document_title: The Secret Life of Pets

question_text: name of the dogs in secret life of pets

question_tokens: name of the dogs in secret life of pets

text: The Secret Life of Pets

text: Plot[edit]

text: A Jack Russell Terrier named Max lives with his owner Katie in a Manhattan apartment. While she is at work during the day, he hangs out with other pets in the building: tabby cat Chloe, pug Mel, dachshund Buddy, and budgerigar Sweet Pea. One day, Katie adopts Duke, a large mongrel from the pound, leaving Max jealous because of her divided focus on Duke. Enraged by Max's attitude towards him, Duke tries to abandon Max in an alley, but they are both attacked by cats led by Sphynx cat Ozone who removed both dogs' collars and leave them to be caught by Animal Control. Duke fears that he will be put down if he goes back to the pound. To her desperation, Gidget, a white Pomeranian actually discovers that Max is missing.

text: Meanwhile, he and Duke are rescued by a white rabbit named Snowball, the leader of "The Flushed Pets" – a gang of sewer-dwelling animals who hate humans because their owners mistreated them. After Max and Duke pretend to despise humans as much as they do by saying they killed their owners, the Flushed Pets invite them to join. Before they can prove their loyalty by allowing a one-fanged viper to bite them, Snowball learns from the cats that Max and Duke are domesticated. The two dogs escape the sewers and board a ferry to Brooklyn, inadvertently killing the viper in the process; Snowball vows to kill them and leads the Flushed Pets after them.

text: Meanwhile, Gidget recruits a red-tailed hawk named Tiberius to find him, but mistakenly returns carrying Ozone, whom Gidget coerces into telling what he knows about the dogs. They then enlisted Mel, Buddy, Chloe, guinea pig Norman and Sweet Pea. On the way, they meet Pops, an old Basset Hound, who helps Gidget and the pets find Max. Meanwhile, Max and Duke raid a sausage factory for food. Gidget and co. encounter Snowball, who vows to kill them as well, and Norman is captured as the rest of Gidget's team flees.

text: Meanwhile, Duke tells Max about his previous owner, Fred, an elderly man who adopted him as a puppy and loved spending time with him. One day, Duke got lost while chasing a butterfly and was caught by Animal Control, but Fred never came to claim him. Max convinces him to visit Fred's house in a nearby neighborhood, confident Fred will still love him and take him back. When they arrive at Fred's house, they learn from the resident cat Reginald that Fred has died. Heartbroken, Duke accuses Max of attempting to get rid of him and barks at the new homeowners who had just returned to the house, who call Animal Control. The handlers catch Max, but Duke interferes long enough for Max to escape and ends up being captured instead.

text: While trying to rescue Duke as he follows the Animal Control van, Max is attacked by Snowball who tries to kill him. However, when his gang is captured, Snowball realizes that he and Max must work together to rescue them. They drive a city bus into the van on the Brooklyn Bridge, stopping traffic. The Flushed Pets encircle Max, unaware of his partnership with Snowball, but Gidget and her team save him. When Gidget is using her kung-fu fighting skills, Max starts to fall in love with her. The van gets stuck in scaffolding and the Flushed Pets escape. Once Max got the keys to Duke's cage, the van plummets into the East River with him inside. Max is unable to free Duke, so Snowball jumps into the river to retrieve the keys, allowing them to escape the sinking van. Once out of the river, Snowball realizes how good being heroic feels.

text: The entire group returns to the apartment block by pig-driven taxi. Max expresses his love for Gidget, who returns his affection. Snowball and the Flushed Pets then come up with a new plan to annihilate all humans, but a little girl named Molly arrives to adopt Snowball and the remaining Flushed Pets return to the sewers. At first, Snowball resists, but gives in and lets himself become a domesticated pet. The domesticated pets return to their homes and embrace their owners, and Max and Duke finally reunite with Katie, sparking a true friendship.

text: In a mid-credits scene, Buddy and Mel show up in costume at a party in poodle Leonard's apartment. Leonard's owner returns and Tattoo crashes to the floor on the chandelier.

text: Cast[edit]

text:
Louis C.K. as Max, a Jack Russell Terrier.[8]
Eric Stonestreet as Duke, a large, brown, shaggy mongrel.[8]
Kevin Hart as Snowball, a white rabbit.[8]
Jenny Slate as Gidget, a white Pomeranian and Max's love interest.
Ellie Kemper as Katie, Max and Duke's friendly owner.[1][9]
Lake Bell as Chloe, an obese and apathetic grey tabby cat[9] who is one of Max's friends.
Dana Carvey as Pops, an elderly Basset Hound,[10] who is paralyzed in his back legs.
Hannibal Buress as Buddy, a laid-back dachshund[9][11] and one of Max's friends.
Bobby Moynihan as Mel, an hyperactive pug and one of Max's friends.
Steve Coogan as Ozone, a Cockney-accented Sphynx cat.

Steve Coogan also voices Reginald, a malevolent Himalayan cat.

Albert Brooks as Tiberius, a curmudgeonly red-tailed hawk.[9][11]
Chris Renaud as Norman, a guinea pig who keeps getting lost trying to find his apartment.
Michael Beattie as Tattoo, a slow-witted tattooed pig who is one of the Flushed Pets.
Sandra Echeverría as Maria, a character from the La Pasión de la Pasión telenovela that Gidget watches.
Jaime Camil as Fernando, a character from the La Pasión de la Pasión telenovela that Gidget watches.
Kiely Renaud as Molly, a little girl who adopts Snowball.

text: Production[edit]

text: In January 2014, it was announced that Louis C.K., Eric Stonestreet, and Kevin Hart had joined the cast of the film, with Chris Renaud set to direct from a screenplay written by Brian Lynch, Cinco Paul and Ken Daurio, while Illumination Entertainment would produce, and Universal Pictures would distribute the film and was originally titled Max.[12][8] In June 2014, Albert Brooks, Hannibal Buress, Bobby Moynihan, Lake Bell and Ellie Kemper joined the cast.[9] The animation was created entirely in France by Illumination MacGuff.[13]

text: Music and soundtrack[edit]

text:
The Secret Life of Pets (Original Motion Picture Soundtrack)

Soundtrack album by Alexandre Desplat

Released
July 1, 2016 (2016-07-01)

Recorded
2016

Genre

Film score
Jazz
Classical

Length
51:31

Label
Back Lot Music

Producer
Dominique Lemonnier

Alexandre Desplat soundtrack chronology

Florence Foster Jenkins
(2016)Florence Foster Jenkins2016
The Secret Life of Pets
(2016)
The Light Between Oceans
(2016)The Light Between Oceans2016

text: The original score for the film was composed by Alexandre Desplat.[14] The soundtrack was released on July 1, 2016 by Back Lot Music.[15]

text: All music composed by Alexandre Desplat, except "We Go Together" by The Sausage Factory Singers.

text:
[show]Track listing

No.
Title
Length

1.
"Meet the Pets"
2:37

2.
"Katie's Leaving"
0:55

3.
"Meet Duke"
3:36

4.
"Fetch Me a Stick"
3:09

5.
"Telenovela Squirrels"
1:24

6.
"Hijack!"
2:00

7.
"Gidget Meets Tiberius"
4:56

8.
"Initiation Time"
1:01

9.
"Rooftop Route"
1:27

10.
"The Viper"
1:49

11.
"You Have an Owner?"
3:04

12.
"Good Morning Max"
1:21

13.
"Sewer Chase"
1:09

14.
"Who's With Me?"
1:21

15.
"Me Like What Me See"
0:54

16.
"Traveling Bossa"
1:56

17.
"Flushed Out To Brooklyn"
2:47

18.
"Sausages!"
1:13

19.
"Duke's Old House / Captured"
3:03

20.
"Brooklyn Bridge Showdown"
2:34

21.
"Rescuing Duke"
2:46

22.
"Wet But Handsome / Blue Taxi"
1:24

23.
"Max and Gidget"
1:36

24.
"Welcome Home"
1:57

25.
"We Go Together" (originally from Grease)
1:24

Total length:
51:31

text: Release[edit]

text: The film was originally scheduled to be released on February 12, 2016, but was pushed back to July 8, 2016.[2] It was released in 2D, 3D, and IMAX 3D using DMR.[16][better source needed] The Secret Life of Pets premiered at the 2016 Annecy International Animated Film Festival on June 16, 2016, and was screened at VidCon.[17] The film is accompanied by a Despicable Me short film entitled Mower Minions.

text: Home media[edit]

text: The Secret Life of Pets was released on Digital HD on November 22, 2016, and on Blu-ray, Blu-ray 3D, Ultra HD Blu-ray, and DVD on December 6, 2016. In addition to the short film Mower Minions, which was released theatrically with the feature film, the releases also include two short films: Norman Television and Weenie.[18]

text: Reception[edit]

text: Box office[edit]

text: The Secret Life of Pets grossed $368.4 million in Canada and the U.S., and $507.1 million in other countries for a worldwide total of $875.5 million, against a production budget of $75 million.[4] Its internationally takings helped push Universal Pictures [International] past $1 billion for the tenth consecutive year,[19] and aided Illumination Entertainment to pass the $4 billion mark for the first time since 2010.[20]

text: Worldwide, it is the fifth highest-grossing original film of all time (behind Avatar, Zootopia, The Lion King, and Finding Nemo),[21] the sixth highest-grossing film of 2016,[22] the 12th highest-grossing animated film of all time, and the 50th highest-grossing film of all time. Deadline.com calculated the net profit of the film to be $374.6 million, when factoring together all expenses and revenues for the film, making it the most profitable release of 2016.[23]

text: North America[edit]

text: In Canada and the United States, the film was projected to gross around $70 million in its opening weekend, with some predictions going as high as $100 million.[24][25][26] It received the widest release for an animated film as well as for a PG-rated film (breaking Shrek Forever After's record) and the seventh widest of all time overall, across 4,370 theaters, the widest release since The Dark Knight Rises.[25][27][28][29] It made $5.3 million from Thursday night previews in 3,009 theaters, which is the second biggest of all time for Illumination, behind only Minions ($6.2 million), and the third biggest of all time for an animated film, behind Finding Dory ($9.2 million) and Minions.[26][30] This was followed by a record-breaking $38.5 million opening day gross (including previews), the biggest for an original film.[31][32] In its opening weekend, it exceeded expectations and grossed $104.4 million, finishing first at the box office.[33] Its debut is the sixth biggest for an animated film, the sixth highest weekend debut in July, and the fourth biggest opening weekend for Universal Pictures.[34] It also set the record for largest opening for an original film, eclipsing the $90.4 million debut of Inside Out.[35][36]

text: In its second weekend, the film made $50.8 million (-51.3%) while maintaining the top spot, despite facing stiff competition with newcomer Ghostbusters.[37] Although it slipped into second place when overtaken by Star Trek Beyond in its third weekend, it was still able to fend off the two other new releases—Lights Out and Ice Age: Collision Course—by earning another $29.6 million.[38] It passed $300 million on its twenty-sixth day—the same day it crossed $400 million worldwide[39]—becoming the seventh film of 2016,[40] the seventh Universal Pictures film (the first for the year),[41] and the twelfth animated film overall to reach the benchmark.[42]

text: Outside North America[edit]

text: Internationally, The Secret Life of Pets received a scattered release from June to September in a total of 69 markets, and faced competition from other animated films such as Finding Dory and Ice Age: Collision Course in the course of its release.[43][44][45] In total, it had number-one openings in 45 markets and recorded the biggest opening for an original animated film in 17 territories, and the No. 1 animated film of 2016 to date in 13 markets.[20][46] It topped the international box office in its ninth weekend after earning $43.8 million in 53 markets (dethroning Suicide Squad).[47] However, this wasn't the film's biggest single weekend gross. Its biggest weekend gross was actually in its seventh weekend when it grossed $69.3 million. That weekend it was No. 3 behind Suicide Squad ($133.3 million) and Chinese film Time Raiders ($64.6 million).[48]

text: It was released in the United Kingdom and Norway two weeks ahead of its U.S. release, where it grossed a combined total of $14.1 million in the two markets.[43] Prior to its official debut in the UK and Ireland on June 24, the film had two days worth of previews on June 18 and 19, from which it made £3.63 million ($5.2 million) in two days. This figure was later counted in the film's official opening weekend.[49] It went on to score the fourth biggest original animation opening weekend ever there, with £9.58 million ($13.4 million) from 592 theaters, trailing behind Inside Out, Monsters, Inc., and Up. Excluding previews, its total three-day opening was $8.1 million.[50][51] This was amidst Brexit and despite facing competition from Independence Day: Resurgence.[43][52] The film held off extremely well in its second and third weekend falling just 20% and 24%, earning £4.47 million ($6.1 million) and £3.62 million ($4.8 million) respectively despite facing some competitions.[53][54] It has topped the box office for three consecutive weekends and has grossed total of $41 million there.[55] In Norway, it took the number one spot as well, with $778,000.[43] It scored the biggest animation opening of 2016 and the second biggest of all time overall in both Taiwan ($2 million)—behind Madagascar, and Hong Kong ($1.9 million)—behind Inside Out.[44]

text: In Argentina, the film had a record-breaking number-one debut with $4 million from 195 screens — the biggest opening weekend ever for an animated film and the second biggest opening ever for any film (behind Furious 7). Similarly, Chile also recorded the second biggest animated opening ever with $1.7 million (behind Minions). Colombia also opened at No. 1 with a huge $1.5 million.[56] In Mexico, other than opening at No. 1, it posted the biggest original animated opening of all time with $7.6 million, surpassing the studio's own previous record holder Inside Out.[45] Also in Russia, it scored the biggest original animated opening and the second biggest ever for an animated film, behind Minions with $16.3 million, although this included $6.4 million five days worth of previews.[47] It went on to become the highest-grossing film of the summer that year.[57] In Japan, the film opened on August 11—the date of Japan's first Mountain Day national holiday—with Toho-Towa distributing, and earned $4.6 million on 370,000 admissions for its two-day opening weekend dethroning Shin Godzilla off the top spot. The cumulative total for the four-day holiday (August 11–14) was $9.7 million.[19][58] Similarly, it had number-one debuts in Germany ($7.1 million) and France ($5.9 million), Australia ($5.6 million), Italy ($5.1 million), Brazil ($4.4 million), Spain ($4.5 million), Poland ($1.7 million), Singapore ($1.3 million), Belgium ($1.2 million) and Denmark ($1.2 million), Indonesia ($1.1 million), the Philippines ($1 million), and South Africa ($463,000). In Germany, it scored the best animation opening since Minions.[45][59][60][61][62][63][64][65] It topped the box office in Israel for five straight weekends and in Australia, Russia and Spain for three weekends.[45][47][61][63] In Japan, the film was the highest-grossing foreign release of the weekend, holding the top spot for four consecutive weekends.[60]

text: In China—the world's second biggest film market—the film opened on a Tuesday, earning a modest $5.2 million on its opening day. To Sunday, it achieved a six-day opening of $29.3 million and $15.5 million for the weekend alone (Friday to Sunday), with per screen averages over $500 per screen per day. In both instances, it was behind the local film Time Raiders.[59][66][67] In South Korea, despite debuting in fifth place, it had an opening of $6.9 million.[59] Elsewhere, it grossed $2.7 million in the Netherlands, $1.2 million in Sweden and in Hungary it scored the biggest opening of the year with $738,000,[59] and also in Portugal ($965,000), Latvia ($224,000),[47] and South Africa.[64]

text: In terms of total earnings, its biggest markets are China ($58 million), United Kingdom and Ireland ($47.2 million), Japan ($41.6 million) and Russia ($34.2 million).[19][47][68] It became the third highest-grossing film of all time in Russia (behind Avatar and Zootopia)[61] and the highest-grossing film of 2016 in Spain.[20][62]

text: Critical response[edit]

text: On Rotten Tomatoes, the film has an approval rating of 74% based on 200 reviews with an average rating of 6.2/10. The site's critical consensus reads, "Fast-paced, funny, and blessed with a talented voice cast, The Secret Life of Pets offers a beautifully animated, cheerfully undemanding family-friendly diversion."[69] On Metacritic, the film has a score of 61 out of 100 based on 39 critics, indicating "generally favorable reviews."[70] Audiences polled by CinemaScore gave the film an average grade of "A–" on an A+ to F scale.[71]

text: Peter Travers of Rolling Stone gave the film three out of four stars and called it "an animated fluffball-a sort of Toy Story with pets does everything to drive you crazy and ends up being totally irresistible."[72] Scott Tobias, writing for NPR, characterized the film's concept as "Toy Story but with house pets," highlighting the film's "thinly distinguished characters" and "gloppy sentimentality."[73] Steven Rea of The Philadelphia Inquirer gave the film three out of four stars and said "In much the same way that the smash Zootopia demonstrated that creatures of different culture and class and species are better off when they come together, The Secret Life of Pets is a testament to teamwork and friendship and fixing the rifts that divide us. Let the fur – and the warm, fuzzy feelings – fly."[74]

text: However, some critics disliked the film's unexpected similarities with Pixar's Toy Story.[75] Matthew Parkinson of The Escapist compared the plots of both films and wrote that "The Secret Life of Pets feels like one of those hour-long ripoffs you'd see on a children's television network."[76] Ethan Anderton of the website /Film criticised the film's characterisation, and noted that the relationship between Max and Duke was akin to "Woody and Buzz Lightyear all over again."[77]

text: Accolades[edit]

text:
List of awards and nominations

Award
Date of ceremony
Category
Recipient(s)
Result
Ref(s)

Annie Awards
February 4, 2017
Outstanding Achievement, Character Design in an Animated Feature Production
Eric Guillon
Nominated
[78]

Outstanding Achievement, Music in an Animated Feature Production
Alexandre Desplat

British Academy Children's Awards
November 20, 2016
BAFTA Kids' Vote
The Secret Life of Pets
[79][80]

Cinema Audio Society
February 18, 2017
Outstanding Achievement in Sound Mixing for a Motion Picture – Animated
David Accord, Jason Butler, Gary A. Rizzo, Carlos Sotolongo and Frank Wolf
[81]

Hollywood Music in Media Awards
November 17, 2016
Best Original Score – Animated Film
Alexandre Desplat
Won
[82][83]

2017 Kids' Choice Awards
March 11, 2017
Favorite Animated Movie
The Secret Life of Pets
Nominated
[84]

Favorite Voice From an Animated Movie
Kevin Hart

Favorite Villain
Won

Most Wanted Pet

NAACP Image Awards
February 11, 2017
Outstanding Character Voice-Over Performance
Nominated
[85]

People's Choice Awards
January 18, 2017
Favorite Family Movie
The Secret Life of Pets
[86]

Favorite Animated Movie Voice
Kevin Hart

Producers Guild of America
January 28, 2017
Best Animated Motion Picture
Janet Healy and Chris Meledandri
[87]

text: Sequel[edit]

text: In August 2016 it was announced that there would be a sequel, with director Chris Renaud, producers Chris Meledandri and Janet Healy, and writer Brian Lynch, with a release date of July 13, 2018.[88] On January 25, 2017, its release date was pushed back to July 3, 2019.[7] On August 15, 2017, its release date was moved forward to June 7, 2019.


document_title: Alcoholic spirits measure

question_text: how much is a single measure of spirit

question_tokens: how much is a single measure of spirit

root

text: Alcoholic spirits measure

text: Optic measure[edit]

text: The optic or non-drip measure is mounted beneath an inverted spirit bottle, so that a pre-defined volume of the bottle's contents drains into the measure. Lifting a lever on the side of the measure first closes off the measure from the bottle, then dispenses the measured liquid into the glass or mixing vessel held underneath. This mechanism ensures that a correct spirit measure can be dispensed each time, as the inverted bottle allows the measure to be replenished in the optic after each shot has been dispensed. These types of measures are commonly used for highly demanded drinks in professional bar settings, and are often incorporated into visually-appealing machines that serve both functional and marketing purposes for the liquor being poured. Bottles are available to the trade with their labels affixed upside down so that they will be the right way up when mounted on the optic.

tag: p

text: Thimble measure[edit]

text: The thimble measure is a stainless steel vessel, like a shot glass, either with predefined measuring lines etched or stamped into the sides, or else pre-sized so that pouring up to the brim of the measure yields the correct volume. This second variation is commonly seen in a double-thimble or "hourglass" form, with two metal cups of different volumes (often in a 3:2 or 2:1 ratio, like a U.S. standard 1.5 fl oz "jigger" and 1 fl oz "pony", or UK standard 25/50mL or 35/70mL combos) spot-welded to each other at their relative bottom surfaces, possibly with a handle between them, allowing one unit to easily measure two common volumes.

text: Thimble measures are popular for home use, as they allow a relatively untrained person to pour quickly and accurately from several bottles without risking cross-contamination between them, or requiring the purchase, use and cleaning of a pour spout for each bottle as would be seen in a professional bar.

text: Free-flow pour spout[edit]

text: Common in U.S. bars, these devices consist of a simple rubber or plastic stopper with a metal or plastic tube fitted into it, and often a second smaller tube extending down into the bottle, designed to replace the cap or cork on a bottle of liquor. The spout, in the U.S., is usually calibrated to allow a flow of 1 fluid ounce per second, so that a bartender can measure accurate and consistent shots of liquor or portions for cocktails based on timed pours, without needing to use a jigger or other measuring device.[3]

text: Measured pour spout[edit]

text: Common in U.S. bars, these devices appear visually to be the same as a free-flow pour spout, but have an internal mechanism to block the tube after a pre-defined and calibrated volume of liquid has passed, oftentimes a set of 2 or 3 balls. Because of the geometry, the bottle with one of these spouts needs to be held at a specific angle, typically 45 degrees, otherwise the volume poured will be under or over the stated calibration.

text: Ball-measuring cap[edit]

text: Common in France, where it is called a bouchon doseur boule, this device consists of a transparent T-shaped glass tube arrangement,[4] with a ball on one end of the horizontal section, a cap or cork on the other end, and a cork or plastic bottle stopper on the bottom of the T, allowing the measure to replace the cap of a liquor bottle. In use, the bottle is inverted until the ball fills with liquor, and then tilted in the other direction to let the liquor pour out the spout in a manner that keeps additional liquor entering the measure from the bottle. A promotional video available on YouTube includes a cutaway rendered animation beginning at 2:15 that demonstrates the 3-ball mechanical arrangement.[5]

text: United Kingdom[edit]

text: The Weights and Measures Act of 1963 made it illegal in Britain for businesses to give short weights or short measures to consumers. Before this there was no legislation, only guidelines as to the correct weight of an alcoholic spirit measure, and if spirit measures or optics were used, they required a government stamp to certify that the measure was accurate.

text: The 1963 Act formalized the legal measures by which spirits and other alcoholic beverages should be dispensed, namely 1/4 gill (35.5 ml), 1/5 gill (28.4 ml) or 1/6 gill (23.7 ml), but this was replaced in 1985 by 25ml and 35ml for single measures, with double measures of 50ml or 70ml being permitted in 2001. Landlords have the option to decide which quantities they sell, with the difference being caused by historically larger measures being used in Scotland and Northern Ireland.

text: Thimble measures are also used in 175ml and 250ml volumes for measuring wine. Although government stamped for the correct volume, the thimble measure does rely on the user measuring the wine out manually into the thimble.


document_title: Blue Lagoon (geothermal spa)

question_text: what makes the blue lagoon in iceland blue

text: Blue Lagoon (geothermal spa)

text: Description[edit]

text: The warm waters are rich in minerals like silica and sulfur and bathing in the Blue Lagoon is reputed to help some people suffering from skin diseases such as psoriasis.[2] The water temperature in the bathing and swimming area of the lagoon averages 37–39 °C (99–102 °F). The Blue Lagoon also operates a research and development facility to help find cures for other skin ailments using the mineral-rich water.

text: The lagoon is a man-made lagoon which is fed by the water output of the nearby geothermal power plant Svartsengi and is renewed every two days. It is the largest in the world. Superheated water is vented from the ground near a lava flow and used to run turbines that generate electricity. After going through the turbines, the steam and hot water passes through a heat exchanger to provide heat for a municipal water heating system. Then the water is fed into the lagoon for recreational and medicinal users to bathe in.

text: Iceland has a strict code of hygiene and guests are required to shower prior to enjoying the geothermal spa. The communal showers are split up by gender[3].

text: Children age 8 and under are only allowed entry with the use of arm floaters, provided free of charge. The lagoon is not suitable for children under the age of 2 years.

text: The Blue Lagoon is accessible for wheelchair users with a ramp that extends into the water and a shower chair. There is also a private changing room available for those with special needs, complete with a roll-in shower.

text: The rich mineral content is provided by the underground geological layers and pushed up to the surface by the hot water (at about 1.2 MPa (170 psi) pressure and 240 °C (464 °F) temperature) used by the plant. Because of its mineral concentration, water cannot be recycled and must be disposed of in the nearby landscape, a permeable lava field that varies in thickness from 50 cm (20 in) to 1 m (3.3 ft). The silicate minerals is the primary cause of that water's milky blue shade. After the minerals have formed a deposit, the water reinfiltrates the ground, but the deposit renders it impermeable over time, hence the necessity for the plant to continuously dig new ponds in the nearby lava field.

text: A small experimental facility is still visible near the plant, where the engineers made decantation tests to evaluate the speed of mineral deposition, which is clearly a limiting factor both to the plant's rentability and sustainability. Hence, geothermal energy exploitation at this location is not without environmental impact.

text: The Blue Lagoon was used as the pit stop for the first leg of The Amazing Race 6. The Blue Lagoon was used for the thermal spa scenes in the filming of Hostel: Part II. It was also shown in the Incubus documentary Look Alive, when the band visited Iceland, in the fifth cycle of Britain's Next Top Model which used as photoshoot location as well as Keeping up with the Kardashians.

text: The Blue Lagoon is situated close to the world's first renewable methanol plant, which uses Carbon Recycling International's carbon dioxide to methanol fuel process.[4]

text: History[edit]

text: In 1976, a pool formed at the site from the waste water of the geothermal power plant that had just been built there. In 1981, people started bathing in it after its supposed healing powers were popularized. In 1992, the Blue Lagoon company was established, and the bathing facility was opened for the public.

text: In recent years, several cosmetics companies have begun marketing skin care products using mud purportedly extracted from the Blue Lagoon, their effectiveness still subject to debate.


document_title: The Covenant (film)

question_text: where does the movie the covenant take place

text: The Covenant (film)

text: Plot[edit]

text: In the town of Ipswich, four college school boys - Caleb, Pogue, Reid and Tyler, together known as the Sons of Ipswich - are the descendants of colonial witch families and thus wield magical abilities which first manifest on their 13th birthday and grow stronger until they Ascend at 18. The Power is linked to their life force; the more they use, the faster they age. This becomes more dangerous as, upon Ascending, their powers reach their full strength and become so addictive, seductive as they say, that some can't resist. Caleb - who's closest to his 18th birthday - exhibits restraint.

text: While attending a rave, Caleb meets Sarah, a transfer student from a community college in Boston. The Sons also meet Chase Collins, a new student at Spencer Academy. Their meeting is cut short when cops appear to break up the party. The boys escape by using their combined magics. After a student is found mysteriously dead near their campus, various paranormal occurrences take place, with Sarah and her roommate Kate being the focus of it. Upset, Caleb suspects Reid - the most reckless of the warlocks - but he angrily denies any abusive usage of his powers.

text: Caleb and later Pogue see a "darkling," a dead spirit and malicious omen. Meanwhile, Caleb and Sarah quickly become romantically close. During a swim race, Caleb notices Chase, who had befriended the group, displaying magic usage. After researching, Caleb concludes that Chase descends from a fifth family, the bastard son of Goodwin “Goody” Pope, one believed long extinct, and that he is the true perpetrator. As the Sons discuss this revelation, Pogue learns that his girlfriend was rendered comatose by a spell. Enraged, he hastily challenges Chase, who swiftly hospitalizes him.

text: Caleb visits Sarah only to fall into Chase's trap. Chase reveals that he was unaware of his magic’s origin, having been adopted. After locating his biological father, he learned of the price for ascension; but it was too late and he has become too addicted to using magic. His biological father then transferred power to him. Chase wants to force other "ascended" witches to transfer power to him as well, starting with Caleb. Despite Caleb's warning that having more power does not save him from aging to death, Chase ignores him. Before leaving, Chase threatens harm on Caleb's family and friends if he does not get what he wants. Caleb reveals the truth to Sarah and takes her to his father, a man of forty-four years with a decrepit old body, exhausted from magic abuse. When Sarah suggests that one of the other three transfer their power to Caleb so he could evenly match Chase, Caleb immediately refuses, explaining that doing so will cost them their lives.

text: On the night of Caleb’s eighteenth birthday, he leaves to face Chase alone, and has Reid and Tyler safeguard Sarah in public. However, Chase easily kidnaps Sarah. At an old barn, the two confront each other. Chase reveals a spellbound Sarah and gives Caleb an ultimatum of his life for hers. Caleb declares he won’t will his power away or let Sarah come to harm. They duel and Caleb is clearly outmatched. At the exact minute of his birth, he ascends, and his power fully matures, which allows him to mount an offensive. However, as Chase has more than one magical share, he still proves superior and the temporary turn of tide does not last long. Back at home, Evelyn, Caleb's mother, begs her husband to save Caleb. He sacrifices himself and transfers his power remotely to his son. Once his father’s power is infused within him, Caleb hits Chase with a final blow that engulfs him in a ball of flame. Sarah, Kate, and Pogue awaken, freed from their curses.

text: Firefighters arrive on the scene. Caleb and Sarah wait until inspection of the barn’s wreckage is complete; they are informed that a third person was not found, suggesting Chase somehow survived and escaped. The pair get into Caleb’s car, and he casually uses magic to fix the busted windshield, which seems visibly unsettling to Sarah. He reassuringly holds her hand, and they drive off.

text: Cast[edit]

text:
Steven Strait as Caleb Danvers - the main protagonist; one of the four Sons of Ipswich, he attends Spenser Academy along with the other Sons and is the oldest and most responsible of the four. He is very cautious about using the Power, due to the effect its had on his father, and is the first to ascend.
Taylor Kitsch as Pogue Parry - One of the four Sons of Ipswich who is close to Caleb and is dating Kate.
Toby Hemingway as Reid Garwin - One of the four Sons of Ipswich and the most reckless of the four who often butts heads with Caleb.
Chace Crawford as Tyler Simms - One of the four Sons of Ipswich who is close to Reid and usually takes his side.
Sebastian Stan as Chase Collins - A new student at Spenser who befriends the sons of Ipswich. He is later revealed to be the last of the Sons of Ipswich as well as being the one behind the supernatural occurrences, and is the film's main antagonist.
Laura Ramsey as Sarah Wenham - A new student at Spenser who transfers from a Boston public school and Caleb's love interest. She is the only one outside the Covenant who learns about the boys' secret.
Sarah Smyth as Kira Snider - A snobby student and Aaron's girlfriend.
Jessica Lucas as Kate Tunney - Sarah's roommate and Pogue's girlfriend.
Kyle Schmid as Aaron Abbot - A pompous student and jock.
Wendy Crewson as Evelyn Danvers - Caleb's alcoholic mother who worries about her son ascending and becoming like his father.
Stephen McHattie as William Danvers III - Caleb's father.
Kenneth Welsh as Provost Higgins - Head of Spenser Academy.
Jon McLaren as Bordy Becklin
Steven Crowder as Party Kid

tag: ul

text: Release[edit]

text: Despite a popular misconception, The Covenant is not based on a comic book title nor on any other book. The confusion comes from the fact that Sony released a comic book of the same name, written by Aron Coleite, and created for the purposes of promoting the film. Neither the authors of the comic-book miniseries nor Top Cow Comics are mentioned in the films’ credit sequences, so the comic-book miniseries is not regarded as source material by The Covenant's producers. In fact, the film originated from a spec script, and went through a number of drafts, by different writers, before J.S. Cardone eventually submitted the final draft. Cardone received sole screenwriting credit.[4]

text: Reception[edit]

text: The Covenant received extremely negative reviews from critics, with a 3% "Rotten" rating on Rotten Tomatoes, and the site's consensus stating, "The Covenant plays out like a teen soap opera, full of pretty faces, wooden acting, laughable dialogue, and little suspense."[5] The film also holds a place on the site's "Worst of the Worst", ranking 31st.[6] The film received a 19 out of 100 score on Metacritic, indicating "overwhelming dislike".[7]

text: Box office[edit]

text: Upon its release in the United States, the film managed to top the box office charts, with a $8.9 million opening on what was called a "weak" weekend.[8] As of October 15, 2006, The Covenant has earned $23,292,105 in the U.S. ($37,256,954 worldwide).[3] The film cost roughly $20 million to produce, not including marketing.

text: Home media[edit]

text: The Covenant was released on DVD and Blu-ray on January 2, 2007. It went on to sell 1,618,891 units, which translated to revenue of $26,578,576.[9]


document_title: Apoptosis

question_text: apoptotic bodies are phagocytized without the process of

text: Apoptosis

text: Discovery and etymology[edit]

text: German scientist Karl Vogt was first to describe the principle of apoptosis in 1842. In 1885, anatomist Walther Flemming delivered a more precise description of the process of programmed cell death. However, it was not until 1965 that the topic was resurrected. While studying tissues using electron microscopy, John Foxton Ross Kerr at the University of Queensland was able to distinguish apoptosis from traumatic cell death.[7] Following the publication of a paper describing the phenomenon, Kerr was invited to join Alastair R. Currie, as well as Andrew Wyllie, who was Currie's graduate student,[8] at University of Aberdeen. In 1972, the trio published a seminal article in the British Journal of Cancer.[9] Kerr had initially used the term programmed cell necrosis, but in the article, the process of natural cell death was called apoptosis. Kerr, Wyllie and Currie credited James Cormack, a professor of Greek language at University of Aberdeen, with suggesting the term apoptosis. Kerr received the Paul Ehrlich and Ludwig Darmstaedter Prize on March 14, 2000, for his description of apoptosis. He shared the prize with Boston biologist H. Robert Horvitz.[10]

text: For many years, neither "apoptosis" nor  "programmed cell death" was a highly cited term. Two discoveries brought cell death from obscurity to a major field of research: identification of components of the cell death control and effector mechanisms, and linkage of abnormalities in cell death to human disease, in particular cancer.

text: The 2002 Nobel Prize in Medicine was awarded to Sydney Brenner, Horvitz and John E. Sulston for their work identifying genes that control apoptosis.  The genes were identified by studies in the nematode C. elegans and homologues of these genes function in humans to regulate apoptosis.

text: In Greek, apoptosis translates to the "falling off" of leaves from a tree.[11] Cormack, professor of Greek language, reintroduced the term for medical use as it had a medical meaning for the Greeks over two thousand years before. Hippocrates used the term to mean "the falling off of the bones". Galen extended its meaning to "the dropping of the scabs". Cormack was no doubt aware of this usage when he suggested the name. Debate continues over the correct pronunciation, with opinion divided between a pronunciation with the second p silent (/æpəˈtoʊsɪs/ ap-ə-TOH-sis[12][13]) and the second p pronounced (/eɪpəpˈtoʊsɪs/),[12][14] as in the original Greek.[citation needed] In English, the p of the Greek -pt- consonant cluster is typically silent at the beginning of a word (e.g. pterodactyl, Ptolemy), but articulated when used in combining forms preceded by a vowel, as in helicopter or the orders of insects: diptera, lepidoptera, etc.

text: In the original Kerr, Wyllie & Currie paper,[9] there is a footnote regarding the pronunciation:

text: "We are most grateful to Professor James Cormack of the Department of Greek, University of Aberdeen, for suggesting this term. The word "apoptosis" (ἀπόπτωσις) is used in Greek to describe the "dropping off" or "falling off" of petals from flowers, or leaves from trees. To show the derivation clearly, we propose that the stress should be on the penultimate syllable, the second half of the word being pronounced like "ptosis" (with the "p" silent), which comes from the same root "to fall", and is already used to describe the drooping of the upper eyelid."

text: Activation mechanisms[edit]

text: The initiation of apoptosis is tightly regulated by activation mechanisms, because once apoptosis has begun, it inevitably leads to the death of the cell.[15][16] The two best-understood activation mechanisms are the intrinsic pathway (also called the mitochondrial pathway) and the extrinsic pathway.[17] The intrinsic pathway is activated by intracellular signals generated when cells are stressed and depends on the release of proteins from the intermembrane space of mitochondria.[18] The extrinsic pathway is activated by extracellular ligands binding to cell-surface death receptors, which leads to the formation of the death-inducing signaling complex (DISC).[19]

text: A cell initiates intracellular apoptotic signaling in response to a stress, which may bring about cell suicide. The binding of nuclear receptors by glucocorticoids,[20] heat,[20] radiation,[20] nutrient deprivation,[20] viral infection,[20] hypoxia[20], increased intracellular concentration of free fatty acids[21] and increased intracellular calcium concentration,[22][23] for example, by damage to the membrane, can all trigger the release of intracellular apoptotic signals by a damaged cell. A number of cellular components, such as poly ADP ribose polymerase, may also help regulate apoptosis.[24] Single cell fluctuations have been observed in experimental studies of stress induced apoptosis.[25][26]

text: Before the actual process of cell death is precipitated by enzymes, apoptotic signals must cause regulatory proteins to initiate the apoptosis pathway. This step allows those signals to cause cell death, or the process to be stopped, should the cell no longer need to die. Several proteins are involved, but two main methods of regulation have been identified: the targeting of mitochondria functionality,[27] or directly transducing the signal via adaptor proteins to the apoptotic mechanisms. An extrinsic pathway for initiation identified in several toxin studies is an increase in calcium concentration within a cell caused by drug activity, which also can cause apoptosis via a calcium binding protease calpain.

text: Intrinsic pathway[edit]

text: The mitochondria are essential to multicellular life.  Without them, a cell ceases to respire aerobically and quickly dies. This fact forms the basis for some apoptotic pathways.  Apoptotic proteins that target mitochondria affect them in different ways.  They may cause mitochondrial swelling through the formation of membrane pores, or they may increase the permeability of the mitochondrial membrane and cause apoptotic effectors to leak out.[20][28]  They are very closely related to intrinsic pathway, and tumors arise more frequently through intrinsic pathway than the extrinsic pathway because of sensitivity.[29] There is also a growing body of evidence indicating that nitric oxide is able to induce apoptosis by helping to dissipate the membrane potential of mitochondria and therefore make it more permeable.[30] Nitric oxide has been implicated in initiating and inhibiting apoptosis through its possible action as a signal molecule of subsequent pathways that activate apoptosis.[31][citation needed]

text: Mitochondrial proteins known as SMACs (second mitochondria-derived activator of caspases) are released into the cell's cytosol following the increase in permeability of the mitochondria membranes.  SMAC binds to proteins that inhibit apoptosis (IAPs) thereby deactivating them, and preventing the IAPs from arresting the process and therefore allowing apoptosis to proceed.  IAP also normally suppresses the activity of a group of cysteine proteases called caspases,[32] which carry out the degradation of the cell. Therefore, the actual degradation enzymes can be seen to be indirectly regulated by mitochondrial permeability.

text: Cytochrome c is also released from mitochondria due to formation of a channel, the mitochondrial apoptosis-induced channel (MAC), in the outer mitochondrial membrane,[33] and serves a regulatory function as it precedes morphological change associated with apoptosis.[20]  Once cytochrome c is released it binds with Apoptotic protease activating factor – 1 (Apaf-1) and ATP, which then bind to pro-caspase-9 to create a protein complex known as an apoptosome.  The apoptosome cleaves the pro-caspase to its active form of caspase-9, which in turn activates the effector caspase-3.

text: MAC (not to be confused with the membrane attack complex formed by complement activation, also commonly denoted as MAC), also called "Mitochondrial Outer Membrane Permeabilization Pore" is regulated by various proteins, such as those encoded by the mammalian Bcl-2 family of anti-apoptopic genes, the homologs of the ced-9 gene found in C. elegans.[34][35] Bcl-2 proteins are able to promote or inhibit apoptosis by direct action on MAC/MOMPP.  Bax and/or Bak form the pore, while Bcl-2, Bcl-xL or Mcl-1 inhibit its formation.

text: Extrinsic pathway[edit]

text: Two theories of the direct initiation of apoptotic mechanisms in mammals have been suggested: the TNF-induced (tumor necrosis factor) model and the  Fas-Fas ligand-mediated model, both involving receptors of the TNF receptor (TNFR) family[36] coupled to extrinsic signals.

text: TNF path

text: TNF-alpha is a cytokine produced mainly by activated macrophages, and is the major extrinsic mediator of apoptosis. Most cells in the human body have two receptors for TNF-alpha: TNFR1 and TNFR2. The binding of TNF-alpha to TNFR1 has been shown to initiate the pathway that leads to caspase activation via the intermediate membrane proteins TNF receptor-associated death domain (TRADD) and Fas-associated death domain protein (FADD). cIAP1/2 can inhibit TNF-α signaling by binding to TRAF2. FLIP inhibits the activation of caspase-8.[37] Binding of this receptor can also indirectly lead to the activation of transcription factors involved in cell survival and inflammatory responses.[38] However, signalling through TNFR1 might also induce apoptosis in a caspase-independent manner.[39] The link between TNF-alpha and apoptosis shows why an abnormal production of TNF-alpha plays a fundamental role in several human diseases, especially in autoimmune diseases.

text: Fas path

text: The fas receptor  (First apoptosis signal) – (also known as Apo-1 or CD95) is a transmembrane protein of the TNF family which binds the Fas ligand (FasL).[36] The interaction between Fas and FasL results in the formation of the death-inducing signaling complex (DISC), which contains the FADD, caspase-8 and caspase-10. In some types of cells (type I), processed caspase-8 directly activates other members of the caspase family, and triggers the execution of apoptosis of the cell. In other types of cells (type II), the Fas-DISC starts a feedback loop that spirals into increasing release of proapoptotic factors from mitochondria and the amplified activation of caspase-8.[40]

text: Common components

text: Following TNF-R1 and Fas activation in mammalian cells a balance between proapoptotic (BAX,[41] BID, BAK, or BAD) and anti-apoptotic (Bcl-Xl and Bcl-2) members of the Bcl-2 family are established. This balance is the proportion of proapoptotic homodimers that form in the outer-membrane of the mitochondrion. The proapoptotic homodimers are required to make the mitochondrial membrane permeable for the release of caspase activators such as cytochrome c and SMAC. Control of proapoptotic proteins under normal cell conditions of nonapoptotic cells is incompletely understood, but in general, Bax or Bak are activated by the activation of BH3-only proteins, part of the Bcl-2 family.

text:
Caspases
Caspases  play the central role in the transduction of ER apoptotic signals. Caspases are proteins that are highly conserved, cysteine-dependent aspartate-specific proteases. There are two types of caspases: initiator caspases, caspase 2,8,9,10,11,12, and effector caspases, caspase 3,6,7. The activation of initiator caspases requires binding to specific oligomeric activator protein. Effector caspases are then activated by these active initiator caspases through proteolytic cleavage. The active effector caspases then proteolytically degrade a host of intracellular proteins to carry out the cell death program.

text:
Caspase-independent apoptotic pathway
There also exists a caspase-independent apoptotic pathway that is mediated by AIF (apoptosis-inducing factor).[42]

text: Apoptosis model in amphibians[edit]

text: Amphibian frog Xenopus laevis serves as an ideal model system for the study of the mechanisms of apoptosis. In fact, iodine and thyroxine also stimulate the spectacular apoptosis of the cells of the larval gills, tail and fins in amphibians metamorphosis, and stimulate the evolution of their nervous system transforming the aquatic, vegetarian tadpole into the terrestrial, carnivorous frog.[43][44][45][46]

text: Negative regulators of apoptosis[edit]

text: Negative regulation of apoptosis inhibits cell death signaling pathways, helping tumors to evade cell death and developing drug resistance. Many families of proteins act as negative regulators categorized into either antiapoptotic factors, such as IAPs and Bcl-2 proteins or prosurvival factors like cFLIP, BNIP3, FADD, Akt,  and NF-κB.[47]

text: Proteolytic caspase cascade: Killing the cell[edit]

text: Many pathways and signals lead to apoptosis, but these converge on a single mechanism that actually causes the death of the cell. After a cell receives stimulus, it undergoes organized degradation of cellular organelles by activated proteolytic caspases. In addition to the destruction of cellular organelles, mRNA is rapidly and globally degraded by a mechanism that is not yet fully characterized.[48] mRNA decay is triggered very early in apoptosis.

text: A cell undergoing apoptosis shows a series of characteristic morphological changes. Early alterations include:

text:
Cell shrinkage and rounding occur because of the retraction lamellipodia and the breakdown of the proteinaceous cytoskeleton by caspases.[49]
The cytoplasm appears dense, and the organelles appear tightly packed.
Chromatin undergoes condensation into compact patches against the nuclear envelope (also known as the perinuclear envelope) in a process known as pyknosis, a hallmark of apoptosis.[50][51]
The nuclear envelope becomes discontinuous and the DNA inside it is fragmented in a process referred to as karyorrhexis. The nucleus breaks into several discrete chromatin bodies or nucleosomal units due to the degradation of DNA.[52]

tag: ol

text: Apoptosis progresses quickly and its products are quickly removed, making it difficult to detect or visualize on classical histology sections. During karyorrhexis, endonuclease activation leaves short DNA fragments, regularly spaced in size. These give a characteristic "laddered" appearance on agar gel after electrophoresis.[53] Tests for DNA laddering differentiate apoptosis from ischemic or toxic cell death.[54]

text: Apoptotic cell disassembly[edit]

text: Before the apoptotic cell is disposed of, there is a process of disassembly. There are three recognized steps in apoptotic cell disassembly:[56]

text:
Membrane blebbing: The cell membrane shows irregular buds known as blebs. Initially these are smaller surface blebs. Later these can grow into larger so-called dynamic membrane blebs.[56] An important regulator of apoptotic cell membrane blebbing is ROCK1 (rho associated coiled-coil-containing protein kinase 1).[57][58]
Formation of membrane protrusions: Some cell types, under specific conditions, may develop different types of long, thin extensions of the cell membrane called membrane protrusions. Three types have been described: microtubule spikes, apoptopodia (feet of death), and beaded apoptopodia (the latter having a beads-on-a-string appearance).[59][60][61] Pannexin 1 is an important component of membrane channels involved in the formation of apoptopodia and beaded apoptopodia.[60]
Fragmentation: The cell breaks apart into multiple vesicles called apoptotic bodies, which undergo phagocytosis. The plasma membrane protrusions may help bring apoptotic bodies closer to phagocytes.

text: Removal of dead cells[edit]

text:
The removal of dead cells by neighboring phagocytic cells has been termed efferocytosis.[62]
Dying cells that undergo the final stages of apoptosis display phagocytotic molecules, such as phosphatidylserine, on their cell surface.[63] Phosphatidylserine is normally found on the inner leaflet surface of the plasma membrane, but is redistributed during apoptosis to the extracellular surface by a protein known as scramblase.[64] These molecules mark the cell for phagocytosis by cells possessing the appropriate receptors, such as macrophages.[65] The removal of dying cells by phagocytes occurs in an orderly manner without eliciting an inflammatory response.[66] During apoptosis cellular RNA and DNA are separated from each other and sorted to different apoptotic bodies; separation of RNA is initiated as nucleolar segregation.[67]

text: Pathway knock-outs[edit]

text: Many knock-outs have been made in the apoptosis pathways to test the function of each of the proteins.  Several caspases, in addition to APAF1 and FADD, have been mutated to determine the new phenotype.  In order to create a tumor necrosis factor (TNF) knockout, an exon containing the nucleotides 3704–5364 was removed from the gene.  This exon encodes a portion of the mature TNF domain, as well as the leader sequence, which is a highly conserved region necessary for proper intracellular processing.  TNF-/- mice develop normally and have no gross structural or morphological abnormalities.  However, upon immunization with SRBC (sheep red blood cells), these mice demonstrated a deficiency in the maturation of an antibody response; they were able to generate normal levels of IgM, but could not develop specific IgG levels.  Apaf-1 is the protein that turns on caspase 9 by cleavage to begin the caspase cascade that leads to apoptosis.  Since a -/- mutation in the APAF-1 gene is embryonic lethal, a gene trap strategy was used in order to generate an APAF-1 -/- mouse.  This assay is used to disrupt gene function by creating an intragenic gene fusion.  When an APAF-1 gene trap is introduced into cells, many morphological changes occur, such as spina bifida, the persistence of interdigital webs, and open brain.  In addition, after embryonic day 12.5, the brain of the embryos showed several structural changes.  APAF-1 cells are protected from apoptosis stimuli such as irradiation.  A BAX-1 knock-out mouse exhibits normal forebrain formation and a decreased programmed cell death in some neuronal populations and in the spinal cord, leading to an increase in motor neurons.

text: The caspase proteins are integral parts of the apoptosis pathway, so it follows that knock-outs made have varying damaging results.  A caspase 9 knock-out leads to a severe brain malformation.  A caspase 8 knock-out leads to cardiac failure and thus embryonic lethality.  However, with the use of cre-lox technology, a caspase 8 knock-out has been created that exhibits an increase in peripheral T cells, an impaired T cell response, and a defect in neural tube closure.  These mice were found to be resistant to apoptosis mediated by CD95, TNFR, etc. but not resistant to apoptosis caused by UV irradiation, chemotherapeutic drugs, and other stimuli.  Finally, a caspase 3 knock-out was characterized by ectopic cell masses in the brain and abnormal apoptotic features such as membrane blebbing or nuclear fragmentation.  A remarkable feature of these KO mice is that they have a very restricted phenotype: Casp3, 9, APAF-1 KO mice have deformations of neural tissue and FADD and Casp 8 KO showed defective heart development, however in both types of KO other organs developed normally and some cell types were still sensitive to apoptotic stimuli suggesting that unknown proapoptotic pathways exist.

text: Methods for distinguishing apoptotic from necrotic (necroptotic) cells[edit]

text: In order to perform analysis of apoptotic versus necrotic (necroptotic) cells, one can do analysis of morphology by time-lapse microscopy, flow fluorocytometry, and transmission electron microscopy. There are  also various biochemical techniques for analysis of cell surface markers (phosphatidylserine exposure versus cell permeability by flow cytometry), cellular markers such as DNA fragmentation[68] (flow cytometry),[69] caspase activation, Bid cleavage, and cytochrome c release (Western blotting). It is important to know how primary and secondary necrotic cells can be distinguished by analysis of supernatant for caspases, HMGB1, and release of cytokeratin 18. However, no distinct surface or biochemical markers of necrotic cell death have been identified yet, and only negative markers are available. These include absence of apoptotic markers (caspase activation, cytochrome c release, and oligonucleosomal DNA fragmentation) and differential kinetics of cell death markers (phosphatidylserine exposure and cell membrane permeabilization). A selection of techniques that can be used to distinguish apoptosis from necroptotic cells could be found in these references.[70][71][72][73]

text: Implication in disease[edit]

text: Defective pathways[edit]

text: The many different types of apoptotic pathways contain a multitude of different biochemical components, many of them not yet understood.[74] As a pathway is more or less sequential in nature, removing or modifying one component leads to an effect in another. In a living organism, this can have disastrous effects, often in the form of disease or disorder. A discussion of every disease caused by modification of the various apoptotic pathways would be impractical, but the concept overlying each one is the same: The normal functioning of the pathway has been disrupted in such a way as to impair the ability of the cell to undergo normal apoptosis. This results in a cell that lives past its "use-by-date" and is able to replicate and pass on any faulty machinery to its progeny, increasing the likelihood of the cell's becoming cancerous or diseased.

text: A recently described example of this concept in action can be seen in the development of a lung cancer called NCI-H460.[75] The X-linked inhibitor of apoptosis protein (XIAP) is overexpressed in cells of the H460 cell line. XIAPs bind to the processed form of caspase-9, and suppress the activity of apoptotic activator cytochrome c, therefore overexpression leads to a decrease in the amount of proapoptotic agonists. As a consequence, the balance of anti-apoptotic and proapoptotic effectors is upset in favour of the former, and the damaged cells continue to replicate despite being directed to die. Defects in regulation of apoptosis in cancer cells occur often at the level of control of transcription factors. As a particular example, defects in molecules that control transcription factor NF-κB in cancer change the mode of transcriptional regulation and the response to apoptotic signals, to curtail dependence on the tissue that the cell belongs. This degree of independence from external survival signals, can enable cancer metastasis.[76]

text: Dysregulation of p53[edit]

text: The tumor-suppressor protein p53 accumulates when DNA is damaged due to a chain of biochemical factors. Part of this pathway includes alpha-interferon and beta-interferon, which induce transcription of the p53 gene, resulting in the increase of p53 protein level and enhancement of cancer cell-apoptosis.[77] p53 prevents the cell from replicating by stopping the cell cycle at G1, or interphase, to give the cell time to repair, however it will induce apoptosis if damage is extensive and repair efforts fail.[78] Any disruption to the regulation of the p53 or interferon genes will result in impaired apoptosis and the possible formation of tumors.

text: Inhibition[edit]

text: Inhibition of apoptosis can result in a number of cancers, autoimmune diseases, inflammatory diseases, and viral infections. It was originally believed that the associated accumulation of cells was due to an increase in cellular proliferation, but it is now known that it is also due to a decrease in cell death. The most common of these diseases is cancer, the disease of excessive cellular proliferation, which is often characterized by an overexpression of IAP family members. As a result, the malignant cells experience an abnormal response to apoptosis induction: Cycle-regulating genes (such as p53, ras or c-myc) are mutated or inactivated in diseased cells, and further genes (such as bcl-2) also modify their expression in tumors. Some apoptotic factors are vital during mitochondrial respiration e.g. cytochrome C.[79] Pathological inactivation of apoptosis in cancer cells is correlated with frequent respiratory metabolic shifts toward glycolysis (an observation known as the “Warburg hypothesis”.[80]

text: HeLa cell[edit]

text: Apoptosis in HeLa[b] cells is inhibited by proteins produced by the cell; these inhibitory proteins target retinoblastoma tumor-suppressing proteins.[81] These tumor-suppressing proteins regulate the cell cycle, but are rendered inactive when bound to an inhibitory protein.[81] HPV E6 and E7 are inhibitory proteins expressed by the human papillomavirus, HPV being responsible for the formation of the cervical tumor from which HeLa cells are derived.[82] HPV E6 causes p53, which regulates the cell cycle, to become inactive.[83] HPV E7 binds to retinoblastoma tumor suppressing proteins and limits its ability to control cell division.[83]  These two inhibitory proteins are partially responsible for HeLa cells' immortality by inhibiting apoptosis to occur.[84] CDV (Canine Distemper Virus) is able to induce apoptosis despite the presence of these inhibitory proteins. This is an important oncolytic property of CDV: this virus is capable of killing canine lymphoma cells. Oncoproteins E6 and E7 still leave p53 inactive, but they are not able to avoid the activation of caspases induced from the stress of viral infection. These oncolytic properties provided a promising link between CDV and lymphoma apoptosis, which can lead to development of alternative treatment methods for both canine lymphoma and human non-Hodgkin lymphoma. Defects in the cell cycle are thought to be responsible for the resistance to chemotherapy or radiation by certain tumor cells, so a virus that can induce apoptosis despite defects in the cell cycle is useful for cancer treatment.[84]

text: Treatments[edit]

text: The main method of treatment for death from signaling-related diseases involves either increasing or decreasing the susceptibility of apoptosis in diseased cells, depending on whether the disease is caused by either the inhibition of or excess apoptosis. For instance, treatments aim to restore apoptosis to treat diseases with deficient cell death, and to increase the apoptotic threshold to treat diseases involved with excessive cell death. To stimulate apoptosis, one can increase the number of death receptor ligands (such as TNF or TRAIL), antagonize the anti-apoptotic Bcl-2 pathway, or introduce Smac mimetics to inhibit the inhibitor (IAPs). The addition of agents such as Herceptin, Iressa, or Gleevec works to stop cells from cycling and causes apoptosis activation by blocking growth and survival signaling further upstream. Finally, adding p53-MDM2 complexes displaces p53 and activates the p53 pathway, leading to cell cycle arrest and apoptosis. Many different methods can be used either to stimulate or to inhibit apoptosis in various places along the death signaling pathway.[85]

text: Apoptosis is a multi-step, multi-pathway cell-death programme that is inherent in every cell of the body. In cancer, the apoptosis cell-division ratio is altered. Cancer treatment by chemotherapy and  irradiation kills target cells primarily by inducing apoptosis.

text: Hyperactive apoptosis[edit]

text:
On the other hand, loss of control of cell death (resulting in excess apoptosis) can lead to neurodegenerative diseases, hematologic diseases, and tissue damage. It is to note that neurons that rely on mitochondrial respiration undergo apoptosis in neurodegenerative diseases such as Alzheimer’s [86] and Parkinson’s [87] (an observation known as the “Inverse Warburg hypothesis” [88][79] ).Moreover, there is an inverse epidemiological comorbidity between neurodegenerative diseases and cancer.[89] The progression of HIV is directly linked to excess, unregulated apoptosis.  In a healthy individual, the number of CD4+ lymphocytes is in balance with the cells generated by the bone marrow; however, in HIV-positive patients, this balance is lost due to an inability of the bone marrow to regenerate CD4+ cells.  In the case of HIV, CD4+ lymphocytes die at an accelerated rate through uncontrolled apoptosis, when stimulated.
At the molecular level, hyperactive apoptosis can be caused by defects in signaling pathways that regulate the Bcl-2 family proteins. Increased expression of apoptotic proteins such as BIM, or their decreased proteolysis, leads to cell death, and can cause a number of pathologies, depending on the cells where excessive activity of BIM occurs. Cancer cells can escape apoptosis through mechanisms that suppress BIM expression or by increased proteolysis of BIM.[90]

text: Treatments[edit]

text: Treatments aiming to inhibit works to block specific caspases. Finally, the Akt protein kinase promotes cell survival through two pathways. Akt phosphorylates and inhibits Bad (a Bcl-2 family member), causing Bad to interact with the 14-3-3 scaffold, resulting in Bcl dissociation and thus cell survival. Akt also activates IKKα, which leads to NF-κB activation and cell survival. Active NF-κB induces the expression of anti-apoptotic genes such as Bcl-2, resulting in inhibition of apoptosis. NF-κB has been found to play both an antiapoptotic role and a proapoptotic role depending on the stimuli utilized and the cell type.[91]

text: HIV progression[edit]

text: The progression of the human immunodeficiency virus infection into AIDS is due primarily to the depletion of CD4+ T-helper lymphocytes in a manner that is too rapid for the body's bone marrow to replenish the cells, leading to a compromised immune system. One of the mechanisms by which T-helper cells are depleted is apoptosis, which results from a series of biochemical pathways:[92]

text:
HIV enzymes deactivate anti-apoptotic Bcl-2. This does not directly cause cell death but primes the cell for apoptosis should the appropriate signal be received.  In parallel, these enzymes activate proapoptotic procaspase-8, which does directly activate the mitochondrial events of apoptosis.
HIV may increase the level of cellular proteins that prompt Fas-mediated apoptosis.
HIV proteins decrease the amount of CD4 glycoprotein marker present on the cell membrane.
Released viral particles and proteins present in extracellular fluid are able to induce apoptosis in nearby "bystander" T helper cells.
HIV decreases the production of molecules involved in marking the cell for apoptosis, giving the virus time to replicate and continue releasing apoptotic agents and virions into the surrounding tissue.
The infected CD4+ cell may also receive the death signal from a cytotoxic T cell.

text: Cells may also die as direct consequences of viral infections. HIV-1 expression induces tubular cell G2/M arrest and apoptosis.[93]  The progression from HIV to AIDS is not immediate or even necessarily rapid; HIV's cytotoxic activity toward CD4+ lymphocytes is classified as AIDS once a given patient's CD4+ cell count falls below 200.[94]

text: Researchers from Kumamoto University in Japan have developed a new method to eradicate HIV in viral reservoir cells, named "Lock-in and apoptosis." Using the synthesized compound Heptanoylphosphatidyl L-Inositol Pentakisphophate (or L-Hippo) to bind strongly to the HIV protein PR55Gag, they were able to suppress viral budding. By suppressing viral budding, the researchers were able to trap the HIV virus in the cell and allow for the cell to undergo apoptosis (natural cell death). Associate Professor Mikako Fujita has stated that the approach is not yet available to HIV patients because the research team has to conduct further research on combining the drug therapy that currently exists with this "Lock-in and apoptosis" approach to lead to complete recovery from HIV.[95]

text: Viral infection[edit]

text: Viral induction of apoptosis occurs when one or several cells of a living organism are infected with a virus, leading to cell death. Cell death in organisms is necessary for the normal development of cells and the cell cycle maturation.[96] It is also important in maintaining the regular functions and activities of cells.

text: Viruses can trigger apoptosis of infected cells via a range of mechanisms including:

text:
Receptor binding
Activation of protein kinase R (PKR)
Interaction with p53
Expression of viral proteins coupled to MHC proteins on the surface of the infected cell, allowing recognition by cells of the immune system (such as Natural Killer and cytotoxic T cells) that then induce the infected cell to undergo apoptosis.[97]

text:
Canine distemper virus (CDV) is known to cause apoptosis in central nervous system and lymphoid tissue of infected dogs in vivo and in vitro.[98]
Apoptosis caused by CDV is typically induced via the extrinsic pathway, which activates caspases that disrupt cellular function and eventually leads to the cells death.[81] In normal cells, CDV activates caspase-8 first, which works as the initiator protein followed by the executioner protein caspase-3.[81]  However, apoptosis induced by CDV in HeLa cells does not involve the initiator protein caspase-8. HeLa cell apoptosis caused by CDV follows a different mechanism than that in vero cell lines.[81]  This change in the caspase cascade suggests CDV induces apoptosis via the intrinsic pathway, excluding the need for the initiator caspase-8. The executioner protein is instead activated by the internal stimuli caused by viral infection not a caspase cascade.[81]

text: The Oropouche virus (OROV) is found in the family Bunyaviridae. The study of apoptosis brought on by Bunyaviridae was initiated in 1996, when it was observed that apoptosis was induced by the La Crosse virus into the kidney cells of baby hamsters and into the brains of baby mice.[99]

text: OROV is a disease that is transmitted between humans by the biting midge (Culicoides paraensis).[100] It is referred to as a zoonotic arbovirus and causes febrile illness, characterized by the onset of a sudden fever known as Oropouche fever.[101]

text: The Oropouche virus also causes disruption in cultured cells – cells that are cultivated in distinct and specific conditions. An example of this can be seen in HeLa cells, whereby the cells begin to degenerate shortly after they are infected.[99]

text: With the use of gel electrophoresis, it can be observed that OROV causes DNA fragmentation in HeLa cells. It can be interpreted by counting, measuring, and analyzing the cells of the Sub/G1 cell population.[99] When HeLA cells are infected with OROV, the cytochrome C is released from the membrane of the mitochondria, into the cytosol of the cells. This type of interaction shows that apoptosis is activated via an intrinsic pathway.[96]

text: In order for apoptosis to occur within OROV, viral uncoating, viral internalization, along with the replication of cells is necessary. Apoptosis in some viruses is activated by extracellular stimuli. However, studies have demonstrated that the OROV infection causes apoptosis to be activated through intracellular stimuli and involves the mitochondria.[99]

text: Many viruses encode proteins that can inhibit apoptosis.[102] Several viruses encode viral homologs of Bcl-2. These homologs can inhibit proapoptotic proteins such as BAX and BAK, which are essential for the activation of apoptosis. Examples of viral Bcl-2 proteins include the Epstein-Barr virus BHRF1 protein and the adenovirus E1B 19K protein.[103] Some viruses express caspase inhibitors that inhibit caspase activity and an example is the CrmA protein of cowpox viruses. Whilst a number of viruses can block the effects of TNF and Fas. For example, the M-T2 protein of myxoma viruses can bind TNF preventing it from binding the TNF receptor and inducing a response.[104] Furthermore, many viruses express p53 inhibitors that can bind p53 and inhibit its transcriptional transactivation activity. As a consequence, p53 cannot induce apoptosis, since it cannot induce the expression of proapoptotic proteins. The adenovirus E1B-55K protein and the hepatitis B virus HBx protein are examples of viral proteins that can perform such a function.[105]

text: Viruses can remain intact from apoptosis in particular in the latter stages of infection. They can be exported in the apoptotic bodies that pinch off from the surface of the dying cell, and the fact that they are engulfed by phagocytes prevents the initiation of a host response. This favours the spread of the virus.[104]

text: Plants[edit]

text: Programmed cell death in plants has a number of molecular similarities to that of animal apoptosis, but it also has differences, notable ones being the presence of a cell wall and the lack of an immune system that removes the pieces of the dead cell.  Instead of an immune response, the dying cell synthesizes substances to break itself down and places them in a vacuole that ruptures as the cell dies.  Whether this whole process resembles animal apoptosis closely enough to warrant using the name apoptosis (as opposed to the more general programmed cell death) is unclear.[106]

text: Caspase-independent apoptosis[edit]

text: The characterization of the caspases allowed the development of caspase inhibitors, which can be used to determine whether a cellular process involves active caspases. Using these inhibitors it was discovered that cells can die while displaying a morphology similar to apoptosis without caspase activation.[107] Later studies linked this phenomenon to the release of AIF (apoptosis-inducing factor) from the mitochondria and its translocation into the nucleus mediated by its NLS (nuclear localization signal). Inside the mitochondria, AIF is anchored to the inner membrane. In order to be released, the protein is cleaved by a calcium-dependent calpain protease.