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37320387 | Wilson's & North Eastern Railway Shipping Co. Ltd | https://en.wikipedia.org/w/index.php?title=Wilson's%20&%20North%20Eastern%20Railway%20Shipping%20Co.%20Ltd | Wilson's & North Eastern Railway Shipping Co. Ltd
Wilsons sought a solution to the Hull services problem and a joint venture was proposed. The negotiations were led by Charles Wilson (Lord Nunburnholme) and his brother Arthur Wilson who was also a Director of the N.E.R.
- 1906 – Joint company formed in March to operate services from Hull. In the same year Thomas Wilson Sons & Co. took control of local shipbuilders Earle's Shipbuilding.
- 1923 – January – The company came under control of the London and North Eastern Railway.
- 1935 – May – the company was merged with other interests to form the Associated Humber Lines.
- 1957 – Final two ships "Selby" and "Harrogate" (1925) were sold and the company ceased to trade.
# Routes.
Hull to | 6,131,600 |
37320387 | Wilson's & North Eastern Railway Shipping Co. Ltd | https://en.wikipedia.org/w/index.php?title=Wilson's%20&%20North%20Eastern%20Railway%20Shipping%20Co.%20Ltd | Wilson's & North Eastern Railway Shipping Co. Ltd
brother Arthur Wilson who was also a Director of the N.E.R.
- 1906 – Joint company formed in March to operate services from Hull. In the same year Thomas Wilson Sons & Co. took control of local shipbuilders Earle's Shipbuilding.
- 1923 – January – The company came under control of the London and North Eastern Railway.
- 1935 – May – the company was merged with other interests to form the Associated Humber Lines.
- 1957 – Final two ships "Selby" and "Harrogate" (1925) were sold and the company ceased to trade.
# Routes.
Hull to Hamburg ; Antwerp / Ghent ; and Dunkirk.
# Livery.
Funnel : Red with black top with thin white dividing line.
Hull : Wilson dark green with red boot topping. | 6,131,601 |
37320399 | Accommodation bridge | https://en.wikipedia.org/w/index.php?title=Accommodation%20bridge | Accommodation bridge
Accommodation bridge
An accommodation bridge or occupation bridge in the United Kingdom preserves a pre-existing private road, path or right of access when a major transport route is built across it. Without the bridge, access would be disrupted. Accommodation bridges are usually built at the cost of the route developer, as part of the conditions for obtaining the land for building the new route.
The term is not applied where the new route crosses an existing public highway.
# Canals.
The first accommodation bridges were built as part of 18th-century canal building. Most were provided for farmers, whose lands and grazing were separated by the canal. The first canals developed from rivers, | 6,131,602 |
37320399 | Accommodation bridge | https://en.wikipedia.org/w/index.php?title=Accommodation%20bridge | Accommodation bridge
with short lengths of canal built to bypass obstacles, such as weirs and millponds. The river represented a long-established and accepted boundary, but these new sections were resented by landlords.
Unlike turnpike roads, drovers could not simply cross the new road but now needed a bridge. These bridges also needed to meet the standards of the canal builder, allowing the towpath through beneath, sufficient clearance for passing boats and being adequately constructed to be robust, without risk of a collapse blocking the canal. To save costs, the Kennet Navigation, in flat country, used swing bridges rather than arches, Other canals such as the Oxford Canal, also used lifting bridges.
# Railways.
Most | 6,131,603 |
37320399 | Accommodation bridge | https://en.wikipedia.org/w/index.php?title=Accommodation%20bridge | Accommodation bridge
accommodation bridges, in the UK at least, were constructed during the railway building boom of the mid-19th century. British practice avoided level crossings wherever possible, except in the flat parts of the country where building a raised approach to a bridge would be more costly. Nevertheless the term is still more usually applied to canal bridges.
As the load carried by the new railway was almost always greater than the old path, nearly all accommodation bridges are overbridges, carrying the old track over the new railway. Underpasses were relatively rare, except where more convenient in hilly country, as the old track may have followed the land more closely than a gradient-sensitive railway.
In | 6,131,604 |
37320399 | Accommodation bridge | https://en.wikipedia.org/w/index.php?title=Accommodation%20bridge | Accommodation bridge
Britain an accommodation bridge is the name for a bridge connecting land that belonged to one owner severed by the railway. An occupation bridge is the name for a bridge that carried an existing private road or took the railway over an existing private road. Similar terms were applied to level crossings.
# Motorways.
Accommodation bridges were rarely needed across roads, as a road junction would be provided instead. This changed in the 1960s, with the development of the motorway network, where slow-speed local traffic is segregated from the high-speed traffic.
# See also.
- Cattle creep
# External links.
- Canal restoration project involving the construction of a new accommodation bridge | 6,131,605 |
37320399 | Accommodation bridge | https://en.wikipedia.org/w/index.php?title=Accommodation%20bridge | Accommodation bridge
e for a bridge connecting land that belonged to one owner severed by the railway. An occupation bridge is the name for a bridge that carried an existing private road or took the railway over an existing private road. Similar terms were applied to level crossings.
# Motorways.
Accommodation bridges were rarely needed across roads, as a road junction would be provided instead. This changed in the 1960s, with the development of the motorway network, where slow-speed local traffic is segregated from the high-speed traffic.
# See also.
- Cattle creep
# External links.
- Canal restoration project involving the construction of a new accommodation bridge to maintain a previous right of access. | 6,131,606 |
37320434 | Oh Look Out | https://en.wikipedia.org/w/index.php?title=Oh%20Look%20Out | Oh Look Out
Oh Look Out
Oh Look Out is an American rock band based in Austin, TX created by singer/songwriter JP Pfertner in 2011. To date, Oh Look Out has released two full-length albums, and is described as "Catchy indie rock/pop with waves of fuzzy guitars and Casio keyboards that hit your brain like an Atari blasting out of a jambox bazooka."
# History.
In 2011, following five years of writing and performing with the band Built By Snow lead singer and songwriter JP Pfertner started releasing home recordings with hand drawn art under the name "Oh Look Out." The first release in fall 2011, "Alright Alright Alright Alright Alright" is a collection of songs selected from recordings made in his bedroom/makeshift | 6,131,607 |
37320434 | Oh Look Out | https://en.wikipedia.org/w/index.php?title=Oh%20Look%20Out | Oh Look Out
studio. The result was a mix of energetic indie rock/pop songs, drawing on inspiration from The Cars, The Rentals, and DEVO, as well as melancholy keyboard pop songs that draw comparisons to the bedroom recording legend, Daniel Johnston.
The day after "Alright Alright Alright Alright Alright" was released, JP pulled out some small amps and guitars, an old drum kit, a cassette tape recorder, a battery powered Casiotone keyboard, and began recording a new album in his garage. These recordings became the second Oh Look Out album, titled "Orchestrated Fuzz." Released in Fall 2012, Orchestrated Fuzz is 9 songs that all connect together with waves of crunchy guitars, punchy keys, big drums and lyrics | 6,131,608 |
37320434 | Oh Look Out | https://en.wikipedia.org/w/index.php?title=Oh%20Look%20Out | Oh Look Out
ongs that all connect together with waves of crunchy guitars, punchy keys, big drums and lyrics about losing your mind, exploding, reliving better years, and wearing out everything that you love. Songs spin backwards, Mellotrons play alongside moogs, cassette tape samples blast through jambox speakers, and songs smash into one another keeping the album moving with no gaps from start to end.
# Discography.
- "Orchestrated Fuzz" (2012) self released
- "Alright Alright Alright Alright Alright" (2011) self released
# Side Projects.
JP Pfertner is also the lead singer/songwriter for the American indie rock band Built By Snow.
# External links.
- Oh Look Out (Official Site)
- Built By Snow | 6,131,609 |
37320432 | Hulk: Gray | https://en.wikipedia.org/w/index.php?title=Hulk:%20Gray | Hulk: Gray
Hulk: Gray
Hulk: Gray is a comic book limited series written by Jeph Loeb and illustrated by Tim Sale.
# Publication history.
The series ran for a total of six issues which followed the early years of Bruce Banner and his problems as the Hulk. Jeph Loeb and Tim Sale also collaborated on other limited series such as: "", "", and "". Each series, like "Hulk: Gray", followed the early years of the different Marvel Comics heroes.
# Plot.
Bruce Banner's life was torn apart by the explosion of the Gamma Bomb. From that moment on, he unleashed the strongest creature on Earth, The Incredible Hulk. No matter how powerful he became, his heart could still be shattered by Betty Ross, the daughter of | 6,131,610 |
37320432 | Hulk: Gray | https://en.wikipedia.org/w/index.php?title=Hulk:%20Gray | Hulk: Gray
# Publication history.
The series ran for a total of six issues which followed the early years of Bruce Banner and his problems as the Hulk. Jeph Loeb and Tim Sale also collaborated on other limited series such as: "", "", and "". Each series, like "Hulk: Gray", followed the early years of the different Marvel Comics heroes.
# Plot.
Bruce Banner's life was torn apart by the explosion of the Gamma Bomb. From that moment on, he unleashed the strongest creature on Earth, The Incredible Hulk. No matter how powerful he became, his heart could still be shattered by Betty Ross, the daughter of General "Thunderbolt" Ross.
# Collected editions.
The series has been collected into a single volume. | 6,131,611 |
37320441 | 2012 Lillestrøm SK season | https://en.wikipedia.org/w/index.php?title=2012%20Lillestrøm%20SK%20season | 2012 Lillestrøm SK season
2012 Lillestrøm SK season
The 2012 season saw Lillestrøm compete in the Tippeligaen as well as the 2012 Norwegian Football Cup. They finished the season in 9th in the Tippeligaen and they were knocked out of in the Fourth Round by Bodø/Glimt. It was the club's first season with Magnus Haglund as their manager.
# Squad.
## Transfers.
### Winter.
In:
Out:
### Summer.
In:
Out: | 6,131,612 |
37320379 | Börje Langefors Best Doctoral Dissertation Award | https://en.wikipedia.org/w/index.php?title=Börje%20Langefors%20Best%20Doctoral%20Dissertation%20Award | Börje Langefors Best Doctoral Dissertation Award
Börje Langefors Best Doctoral Dissertation Award
The Börje Langefors Award (‘Börje Langeforspriset’ in Swedish) is an academic prize awarded each year by the Swedish Information Systems Academy (Svenska informationssystemakademin or SISA) for the best doctoral dissertation in Sweden in the subject areas - informatics, information systems, data and information science or equivalent. The prize aims to reward and encourage development of high standard research in Sweden, and to demonstrate exemplary research in informatics.
# Origin.
The award has been named after Professor Börje Langefors (1915–2009), one of those who made systems development a science. professor Börje was a Swedish engineer | 6,131,613 |
37320379 | Börje Langefors Best Doctoral Dissertation Award | https://en.wikipedia.org/w/index.php?title=Börje%20Langefors%20Best%20Doctoral%20Dissertation%20Award | Börje Langefors Best Doctoral Dissertation Award
and computer scientist, and emeritus professor of business information systems at the Department of Computer and Systems Science, Stockholm University and Royal Institute of Technology, Stockholm. Börje Langefors was a pioneer of IT and one of the initiators of "informatics" as an academic area of study. He was the first IT professor in Sweden and one of the first in the world. Börje contributed strongly to put Sweden on the international IT map and brought into a focus in particular to the user's role in data processing. Börje Langefors brought more than 20 graduate students to degree most of which today are professors who in turn have brought their students to graduates.
# Award criteria.
The | 6,131,614 |
37320379 | Börje Langefors Best Doctoral Dissertation Award | https://en.wikipedia.org/w/index.php?title=Börje%20Langefors%20Best%20Doctoral%20Dissertation%20Award | Börje Langefors Best Doctoral Dissertation Award
following quality criteria are applied for the evaluation of individual doctoral thesis:
- Relevance: Articulate, well-defined and well-motivated research question(s)
- Articulate and well-reflected research design
- Comprehensiveness: Chosen and used well described theory base
- Well described empirical base
- Validity of knowledge (empirically and theoretically well-grounded)
- Contribution validity and durability (abstraction) to further research
- Innovative value in knowledge contributions
- Independence (of author's own contribution)
- Communicability: Clarity, transparency and conceptual clarity
- Internal coherence: holistic and coherent argument
- Subject (IS field) congruency
- | 6,131,615 |
37320379 | Börje Langefors Best Doctoral Dissertation Award | https://en.wikipedia.org/w/index.php?title=Börje%20Langefors%20Best%20Doctoral%20Dissertation%20Award | Börje Langefors Best Doctoral Dissertation Award
Ability to serve as a "Role model"
- International exposure /review
# Prize committee.
Every year in spring (usually in May), a prize committee assesses the theses submitted by the universities/institutions in Sweden and nominates the best dissertation, which is finally announced in connection with SISA's annual conference. Information about this conference can be found here.
The members of Committee for Börje Langefors Award in 2016 are:
- Professor Karin Hedström , Örebro University
- Professor Tero Päivärinta, Luleå University
- Professor Jan Ljungberg, University of Gothenburg
- Professor Jeremy Rose, University of Skövde
- Professor Vivian Vimarlund, Linköping University
Börje | 6,131,616 |
37320379 | Börje Langefors Best Doctoral Dissertation Award | https://en.wikipedia.org/w/index.php?title=Börje%20Langefors%20Best%20Doctoral%20Dissertation%20Award | Börje Langefors Best Doctoral Dissertation Award
Langefors Awards during the period 2011-2015 was sponsored by Nethouse and Sitevision
# The recipients.
## 2015.
First prize
"Johan Sandberg", Postdoctoral Researcher at the Umeå University awarded the first best for his doctoral dissertation entitled "Digital Capability: Investigating Coevolution of IT and Business Strategies"
## 2014.
First prize
"Mathias Hatakka", Senior Lecturer at the Örebro University awarded the first best for his doctoral dissertation entitled "The capability approach in ict4d research".
## 2013.
First prize
"Anders Olof Larsson" at the Department of Informatics and Media of Uppsala University for his thesis "Doing Things in Relation to Machines –
Studies | 6,131,617 |
37320379 | Börje Langefors Best Doctoral Dissertation Award | https://en.wikipedia.org/w/index.php?title=Börje%20Langefors%20Best%20Doctoral%20Dissertation%20Award | Börje Langefors Best Doctoral Dissertation Award
on Online Interactivity". The motivation for the award as described by the SISA Börje Langefors Prize Committee : "The thesis is based on a socially relevant contemporary topic, well-designed and well-defined subject area with contrasting perspectives based on exceptional and interesting empirical material. The thesis is easy to read and well structured with well linked articles. It has a very good international exposure". Thesis download link is here.
## 2012.
First best
"Henrik Wimelius", Assistant professor at the Umeå University awarded the first best for his doctoral dissertation entitled “Duplicate Systems: Investigating Unintended Consequences of Information Technology in Organizations”. | 6,131,618 |
37320379 | Börje Langefors Best Doctoral Dissertation Award | https://en.wikipedia.org/w/index.php?title=Börje%20Langefors%20Best%20Doctoral%20Dissertation%20Award | Börje Langefors Best Doctoral Dissertation Award
The motivation for selected him as the first is, as stated by SISA is “Henrik Wimelius dissertation is well written and clearly positioned against existing literature. The question that is addressed both theoretically and practically interesting. Methodologically the research is based on a rigorous process, presented in a reflexive manner. Furthermore, logic and structure of the thesis is well thought. The existence of parallel, competing IT systems in organizations and activities tend to help Henrik with valuable insights and lessons learned. His thesis is an excellent knowledge base which can advantageously be further exploited.” Thesis download link is here.
Second best
"M. Sirajul Islam", | 6,131,619 |
37320379 | Börje Langefors Best Doctoral Dissertation Award | https://en.wikipedia.org/w/index.php?title=Börje%20Langefors%20Best%20Doctoral%20Dissertation%20Award | Börje Langefors Best Doctoral Dissertation Award
Assistant Professor at the School of Business (Informatics) of Örebro University received the second best award for his dissertation – “Creating Opportunity by Connecting the Unconnected: Deploying Mobile Phone based Agriculture Market Information Service for Farmers in Bangladesh” . The motivation for the award conferred to Sirajul as stated: “Sirajul Islam‘s dissertation reports a design-oriented action research project that sought to create sustainable societal effects by facilitating mobile technology adoption. One interesting aspect of this change effort is that it nicely illustrates how informatics research can help underprivileged groups to strengthen their positions through innovative | 6,131,620 |
37320379 | Börje Langefors Best Doctoral Dissertation Award | https://en.wikipedia.org/w/index.php?title=Börje%20Langefors%20Best%20Doctoral%20Dissertation%20Award | Börje Langefors Best Doctoral Dissertation Award
IT use. The research project was executed through a well-designed process that is presented in a comprehensive yet detailed way. In particular, it reveals how and why certain practical and theoretical issues were tackled to push the project forward. Constituting the core of the thesis, the set of articles suggests that the result produced were not only locally relevant but also globally impactful.” Thesis download link is here. A brief interview with Sirajul about this Award is available here (in Swedish).
## 2011.
First prize
"Annika Andersson" at the Informatics department of Örebro University for her thesis " Learning to learn in e-Learning: constructive practices for development ". The | 6,131,621 |
37320379 | Börje Langefors Best Doctoral Dissertation Award | https://en.wikipedia.org/w/index.php?title=Börje%20Langefors%20Best%20Doctoral%20Dissertation%20Award | Börje Langefors Best Doctoral Dissertation Award
motivation for the award as described by the SISA Börje Langefors Prize Committee : "Annika Andersson is awarded the prize for best dissertation for the following reasons: socially relevant subject matter, well formulated and relevant theoretical basis, proper research design with appropriate method triangulation and sequencing of sub-studies, comprehensive and interesting empirical work, good incremental cumulative knowledge, clear and well-structured presentation, good design of compilation thesis with a well-developed cover paper, a good international exposure.". Thesis download link is here.
Runner-up
"Jonas Sjöström", Senior Lecturer and Researcher at the Department of Informatics and | 6,131,622 |
37320379 | Börje Langefors Best Doctoral Dissertation Award | https://en.wikipedia.org/w/index.php?title=Börje%20Langefors%20Best%20Doctoral%20Dissertation%20Award | Börje Langefors Best Doctoral Dissertation Award
Senior Lecturer and Researcher at the Department of Informatics and Media, Uppsala University stood runner-up for the Börje Langefors Award for best Information Systems (IS) thesis in Sweden 2009-2010. The title of his thesis is Designing Information Systems - A Pragmatic Account. The motivation was : “ Topical subject, well grounded in an interesting and well-reflected theoretical perspective that integrate essential national and international theory within as well as outside of the IS discipline, important contributions to the theorization of the IT artifact, solid knowledge contributions as a foundation for future research, and a good international exposure.” Thesis download link is here. | 6,131,623 |
37320488 | NA-17 (Haripur) | https://en.wikipedia.org/w/index.php?title=NA-17%20(Haripur) | NA-17 (Haripur)
NA-17 (Haripur)
NA-17 (Haripur) () is a constituency for the National Assembly of Pakistan. It covers tho whole of district Haripur. The constituency was formerly known as NA-19 (Haripur) from 1977 to 2018. The name changed to NA-17 (Haripur) after the delimitation in 2018.
# Detailed results.
## 2002 General Election.
"A total of 6,277 votes were rejected."
## 2008 General Election.
"A total of 5,757 votes were rejected."
## 2013 General Election.
"A total of 8,467 votes were rejected."
## 2015 By-election.
A by-election took place on 16 August 2015.
"A total of 3,918 votes were rejected."
## 2018 General Election.
General elections were held on 25 July 2018. The constituency got | 6,131,624 |
37320488 | NA-17 (Haripur) | https://en.wikipedia.org/w/index.php?title=NA-17%20(Haripur) | NA-17 (Haripur)
tailed results.
## 2002 General Election.
"A total of 6,277 votes were rejected."
## 2008 General Election.
"A total of 5,757 votes were rejected."
## 2013 General Election.
"A total of 8,467 votes were rejected."
## 2015 By-election.
A by-election took place on 16 August 2015.
"A total of 3,918 votes were rejected."
## 2018 General Election.
General elections were held on 25 July 2018. The constituency got the third most highest total votes polled in all of Pakistan. Omar Ayub Khan of Pakistan Tehreek-e-Insaf won getting most votes by any candidate in all of Pakistan.
# See also.
- NA-16 (Abbottabad-II)
- NA-18 (Swabi-I)
# External links.
- Election result's official website | 6,131,625 |
37320419 | François Gernelle | https://en.wikipedia.org/w/index.php?title=François%20Gernelle | François Gernelle
François Gernelle
François Gernelle (born December 20, 1944) is a French engineer, computer scientist and entrepreneur famous for inventing the first micro-computer using a micro-processor, the Micral N.
# Education.
In the late 1960s, Gernelle earned an engineering degree at the Conservatoire National des Arts et Métiers. In 1978, he earned a Ph.D in computer science at the Pierre Mendès-France University of Grenoble.
# Career.
## Intertechnique and R2E.
In 1968, he was hired by Intertechnique, a company specialized in electronic measurement for aviation. There he discovered the Intel 8008 microprocessor and imagined all its potential applications. As his hierarchy didn't share his views | 6,131,626 |
37320419 | François Gernelle | https://en.wikipedia.org/w/index.php?title=François%20Gernelle | François Gernelle
on the i8008 development capacity, he resigned in 1972 and joined , a company created and led by Paul Magneron. He designed a micro-computer to answer a request of INRA to measure agricultural hygrometry. During this project, he granted two patents.
In 1973 he supported the Micral N and, with the company's growth, helped to design 20 other multi-user microcomputers for some were multi-processor one.
## Bull Micral.
In 1981, the Bull company acquired R2E and he then joined the new entity Bull Micral. But the company wanted him to design IBM PC compatible machines and François Gernelle didn't agree because he thought this machine was bad designed, using an 8 bit mono-tasking mono-user i8088 | 6,131,627 |
37320419 | François Gernelle | https://en.wikipedia.org/w/index.php?title=François%20Gernelle | François Gernelle
powered by a poor operating system. In his mind, this kind of poor computer design was a dead-end at a time where really good micro-processors existed and offered capacity to design powerful multi-user and multi-tasking systems at medium and even low cost.
## FORUM International.
In 1983, he resigned from Bull and founded a new company named which will create professional computers powered by .
# External References.
- This article is based on the .
- Interview with F. Gernelle issued in Le Choc du Mois issue #18 of december 2007.
- «La naissance du premier micro-ordinateur: le Micral N» ("Birth of the first micro-computer: the Micral N") article written by F.Gernelle where he talks about | 6,131,628 |
37320419 | François Gernelle | https://en.wikipedia.org/w/index.php?title=François%20Gernelle | François Gernelle
existed and offered capacity to design powerful multi-user and multi-tasking systems at medium and even low cost.
## FORUM International.
In 1983, he resigned from Bull and founded a new company named which will create professional computers powered by .
# External References.
- This article is based on the .
- Interview with F. Gernelle issued in Le Choc du Mois issue #18 of december 2007.
- «La naissance du premier micro-ordinateur: le Micral N» ("Birth of the first micro-computer: the Micral N") article written by F.Gernelle where he talks about his career at R2E, Bull and the creation of FORUM International.
- History of Computer Graphics by Dan Ryan, page 336. 2011, AuthorHouse. | 6,131,629 |
37319988 | The Concert for Bangladesh (film) | https://en.wikipedia.org/w/index.php?title=The%20Concert%20for%20Bangladesh%20(film) | The Concert for Bangladesh (film)
The Concert for Bangladesh (film)
The Concert for Bangladesh is a film directed by Saul Swimmer and released in 1972. The film documents the two benefit concerts that were organised by George Harrison and Ravi Shankar to raise funds for refugees of the Bangladesh Liberation War, and were held on Sunday, 1 August 1971 at Madison Square Garden in New York City. As well as notable performances from Harrison and Shankar, the film includes "main performer" contributions from Harrison's fellow ex-Beatle Ringo Starr, Billy Preston and Leon Russell, and a surprise walk-on from Bob Dylan. Other contributing musicians include Ali Akbar Khan, Eric Clapton, the band Badfinger, Klaus Voormann, Jesse Ed Davis, | 6,131,630 |
37319988 | The Concert for Bangladesh (film) | https://en.wikipedia.org/w/index.php?title=The%20Concert%20for%20Bangladesh%20(film) | The Concert for Bangladesh (film)
Jim Horn and Jim Keltner.
The film was the final part of Harrison's "pioneering" aid project for the people of former East Pakistan, following his "Bangla Desh" charity single, the UNICEF benefit concerts, and a triple live album of the event credited to "George Harrison and Friends". "The Concert for Bangladesh" was produced by The Beatles' Apple Films; after delays caused by problems with inadequate footage from the event, it opened in US cinemas in the spring of 1972. The film was released on DVD in 2005 accompanied by a newly created documentary feature, "The Concert for Bangladesh Revisited with George Harrison and Friends", which included recollections from many of the project's participants | 6,131,631 |
37319988 | The Concert for Bangladesh (film) | https://en.wikipedia.org/w/index.php?title=The%20Concert%20for%20Bangladesh%20(film) | The Concert for Bangladesh (film)
and contextual input from then UN secretary-general Kofi Annan, US Fund for UNICEF president Charles Lyons and Live Aid founder Bob Geldof.
As with the live album, sales of the DVD release of the film continue to benefit the George Harrison Fund for UNICEF.
# Production.
Saul Swimmer's "Concert for Bangladesh" documentary combined footage from both of the Madison Square Garden shows held on 1 August 1971, using George Harrison's preference of the performances of the songs. Harrison later explained that much of the concert footage was unusable, as a camera on the right-hand side of the venue was faulty and out of focus throughout, while the one opposite, down the left side, had cables hanging | 6,131,632 |
37319988 | The Concert for Bangladesh (film) | https://en.wikipedia.org/w/index.php?title=The%20Concert%20for%20Bangladesh%20(film) | The Concert for Bangladesh (film)
down in front of it. The compromised quality would result in some brutal edits in the released movie – Eric Clapton, for instance, appears to change jackets and guitar part-way through a song (Leon Russell's medley in particular). In an interview accompanying the 2005 DVD release of the film, Swimmer would cite the audio syncing and the frame-by-frame conversion to 70mm format (from the original 16mm) as other challenging, labour-intensive tasks. With work almost completed on the "Concert for Bangladesh" live album, Harrison is said to have begun editing the footage on 6 September; at some stage during the next few months, he was joined in this lengthy process by Bob Dylan.
A clip of Harrison's | 6,131,633 |
37319988 | The Concert for Bangladesh (film) | https://en.wikipedia.org/w/index.php?title=The%20Concert%20for%20Bangladesh%20(film) | The Concert for Bangladesh (film)
performance of "My Sweet Lord" was previewed during his appearance on ABC-TV's "The Dick Cavett Show" on 23 November, but the movie would not be ready for release until the following spring.
# Synopsis.
The opening of the movie features footage from the New York press conference, held at Allen Klein's ABKCO offices five days before the concerts, during which Harrison and Ravi Shankar discuss the upcoming shows. Harrison is asked by a reporter: "With all the enormous problems in the world, how did you happen to choose this one to do something about?" "Because I was asked by a friend if I would help, you know – that's all," is his reply.
The scene then shifts to inside Madison Square Garden, | 6,131,634 |
37319988 | The Concert for Bangladesh (film) | https://en.wikipedia.org/w/index.php?title=The%20Concert%20for%20Bangladesh%20(film) | The Concert for Bangladesh (film)
showing musicians and support crew preparing for the first show. In voiceover, Harrison provides a brief explanation of how the project came together.
The concert begins with Harrison taking to the stage alone and addressing the audience, his comment "We've got a good show lined up – well, I hope so anyway ..." alluding to the speed with which the event was organised. He then introduces the first group of musicians, led by Shankar, who, like Harrison, attempts to convey the intricacies of Indian classical music to the audience, as well as outlining the reason for this "special benefit concert". Shankar and Ali Akbar Khan proceed to tune their instruments and then stop after about 90 seconds. | 6,131,635 |
37319988 | The Concert for Bangladesh (film) | https://en.wikipedia.org/w/index.php?title=The%20Concert%20for%20Bangladesh%20(film) | The Concert for Bangladesh (film)
The audience, apparently believing they have heard an entire piece, respond with enthusiastic applause, to which Shankar replies: "Thank you, if you appreciate the tuning so much, I hope you will enjoy the playing more." The four musicians then launch into a fifteen-minute dhun.
A brief interlude ensues, consisting of behind-the-scenes footage that shows Phil Spector, Harrison and other performers making their way to the stage. Harrison starts off the rock portion of the concert with a string of songs from his hit album "All Things Must Pass". He is backed by a large band, including two drummers, Ringo Starr and Jim Keltner, pianist Leon Russell, organist Billy Preston, Klaus Voormann on bass, | 6,131,636 |
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two other lead guitarists, Eric Clapton and Jesse Ed Davis, the four members of Badfinger on rhythm guitars and tambourine, a six-piece horn section in matching blue patterned shirts, and a small choir of backing vocalists, a few of whom are also playing percussion. Harrison then turns the concert over to his friends briefly.
Towards the end of Billy Preston's song, "That's the Way God Planned It", Preston gets up from his stool and dances across the stage and back again – much to Harrison's delight. (Like most of the concert visuals and audio, this footage is taken from the evening performance on 1 August.) Starr sings his hit song "It Don't Come Easy" and appears flustered as he forgets some | 6,131,637 |
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of the words. Harrison then returns to the spotlight for two of his own numbers, the second being "While My Guitar Gently Weeps", at the end of which Clapton clearly struggles on lead guitar next to Harrison's more measured soloing. Before this, Harrison pauses for a few minutes to introduce the many musicians around him, and to his own amazement misses out one of the key performers so far – "We've forgotten Billy Preston!" he declares eventually.
With a slight change in personnel, Russell delivers a rock and roll medley, before Harrison performs an intimate version of another Beatles-era song, "Here Comes the Sun". Harrison now introduces a newcomer to the stage, as Bob Dylan appears for a | 6,131,638 |
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semi-acoustic set of four of his songs.
Two more numbers from Harrison close the show. When he apparently forgets part of the second verse in "Something", he looks around, seemingly at Russell or Clapton, and grins his way through it. The film concludes with a spirited version of Harrison's then-current single, "Bangla Desh", intercut at first with footage of the suffering refugees the concert was aiming to provide aid for. Towards the end of the song, Harrison exits the stage while the rest of the band plays on.
# Performances in the film.
"All songs composed and performed by George Harrison, unless otherwise noted."
## Ravi Shankar.
Performers: Ravi Shankar (sitar), Ali Akbar Khan (sarod), | 6,131,639 |
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Alla Rakha (tabla), Kamala Chakravarty (tambura)
- "Bangla Dhun" (P.D.) – traditional dhun
## George Harrison and band.
Performers: George Harrison (vocals, electric and acoustic guitars), Ringo Starr (vocals, drums), Leon Russell (vocals, piano), Billy Preston (vocals, organ), Eric Clapton (electric guitar, slide guitar), Jesse Ed Davis (electric guitar, slide guitar), Klaus Voormann (bass), Jim Keltner (drums); with Badfinger: Pete Ham (acoustic guitar), Tom Evans (12-string acoustic guitar), Joey Molland (acoustic guitar), Mike Gibbins (percussion); with The Hollywood Horns: Jim Horn, Chuck Findley, Jackie Kelso, Allan Beutler, Lou McCreary, Ollie Mitchell; and The Soul Choir: Don Nix, | 6,131,640 |
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Claudia Linnear, Joe Greene, Dolores Hall, Jeanie Greene, Marlin Greene, Don Preston
- "Wah-Wah"
- "My Sweet Lord"
- "Awaiting on You All"
- "That's the Way God Planned It" (Preston) – performed by Billy Preston
- "It Don't Come Easy" (Starkey) – performed by Ringo Starr
- "Beware of Darkness" – featuring Leon Russell on second lead vocal
- "While My Guitar Gently Weeps" – featuring Eric Clapton on lead guitar
- "Jumpin' Jack Flash"/"Young Blood" (Jagger–Richards/Leiber–Stoller–Pomus) – performed by Leon Russell, with additional lead vocals on "Young Blood" by Don Preston and George Harrison; featuring Preston on lead guitar and Carl Radle on bass (in place of Voormann)
- "Here Comes | 6,131,641 |
37319988 | The Concert for Bangladesh (film) | https://en.wikipedia.org/w/index.php?title=The%20Concert%20for%20Bangladesh%20(film) | The Concert for Bangladesh (film)
the Sun" – featuring Pete Ham on second acoustic guitar
## Bob Dylan.
Performers: Bob Dylan (vocals, acoustic guitar, harmonica), George Harrison (electric guitar, slide guitar, backing vocals), Leon Russell (bass, backing vocals), Ringo Starr (tambourine)
- "A Hard Rain's a-Gonna Fall" (Dylan)
- "It Takes a Lot to Laugh, It Takes a Train to Cry" (Dylan)
- "Blowin' in the Wind" (Dylan)
- "Just Like a Woman" (Dylan)
### encore.
Performers: as for George Harrison and band (above), but with Don Preston on electric guitar on "Bangla Desh"
- "Something"
- "Bangla Desh"
# Songs not in the film.
- "Mr. Tambourine Man" – written and performed by Bob Dylan; from the evening show and included | 6,131,642 |
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only on the "Concert for Bangladesh" album.
- "Love Minus Zero/No Limit" – written and performed by Dylan; recorded during the afternoon show and included as an extra on the 2005 DVD.
- "Hear Me Lord" – written and performed by George Harrison; played following Dylan's five-song set during the afternoon show but not issued on either the 1971–72 releases or the 2005 reissues.
- "If Not for You" – written and performed by Dylan, with George Harrison on harmony vocals and acoustic guitar, and Klaus Voormann on electric bass; recorded during the soundcheck on 31 July and later included as an extra on the 2005 DVD, after a very brief portion had been aired in the original film.
- "Come on in | 6,131,643 |
37319988 | The Concert for Bangladesh (film) | https://en.wikipedia.org/w/index.php?title=The%20Concert%20for%20Bangladesh%20(film) | The Concert for Bangladesh (film)
My Kitchen" – composed by Robert Johnson and performed by Leon Russell (vocals and piano) with George Harrison (lead guitar and backing vocals), Eric Clapton (second lead guitar), Billy Preston (organ), Carl Radle (bass), and Ringo Starr and Jim Keltner (both on drums); recorded during the soundcheck and included as an extra on the 2005 DVD.
# Release and reception.
Marketed with the tagline "The greatest concert of the decade. Now you can see it and hear it ... as if you were there!" together with Tom Wilkes's confronting poster image, "The Concert for Bangla Desh" received a preview screening on 22 March 1972 at New York's DeMille Theater before its official premiere the following day. John | 6,131,644 |
37319988 | The Concert for Bangladesh (film) | https://en.wikipedia.org/w/index.php?title=The%20Concert%20for%20Bangladesh%20(film) | The Concert for Bangladesh (film)
Lennon was among those attending the preview, along with wife Yoko Ono, although he left the cinema during Dylan's segment. The UK release was delayed until 27 July, however,John Pidgeon, ""Bangla Desh"", "NME", 15 July 1972, p. 24; available at Rock's Back Pages (retrieved 15 July 2012). preceded by a screening at the Rialto Cinema in central London. "The Concert for Bangladesh" was an instant commercial success internationally, breaking previous records for daily box-office takings in London. In his review for "NME", John Pidgeon concluded: "The film tries to be no more than a visual album, an aim which it pursues even as far as separating the 'tracks' with darkness, and in which it undisputedly | 6,131,645 |
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succeeds." Reviewing the 2005 reissue for BBC Online, Chris Jones described the concert as a "masterclass" in how to carry out a superstar charity benefit.
# 2005 DVD release.
A two-disc special edition DVD of "The Concert for Bangladesh" was issued in October 2005, with the concert on disc one – at 99 minutes, slightly shorter than the original film – and an all-new documentary, "The Concert for Bangladesh Revisited with George Harrison and Friends", on the second disc.
Performers interviewed for the documentary include Ravi Shankar, Eric Clapton, Ringo Starr, Billy Preston, Jim Keltner, Jim Horn, Leon Russell and Klaus Voormann, who offer their recollections of the concert. George Harrison | 6,131,646 |
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talks about organising the concert in voiceovers only. Other interviews are with "Rolling Stone" founder Jann Wenner and Live Aid organiser Bob Geldof, both of whom talk of the historic importance of the event, as well as Apple Corps executive Neil Aspinall. Charles Lyons, then president of the US Fund for UNICEF, discusses the immediate benefits of the concerts in providing funds to treat cholera among the refugees, as well as the longer-term influence of the Bangladesh relief project, as the success of the live album and film forced foreign governments to ask themselves, "Are we on the right side on this Bangladesh issue?"
Regarding Clapton's absence from the preparations, Harrison explains | 6,131,647 |
37319988 | The Concert for Bangladesh (film) | https://en.wikipedia.org/w/index.php?title=The%20Concert%20for%20Bangladesh%20(film) | The Concert for Bangladesh (film)
that he was booked on every airline flight from London to New York City for a week before the shows. After finally boarding a plane, Clapton performed without the benefit of a rehearsal, and "he was brilliant," Harrison says, somewhat generously. Clapton, for his part, recalls the time as a period of "retirement" and states that he "really made it hard" for himself in the concerts, choosing to play a hollow-body Gibson Byrdland for his solo on "While My Guitar Gently Weeps", when a solid-body would have been more appropriate.
There are also short features on the making of Saul Swimmer's film, the release of the live album, movie poster and album artwork, concert photography, and personal recollections | 6,131,648 |
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of the historic day, Sunday, 1 August 1971. Participants in these features include Swimmer, A&M Studios engineer Norm Kinney, former Capitol Records boss Bhaskar Menon, designer Tom Wilkes, photographer Barry Feinstein, and British film mogul David Puttnam. Menon is noticeably more contrite when discussing Capitol's controversial delaying of the album than his comments at the time suggested. In the main documentary, Leon Russell describes the day of the concerts as "one high-level experience from beginning to end".
# References.
- Keith Badman, "The Beatles Diary Volume 2: After the Break-Up 1970–2001", Omnibus Press (London, 2001; ).
- Harry Castleman & Walter J. Podrazik, "All Together | 6,131,649 |
37319988 | The Concert for Bangladesh (film) | https://en.wikipedia.org/w/index.php?title=The%20Concert%20for%20Bangladesh%20(film) | The Concert for Bangladesh (film)
Now: The First Complete Beatles Discography 1961–1975", Ballantine Books (New York, NY, 1976; ).
- Alan Clayson, "George Harrison", Sanctuary (London, 2003; ).
- "The Concert for Bangladesh Revisited with George Harrison and Friends" DVD, Apple Corps, 2005 (directed by Claire Ferguson; produced by Olivia Harrison, Jonathan Clyde & Jo Human).
- The Editors of "Rolling Stone", "Harrison", Rolling Stone Press/Simon & Schuster (New York, NY, 2002; ).
- George Harrison, "I Me Mine", Chronicle Books (San Francisco, CA, 2002; ).
- Nicholas Schaffner, "The Beatles Forever", McGraw-Hill (New York, NY, 1978; ).
- Bruce Spizer, "The Beatles Solo on Apple Records", 498 Productions (New Orleans, LA, | 6,131,650 |
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61–1975", Ballantine Books (New York, NY, 1976; ).
- Alan Clayson, "George Harrison", Sanctuary (London, 2003; ).
- "The Concert for Bangladesh Revisited with George Harrison and Friends" DVD, Apple Corps, 2005 (directed by Claire Ferguson; produced by Olivia Harrison, Jonathan Clyde & Jo Human).
- The Editors of "Rolling Stone", "Harrison", Rolling Stone Press/Simon & Schuster (New York, NY, 2002; ).
- George Harrison, "I Me Mine", Chronicle Books (San Francisco, CA, 2002; ).
- Nicholas Schaffner, "The Beatles Forever", McGraw-Hill (New York, NY, 1978; ).
- Bruce Spizer, "The Beatles Solo on Apple Records", 498 Productions (New Orleans, LA, 2005; ).
# External links.
- Official site | 6,131,651 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
Genetic engineering techniques
Genetic engineering can be accomplished using multiple techniques. There are a number of steps that are followed before a genetically modified organism (GMO) is created. Genetic engineers must first choose what gene they wish to insert, modify or delete. The gene must then be isolated and incorporated, along with other genetic elements, into a suitable vector. This vector is then used to insert the gene into the host organism, creating the GMO.
The ability to genetically engineer organisms is built on years of research and discovery on how genes function and how we can manipulate them. Following the discovery of genes by Gregor Mendel and the proof that they were | 6,131,652 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
involved in inheritance tools were developed that allowed their direct manipulation. Important advances included the discovery of restriction enzymes and DNA ligases and the development of polymerase chain reaction and sequencing.
This allowed the gene of interest to be isolated and then incorporated into a vector. Often a promoter and terminator region was added as well as a selectable marker gene. The gene may be modified further at this point to make it express more efficiently. This vector is then inserted into the host organism's genome. For animals, the gene is typically inserted into embryonic stem cells, while in plants it can be inserted into any tissue that can be cultured into a | 6,131,653 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
fully developed plant. Common techniques include microinjection, virus-mediated, "Agrobacterium"-mediated or biolistics. Further tests are carried out on the resulting organism to ensure stable integration, inheritance and expression. First generation offspring are heterozygous, requiring them to be inbred to create the homozygous pattern necessary for stable inheritance. Homozygosity must be confirmed in second generation specimens.
Traditional techniques inserted the genes randomly into the hosts genome. Advances have allowed genes to be inserted at specific locations within a genome, which reduces the unintended side effects of random insertion. Early targeting systems relied on meganucleases | 6,131,654 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
and zinc finger nucleases. Since 2009 more accurate and easier systems to implement have been developed. Transcription activator-like effector nucleases (TALENs) and the Cas9-guideRNA system (adapted from CRISPR) are the two most commonly used. They may potentially be useful in gene therapy and other procedures that require accurate or high through put targeting.
# History.
Many different discoveries and advancements lead to the development of genetic engineering. Human-directed genetic manipulation began with the domestication of plants and animals through artificial selection in about 12,000 BC. Various techniques were developed to aid in breeding and selection. Hybridization was one way | 6,131,655 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
rapid changes in an organisms makeup could be introduced. Hybridization most likely first occurred when humans first grew similar, yet slightly different plants in close proximity. Some plants were able to be propagated by vegetative cloning.
Genetic inheritance was first discovered by Gregor Mendel in 1865, following experiments crossing peas. In 1928 Frederick Griffith proved the existence of a "transforming principle" involved in inheritance, which was identified as DNA in 1944 by Oswald Avery, Colin MacLeod, and Maclyn McCarty. Frederick Sanger developed a method for sequencing DNA in 1977, greatly increasing the genetic information available to researchers.
After discovering the existence | 6,131,656 |
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and properties of DNA, tools had to be developed that allowed it to be manipulated. In 1970 Hamilton Smiths lab discovered restriction enzymes, enabling scientists to isolate genes from an organism's genome. DNA ligases, which join broken DNA together, were discovered earlier in 1967. By combining the two enzymes it became possible to "cut and paste" DNA sequences to create recombinant DNA. Plasmids, discovered in 1952, became important tools for transferring information between cells and replicating DNA sequences. Polymerase chain reaction (PCR), developed by Kary Mullis in 1983, allowed small sections of DNA to be amplified (replicated) and aided identification and isolation of genetic material.
As | 6,131,657 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
well as manipulating DNA, techniques had to be developed for its insertion into an organism's genome. Griffith's experiment had already shown that some bacteria had the ability to naturally uptake and express foreign DNA. Artificial competence was induced in "Escherichia coli" in 1970 by treating them with calcium chloride solution (CaCl). Transformation using electroporation was developed in the late 1980s, increasing the efficiency and bacterial range. In 1907 a bacterium that caused plant tumors, "Agrobacterium tumefaciens", had been discovered. In the early 1970s it was found that this bacteria inserted its DNA into plants using a Ti plasmid. By removing the genes in the plasmid that caused | 6,131,658 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
the tumor and adding in novel genes, researchers were able to infect plants with "A. tumefaciens" and let the bacteria insert their chosen DNA into the genomes of the plants.
# Choosing target genes.
The first step is to identify the target gene or genes to insert into the host organism. This is driven by the goal for the resultant organism. In some cases only one or two genes are affected. For more complex objectives entire biosynthetic pathways involving multiple genes may be involved. Once found genes and other genetic information from a wide range of organisms can be inserted into bacteria for storage and modification, creating genetically modified bacteria in the process. Bacteria are | 6,131,659 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
cheap, easy to grow, clonal, multiply quickly, relatively easy to transform and can be stored at -80 °C almost indefinitely. Once a gene is isolated it can be stored inside the bacteria providing an unlimited supply for research.
Genetic screens can be carried out to determine potential genes followed by other tests identify the best candidates. A simple screen involves randomly mutating DNA with chemicals or radiation and then selecting those that display the desired trait. For organisms where mutation is not practical, scientist instead look for individuals among the population who present the characteristic through naturally-occurring mutations. Processes that look at a phenotype and then | 6,131,660 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
try and identify the gene responsible are called forward genetics. The gene then needs to be mapped by comparing the inheritance of the phenotype with known genetic markers. Genes that are close together are likely to be inherited together.
Another option is reverse genetics. This approach involves targeting a specific gene with a mutation and then observing what phenotype develops. The mutation can be designed to inactivate the gene or only allow it to become active only under certain conditions. Conditional mutations are useful for identifying genes that are normally lethal if non-functional. As genes with similar functions share similar sequences (homologous) it is possible to predict the | 6,131,661 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
likely function of a gene by comparing its sequence to that of well-studied genes from model organisms. The development of microarrays, transcriptomes and genome sequencing has made it much easier to find desirable genes.
The bacteria "Bacillus thuringiensis" was first discovered in 1901 as the causative agent in the death of silkworms. Due to these insecticidal properties the bacteria was used as a biological insecticide, developed commercially in 1938. The cry proteins were discovered to provide the insecticidal activity in 1956 and by the 1980s scientists had successfully cloned the gene that codes for this protein and expressed it in plants. Luck may also be involved. The gene that provides | 6,131,662 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
resistance to the glyphosate herbicide was found, after seven years searching, in bacteria that were surviving in the outflow pipe of a Monsanto RoundUp manufacturing facility. In animals the majority of genes used are growth hormone genes.
# Gene manipulation.
All genetic engineering processes involve the modification of DNA. Traditionally DNA was isolated from the cells of organisms. Later, genes came to be cloned from a DNA segment after the creation of a DNA library or artificially synthesised. Once isolated, additional genetic elements are added to the gene to allow it to be expressed in the host organism and to aid selection.
## Extraction from cells.
First the cell must be gently | 6,131,663 |
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opened, exposing the DNA without causing too much damage to it. The methods used vary depending on the type of cell. Once open, the DNA must be separated from the other cellular components. A ruptured cell contains proteins and other cell debris. By mixing with phenol and/or chloroform, followed by centrifuging, the nucleic acids can be separated from this debris into an upper aqueous phase. This aqueous phase can be removed and further purified if necessary by repeating the phenol-chloroform steps. The nucleic acids can then be precipitated from the aqueous solution using ethanol or isopropanol. Any RNA can be removed by adding a ribonuclease that will degrade it. Many companies now sell kits | 6,131,664 |
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that simplify the process.
## Gene isolation.
The gene of interest must be separated from the extracted DNA. If the sequence is not known then a common method is to break the DNA up with a random digestion method. This is usually accomplished using restriction enzymes (enzymes that cut DNA). A partial restriction digest cuts only some of the restriction sites, resulting in overlapping DNA fragment segments. The DNA fragments are put into individual plasmid vectors and grown inside bacteria. Once in the bacteria the plasmid is copied as the bacteria divides. To determine if a useful gene is present on a particular fragment the DNA library is screened for the desired phenotype. If the phenotype | 6,131,665 |
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is detected then it is possible that the bacteria contains the target gene.
If the gene does not have a detectable phenotype or a DNA library does not contain the correct gene, other methods must be used to isolate it. If the position of the gene can be determined using molecular markers then chromosome walking is one way to isolate the correct DNA fragment. If the gene expresses close homology to a known gene in another species, then it could be isolated by searching for genes in the library that closely match the known gene.
For known DNA sequences, restriction enzymes that cut the DNA on either side of the gene can be used. Gel electrophoresis then sorts the fragments according to length. | 6,131,666 |
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Some gels can separate sequences that differ by a single base-pair. The DNA can be visualised by staining it with ethidium bromide and photographing under UV light. A marker with fragments of known lengths can be laid alongside the DNA to estimate the size of each band. The DNA band at the correct size should contain the gene, where it can be excised from the gel. Another technique to isolate genes of known sequences involves polymerase chain reaction (PCR). PCR is a powerful tool that can amplify a given sequence, which can then be isolated through gel electrophoresis. Its effectiveness drops with larger genes and it has the potential to introduce errors into the sequence.
It is possible to | 6,131,667 |
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artificially synthesise genes. Some synthetic sequences are available commercially, forgoing many of these early steps.
## Modification.
The gene to be inserted must be combined with other genetic elements in order for it to work properly. The gene can be modified at this stage for better expression or effectiveness. As well as the gene to be inserted most constructs contain a promoter and terminator region as well as a selectable marker gene. The promoter region initiates transcription of the gene and can be used to control the location and level of gene expression, while the terminator region ends transcription. A selectable marker, which in most cases confers antibiotic resistance to the | 6,131,668 |
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organism it is expressed in, is used to determine which cells are transformed with the new gene. The constructs are made using recombinant DNA techniques, such as restriction digests, ligations and molecular cloning.
# Inserting DNA into the host genome.
Once the gene is constructed it must be stably integrated into the target organisms genome or exist as extrachromosomal DNA. There are a number of techniques available for inserting the gene into the host genome and they vary depending on the type of organism targeted. In multicellular eukaryotes, if the transgene is incorporated into the host's germline cells, the resulting host cell can pass the transgene to its progeny. If the transgene | 6,131,669 |
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is incorporated into somatic cells, the transgene can not be inherited.
## Transformation.
Transformation is the direct alteration of a cells genetic components by passing the genetic material through the cell membrane. About 1% of bacteria are naturally able to take up foreign DNA, but this ability can be induced in other bacteria. Stressing the bacteria with a heat shock or electroporation can make the cell membrane permeable to DNA that may then incorporate into the genome or exist as extrachromosomal DNA. Typically the cells are incubated in a solution containing divalent cations (often calcium chloride) under cold conditions, before being exposed to a heat pulse (heat shock). Calcium | 6,131,670 |
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chloride partially disrupts the cell membrane, which allows the recombinant DNA to enter the host cell. It is suggested that exposing the cells to divalent cations in cold condition may change or weaken the cell surface structure, making it more permeable to DNA. The heat-pulse is thought to create a thermal imbalance across the cell membrane, which forces the DNA to enter the cells through either cell pores or the damaged cell wall. Electroporation is another method of promoting competence. In this method the cells are briefly shocked with an electric field of 10-20 kV/cm, which is thought to create holes in the cell membrane through which the plasmid DNA may enter. After the electric shock, | 6,131,671 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
the holes are rapidly closed by the cell's membrane-repair mechanisms. Up-taken DNA can either integrate with the bacterials genome or, more commonly, exist as extrachromosomal DNA.
In plants the DNA is often inserted using "Agrobacterium"-mediated recombination, taking advantage of the "Agrobacterium"s T-DNA sequence that allows natural insertion of genetic material into plant cells. Plant tissue are cut into small pieces and soaked in a fluid containing suspended "Agrobacterium". The bacteria will attach to many of the plant cells exposed by the cuts. The bacteria uses conjugation to transfer a DNA segment called T-DNA from its plasmid into the plant. The transferred DNA is piloted to the | 6,131,672 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
plant cell nucleus and integrated into the host plants genomic DNA.The plasmid T-DNA is integrated semi-randomly into the genome of the host cell.
By modifying the plasmid to express the gene of interest, researchers can insert their chosen gene stably into the plants genome. The only essential parts of the T-DNA are its two small (25 base pair) border repeats, at least one of which is needed for plant transformation. The genes to be introduced into the plant are cloned into a plant transformation vector that contains the T-DNA region of the plasmid. An alternative method is agroinfiltration.
Another method used to transform plant cells is biolistics, where particles of gold or tungsten are | 6,131,673 |
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coated with DNA and then shot into young plant cells or plant embryos. Some genetic material enters the cells and transforms them. This method can be used on plants that are not susceptible to "Agrobacterium" infection and also allows transformation of plant plastids. Plants cells can also be transformed using electroporation, which uses an electric shock to make the cell membrane permeable to plasmid DNA. Due to the damage caused to the cells and DNA the transformation efficiency of biolistics and electroporation is lower than agrobacterial transformation.
## Transfection.
Transformation has a different meaning in relation to animals, indicating progression to a cancerous state, so the process | 6,131,674 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
used to insert foreign DNA into animal cells is usually called transfection. There are many ways to directly introduce DNA into animal cells "in vitro". Often these cells are stem cells that are used for gene therapy. Chemical based methods uses natural or synthetic compounds to form particles that facilitate the transfer of genes into cells. These synthetic vectors have the ability to bind DNA and accommodate large genetic transfers. One of the simplest methods involves using calcium phosphate to bind the DNA and then exposing it to cultured cells. The solution, along with the DNA, is encaspulated by the cells and a small amount of DNA can be integrated into the genome. Liposomes and polymers | 6,131,675 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
can be used as vectors to deliver DNA into cultured animal cells. Positively charged liposomes bind with DNA, while polymers can designed that interact with DNA. They form lipoplexes and polyplexes respectively, which are then up-taken by the cells. Other techniques include using electroporation and biolistics.
To create transgenic animals the DNA must be inserted into viable embryos or eggs. This is usually accomplished using microinjection, where DNA is injected through the cell's nuclear envelope directly into the nucleus. Superovulated fertilised eggs are collected at the single cell stage and cultured "in vitro". When the pronuclei from the sperm head and egg are visible through the protoplasm | 6,131,676 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
the genetic material is injected into one of them. The oocyte is then implanted in the oviduct of a pseudopregnant animal. Another method is Embryonic Stem Cell-Mediated Gene Transfer. The gene is transfected into embryonic stem cells and then they are inserted into mouse blastocysts that are then implanted into foster mothers. The resulting offspring are chimeric, and further mating can produce mice fully transgenic with the gene of interest.
## Transduction.
Transduction is the process by which foreign DNA is introduced into a cell by a virus or viral vector. Genetically modified viruses can be used as viral vectors to transfer target genes to another organism in gene therapy. First the | 6,131,677 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
virulent genes are removed from the virus and the target genes are inserted instead. The sequences that allow the virus to insert the genes into the host organism must be left intact. Popular virus vectors are developed from retroviruses or adenoviruses. Other viruses used as vectors include, lentiviruses, pox viruses and herpes viruses. The type of virus used will depend on the cells targeted and whether the DNA is to be altered permanently or temporarily.
## Regeneration.
As often only a single cell is transformed with genetic material, the organism must be regenerated from that single cell. In plants this is accomplished through the use of tissue culture. Each plant species has different | 6,131,678 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
requirements for successful regeneration. If successful, the technique produces an adult plant that contains the transgene in every cell. In animals it is necessary to ensure that the inserted DNA is present in the embryonic stem cells. Offspring can be screened for the gene. All offspring from the first generation are heterozygous for the inserted gene and must be inbred to produce a homozygous specimen. Bacteria consist of a single cell and reproduce clonally so regeneration is not necessary. Selectable markers are used to easily differentiate transformed from untransformed cells.
Cells that have been successfully transformed with the DNA contain the marker gene, while those not transformed | 6,131,679 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
will not. By growing the cells in the presence of an antibiotic or chemical that selects or marks the cells expressing that gene, it is possible to separate modified from unmodified cells. Another screening method involves a DNA probe that sticks only to the inserted gene. These markers are usually present in the transgenic organism, although a number of strategies have been developed that can remove the selectable marker from the mature transgenic plant.
## Confirmation.
Finding that a recombinant organism contains the inserted genes is not usually sufficient to ensure that they will be appropriately expressed in the intended tissues. Further testing using PCR, Southern hybridization, and | 6,131,680 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
DNA sequencing is conducted to confirm that an organism contains the new gene. These tests can also confirm the chromosomal location and copy number of the inserted gene. Once confirmed methods that look for and measure the gene products (RNA and protein) are also used to assess gene expression, transcription, RNA processing patterns and expression and localization of protein product(s). These include northern hybridisation, quantitative RT-PCR, Western blot, immunofluorescence, ELISA and phenotypic analysis. When appropriate, the organism's offspring are studied to confirm that the transgene and associated phenotype are stably inherited.
# Gene targeting.
Traditional methods of genetic engineering | 6,131,681 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
generally insert the new genetic material randomly within the host genome. This can impair or alter other genes within the organism. Methods were developed that inserted the new genetic material into specific sites within an organism genome. Early methods that targeted genes at certain sites within a genome relied on homologous recombination. By creating DNA constructs that contain a template that matches the targeted genome sequence, it is possible that the HR processes within the cell will insert the construct at the desired location. Using this method on embryonic stem cells led to the development of transgenic mice with targeted knocked out. It has also been possible to knock in genes or | 6,131,682 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
alter gene expression patterns.
If a vital gene is knocked out it can prove lethal to the organism. In order to study the function of these genes, site specific recombinases (SSR) were used. The two most common types are the Cre-LoxP and Flp-FRT systems. Cre recombinase is an enzyme that removes DNA by homologous recombination between binding sequences known as Lox-P sites. The Flip-FRT system operates in a similar way, with the Flip recombinase recognizing FRT sequences. By crossing an organism containing the recombinase sites flanking the gene of interest with an organism that expresses the SSR under control of tissue specific promoters, it is possible to knock out or switch on genes only | 6,131,683 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
in certain cells. This has also been used to remove marker genes from transgenic animals. Further modifications of these systems allowed researchers to induce recombination only under certain conditions, allowing genes to be knocked out or expressed at desired times or stages of development.
Genome editing uses artificially engineered nucleases that create specific double-stranded breaks at desired locations in the genome. The breaks are subject to cellular DNA repair processes that can be exploited for targeted gene knock-out, correction or insertion at high frequencies. If a donor DNA containing the appropriate sequence (homologies) is present, then new genetic material containing the transgene | 6,131,684 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
will be integrated at the targeted site with high efficiency by homologous recombination. There are four families of engineered nucleases: meganucleases, ZFNs, transcription activator-like effector nucleases (TALEN), the CRISPR/Cas (clustered regularly interspaced short palindromic repeat/CRISPRassociated protein (e.g. CRISPR/Cas9). Among the four types, TALEN and CRISPR/Cas are the two most commonly used. Recent advances have looked at combining multiple systems to exploit the best features of both (e.g. megaTAL that are a fusion of a TALE DNA binding domain and a meganuclease). Recent research has also focused on developing strategies to create gene knock-out or corrections without creating | 6,131,685 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
double stranded breaks (base editors).
## Meganucleases and Zinc finger nucleases.
Meganucleases were first used in 1988 in mammalian cells. Meganucleases are endodeoxyribonucleases that function as restriction enzymes with long recognition sites, making them are more specific to their target site than other restriction enzymes. This increases their specificity and reduces their toxicity as they will not target as many sites within a genome. The most studied meganucleases are the LAGLIDADG family. While meganucleases are still quite susceptible to off-target binding, which makes them less attractive than other gene editing tools, their smaller size still makes them attractive particularly | 6,131,686 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
for viral vectorization perspectives.
Zinc-finger nucleases (ZFNs), used for the first time in 1996, are typically created through the fusion of Zinc-finger domains and the "Fok"I nuclease domain. ZFNs have thus the ability to cleave DNA at target sites. By engineering the zinc finger domain to target a specific site within the genome, it is possible to edit the genomic sequence at the desired location. ZFNs have a greater specificity, but still hold the potential to bind to non-specific sequences.. While a certain amount of off-target cleavage is acceptable for creating transgenic model organisms, they might not be optimal for all human gene therapy treatments.
## TALEN and CRISPR.
Access | 6,131,687 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
to the code governing the DNA recognition by transcription activator-like effectors (TALE) in 2009 opened the way to the development of a new class of efficient TAL-based gene editing tools. TALE, proteins secreted by the Xanthomonas plant pathogen, bind with great specificity to genes within the plant host and initiate transcription of the genes helping infection. Engineering TALE by fusing the DNA binding core to the "Fok"I nuclease catalytic domain allowed creation of a new tool of designer nucleases, the TALE nuclease (TALEN). They have one of the greatest specificities of all the current engineered nucleases. Due to the presence of repeat sequences, they are difficult to construct through | 6,131,688 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
standard molecular biology procedure and rely on more complicated method of such as Golden gate cloning.
In 2011, another major breakthrough technology was developed based on CRISPR/Cas (clustered regularly interspaced short palindromic repeat / CRISPR associated protein) systems that function as an adaptive immune system in bacteria and archaea. The CRISPR/Cas system allows bacteria and archaea to fight against invading viruses by cleaving viral DNA and inserting pieces of that DNA into their own genome. The organism then transcribed this DNA into RNA and combined with Cas9 proteins to make double-stranded breaks in the invading viral DNA. By pairing Cas proteins with a designed guide RNA | 6,131,689 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
CRISPR/Cas9 could be used to induce double-stranded breaks at specific points within DNA sequences. It was later demonstrating that CRISPR/Cas9 could edit human cells in a dish. Although the early generation lacks the specificity of TALEN, the major advantage of this technology is the simplicity of the design. It also allows multiple sites to be targeted simultaneously, allowing the editing of multiple genes at once. CRISPR/Cpf1 is a more recently discovered system that requires a different guide RNA to create particular double-stranded breaks (leaves overhangs when cleaving the DNA) when compared to CRISPR/Cas9.
# See also.
- List of genetic engineering software: software to code the genetic | 6,131,690 |
37319629 | Genetic engineering techniques | https://en.wikipedia.org/w/index.php?title=Genetic%20engineering%20techniques | Genetic engineering techniques
be used to induce double-stranded breaks at specific points within DNA sequences. It was later demonstrating that CRISPR/Cas9 could edit human cells in a dish. Although the early generation lacks the specificity of TALEN, the major advantage of this technology is the simplicity of the design. It also allows multiple sites to be targeted simultaneously, allowing the editing of multiple genes at once. CRISPR/Cpf1 is a more recently discovered system that requires a different guide RNA to create particular double-stranded breaks (leaves overhangs when cleaving the DNA) when compared to CRISPR/Cas9.
# See also.
- List of genetic engineering software: software to code the genetic modifications | 6,131,691 |
37320563 | Simon Walker | https://en.wikipedia.org/w/index.php?title=Simon%20Walker | Simon Walker
Simon Walker
Simon Walker may refer to:
- Simon Walker (business) (born 1953), Director General of the Institute of Directors
- Simon Walker (composer) (born 1961), Australian composer
- Simon Walker (sailor), English yachtsman, adventurer and author
- Simon Walker (Hollyoaks), a character on the British soap opera "Hollyoaks"
- Simon Walker (historian) (1958–2004), English medievalist | 6,131,692 |
37320543 | Cardiff West Yard Locomotive Works | https://en.wikipedia.org/w/index.php?title=Cardiff%20West%20Yard%20Locomotive%20Works | Cardiff West Yard Locomotive Works
Cardiff West Yard Locomotive Works
West Yard Works was the Taff Vale Railway's locomotive repair and construction factory. It was located in Cardiff between Bute Street and the Glamorganshire Canal.
A small engine shed with room for one locomotive and a repair shop was built there when the railway was first constructed in 1839, but much of the work had to be carried out in the open air.
In 1846 Henry Clement was appointed as the railway's Resident Engineer and one of the first things he did was to have a locomotive works built there. In 1857 the first locomotive was built at the works, 'Venus' a small 2-4-0 passenger loco.
When the Taff Vale introduced the 0-6-2T type, which was to become | 6,131,693 |
37320543 | Cardiff West Yard Locomotive Works | https://en.wikipedia.org/w/index.php?title=Cardiff%20West%20Yard%20Locomotive%20Works | Cardiff West Yard Locomotive Works
ubiquitous across South Wales, the works traverser could not accommodate the longer wheelbase so locomotives had to have their trailing radial wheels removed while within the works.
The works could only be accessed by level crossings accessed by turntables on the main line near the railway's terminus at Bute Road station.
Shortage of space finally led Tom Hurry Riches, the railway's Locomotive Superintendent to suspend locomotive building by the company itself after the completion of the O1 class in 1897. However in 1903 a small steam engine unit for the company's first rail motor was built there, the carriage part being built at their Cathays Carriage and Wagon Works, however further rail | 6,131,694 |
37320543 | Cardiff West Yard Locomotive Works | https://en.wikipedia.org/w/index.php?title=Cardiff%20West%20Yard%20Locomotive%20Works | Cardiff West Yard Locomotive Works
motors were all built by outside contractors.
After the Great War there were plans to build a new works at Radyr but as the company was to amalgamate with the Great Western Railway and other South Wales companies in 1922 that plan was abandoned and the Great Western subsequently concentrated all major locomotive repair work in South Wales at the former Rhymney Railway's Caerphilly Works.
New workshops were constructed at Caerphilly and after their opening West Yard Works finally closed on 28 August 1926, the remaining workforce transferring to Caerphilly.
One locomotive built at West Yard has survived, as the last standard gauge loco built in Wales. No 28, an O1 class 0-6-2T is now a part | 6,131,695 |
37320543 | Cardiff West Yard Locomotive Works | https://en.wikipedia.org/w/index.php?title=Cardiff%20West%20Yard%20Locomotive%20Works | Cardiff West Yard Locomotive Works
plans to build a new works at Radyr but as the company was to amalgamate with the Great Western Railway and other South Wales companies in 1922 that plan was abandoned and the Great Western subsequently concentrated all major locomotive repair work in South Wales at the former Rhymney Railway's Caerphilly Works.
New workshops were constructed at Caerphilly and after their opening West Yard Works finally closed on 28 August 1926, the remaining workforce transferring to Caerphilly.
One locomotive built at West Yard has survived, as the last standard gauge loco built in Wales. No 28, an O1 class 0-6-2T is now a part of the National Collection, currently under restoration at the Gwili Railway. | 6,131,696 |
37320561 | Tales for the Midnight Hour | https://en.wikipedia.org/w/index.php?title=Tales%20for%20the%20Midnight%20Hour | Tales for the Midnight Hour
Tales for the Midnight Hour
Tales for the Midnight Hour is a series of scary children's books written by Judith Bauer Stamper. This anthology horror series served as the precursor to various other similar works, including "Scary Stories to Tell in the Dark" and "Scary Stories for Sleep-overs". Published by Scholastic's Point Horror banner, this popular series spawned 3 sequels and lasted from 1977-1991.
# Overview.
The series was written for a younger audience, but was told much darker than many books of the time. With the exception of the first volume, each book contained 13 stories, usually involving youths trying to find their way out of spooky/paranormal situations.
In 1992, the first | 6,131,697 |
37320561 | Tales for the Midnight Hour | https://en.wikipedia.org/w/index.php?title=Tales%20for%20the%20Midnight%20Hour | Tales for the Midnight Hour
Published by Scholastic's Point Horror banner, this popular series spawned 3 sequels and lasted from 1977-1991.
# Overview.
The series was written for a younger audience, but was told much darker than many books of the time. With the exception of the first volume, each book contained 13 stories, usually involving youths trying to find their way out of spooky/paranormal situations.
In 1992, the first book in the series was released on audiobook in cassette form.
The series has since been republished in 2005, with new cover art, in a 2-volume collection.
# Books.
# See also.
- Scholastic Books
- "Scary Stories to Tell in the Dark"
- "Scary Stories for Sleep-overs"
- "Short & Shivery" | 6,131,698 |
37320554 | Arthur Barnwell House | https://en.wikipedia.org/w/index.php?title=Arthur%20Barnwell%20House | Arthur Barnwell House
Arthur Barnwell House
The Arthur Barnwell House, which is also known as the Barnwell-DeCamps House, is a Queen Anne house in Greer, South Carolina that was built in the period 1880-1900. It was named to the National Register of Historic Places on 1992. As of 2013, the house was in the process of being moved and rebuilt at a new location.
# History.
The house was built for the first president of Pelham Manufacturing Company, Arthur Barnwell, on the banks of the Enoree River. The ruins of Pelham Mill and the Pelham mill village are on the opposite side of the river. It was believed to be built some time between 1880 and 1900.
It has been disassembled in ca 2015 and the parts are now forgotten | 6,131,699 |
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