prompt_front: Now, answer the following question based on the text below, only give me the answer and do not output any other words.

Question: When did KSTP switch to a sports radio format?

Read the following text and answer briefly.

Context 1: KSTP (1500 AM; SKOR North) is a commercial AM radio station licensed to Saint Paul, Minnesota  It is the flagship AM radio station of Hubbard Broadcasting, which also owns several other television and radio stations across the United States   KSTP has a sports radio format and is the ESPN Radio Network affiliate for Minneapolis-St  Paul   The radio studios are  on University Avenue in Minneapolis, shared with sister stations KSTP-FM, KSTP-TV, KTMY, and KSTC-TV  On weekdays, KSTP airs local sports shows from 9 a m  to 9 p m  and carries ESPN programming weekday mornings, late nights and weekends  Some KSTP shows are simulcast on other sports radio stations in the region 

KSTP runs the maximum power for AM stations, 50,000 watts   It shares clear-channel, Class A status on 1500 AM with WFED in Washington, D C   KSTP broadcasts a directional signal at night, using a three-tower array, with its transmitter on U S  Route 61 at Beam Avenue in Maplewood   Programming is also heard on 250 watt FM translator K235BP at 94 9 MHz in Bemidji 

History

WAMD and KFOY
KSTP's start in 1928 was the product of a merger between two pioneering Twin Cities stations: WAMD ("Where All Minneapolis Dances") in Minneapolis, first licensed on February 16, 1925 to Stanley E  Hubbard, and KFOY in St  Paul, first licensed on March 12, 1924 to the Beacon Radio Service in St  Paul 

Following a few test transmissions, WAMD made its formal debut broadcast on February 22, 1925  (In later interviews Stanley Hubbard traced WAMD's start to April 1924 ) It was located at the Marigold Dance Garden, and featured nightly "Midnight Frolics" broadcasts by the ballroom's orchestra  It is claimed that WAMD was the first radio station to be completely supported by running paid advertisements  Effective June 15, 1927, WAMD was assigned to 1330 kHz 

On November 11, 1927 WAMD's transmitter site at Oxboro Heath on Lyndale Avenue South burned down, two weeks after the station had been sold to the National Battery Company  An initial arrangement was made to carry WAMD's programs over WRHM (now WWTC), transmitting on WAMD's 1330 kHz frequency  Beginning on November 24, 1927 the WAMD broadcasts, still on 1330 kHz, were shifted to KFOY's facility in St  Paul  (At this time KFOY was assigned to 1050 kHz)  The next day it was announced that National Battery had purchased KFOY, and as of December 1, 1927 both KFOY and WAMD were reassigned to 1350 kHz  WAMD continued making regular broadcasts until the end of March 1928, while KFOY, although it continued to be licensed for a few more months on a time-sharing basis with WAMD, ceased operations at this point 

National Battery Company
In mid-December 1927, the National Battery Company announced it had received permission from the Federal Radio Commission (FRC) to build a new station, with the call letters KSTP, operating from a transmitter site to be constructed three miles south of Wescott  The next month it was reported that the new station, still under construction, had been assigned to 1360 kHz  KSTP made its debut broadcast on March 29, 1928  Although technically it was a separate station from WAMD and KFOY, both of which were formally deleted on April 30, 1928, overall KSTP was treated as the direct successor to a consolidated WAMD and KFOY 

Hubbard became the merged station's general manager, acquiring controlling interest in 1941  A month after the merger, KSTP became an affiliate for the NBC Red Network   It remained with NBC for 46 years  On November 11, 1928, under the provisions of the FRC's General Order 40, KSTP was assigned to a "high-powered regional" frequency of 1460 kHz  The only other station assigned to this frequency was WTFF in Mount Vernon Hills, Virginia (later WJSV, now WFED, Washington, D C )  On February 7, 1933, the FRC authorized KSTP to increase its daytime power to 25 KW  In 1938 and 1939 KSTP also operated a high-fidelity AM "experimental audio broadcasting station" Apex station, W9XUP, originally on 25,950 kHz and later on 26,150 kHz  In 1941, as part of the implementation of the North American Regional Broadcasting Agreement, KSTP was assigned to its current "clear channel" frequency of 1500 kHz, with the provision that it and WJSV, as "Class I-B" stations, had to maintain directional antennas at night in order to mutually protect each other from interference  An FM station, KSTP-FM, was founded in 1946 but shut down in 1952 

Hubbard reportedly acquired an RCA TV camera in 1939, and started experimenting with television broadcasts   But World War II put a hold on the development of television   In 1948, with the war over, KSTP-TV became the first television station in Minnesota   With KSTP 1500 already associated with NBC Radio, KSTP-TV became an NBC Television Network affiliate   From 1946 to 1952, KSTP also had an FM counterpart   KSTP-FM 102 1 was only on the air four years   There were few radios equipped to receive FM signals in that era, and management decided to discontinue FM broadcasts 

MOR and Top 40
As network programming moved from radio to television, KSTP programmed a full service Middle of the Road (MOR) radio format, in the shadow of its chief competitor, CBS Radio affiliate 830 WCCO  In 1965, a new FM station, reviving the KSTP-FM call sign, was put on the air, largely simulcasting the AM station   But by the late 1960s, KSTP-FM began a separate format of beautiful music   KSTP was the radio home of the Minnesota Vikings football team from 1970 to 1975  

In 1973, KSTP broke away from its longtime adult MOR sound and became one of four area stations at the time to program a Top 40 format  "15 KSTP, The Music Station" competed with Top 40 AM rivals WDGY, KDWB and later, WYOO  The competition would eventually shake itself out, with outrageous rocker WYOO dropping out after being sold in 1976, and then the staid WDGY switching to country music the following year   As for uptempo hits station 15 KSTP, it went from a tight Top 40 format to leaning adult rock in 1978, to leaning adult contemporary in 1979, to evolving into adult contemporary/talk by 1980   In 1982, it officially shifted to talk   Most Top 40 rock music, by this time, had moved to the FM band 

Past Personalities

Notable hosts who have been on KSTP include John Hines, Jesse Ventura, Larry Carolla, Tom Barnard, Big Al Davis, Don Vogel, John MacDougall, Griff, Mike Edwards, Geoff Charles, Joe Soucheray, James Lileks, Leigh Kamman, Barbara Carlson, Peter Thiele, Tom Mischke, Jason Lewis, Chuck Knapp, Machine Gun Kelly, Charle Bush, Mark O'Connell and Paul Brand  These broadcasters were supported by producers such as Bruce Huff, Rob Pendleton, Alison Brown, Jean Bjorgen, David Elvin (who Vogel dubbed the "Steven Spielberg of Talk Radio"), Mitch Berg and others 

The station has, for the most part, emphasized local hosts over the years   But in 1988, KSTP was one of Rush Limbaugh's first affiliates when his conservative talk show was rolled out for national syndication   (Clear Channel-owned KTLK-FM took over rights to Limbaugh's show in January 2006)  Other syndicated hosts previously heard on KSTP include Sean Hannity, Bruce Williams, Larry King, and Owen Spann 

Sports Radio
KSTP switched to Sports Radio on February 15, 2010   As the station had to wait for ESPN's contract with rival KFAN and its sister station KFXN to expire, it did not become an ESPN Radio affiliate until April 12, the same day that the Minnesota Twins were scheduled to play the first game in their new ball park, Target Field, against the Boston Red Sox   As a result Coast to Coast AM and Live on Sunday Night, it's Bill Cunningham were retained during this period   One ESPN Radio network program, The Herd with Colin Cowherd, was picked up by KSTP immediately following the format change 

In 2018, the station was approved for an FM translator on 94 1 FM, broadcasting from a transmitter atop the IDS Center in downtown Minneapolis  The two-watt signal threw most of its power to the west, preventing interference to low powered FM stations on the same channel including WFNU-LP in St  Paul  With only two watts of power, however, the signal was limited to the immediate downtown area surrounding the IDS Center   It later acquired a 250 watt translator, K235BP at 94 9 MHz   The original translator was discontinued 

On January 15, 2019, KSTP rebranded as "SKOR North" (a reference to the Vikings team song/chant, "Skol, Vikings"), with local programming between 12 noon and 7 pm  About a year later, in May of 2020, KSTP suspended most of its local programming and laid off nearly all of its local staff  Station management cited the economic toll of the coronavirus for the changes  Sports broadcasting continues, primarily composed of ESPN radio network broadcasts 

Sports Teams

KSTP-AM served as the radio flagship for the Minnesota Vikings football team from 1970 to 1975 

On August 1, 2006, the station announced that it would be the new flagship station for the Minnesota Twins baseball team, effective with the start of the 2007 season  The Twins had been on rival WCCO since arriving in Minnesota in 1961   KSTP served as the flagship for the Twins until the end of the 2012 season, when games moved to 96 3 KTWN-FM (now KMWA)   The Twins have since returned to WCCO 830 

The switch to a fairly weak FM station caused dissent among some listeners, particularly in communities that had trouble picking up KSTP 1500   Although KSTP is the state's second most powerful AM station, it must operate directionally at night, delivering a reduced signal to parts of the market  WCCO, by comparison, offers a signal with a wider coverage area during the day than KSTP does, with WCCO's non-directional 50,000 watt signal  In response, the Twins have expanded the number of affiliates 

On March 9, 2011, KSTP announced it would be the new flagship for the University of Minnesota Golden Gophers men's and women's basketball and men's ice hockey, ending a 68-year run on WCCO  The rights have since moved to KFXN-FM, which already aired Gopher football 

On March 2, 2017, KSTP announced it would be the first radio broadcaster for Minnesota United FC   The move brings live soccer action to 1500 AM 

Previous logos

References

External links
KSTP website

FCC History Cards for KSTP (covering 1928-1980)
Radiotapes com Historic Minneapolis/St  Paul airchecks dating back to 1924 including KSTP and other Twin Cities radio stations 
Rick Burnett's TwinCitiesRadioAirchecks com has additional airchecks of KSTP and other Twin Cities radio stations from the '60s and '70s, including Chuck Knapp's 2nd show on KSTP 

Hubbard Broadcasting
ESPN Radio stations
Peabody Award winners
Radio stations in Minneapolis–Saint Paul
Radio stations established in 1925
1925 establishments in Minnesota
Minnesota Kicks
Sports radio stations in the United States
Clear-channel radio stations
Context 3: PROBLEM TO BE SOLVED: To provide a vacuum processor which can suppress the rise of manufacture cost while coping with the enlargement of diameter of a sample, and besides is excellent in maintainability  SOLUTION: This vacuum processor comprises an air loader 1 which is equipped with a plurality of juxtaposed cassette stands 2a and 2b and a carrier 13 for carrying a wafer 20 from or to the cassette stands, a vacuum loader 7 which is equipped with etching treatment chambers 11 and 11b for processing the wafer and a vacuum carriage chamber 16 connected to them through gate valves 15a and 15b, and a lock device 6 consisting of a load lock chamber 6b and an unload lock chamber 6b equipped with gate valves 12a, 12b, 12c, and 12d for connecting the said carrier 13 with the vacuum carriage chamber 16  For the etching treatment chambers, two are provided only on the opposite side of the vacuum carriage chamber symmetrically about the axis line A passing the center, and besides the arrangement positions of the two etching treatment chambers are at an acute angle on the opposite side of the vacuum carriage chamber 
エッチング，ＣＶＤ（化学的気相成長），スパッタリング，アッシング，リンサ（水洗）等の枚葉処理をするのに好適な真空処理装置とそれを用いて半導体デバイスを製造する半導体製造ラインに関するものである。 Those etching, CVD (chemical vapor deposition), sputtering, ashing, to rinser (water washing) suitable vacuum processing apparatus for single wafer processing, such as a semiconductor manufacturing line for manufacturing a semiconductor device using the same  
【従来の技術】試料を処理する真空処理装置は、大別すると、カセットブロックと真空処理ブロックから構成されており、カセットブロックは、半導体製造ラインのベイ通路に面して長手方向に伸びるフロントを有し、試料用のカセットや試料のオリエンテーションを合わせるアライメントユニットと、大気ロボットがある。 BACKGROUND OF THE INVENTION Vacuum processing apparatus for processing a sample, the roughly is constituted by a cassette block and a vacuum processing block, the cassette block, a front extending in the longitudinal direction facing the bay aisle of a semiconductor manufacturing line a, an alignment unit for aligning the cassette and sample orientation for sample, there is an atmospheric robot  真空処理ブロックには、ロード側ロードロック室，アンロード側ロードロック室，真空処理室，後真空処理室，真空ポンプ及び真空ロボット等が設けられている。 The vacuum processing block, the load-side load lock chamber, the unload side load lock chamber, the vacuum processing chamber, a rear vacuum processing chamber, a vacuum pump and a vacuum robot and the like 
【０００３】これらの真空処理装置では、カセットブロックのカセットから取り出された試料が、大気ロボットにより真空処理ブロックのロードロック室まで搬送される。  In these vacuum processing apparatus, a sample taken from the cassette in the cassette block is transported to the load lock chamber in the vacuum processing block by the atmospheric robot  ロードロック室から真空ロボットによりさらに処理室に搬送され、電極構造体上にセットされた試料は、プラズマエッチング等の処理がなされる。 Is conveyed from the load lock chamber to the further processing chamber by the vacuum robot, the sample is set on an electrode structure, processing such as plasma etching is performed  その後、必要に応じて後真空処理室に搬送，処理される。 Thereafter, the conveyance to the rear vacuum processing chamber as necessary and processed  処理済みの試料は、真空ロボット及び大気ロボットによりカセットブロックのカセットに搬送される。 Processed sample is conveyed to the cassette of the cassette block by the vacuum robot and the atmospheric robot 
【０００４】試料をプラズマエッチング処理する真空処理装置の例としては、例えば特公昭61−8153号公報，特開昭63−133532号公報，特公平6−30369号公報，特開平  Examples of the sample vacuum processing apparatus for plasma etching treatment, for example Japanese Patent Publication 61-8153, JP-Sho 63-133532 and JP Kokoku 6-30369, JP-A No 
6−314729号公報，特開平6−314730号公報，米国特許第 6-314729, JP-A No  6-314730, JP-U S  Patent No 
5,314,509号明細書および5,784,799号明細書に記載されたようなものがある。 There are such as described in Pat and 5,784,799 Pat 5,314,509 
509号明細書に記載された装置は、真空処理ブロックの中央付近に真空ロボット、その周囲に３個の処理室が同心状に配置され、真空ロボットとカセットブロックの間に、ロード側ロードロック室，アンロード側ロードロック室が設けられている。 Device described in 509 Pat are vacuum robot in the vicinity of the center of the vacuum processing block, three process chambers around it are arranged concentrically, between the vacuum robot and the cassette block, the load-side load-lock chamber , unload side load lock chamber is provided  これらの装置では、大気ロボットや真空ロボットの搬送アームの回転角度が大きく従って装置全体の必要床面積が大きいという問題がある。 In these devices, there is a problem that needs floor space for the entire rotation angle is large therefore device of the transfer arm of the atmospheric robot and the vacuum robot is large 
【０００６】一方、真空処理装置の真空処理ブロック内の処理室や真空ポンプその他各種の配管機器については、定期，不定期に点検修理等のメンテナンスを行うことが必要である。  On the other hand, the processing chamber and the vacuum pump and other various piping components of the vacuum processing block of the vacuum processing apparatus, periodically, it is necessary to perform maintenance such as inspection and repair irregularly  そのため、一般に、真空処理ブロックの周囲には、扉が設けられており、この扉を開けることにより、ロードロック室，アンロードロック室，処理室，真空ロボット及び各種の配管機器の点検修理ができるようになっている。 Therefore, in general, around the vacuum processing block, the door is provided by opening the door, load lock chambers, unload lock chambers, processing chambers, the servicing of the vacuum robot and various piping devices It has become way 
キャリアポッドが必要となるために、約３５０mm程度と大きくなり、複数のキャリアポッドを収納するカセットブロックの幅も大きくなる。 For carrier pod required, large as about 350 mm, the greater width of the cassette block for housing a plurality of carrier pods  この幅に合わせて真空処理ブロックの幅を決定すると、真空処理装置全体が大きなスペースを必要とすることになる。 When determining the width of the vacuum processing block in accordance with the this width, the entire vacuum processing apparatus requires a large space  一例として、４個のキャリアポッドを収納するカセットブロックについて考えると、試料の直径ｄが８インチから１２インチになった場合、カセットの幅は少なくとも約４０cm以上大きくならざるを得ない。 As an example, considering the cassette block for accommodating four carriers pods, if the diameter d of the sample was a 12-inch 8 inch wide cassette inevitably increases at least about 40cm or more 
【０００８】一方、試料に各種の処理を行いながら大量の処理を行うために、一般の半導体製造ラインでは、同じ処理を行う複数の真空処理装置を同じベイに集め、各ベイ間の搬送を自動またはマニュアルで行っている。 On the other hand, in order to perform a lot of processing while performing various processes in the sample, in a general semiconductor manufacturing lines, gathering a plurality of vacuum processing apparatus for performing the same processing in the same bay, automatic conveyance between the bays or it is carried out manually  このような半導体製造ラインは、高いクリーン度を必要とするため、半導体製造ライン全体が大きなクリーンルーム内に設置される。 Such a semiconductor manufacturing line, requires a high degree of cleanliness, the whole semiconductor manufacturing line is placed in a large clean room  試料の大口径化に伴う真空処理装置の大型化は、クリーンルーム占有面積の大型化を伴うが、これはもともと建設コストの高いクリーンルームの建設コストを一層増加させることになる。 Size of the vacuum processing apparatus due to the large diameter of the sample is accompanied by a large clean room area occupied, which will be further increased construction costs of the high construction cost clean room originally  もし、同じ面積のクリーンルームに占有面積の大きな真空処理装置を設置するとすれば、真空処理装置の全体の台数を減らすか、あるいは各真空処理装置間の間隔を狭くせざるを得ない。 If, if the clean room of the same area to install a large vacuum processing apparatus of the occupied area, reduce the overall number of the vacuum processing apparatus, or interval narrower forced between the vacuum processing apparatus  同じ面積のクリーンルームにおける真空処理装置の設置台数減少は、必然的に半導体の製造ラインの生産性の低下ひいては半導体の製造コストの上昇を伴う。 Installed base reduction in the vacuum processing apparatus in a clean room having the same area is accompanied inevitably rise of the semiconductor decrease and thus the semiconductor manufacturing cost of productivity of the production line  他方、各真空処理装置間の間隔を狭くすることは、点検修理のためのメンテナンススペースが不足し、真空処理装置のメンテナンス性を著しく阻害する。 On the other hand, to reduce the distance between each of the vacuum processing apparatus, the maintenance space for inspection and repair is insufficient to significantly inhibit the maintenance of the vacuum processing apparatus 
【０００９】本発明の目的は、試料の大口径化に対応しつつ、製造コストの上昇を抑えることのできる真空処理装置を提供することにある。 An object of the present invention, while corresponding to the large diameter of the sample, is to provide a vacuum processing apparatus capable of suppressing an increase in manufacturing cost 
【００１０】本発明の他の目的は、試料の大口径化に対応しつつ、メンテナンス性に優れた真空処理装置を提供することにある。 Another object of the present invention, while corresponding to the large diameter of the sample is to provide a vacuum processing apparatus having excellent maintainability 
【００１１】本発明の他の目的は、試料の大口径化に対応しつつ、真空処理装置の必要設置台数を確保して製造コストの上昇を抑え、かつ、メンテナンス性も損なわない半導体製造ラインを提供することにある。 Another object of the present invention, while corresponding to the large diameter of the sample, to ensure the necessary number of installed vacuum processing apparatus suppressing an increase in manufacturing cost, and a semiconductor manufacturing line is not impaired maintainability It is to provide 
【課題を解決するための手段】本発明は、並設した複数のカセット台およびカセット台から、あるいはカセット台へウエハを搬送するための搬送装置を備えた大気ローダと、ウエハを処理するための真空処理室およびこれにゲート弁を介して連接された真空搬送室を備えた真空ローダと、前記搬送装置と前記真空搬送室とを連接するためのゲート弁を備えたロードロック室およびアンロードロック室からなるロック装置とを含んで構成される真空処理装置において、ウエハを処理するための真空処理室は、有磁場ＵＨＦ帯電磁波放射放電方式リアクタ（以下、ＵＨＦ−ＥＣＲリアクタという。）によって形成される真空処理室であり、該真空処理室には、分解可能な側壁インナーユニットおよびアンテナが設けられ、該真空処理室は、真空搬 The present invention SUMMARY OF THE INVENTION from a plurality of cassette tables and cassette stand juxtaposed, or the atmosphere loader having a conveying device for conveying the wafer to the cassette table, for processing a wafer the load lock chamber and the unload lock having a gate valve for connecting the vacuum loader vacuum processing chamber and having a vacuum transfer chamber which is connected via a gate valve to the said vacuum transfer chamber and the conveying device in the vacuum processing apparatus configured to include a lock device comprising a chamber, a vacuum processing chamber for processing a wafer it is formed by a magnetic field UHF band electromagnetic wave radiation discharge type reactor (hereinafter  referred UHF-ECR reactor) that a vacuum processing chamber, the vacuum processing chamber, degradable sidewall inner unit and an antenna are provided, the vacuum processing chamber, a vacuum transportable 室およびロック装置の中央を通る軸線に対して対称にして、かつ真空搬送室を中心にしてロック装置の反対側のみに２つ設けられ、かつ真空搬送室に対して前記２つの真空処理室の配置位置は鋭角をなしている真空処理装置を提供する。 And symmetrically with respect to the axis passing through the center of the chamber and the locking device, and the opposite side only two provided for to lock device around the vacuum transfer chamber, and the two vacuum processing chamber to the vacuum transfer chamber position is to provide a vacuum processing apparatus which forms an acute angle 
ＣＲリアクタによって形成される真空処理室であり、該真空処理室は、真空搬送室およびロック装置の中央を通る軸線に対して対称にして、かつ真空搬送室を中心にしてロック装置の反対側のみに２つ設けられ、かつ真空搬送室に対して前記２つの真空処理室の配置位置は鋭角をなしており、ＵＨＦ−ＥＣＲのアンテナは、前記軸線に対して平行で、かつ前記真空搬送室とは反対側に開放される真空処理装置を提供する。 A vacuum processing chamber formed by a CR reactor, vacuum processing chamber, and symmetrically with respect to the axis passing through the center of the vacuum transfer chamber and the locking device, and the opposite side of the locking device around the vacuum transfer chamber only two provided, and positions of the two vacuum processing chamber to the vacuum transfer chamber is an acute angle, the UHF-ECR antennas, and parallel, and the vacuum transfer chamber with respect to said axis to provides a vacuum processing apparatus is opened to the opposite side 
かつ真空搬送室を中心にしてロック装置の反対側のみに２つ設けられ、かつ真空搬送室に対して前記２つの真空処理室の配置位置は鋭角をなしており、大気ローダ，真空ローダおよびロック装置はＴ字配置とされた真空処理方法を提供する。 And around the vacuum transfer chamber opposite only two provided a locking device, and positions of the two vacuum processing chamber to the vacuum transfer chamber is an acute angle, atmospheric loader, vacuum loader and locking apparatus to provide a vacuum processing method which is a T-arrangement 
【００１５】本発明は、並設した複数のカセット台およびカセット台から、あるいはカセット台へウエハを搬送するための搬送装置を備えた大気ローダと、ウエハを処理するための真空処理室およびこれにゲート弁を介して連接された真空搬送室を備えた真空ローダと、前記搬送装置と前記真空搬送室とを連接するためのゲート弁を備えたロードロック室およびアンロードロック室からなるロック装置とを含んで構成される真空処理装置が平行に複数台並設された真空処理システムにおいて、ウエハを処理するための真空処理室は、ＵＨＦ−ＥＣＲリアクタによって形成される真空処理室であり、該真空処理室は、真空搬送室およびロック装置の中央を通る軸線に対して対称にして、かつ真空搬送室を中心にしてロック装置の反対側のみに２ The present invention, a plurality of cassette tables and cassette stand juxtaposed, or the atmosphere loader having a conveying device for conveying the wafer to the cassette base, the vacuum processing chamber for processing the wafer and to a vacuum loader having a vacuum transfer chamber which is connected via a gate valve, the locking consisting of the conveying device and the load lock chamber and the unload lock chamber with gate valves for connecting the said vacuum transfer chamber apparatus and in the vacuum processing system vacuum processing apparatus is constituted with a plurality Tainami set in parallel include vacuum processing chamber for processing a wafer is vacuum processing chamber formed by the UHF-ECR reactor, vacuum processing chamber, and symmetrically with respect to the axis passing through the center of the vacuum transfer chamber and the locking device, and 2 only on the opposite side of the locking device around the vacuum transfer chamber 設けられ、かつ真空搬送室に対して前記２つの真空処理室の配置位置は鋭角をなしており、並設された複数の真空処理装置のすべての真空処理室に一直線上に配列される真空処理システムを提供する。 Provided, and positions of the two vacuum processing chamber to the vacuum transfer chamber is an acute angle, a vacuum process that is arranged in alignment with all of the vacuum processing chamber of a plurality of vacuum processing apparatus are arranged in parallel to provide a system 
【発明の実施の形態】以下、本発明にかかる一実施例を図面に基づいて説明する。 BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, will be explained based on an embodiment according to the present invention with reference to the accompanying drawings 
１０Ｂ，１０Ｃで示す。 10B, shown by 10C 
【００１８】図１に示す真空処理システムを説明する前に、図２から図４に基づいて真空処理装置を説明する。  Before describing the vacuum processing system shown in FIG  1, illustrating a vacuum processing apparatus on the basis of FIGS  2-4 
５ｂを介して連接された真空搬送室１６を備えた真空ローダ７と、前記搬送装置と前記真空搬送室とを連接するためのゲート弁を備えたロードロック室６ａおよびアンロードロック室６ｂからなるロック装置６とを含んで構成される。 5b a vacuum loader 7 having a vacuum transfer chamber 16 which is connected via consist load lock chamber 6a and the unload lock chamber 6b has a gate valve for connecting and said vacuum transfer chamber and the conveying device configured to include a locking device 6 
平行形に配置され、その位置および姿勢を変えることなく、装置への導入／払出しが可能な位置、すなわち、カセット１ａないし１ｃを略水平の平面上で常に一定の位置に固定される。 Arranged parallel type, without changing its position and orientation, the introduction / dispensing of the locations of the device, i e , always fixed to a constant position to free the cassette 1a 1c on a substantially horizontal plane  カセット台２ａおよび２ｂは、平行に隣合わせて配置してある。 Cassette tables 2a and 2b, are arranged side by side in parallel  カセット台２ｃは、最右端に配置してある。 Cassette table 2c is, are arranged on the top right edge  カセット１ａおよび１ｂは、処理を行うための末処理ウエハを収納したり、処理済みのウエハを回収するためのもので、複数枚（通常２５枚）の被処理基板であるウエハ２０が収納可能となっている。 Cassettes 1a and 1b, or storing the processed wafers end for processing, for recovering the processed wafer, the wafer 20 is a substrate to be processed in a plurality (25 pieces Normal) can be stored going on  カセット１ｃは、この場合、プラズマを用いたドライクリーニング（以下、「プラズマクリーニング」という。）を行うためのダミーウエハを収納したり、プラズマクリーニング後のダミーウエハを回収するためのもので、複数枚（通常２５枚）のダミーウエハ３０が収納可能となっている。 Cassette 1c in this case, dry cleaning using plasma (hereinafter, "plasma cleaning" hereinafter ) Or housing a dummy wafer for performing, for recovering the dummy wafers after plasma cleaning, a plurality (usually dummy wafer 30 of 25 sheets) has become can be stored 
とカセット１ａ，１ｂとの間でウエハ２０を、そしてロードロック室６ａおよびアンロードロック室６ｂとカセット１ｃとの間でダミーウエハ３０を授受可能に動作する。 A cassette 1a, the wafer 20 with the 1b, and operates to allow exchanging dummy wafer 30 between the load lock chamber 6a and the unload lock chamber 6b and the cassette 1c 
ａ，１５ｂを介して真空処理室であるエッチング処理室１１ａ，１１ｂが設けてある。 a, etching chamber is a vacuum processing chamber through a 15b 11a, 11b are provided  以下、エッチング処理室を例に取って説明する。 Hereinafter will be described taking the etching chamber as an example  真空搬送室１６内には、ロードロック室６ａ，アンロード室６ｂおよびエッチング処理室１１ａ，１１ｂとの間でウエハ２０またはダミーウエハ３０を授受可能に動作する搬送装置１４が設けてある。 The vacuum transfer chamber 16, load lock chambers 6a, unload chamber 6b and etching chambers 11a, conveying device 14 is provided that operates to allow transferring the wafer 20 or dummy wafer 30 between 11b  真空搬送室１６は、独立に真空排気可能な真空排気装置１７を装備している。 Vacuum transfer chamber 16 is equipped with a vacuum evacuable vacuum evacuation device 17 independently 
【００２２】ＵＨＦ−ＥＣＲリアクタのエッチング処理室１１ａ，１１ｂは、この場合、同一の構成で対称配置とされてエッチング処理が行われるようになっている。 The UHF-ECR reactor etching chambers 11a, 11b is in this case are symmetrically arranged in the same configuration so that the etching process is performed 
エッチング処理室１１ａを例に説明する。 The etching chamber 11a will be described as an example  エッチング処理室１１ａは、ウエハ２０を配置するための試料台を有し、試料台８ａの上部に放電部７ａを形成するように放電室が設けてある。 Etching chamber 11a has a sample stage for placing the wafer 20, the discharge chamber so as to form a discharge portion 7a at the top of the sample table 8a is provided  エッチング処理室１１ａは、放電部７ａへの処理ガス供給のためのガス導入装置１０ａを有するとともに、エッチング処理室１１ａ内を所定圧力に減圧排気する真空排気装置９ａを有し、放電部７ａの処理ガスをプラズマ化するための、この場合、ＵＨＦ波と磁場の発生手段を有している。 The etching chambers 11a, which has a gas introduction device 10a for processing the gas supply to the discharge portion 7a, having a vacuum exhaust device 9a for evacuating the etching chamber 11a to a predetermined pressure, the process of the discharge portion 7a for plasma gas, in this case, it has a generating means of the UHF wave and magnetic field 
９は、センサ１８からの計測値を所定値と比較して、エッチング処理室内のクリーニング時期を判断する。 9, the measured value from the sensor 18 is compared with a predetermined value to determine the cleaning time of the etching chamber  また、制御装置１９は、真空搬送装置１３および１４を制御して、ダミーウエハ３０をカセット１ｃおよびエッチング処理室１１ａないし１１ｂの間で搬送制御する。 The control device 19 controls the vacuum transfer apparatus 13 and 14, to the dummy wafer 30 to the cassette 1c and the etching process chamber 11a carrying controlled between 11b 
いずれかの方法によりウエハ処理またはプラズマクリーニングを実行する。 It executes the wafer processing or plasma cleaning by any method 
によって、カセット１ａ内のウエハ２０を下から順にエッチング処理室１１ａ，１１ｂに搬入し、それぞれのウエハ２０をエッチング処理する。 Accordingly, the wafer 20 to an etching treatment chamber in order from the bottom 11a of the cassette 1a, and carried into 11b, and each of the wafer 20 is etched  処理されたそれぞれのウエハ２０は、真空搬送装置１４および搬送装置１３によって、カセット１ａ内の元の位置に収納する。 Each wafer 20 processed by vacuum transfer apparatus 14 and the carrier 13 is housed in its original position in the cassette 1a  この場合、運転開始から終了に至る間、カセットの位置および姿勢を変えることなく未処理のウエハを取り出し、そして処理済みのウエハを未処理のウエハが収納されていた元の位置に戻して収納する。 In this case, while leading to completion of the start of the operation, it takes out an unprocessed wafer without changing the position and posture of the cassettes, and the processed wafer unprocessed wafer is accommodated back into position which is housed   このようにすることで、生産ラインの自動化への対応が容易で、且つ、ゴミの発生によるウエハの汚染を低減でき、高い生産効率と高い製品歩留まりを実現できる。 In this way, it is easy to respond to automation of production lines, and can reduce the contamination of the wafer due to the generation of dust can achieve high production efficiency and high product yield 
の処理が全て終り次のカセットの内のウエハ処理に移る前でも良い。 Of the process is good, even before moving on to the wafer processing of all end next cassette 
【００２７】プラズマクリーニングの実施にあたっては、次の順序で行われる。 The practice of plasma cleaning is performed in the following order  この場合、カセット１ｃに収納されたダミーウエハ３０（この



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Question: What is the recommended daily intake of vitamin K for adult women and men?

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Context 2: Vitamin K - Wikipedia
(Redirected from Vitamin k)
This article needs more medical references for verification or relies too heavily on primary sources  Please review the contents of the article and add the appropriate references if you can  Unsourced or poorly sourced material may be challenged and removed  (November 2015)
This article is about the family of vitamers  For vitamin K1 the form usually used as a supplement, see Phytomenadione 
Vitamin K structures  MK-4 and MK-7 are both subtypes of K2 
Vitamin K deficiency, Warfarin overdose
Vitamin K is a group of structurally similar, fat-soluble vitamins the human body requires for complete synthesis of certain proteins that are prerequisites for blood coagulation and which the body also needs for controlling binding of calcium in bones and other tissues  The vitamin K-related modification of the proteins allows them to bind calcium ions, which they cannot do otherwise  Without vitamin K, blood coagulation is seriously impaired, and uncontrolled bleeding occurs  Low levels of vitamin K also weaken bones and promote calcification of arteries and other soft tissues[citation needed] 
Chemically, the vitamin K family comprises 2-methyl-1,4-naphthoquinone (3-) derivatives  Vitamin K includes two natural vitamers: vitamin K1 and vitamin K2 [1] Vitamin K2, in turn, consists of a number of related chemical subtypes, with differing lengths of carbon side chains made of isoprenoid groups of atoms 
Vitamin K1, also known as phylloquinone, is made by plants, and is found in highest amounts in green leafy vegetables because it is directly involved in photosynthesis  It may be thought of as the plant form of vitamin K  It is active as a vitamin in animals and performs the classic functions of vitamin K, including its activity in the production of blood-clotting proteins  Animals may also convert it to vitamin K2 
Bacteria in the gut flora can also convert K1 into vitamin K2  In addition, bacteria typically lengthen the isoprenoid side chain of vitamin K2 to produce a range of vitamin K2 forms, most notably the MK-7 to MK-11 homologues of vitamin K2  All forms of K2 other than MK-4 can only be produced by bacteria, which use these forms in anaerobic respiration  The MK-7 and other bacterially derived forms of vitamin K2 exhibit vitamin K activity in animals, but MK-7's extra utility over MK-4, if any, is unclear and is a matter of investigation 
Three synthetic types of vitamin K are known: vitamins K3, K4, and K5  Although the natural K1 and all K2 homologues and synthetic K4 and K5 have proven nontoxic, the synthetic form K3 (menadione) has shown toxicity [2]
1 2 Cardiovascular health
1 4 Coumarin poisoning
4 1 Conversion of vitamin K1 to vitamin K2
4 2 Vitamin K2
6 Absorption and dietary need
7 Dietary reference intake
10 Biochemistry
10 1 Function in animals
10 2 Gamma-carboxyglutamate proteins
10 3 Methods of assessment
10 4 Function in bacteria
11 Injection in newborns
11 3 Controversy
A review of 2014 concluded that there is positive evidence that monotherapy using MK-4, one of the forms of Vitamin K2, reduces fracture incidence in post-menopausal women with osteoporosis, and suggested further research on the combined use of MK-4 with bisphosphonates  In contrast, an earlier review article of 2013 concluded that there is no good evidence that vitamin K supplementation helps prevent osteoporosis or fractures in postmenopausal women [3]
A Cochrane systematic review of 2006 suggested that supplementation with Vitamin K1 and with MK4 reduces bone loss; in particular, a strong effect of MK-4 on incident fractures among Japanese patients was emphasized [4]
A review article of 2016 suggested to consider, as one of several measures for bone health, increasing the intake of foods rich in vitamins K1 and K2 [5]
Cardiovascular health[edit]
Adequate intake of vitamin K is associated with the inhibition of arterial calcification and stiffening,[6] but there have been few interventional studies and no good evidence that vitamin K supplementation is of any benefit in the primary prevention of cardiovascular disease [7]
One 10-year population study, the Rotterdam Study, did show a clear and significant inverse relationship between the highest intake levels of menaquinone (mainly MK-4 from eggs and meat, and MK-8 and MK-9 from cheese) and cardiovascular disease and all-cause mortality in older men and women [8]
Vitamin K has been promoted in supplement form with claims it can slow tumor growth; there is however no good medical evidence that supports such claims [9]
Coumarin poisoning[edit]
Vitamin K is part of the suggested treatment regime for poisoning by rodenticide (coumarin poisoning) [10]
Although allergic reaction from supplementation is possible, no known toxicity is associated with high doses of the phylloquinone (vitamin K1) or menaquinone (vitamin K2) forms of vitamin K, so no tolerable upper intake level (UL) has been set [11]
Blood clotting (coagulation) studies in humans using 45 mg per day of vitamin K2 (as MK-4)[12] and even up to 135 mg per day (45 mg three times daily) of K2 (as MK-4),[13] showed no increase in blood clot risk  Even doses in rats as high as 250 mg/kg, body weight did not alter the tendency for blood-clot formation to occur [14]
Unlike the safe natural forms of vitamin K1 and vitamin K2 and their various isomers, a synthetic form of vitamin K, vitamin K3 (menadione), is demonstrably toxic at high levels  The U S  FDA has banned this form from over-the-counter sale in the United States because large doses have been shown to cause allergic reactions, hemolytic anemia, and cytotoxicity in liver cells [2]
Phylloquinone (K1)[15][16] or menaquinone (K2) are capable of reversing the anticoagulant activity of the anticoagulant warfarin (tradename Coumadin)  Warfarin works by blocking recycling of vitamin K, so that the body and tissues have lower levels of active vitamin K, and thus a deficiency of vitamin K 
Supplemental vitamin K (for which oral dosing is often more active than injectable dosing in human adults) reverses the vitamin K deficiency caused by warfarin, and therefore reduces the intended anticoagulant action of warfarin and related drugs [17] Sometimes small amounts of vitamin K are given orally to patients taking warfarin so that the action of the drug is more predictable [17] The proper anticoagulant action of the drug is a function of vitamin K intake and drug dose, and due to differing absorption must be individualized for each patient [citation needed] The action of warfarin and vitamin K both require two to five days after dosing to have maximum effect, and neither warfarin or vitamin K shows much effect in the first 24 hours after they are given [18]
The newer anticoagulants dabigatran and rivaroxaban have different mechanisms of action that do not interact with vitamin K, and may be taken with supplemental vitamin K [19][20]
Vitamin K2 (menaquinone)  In menaquinone, the side chain is composed of a varying number of isoprenoid residues  The most common number of these residues is four, since animal enzymes normally produce menaquinone-4 from plant phylloquinone 
A sample of phytomenadione for injection, also called phylloquinone
The three synthetic forms of vitamin K are vitamins K3 (menadione), K4, and K5, which are used in many areas, including the pet food industry (vitamin K3) and to inhibit fungal growth (vitamin K5) [21]
Conversion of vitamin K1 to vitamin K2[edit]
Vitamin K1 (phylloquinone) – both forms of the vitamin contain a functional naphthoquinone ring and an aliphatic side chain  Phylloquinone has a phytyl side chain 
The MK-4 form of vitamin K2 is produced by conversion of vitamin K1 in the testes, pancreas, and arterial walls [22] While major questions still surround the biochemical pathway for this transformation, the conversion is not dependent on gut bacteria, as it occurs in germ-free rats[23][24] and in parenterally-administered K1 in rats [25][26] In fact, tissues that accumulate high amounts of MK-4 have a remarkable capacity to convert up to 90% of the available K1 into MK-4 [27][28] There is evidence that the conversion proceeds by removal of the phytyl tail of K1 to produce menadione as an intermediate, which is then condensed with an activated geranylgeranyl moiety (see also prenylation) to produce vitamin K2 in the MK-4 (menatetrione) form [29]
Vitamin K2[edit]
Main article: Vitamin K2
Vitamin K2 (menaquinone) includes several subtypes  The two subtypes most studied are menaquinone-4 (menatetrenone, MK-4) and menaquinone-7 (MK-7) 
Vitamin K1, the precursor of most vitamin K in nature, is a stereoisomer of phylloquinone, an important chemical in green plants, where it functions as an electron acceptor in photosystem I during photosynthesis  For this reason, vitamin K1 is found in large quantities in the photosynthetic tissues of plants (green leaves, and dark green leafy vegetables such as romaine lettuce, kale and spinach), but it occurs in far smaller quantities in other plant tissues (roots, fruits, etc )  Iceberg lettuce contains relatively little  The function of phylloquinone in plants appears to have no resemblance to its later metabolic and biochemical function (as "vitamin K") in animals, where it performs a completely different biochemical reaction 
Vitamin K (in animals) is involved in the carboxylation of certain glutamate residues in proteins to form gamma-carboxyglutamate (Gla) residues  The modified residues are often (but not always) situated within specific protein domains called Gla domains  Gla residues are usually involved in binding calcium, and are essential for the biological activity of all known Gla proteins [30]
At this time[update], 17 human proteins with Gla domains have been discovered, and they play key roles in the regulation of three physiological processes:
Blood coagulation: prothrombin (factor II), factors VII, IX, and X, and proteins C, S, and Z[31]
Bone metabolism: osteocalcin, also called bone Gla protein (BGP), matrix Gla protein (MGP),[32] periostin,[33] and the recently discovered Gla-rich protein (GRP) [34][35]
Vascular biology: growth arrest-specific protein 6 (Gas6)[36]
Unknown function: proline-rich γ-carboxyglutamyl proteins (PRGPs) 1 and 2, and transmembrane γ-carboxy glutamyl proteins (TMGs) 3 and 4 [37]
Like other lipid-soluble vitamins (A, D and E), vitamin K is stored in the fatty tissue of the human body 
Absorption and dietary need[edit]
Previous theory held that dietary deficiency is extremely rare unless the small intestine was heavily damaged, resulting in malabsorption of the molecule  Another at-risk group for deficiency were those subject to decreased production of K2 by normal intestinal microbiota, as seen in broad spectrum antibiotic use [38] Taking broad-spectrum antibiotics can reduce vitamin K production in the gut by nearly 74% in people compared with those not taking these antibiotics [39] Diets low in vitamin K also decrease the body's vitamin K concentration [40] Those with chronic kidney disease are at risk for vitamin K deficiency, as well as vitamin D deficiency, and particularly those with the apoE4 genotype [41] Additionally, in the elderly there is a reduction in vitamin K2 production [42]
The National Academy of Medicine (NAM) updated an estimate of what constitutes an adequate intake (AI) for vitamin K in 2001  The NAM does not distinguish between K1 and K2 – both are counted as vitamin K  At that time there was not sufficient evidence to set the more rigorous estimated average requirement (EAR) or recommended dietary allowance (RDA) given for most of the essential vitamins and minerals  The current daily AIs for vitamin K for adult women and men are 90 μg and 120 μg respectively  The AI for pregnancy and lactation is 90 μg  For infants up to 12 months the AI is 2–2 5 μg, and for children aged 1 to 18 years the AI increases with age from 30 to 75 μg  As for safety, the FNB also sets tolerable upper intake levels (known as ULs) for vitamins and minerals when evidence is sufficient  In the case of vitamin K no UL is set, as evidence for adverse effects is not sufficient  Collectively EARs, RDAs, AIs and ULs are referred to as dietary reference intakes [43] The European Food Safety Authority reviewed the same safety question and did not set an UL [44]
For U S  food and dietary supplement labeling purposes, the amount in a serving is expressed as a percentage of daily value (%DV)  For vitamin K labeling purposes the daily value was 80 μg, but as of May 2016 it has been revised upwards to 120 μg  A table of the pre-change adult daily values is provided at reference daily intake  Food and supplement companies have until 28 July 2018 to comply with the change 
See also: Vitamin K2 § Dietary sources
K1 (μg)[45]
Kale, cooked
Collards, cooked
Collards, raw
Swiss chard, cooked
Swiss chard, raw
Turnip greens, raw
Romaine lettuce, raw
Table from "Important information to know when you are taking: Warfarin (Coumadin) and Vitamin K", Clinical Center, National Institutes of Health Drug Nutrient Interaction Task Force [46]
Vitamin K1 is found chiefly in leafy green vegetables such as dandelion greens (which contain 778 4 μg per 100 g, or 741% of the recommended daily amount), spinach, swiss chard, lettuce and Brassica vegetables (such as cabbage, kale, cauliflower, broccoli, and brussels sprouts) and often the absorption is greater when accompanied by fats such as butter or oils; some fruits, such as avocados, kiwifruit and grapes, are also high in vitamin K  By way of reference, two tablespoons of parsley contains 153% of the recommended daily amount of vitamin K [47] Some vegetable oils, notably soybean oil, contain vitamin K, but at levels that would require relatively large calorie consumption to meet the USDA-recommended levels [48] colonic bacteria synthesize a significant portion of humans' vitamin K needs; newborns often receive a vitamin K shot at birth to tide them over until their colons become colonized at five to seven days of age from the consumption of breast milk 
The tight binding of vitamin K1 to thylakoid membranes in chloroplasts makes it less bioavailable  For example, cooked spinach has a 5% bioavailability of phylloquinone, however, fat added to it increases bioavailability to 13% due to the increased solubility of vitamin K in fat [49]
Main article: Vitamin K deficiency
Average diets are usually not lacking in vitamin K, and primary deficiency is rare in healthy adults  Newborn infants are at an increased risk of deficiency  Other populations with an increased prevalence of vitamin K deficiency include those who suffer from liver damage or disease (e g  alcoholics), cystic fibrosis, or inflammatory bowel diseases, or have recently had abdominal surgeries  Secondary vitamin K deficiency can occur in people with bulimia, those on stringent diets, and those taking anticoagulants  Other drugs associated with vitamin K deficiency include salicylates, barbiturates, and cefamandole, although the mechanisms are still unknown  Vitamin K1 deficiency can result in coagulopathy, a bleeding disorder [50]Symptoms of K1 deficiency include anemia, bruising, nosebleeds and bleeding of the gums in both sexes, and heavy menstrual bleeding in women 
Osteoporosis[51][52] and coronary heart disease[53][54] are strongly associated with lower levels of K2 (menaquinone)  Vitamin K2 (as menaquinones MK-4 through MK-10) intake level is inversely related to severe aortic calcification and all-cause mortality [8]
Function in animals[edit]
Mechanism of action of vitamin K1 
The function of vitamin K2 in the animal cell is to add a carboxylic acid functional group to a glutamate (Glu) amino acid residue in a protein, to form a gamma-carboxyglutamate (Gla) residue  This is a somewhat uncommon posttranslational modification of the protein, which is then known as a "Gla protein"  The presence of two −COOH (carboxylic acid) groups on the same carbon in the gamma-carboxyglutamate residue allows it to chelate calcium ions  The binding of calcium ions in this way very often triggers the function or binding of Gla-protein enzymes, such as the so-called vitamin K-dependent clotting factors discussed below 
Within the cell, vitamin K undergoes electron reduction to a reduced form called vitamin K hydroquinone, catalyzed by the enzyme vitamin K epoxide reductase (VKOR) [55] Another enzyme then oxidizes vitamin K hydroquinone to allow carboxylation of Glu to Gla; this enzyme is called gamma-glutamyl carboxylase[56][57] or the vitamin K-dependent carboxylase  The carboxylation reaction only proceeds if the carboxylase enzyme is able to oxidize vitamin K hydroquinone to vitamin K epoxide at the same time  The carboxylation and epoxidation reactions are said to be coupled  Vitamin K epoxide is then reconverted to vitamin K by VKOR  The reduction and subsequent reoxidation of vitamin K coupled with carboxylation of Glu is called the vitamin K cycle [58] Humans are rarely deficient in vitamin K1 because, in part, vitamin K1 is continuously recycled in cells [59]
Warfarin and other 4-hydroxycoumarins block the action of VKOR [60] This results in decreased concentrations of vitamin K and vitamin K hydroquinone in tissues, such that the carboxylation reaction catalyzed by the glutamyl carboxylase is inefficient  This results in the production of clotting factors with inadequate Gla  Without Gla on the amino termini of these factors, they no longer bind stably to the blood vessel endothelium and cannot activate clotting to allow formation of a clot during tissue injury  As it is impossible to predict what dose of warfarin will give the desired degree of clotting suppression, warfarin treatment must be carefully monitored to avoid overdose 
Gamma-carboxyglutamate proteins[edit]
Main article: Gla domain
The following human Gla-containing proteins ("Gla proteins") have been characterized to the level of primary structure: blood coagulation factors II (prothrombin), VII, IX, and X, anticoagulant proteins C and S, and the factor X-targeting protein Z  The bone Gla protein osteocalcin, the calcification-inhibiting matrix Gla protein (MGP), the cell growth regulating growth arrest specific gene 6 protein (Gas6), and the four transmembrane Gla proteins (TMGPs), the function of which is at present unknown  Gas6 can function as a growth factor to activate the Axl receptor tyrosine kinase and stimulate cell proliferation or prevent apoptosis in some cells  In all cases in which their function was known, the presence of the Gla residues in these proteins turned out to be essential for functional activity 
Gla proteins are known to occur in a wide variety of vertebrates: mammals, birds, reptiles, and fish  The venom of a number of Australian snakes acts by activating the human blood-clotting system  In some cases, activation is accomplished by snake Gla-containing enzymes that bind to the endothelium of human blood vessels and catalyze the conversion of procoagulant clotting factors into activated ones, leading to unwanted and potentially deadly clotting 
Another interesting class of invertebrate Gla-containing proteins is synthesized by the fish-hunting snail Conus geographus [61] These snails produce a venom containing hundreds of neuroactive peptides, or conotoxins, which is sufficiently toxic to kill an adult human  Several of the conotoxins contain two to five Gla residues [62]
Methods of assessment[edit]
Vitamin K status can be assessed by:
The prothrombin time (PT) test measures the time required for blood to clot  A blood sample is mixed with citric acid and put in a fibrometer; delayed clot formation indicates a deficiency  This test is insensitive to mild deficiency, as the values do not change until the concentration of prothrombin in the blood has declined by at least 50% [63]
Undercarboxylated prothrombin (PIVKA-II); in a study of 53 newborns, found "PT (prothrombin time) is a less sensitive marker than PIVKA II",[64] and as indicated above, PT is unable to detect subclinical deficiencies that can be detected with PIVKA-II testing 
Plasma phylloquinone was found to be positively correlated with phylloquinone intake in elderly British women, but not men,[65] but an article by Schurgers et al  reported no correlation between FFQ[further explanation needed] and plasma phylloquinone [66]
Urinary γ-carboxyglutamic acid responds to changes in dietary vitamin K intake  Several days are required before any change can be observed  In a study by Booth et al , increases of phylloquinone intakes from 100 μg to between 377 and 417 μg for five days did not induce a significant change  Response may be age-specific [67]
Undercarboxylated osteocalcin (UcOc) levels have been inversely correlated with stores of vitamin K[68] and bone strength in developing rat tibiae  Another study following 78 post-menopausal Korean women found a supplement regimen of vitamins K and D, and calcium, but not a regimen of vitamin D and calcium, was inversely correlated with reduced UcOc levels [69]
Function in bacteria[edit]
Many bacteria, such as Escherichia coli found in the large intestine, can synthesize vitamin K2 (menaquinone-7 or MK-7, up to MK-11),[70] but not vitamin K1 (phylloquinone)  In these bacteria, menaquinone transfers two electrons between two different small molecules, during oxygen-independent metabolic energy production processes (anaerobic respiration) [71] For example, a small molecule with an excess of electrons (also called an electron donor) such as lactate, formate, or NADH, with the help of an enzyme, passes two electrons to menaquinone  The menaquinone, with the help of another enzyme, then transfers these two electrons to a suitable oxidant, such fumarate or nitrate (also called an electron acceptor)  Adding two electrons to fumarate or nitrate converts the molecule to succinate or nitrite plus water, respectively 
Some of these reactions generate a cellular energy source, ATP, in a manner similar to eukaryotic cell aerobic respiration, except the final electron acceptor is not molecular oxygen, but fumarate or nitrate  In aerobic respiration, the final oxidant is molecular oxygen (O2), which accepts four electrons from an electron donor such as NADH to be converted to water  E  coli, as facultative anaerobes, can carry out both aerobic respiration and menaquinone-mediated anaerobic respiration 
Injection in newborns[edit]
The blood clotting factors of newborn babies are roughly 30–60% that of adult values; this may be due to the reduced synthesis of precursor proteins and the sterility of their guts  Human milk contains 1–4 μg/L of vitamin K1, while formula-derived milk can contain up to 100 μg/L in supplemented formulas  Vitamin K2 concentrations in human milk appear to be much lower than those of vitamin K1  Occurrence of vitamin K deficiency bleeding in the first week of the infant's life is estimated at 0 25–1 7%, with a prevalence of 2–10 cases per 100,000 births [72] Premature babies have even lower levels of the vitamin, so they are at a higher risk from this deficiency 
Bleeding in infants due to vitamin K deficiency can be severe, leading to hospitalization, blood transfusions, brain damage, and death  Supplementation can prevent most cases of vitamin K deficiency bleeding in the newborn  Intramuscular administration is more effective in preventing late vitamin K deficiency bleeding than oral administration [73][74]
As a result of the occurrences of vitamin K deficiency bleeding, the Committee on Nutrition of the American Academy of Pediatrics has recommended 0 5–1 mg of vitamin K1 be administered to all newborns shortly after birth [74]
In the UK vitamin K supplementation is recommended for all newborns within the first 24 hours [75] This is usually given as a single intramuscular injection of 1 mg shortly after birth but as a second-line option can be given by three oral doses over the first month [76]
Controversy arose in the early 1990s regarding this practice, when two studies suggested a relationship between parenteral administration of vitamin K and childhood cancer,[77] however, poor methods and small sample sizes led to the discrediting of these studies, and a review of the evidence published in 2000 by Ross and Davies found no link between the two [78] Doctors reported emerging concerns in 2013,[79] after treating children for serious bleeding problems  They cited lack-of newborn vitamin K administration, as the reason that the problems occurred, and recommended that breastfed babies could have an increased risk unless they receive a preventative dose 
In the early 1930s, Danish scientist Henrik Dam investigated the role of cholesterol by feeding chickens a cholesterol-depleted diet [80] He initially replicated experiments reported by scientists at the Ontario Agricultural College (OAC) [81] McFarlane, Graham and Richardson, working on the chick feed program at OAC, had used chloroform to remove all fat from chick chow  They noticed that chicks fed only fat-depleted chow developed hemorrhages and started bleeding from tag sites [82] Dam found that these defects could not be restored by adding purified cholesterol to the diet  It appeared that – together with the cholesterol – a second compound had been extracted from the food, and this compound was called the coagulation vitamin  The new vitamin received the letter K because the initial discoveries were reported in a German journal, in which it was designated as Koagulationsvitamin  Edward Adelbert Doisy of Saint Louis University did much of the research that led to the discovery of the structure and chemical nature of vitamin K [83] Dam and Doisy shared the 1943 Nobel Prize for medicine for their work on vitamin K (K1 and K2) published in 1939  Several laboratories synthesized the compound(s) in 1939 [84]
For several decades, the vitamin K-deficient chick model was the only method of quantifying vitamin K in various foods: the chicks were made vitamin K-deficient and subsequently fed with known amounts of vitamin K-containing food  The extent to which blood coagulation was restored by the diet was taken as a measure for its vitamin K content  Three groups of physicians independently found this: Biochemical Institute, University of Copenhagen (Dam and Johannes Glavind), University of Iowa Department of Pathology (Emory Warner, Kenneth Brinkhous, and Harry Pratt Smith), and the Mayo Clinic (Hugh Butt, Albert Snell, and Arnold Osterberg) [85]
The first published report of successful treatment with vitamin K of life-threatening hemorrhage in a jaundiced patient with prothrombin deficiency was made in 1938 by Smith, Warner, and Brinkhous [86]
The precise function of vitamin K was not discovered until 1974, when three laboratories (Stenflo et al ,[87] Nelsestuen et al ,[88] and Magnusson et al [89]) isolated the vitamin K-dependent coagulation factor prothrombin (factor II) from cows that received a high dose of a vitamin K antagonist, warfarin  It was shown that, while warfarin-treated cows had a form of prothrombin that contained 10 glutamate (Glu) amino acid residues near the amino terminus of this protein, the normal (untreated) cows contained 10 unusual residues that were chemically identified as γ-carboxyglutamate (Gla)  The extra carboxyl group in Gla made clear that vitamin K plays a role in a carboxylation reaction during which Glu is converted into Gla 
The biochemistry of how vitamin K is used to convert Glu to Gla has been elucidated over the past thirty years in academic laboratories throughout the world 
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^ Whitlon, D  S ; Sadowski, J  A ; Suttie, J  W  (Apr 1978)  "Mechanism of coumarin action: significance of vitamin K epoxide reductase inhibition"  Biochemistry  17 (8): 1371–1377  doi:10 1021/bi00601a003  PMID 646989  ^ Terlau, H ; Olivera, B  M  (Jan 2004)  "Conus venoms: a rich source of novel ion channel-targeted peptides"  Physiological Reviews  84 (1): 41–68  doi:10 1152/physrev 00020 2003  PMID 14715910  ^ Buczek, O ; Bulaj, G ; Olivera, BM (Dec 2005)  "Conotoxins and the posttranslational modification of secreted gene products"  Cellular and Molecular Life Sciences  62 (24): 3067–3079  doi:10 1007/s00018-005-5283-0  PMID 16314929  ^ "Prothrombin Time"  WebMD  ^ Dituri, F ; Buonocore, G ; Pietravalle, A ; Naddeo, F ; Cortesi, M; Pasqualetti, P; Tataranno M  L ; R , Agostino (Sep 2012)  "PIVKA-II plasma levels as markers of subclinical vitamin K deficiency in term infants"  Journal of Maternal, Fetal & Neonatal Medicine  25 (9): 1660–1663  doi:10 3109/14767058 2012 657273  PMID 22280352  ^ Thane, C  W ; 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Rhéaume-Bleue, Kate (2012)  Vitamin K2 and the Calcium Paradox  John Wiley & Sons, Canada  ISBN 1-118-06572-7  External links[edit]
"Vitamin K: Another Reason to Eat Your Greens"  v
TPP / ThDP (B1)
FMN, FAD (B2)
NAD+, NADH, NADP+, NADPH (B3)
Coenzyme A (B5)
PLP / P5P (B6)
THFA / H4FA, DHFA / H2FA, MTHF (B9)
AdoCbl, MeCbl (B12)
Phylloquinone (K1), Menaquinone (K2)
non-vitamins
Coenzyme B
Heme / Haem (A, B, C, O)
Molybdopterin/Molybdenum cofactor
THMPT / H4MPT
Fe2+, Fe3+
vitamins: see vitamins
Antihemorrhagics (B02)
(coagulation)
Phytomenadione (K1)
Menadione (K3)
intrinsic: IX/Nonacog alfa
VIII/Moroctocog alfa/Turoctocog alfa
extrinsic: VII/Eptacog alfa
common: X
II/Thrombin
I/Fibrinogen
XIII/Catridecacog
combinations: Prothrombin complex concentrate (II, VII, IX, X, protein C and S)
Carbazochrome
thrombopoietin receptor agonist (Romiplostim
Eltrombopag)
Tetragalacturonic acid hydroxymethylester
Epinephrine/Adrenalone
amino acids (Aminocaproic acid
Aminomethylbenzoic acid)
serpins (Aprotinin
Alfa1 antitrypsin
Camostat) 



ground_truth: 90 μg for women and 120 μg for men.

prompt_front: Now, answer the following question based on the text below, only give me the answer and do not output any other words.

Question: How is the vacuum processing system configured in terms of the arrangement of the vacuum processing apparatus?

Read the following text and answer briefly.

Context 4:  Institutions: University of Mons Long-term potentiation (LTP) is a type of synaptic plasticity characterized by an increase in synaptic strength and believed to be involved in memory encoding  LTP elicited in the CA1 region of acute hippocampal slices has been extensively studied  However the molecular mechanisms underlying the maintenance phase of this phenomenon are still poorly understood  This could be partly due to the various experimental conditions used by different laboratories  Indeed, the maintenance phase of LTP is strongly dependent on external parameters like oxygenation, temperature and humidity  It is also dependent on internal parameters like orientation of the slicing plane and slice viability after dissection 
The optimization of all these parameters enables the induction of a very reproducible and very stable long-term potentiation  This methodology offers the possibility to further explore the molecular mechanisms involved in the stable increase in synaptic strength in hippocampal slices  It also highlights the importance of experimental conditions in in vitro investigation of neurophysiological phenomena Neuroscience, Issue 76, Neurobiology, Anatomy, Physiology, Biomedical Engineering, Surgery, Memory Disorders, Learning, Memory, Neurosciences, Neurophysiology, hippocampus, long-term potentiation, mice, acute slices, synaptic plasticity, in vitro, electrophysiology, animal model50483Play ButtonIn Vivo Modeling of the Morbid Human Genome using Danio rerioAuthors: Adrienne R  Niederriter, Erica E  Davis, Christelle Golzio, Edwin C  Oh, I-Chun Tsai, Nicholas Katsanis  Institutions: Duke University Medical Center, Duke University, Duke University Medical Center Here, we present methods for the development of assays to query potentially clinically significant nonsynonymous changes using in vivo complementation in zebrafish  Zebrafish (Danio rerio) are a useful animal system due to their experimental tractability; embryos are transparent to enable facile viewing, undergo rapid development ex vivo, and can be genetically manipulated 1 These aspects have allowed for significant advances in the analysis of embryogenesis, molecular processes, and morphogenetic signaling  Taken together, the advantages of this vertebrate model make zebrafish highly amenable to modeling the developmental defects in pediatric disease, and in some cases, adult-onset disorders  Because the zebrafish genome is highly conserved with that of humans (~70% orthologous), it is possible to recapitulate human disease states in zebrafish  This is accomplished either through the injection of mutant human mRNA to induce dominant negative or gain of function alleles, or utilization of morpholino (MO) antisense oligonucleotides to suppress genes to mimic loss of function variants  Through complementation of MO-induced phenotypes with capped human mRNA, our approach enables the interpretation of the deleterious effect of mutations on human protein sequence based on the ability of mutant mRNA to rescue a measurable, physiologically relevant phenotype  Modeling of the human disease alleles occurs through microinjection of zebrafish embryos with MO and/or human mRNA at the 1-4 cell stage, and phenotyping up to seven days post fertilization (dpf)  This general strategy can be extended to a wide range of disease phenotypes, as demonstrated in the following protocol  We present our established models for morphogenetic signaling, craniofacial, cardiac, vascular integrity, renal function, and skeletal muscle disorder phenotypes, as well as others  Molecular Biology, Issue 78, Genetics, Biomedical Engineering, Medicine, Developmental Biology, Biochemistry, Anatomy, Physiology, Bioengineering, Genomics, Medical, zebrafish, in vivo, morpholino, human disease modeling, transcription, PCR, mRNA, DNA, Danio rerio, animal model50338Play ButtonDirect Imaging of ER Calcium with Targeted-Esterase Induced Dye Loading (TED)Authors: Samira Samtleben, Juliane Jaepel, Caroline Fecher, Thomas Andreska, Markus Rehberg, Robert Blum  Institutions: University of Wuerzburg, Max Planck Institute of Neurobiology, Martinsried, Ludwig-Maximilians University of Munich Visualization of calcium dynamics is important to understand the role of calcium in cell physiology  To examine calcium dynamics, synthetic fluorescent Ca2+ indictors have become popular  Here we demonstrate TED (= targeted-esterase induced dye loading), a method to improve the release of Ca2+ indicator dyes in the ER lumen of different cell types  To date, TED was used in cell lines, glial cells, and neurons in vitro  TED bases on efficient, recombinant targeting of a high carboxylesterase activity to the ER lumen using vector-constructs that express Carboxylesterases (CES)  The latest TED vectors contain a core element of CES2 fused to a red fluorescent protein, thus enabling simultaneous two-color imaging  The dynamics of free calcium in the ER are imaged in one color, while the corresponding ER structure appears in red  At the beginning of the procedure, cells are transduced with a lentivirus  Subsequently, the infected cells are seeded on coverslips to finally enable live cell imaging  Then, living cells are incubated with the acetoxymethyl ester (AM-ester) form of low-affinity Ca2+ indicators, for instance Fluo5N-AM, Mag-Fluo4-AM, or Mag-Fura2-AM  The esterase activity in the ER cleaves off hydrophobic side chains from the AM form of the Ca2+ indicator and a hydrophilic fluorescent dye/Ca2+ complex is formed and trapped in the ER lumen  After dye loading, the cells are analyzed at an inverted confocal laser scanning microscope  Cells are continuously perfused with Ringer-like solutions and the ER calcium dynamics are directly visualized by time-lapse imaging  Calcium release from the ER is identified by a decrease in fluorescence intensity in regions of interest, whereas the refilling of the ER calcium store produces an increase in fluorescence intensity  Finally, the change in fluorescent intensity over time is determined by calculation of ΔF/F0 Cellular Biology, Issue 75, Neurobiology, Neuroscience, Molecular Biology, Biochemistry, Biomedical Engineering, Bioengineering, Virology, Medicine, Anatomy, Physiology, Surgery, Endoplasmic Reticulum, ER, Calcium Signaling, calcium store, calcium imaging, calcium indicator, metabotropic signaling, Ca2+, neurons, cells, mouse, animal model, cell culture, targeted esterase induced dye loading, imaging50317Play ButtonPreparation of Dissociated Mouse Cortical Neuron CulturesAuthors: Lutz G  W  Hilgenberg, Martin A  Smith  Institutions: University of California, Irvine (UCI) This video will guide you through the process for generating cortical neuronal cultures from late embryo and early postnatal mouse brain  These cultures can be used for a variety of applications including immunocytochemistry, biochemistry, electrophysiology, calcium and sodium imaging, protein and/or RNA isolation  These cultures also provide a platform to study the neuronal development of transgenic animals that carry a late embryonic or postnatal lethal gene mutation  The procedure is relatively straight forward, requires some experience in tissue culture technique and should not take longer than two to three hours if you are properly prepared  Careful separation of the cortical rind from the thalamo-cortical fiber tract will reduce the number of unwanted non-neuronal cells  To increase yields of neuronal cells triturate the pieces of the cortical tissue gently after the enzyme incubation step  This is imperative as it prevents unnecessary injury to cells and premature neuronal cell death  Since these cultures are maintained in the absence of glia feeder cells, they also offer an added advantage of growing cultures enriched in neurons Neuroscience, Issue 10, cellular, molecular, neurobiology, neuron, calcium/sodium imaging, primary cultures, mouse562Play ButtonAnalysis of Schwann-astrocyte Interactions Using In Vitro AssaysAuthors: Fardad T  Afshari, Jessica C  Kwok, James W  Fawcett  Institutions: University of Cambridge Schwann cells are one of the commonly used cells in repair strategies following spinal cord injuries  Schwann cells are capable of supporting axonal regeneration and sprouting by secreting growth factors 1,2 and providing growth promoting adhesion molecules 3 and extracellular matrix molecules 4  In addition they myelinate the demyelinated axons at the site of injury 5 
However following transplantation, Schwann cells do not migrate from the site of implant and do not intermingle with the host astrocytes 6,7  This results in formation of a sharp boundary between the Schwann cells and astrocytes, creating an obstacle for growing axons trying to exit the graft back into the host tissue proximally and distally  Astrocytes in contact with Schwann cells also undergo hypertrophy and up-regulate the inhibitory molecules 8-13 
In vitro assays have been used to model Schwann cell-astrocyte interactions and have been important in understanding the mechanism underlying the cellular behaviour 
These in vitro assays include boundary assay, where a co-culture is made using two different cells with each cell type occupying different territories with only a small gap separating the two cell fronts  As the cells divide and migrate, the two cellular fronts get closer to each other and finally collide  This allows the behaviour of the two cellular populations to be analyzed at the boundary  Another variation of the same technique is to mix the two cellular populations in culture and over time the two cell types segregate with Schwann cells clumped together as islands in between astrocytes together creating multiple Schwann-astrocyte boundaries 
The second assay used in studying the interaction of two cell types is the migration assay where cellular movement can be tracked on the surface of the other cell type monolayer 14,15  This assay is commonly known as inverted coverslip assay  Schwann cells are cultured on small glass fragments and they are inverted face down onto the surface of astrocyte monolayers and migration is assessed from the edge of coverslip 
Both assays have been instrumental in studying the underlying mechanisms involved in the cellular exclusion and boundary formation  Some of the molecules identified using these techniques include N-Cadherins 15, Chondroitin Sulphate proteoglycans(CSPGs) 16,17, FGF/Heparin 18, Eph/Ephrins19 
This article intends to describe boundary assay and migration assay in stepwise fashion and elucidate the possible technical problems that might occur Cellular Biology, Issue 47, Schwann cell, astrocyte, boundary, migration, repulsion2214Play ButtonQuantifying Synapses: an Immunocytochemistry-based Assay to Quantify Synapse NumberAuthors: Dominic M  Ippolito, Cagla Eroglu  Institutions: Duke University, Duke University One of the most important goals in neuroscience is to understand the molecular cues that instruct early stages of synapse formation  As such it has become imperative to develop objective approaches to quantify changes in synaptic connectivity  Starting from sample fixation, this protocol details how to quantify synapse number both in dissociated neuronal culture and in brain sections using immunocytochemistry  Using compartment-specific antibodies, we label presynaptic terminals as well as sites of postsynaptic specialization  We define synapses as points of colocalization between the signals generated by these markers  The number of these colocalizations is quantified using a plug in Puncta Analyzer (written by Bary Wark, available upon request, c eroglu@cellbio duke edu) under the ImageJ analysis software platform  The synapse assay described in this protocol can be applied to any neural tissue or culture preparation for which you have selective pre- and postsynaptic markers  This synapse assay is a valuable tool that can be widely utilized in the study of synaptic development Neuroscience, Issue 45, synapse, immunocytochemistry, brain, neuron, astrocyte2270Play ButtonPreparation of Acute Hippocampal Slices from Rats and Transgenic Mice for the Study of Synaptic Alterations during Aging and Amyloid PathologyAuthors: Diana M  Mathis, Jennifer L  Furman, Christopher M  Norris  Institutions: University of Kentucky College of Public Health, University of Kentucky College of Medicine, University of Kentucky College of Medicine The rodent hippocampal slice preparation is perhaps the most broadly used tool for investigating mammalian synaptic function and plasticity  The hippocampus can be extracted quickly and easily from rats and mice and slices remain viable for hours in oxygenated artificial cerebrospinal fluid  Moreover, basic electrophysisologic techniques are easily applied to the investigation of synaptic function in hippocampal slices and have provided some of the best biomarkers for cognitive impairments  The hippocampal slice is especially popular for the study of synaptic plasticity mechanisms involved in learning and memory  Changes in the induction of long-term potentiation and depression (LTP and LTD) of synaptic efficacy in hippocampal slices (or lack thereof) are frequently used to describe the neurologic phenotype of cognitively-impaired animals and/or to evaluate the mechanism of action of nootropic compounds  This article outlines the procedures we use for preparing hippocampal slices from rats and transgenic mice for the study of synaptic alterations associated with brain aging and Alzheimer's disease (AD)1-3  Use of aged rats and AD model mice can present a unique set of challenges to researchers accustomed to using younger rats and/or mice in their research  Aged rats have thicker skulls and tougher connective tissue than younger rats and mice, which can delay brain extraction and/or dissection and consequently negate or exaggerate real age-differences in synaptic function and plasticity  Aging and amyloid pathology may also exacerbate hippocampal damage sustained during the dissection procedure, again complicating any inferences drawn from physiologic assessment  Here, we discuss the steps taken during the dissection procedure to minimize these problems  Examples of synaptic responses acquired in "healthy" and "unhealthy" slices from rats and mice are provided, as well as representative synaptic plasticity experiments  The possible impact of other methodological factors on synaptic function in these animal models (e g  recording solution components, stimulation parameters) are also discussed  While the focus of this article is on the use of aged rats and transgenic mice, novices to slice physiology should find enough detail here to get started on their own studies, using a variety of rodent models Neuroscience, Issue 49, aging, amyloid, hippocampal slice, synaptic plasticity, Ca2+, CA1, electrophysiology2330Play ButtonMesenteric Artery Contraction and Relaxation Studies Using Automated Wire MyographyAuthors: Lakeesha E  Bridges, Cicely L  Williams, Mildred A  Pointer, Emmanuel M  Awumey  Institutions: North Carolina Central University, Durham, North Carolina Central University, Durham, Wake Forest University School of Medicine Proximal resistance vessels, such as the mesenteric arteries, contribute substantially to the peripheral resistance  These small vessels of between 100-400 μm in diameter function primarily in directing blood flow to various organs according to the overall requirements of the body  The rat mesenteric artery has a diameter greater than 100 μm  The myography technique, first described by Mulvay and Halpern1, was based on the method proposed by Bevan and Osher2  The technique provides information about small vessels under isometric conditions, where substantial shortening of the muscle preparation is prevented  Since force production and sensitivity of vessels to different agonists is dependent on the extent of stretch, according to active tension-length relation, it is essential to conduct contraction studies under isometric conditions to prevent compliance of the mounting wires  Stainless steel wires are preferred to tungsten wires because of oxidation of the latter, which affects recorded responses3 The technique allows for the comparison of agonist-induced contractions of mounted vessels to obtain evidence for normal function of vascular smooth muscle cell receptors 
Medicine, Issue 55, cardiovascular, resistant arteries, contraction, relaxation, myography3119Play ButtonVisualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero ElectroporationAuthors: Emilie Pacary, Matilda A  Haas, Hendrik Wildner, Roberta Azzarelli, Donald M  Bell, Djoher Nora Abrous, François Guillemot  Institutions: MRC National Institute for Medical Research, National Institute for Medical Research, Université de Bordeaux In utero electroporation (IUE) has become a powerful technique to study the development of different regions of the embryonic nervous system 1-5  To date this tool has been widely used to study the regulation of cellular proliferation, differentiation and neuronal migration especially in the developing cerebral cortex 6-8  Here we detail our protocol to electroporate in utero the cerebral cortex and the hippocampus and provide evidence that this approach can be used to study dendrites and spines in these two cerebral regions 
Finally, IUE provides a useful tool to identify functional interactions between genes involved in dendrite, spine and/or synapse development  Indeed, in contrast to other gene transfer methods such as virus, it is straightforward to combine multiple RNAi or transgenes in the same population of cells  In summary, IUE is a powerful method that has already contributed to the characterization of molecular mechanisms underlying brain function and disease and it should also be useful in the study of dendrites and spines Neuroscience, Issue 65, Developmental Biology, Molecular Biology, Neuronal development, In utero electroporation, dendrite, spines, hippocampus, cerebral cortex, gain and loss of function4163Play ButtonImaging Analysis of Neuron to Glia Interaction in Microfluidic Culture Platform (MCP)-based Neuronal Axon and Glia Co-culture SystemAuthors: Haruki Higashimori, Yongjie Yang  Institutions: Tufts University, Tufts Sackler School of Graduate Biomedical Sciences Proper neuron to glia interaction is critical to physiological function of the central nervous system (CNS)  This bidirectional communication is sophisticatedly mediated by specific signaling pathways between neuron and glia1,2   Identification and characterization of these signaling pathways is essential to the understanding of how neuron to glia interaction shapes CNS physiology  Previously, neuron and glia mixed cultures have been widely utilized for testing and characterizing signaling pathways between neuron and glia  What we have learned from these preparations and other in vivo tools, however, has suggested that mutual signaling between neuron and glia often occurred in specific compartments within neurons (i e , axon, dendrite, or soma)3  This makes it important to develop a new culture system that allows separation of neuronal compartments and specifically examines the interaction between glia and neuronal axons/dendrites  In addition, the conventional mixed culture system is not capable of differentiating the soluble factors and direct membrane contact signals between neuron and glia  Furthermore, the large quantity of neurons and glial cells in the conventional co-culture system lacks the resolution necessary to observe the interaction between a single axon and a glial cell 
In this study, we describe a novel axon and glia co-culture system with the use of a microfluidic culture platform (MCP)  In this co-culture system, neurons and glial cells are cultured in two separate chambers that are connected through multiple central channels  In this microfluidic culture platform, only neuronal processes (especially axons) can enter the glial side through the central channels  In combination with powerful fluorescent protein labeling, this system allows direct examination of signaling pathways between axonal/dendritic and glial interactions, such as axon-mediated transcriptional regulation in glia, glia-mediated receptor trafficking in neuronal terminals, and glia-mediated axon growth  The narrow diameter of the chamber also significantly prohibits the flow of the neuron-enriched medium into the glial chamber, facilitating probing of the direct membrane-protein interaction between axons/dendrites and glial surfaces Neuroscience, Issue 68, Molecular Biology, Cellular Biology, Biophysics, Microfluidics, Microfluidic culture platform, Compartmented culture, Neuron to glia signaling, neurons, glia, cell culture4448Play ButtonFluorescence Recovery After Photobleaching (FRAP) of Fluorescence Tagged Proteins in Dendritic Spines of Cultured Hippocampal NeuronsAuthors: Chan-Ying Zheng, Ronald S  Petralia, Ya-Xian Wang, Bechara Kachar  Institutions: National Institutes of Health, Bethesda FRAP has been used to quantify the mobility of GFP-tagged proteins  Using a strong excitation laser, the fluorescence of a GFP-tagged protein is bleached in the region of interest  The fluorescence of the region recovers when the unbleached GFP-tagged protein from outside of the region diffuses into the region of interest  The mobility of the protein is then analyzed by measuring the fluorescence recovery rate  This technique could be used to characterize protein mobility and turnover rate 
This FRAP protocol shows how to perform a basic FRAP experiment as well as how to analyze the data Neuroscience, Issue 50, Spine, FRAP, hippocampal neurons, live cell imaging, protein mobility2568Play ButtonPrimary Neuronal Cultures from the Brains of Late Stage Drosophila PupaeAuthors: Beatriz Sicaeros, Jorge M  Campusano, Diane K  O'Dowd  Institutions: University of California, Irvine (UCI) In this video, we demonstrate the preparation of primary neuronal cultures from the brains of late stage Drosophila pupae  The procedure begins with the removal of brains from animals at 70-78 hrs after puparium formation  The isolated brains are shown after brief incubation in papain followed by several washes in serum-free growth medium  The process of mechanical dissociation of each brain in a 5 ul drop of media on a coverslip is illustrated  The axons and dendrites of the post-mitotic neurons are sheered off near the soma during dissociation but the neurons begin to regenerate processes within a few hours of plating  Images show live cultures at 2 days  Neurons continue to elaborate processes during the first week in culture  Specific neuronal populations can be identified in culture using GAL4 lines to drive tissue specific expression of fluorescent markers such as GFP or RFP  Whole cell recordings have demonstrated the cultured neurons form functional, spontaneously active cholinergic and GABAergic synapses  A short video segment illustrates calcium dynamics in the cultured neurons using Fura-2 as a calcium indicator dye to monitor spontaneous calcium transients and nicotine evoked calcium responses in a dish of cultured neurons  These pupal brain cultures are a useful model system in which genetic and pharmacological tools can be used to identify intrinsic and extrinsic factors that influence formation and function of central synapses 



ground_truth: Multiple vacuum processing apparatuses are arranged in parallel.