cookies enabled, you will need to enable them in your internet browser before continuing. Sign up for our newsletter today and you will receive all the latest special offers before anyone else. Sign up now!	{ "pile_set_name": "Pile-CC" }
Is a Memory Foam Mattress Right for My Child? This entry was posted on April 15, 2016 by TCDEV. Selecting the right mattress for your child is no easy task, especially if it is their first ‘grown up’ bed. These days it seems many adults are opting for memory foam beds for themselves, but what about for our little ones? In recent years there has been a lot of talk about the benefits of memory foam over traditional innerspring mattresses - for both our bodies and sleep schedule. With this shift towards memory foam comes questions about the safety - and appropriateness - of a memory foam bed for children. Given the fact that children sleep longer and deeper than adults, it is absolutely imperative that they are resting their little heads on a quality, supportive mattress. Furthermore, a quality mattress is a huge factor in whether a child is getting a good night’s sleep and waking up rested and ready to conquer the day. While there are undoubtedly many different options when it comes to picking out a child’s mattress, today we are going to focus on the benefits of memory foam. While there is some debate as to whether memory foam is good for infants and toddlers, the general consensus is that older children and teens can certainly reap the benefits of memory foam, without cause for concern. What is Memory Foam? Many people do not realize this, but memory foam was actually developed by NASA in response to a need for a foam material that could mold to a person’s body, then bounce back to its original shape. Since first introduced, memory foam has proved to be a viable option when it comes to a mattress, for people of all different ages and occupation. Made from polyurethane foam, memory foam mattresses react to pressure and heat, working to conform to the body when you lie down. While these body-contouring properties are desirable for adults, these characteristics can actually be quite dangerous to infants and toddlers. Benefits of a Memory Foam Mattress If you are looking for a supportive and long-lasting mattress for your older child or teenager, memory foam is a wonderful option. Some of the top benefits of this type of mattress include: Contours to the body - As mentioned earlier, memory foam mattresses actively mold to the body in response to heat and pressure. This offers a lot of support and helps to evenly distribute the body’s weight. Allergy-free - Another reason why memory foam may be the right choice for your child is that it is allergy free. Made from polyurethane foam, this type of mattress is full of fibers that actually prevent allergy-causing dust mites from collecting within the bed. Extremely durable - Finally, memory foam mattresses are one of the most durable on the market. They are actually known to last much longer than average spring mattresses, making them a wonderful option for growing children. If you are in the market for a new children’s mattress, be sure and check out our memory foam options here. As always, if you have any questions or would like additional information about any of our products, please do not hesitate to ask.	{ "pile_set_name": "Pile-CC" }
Hartz -- Szuba November 20, 2006 SOUTH BEND -- Christ the King Catholic Church was the setting for the 3 p.m. July 22 wedding of Rachel L. Szuba and Dave Hartz. They are home in Osceola. Their parents are Mary and Jim Szuba of 52060 Chicory Lane, Granger, and Lois and Chuck Hartz of 14097 Kimberly Lane, Middlebury. The bride graduated from Penn High School, Mishawaka, and Indiana University South Bend with a bachelor's degree in elementary education. She works for South Bend Community School Corp. as a first-grade teacher at Warren Primary Center. The groom graduated from Northridge High School, Middlebury, and IUSB with a bachelor's degree in public affairs. He is a national sales manager for Omega Plastics, Elkhart.	{ "pile_set_name": "Pile-CC" }
[Extraction and analysis of profenofos residue in tomato and cabbage by gas chromatography-flame photometric detector]. A quick and effective extraction and clean-up method of profenofos residue in tomato and cabbage is presented. Tomato and cabbage samples were homogenized with a mixture of acetone-hexane (1:1, V/V) using a mechanical homogenizer. The resultant homogenate was cleaned-up by adding active carbon and then filtered under reduced pressure. The filter cake was extracted twice with the same solvent mixture. The filtrates were combined and transferred into a separatory funnel. The organic layer was separated and evaporated to dryness using a rotary evaporator. The residue was dissolved in 2 mL of acetone and transferred into a small glass vial and then determined by GC on a 5% OV-101 Chromosorb W-HP column with flame photometric detector. The results showed that this analytical method can be used for an accurate determination of profenofos residues in tomato and cabbage. The minimum detectable concentration of profenofos in samples was 0.06 mg/kg. The recoveries of profenofos in tomato and cabbage were in the range of 96.2%-105.9% and 94.7%-102.3%, respectively. The relative standard deviations were in the range of 3.7%-4.9% and 3.7%-5.0%, respectively. The tomato and cabbage samples were collected 3 weeks after applying profenofos in the field, and the contents of profenofos were determined. The average contents of profenofos in tomato and cabbage were (13.8 +/- 0.8) mg/kg and (14.4 +/- 0.7) mg/kg, respectively.	{ "pile_set_name": "PubMed Abstracts" }

Subsets and Splits

No community queries yet

The top public SQL queries from the community will appear here once available.