Scientists decode TREX which could see new treatments for cancer realized

Public release date: 14-Aug-2012
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Contact: Paul Mannion
p.f.mannion@sheffield.ac.uk
01-142-229-851
University of Sheffield

Decoded process could hold the key to future treatments for a wide range of chronic health problems including Motor Neuron Disease, myotonic dystrophy and a wide range of cancers, University of Sheffield scientists have revealed.

Experts from the University's Department of Molecular Biology and Biotechnology, collaborating with scientists from Harvard Medical School in the USA, have revealed how a complicated set of proteins called TREX act as a passport for the transfer of cell blueprints which create proteins that are essential for life.

The researchers believe their better understanding will mean they can ultimately fix problems in the process which cause fatal health conditions like Motor Neuron Disease, myotonic dystrophy and a wide range of cancers.

Professor Stuart Wilson, who led the groundbreaking project, said: "Protein production is an essential part of life for all organisms. This process involves reading the code in genes and converting this to a message which is ultimately decoded to make a protein.

"The message is made of a special molecule called mRNA. In all organisms from yeast through to man the mRNA is made in a compartment in the cell called the nucleus, but then has to be transported to a separate compartment, called the cytoplasm, where it is decoded to make a protein. This process of mRNA transport is essential for life and when it malfunctions in humans it can lead to diseases such as Motor Neuron Disease or cancer."

Researchers have shown how proteins called TREX provide a mark on the mRNA which acts as the passport which unlocks a transporter protein called Nxf1, allowing it to land on the mRNA and transport it to the cytoplasm to create proteins.

Professor Wilson added: "When a car is built in a factory, it goes through various stages of production with parts being added and, hopefully, quality control checks before it is driven out of the factory and into the salesroom.

"Similarly mRNA goes through various modifications in the nucleus, with bits added on and other sections removed. Only when the mRNA reaches the end of the production line in the nucleus and passes all the quality control checks is it given the passport which allows it to be transported to the cytoplasm and then used to make proteins.

"Until now, it has not been clear how the cell knows when the mRNA has reached the end of the production line in the nucleus and thus when it should be given a passport allowing passage to the cytoplasm. Now we have identified how the passport is issued, allowing the mRNA transport and the production of proteins in the cell which is essential for life."

###

Notes to editors:

The University of Sheffield

With nearly 25,000 students from 125 countries, the University of Sheffield is one of the UK's leading and largest universities. A member of the Russell Group, it has a reputation for world-class teaching and research excellence across a wide range of disciplines. The University of Sheffield has been named University of the Year in the Times Higher Education Awards for its exceptional performance in research, teaching, access and business performance. In addition, the University has won four Queen's Anniversary Prizes (1998, 2000, 2002, and 2007).

These prestigious awards recognise outstanding contributions by universities and colleges to the United Kingdom's intellectual, economic, cultural and social life. Sheffield also boasts five Nobel Prize winners among former staff and students and many of its alumni have gone on to hold positions of great responsibility and influence around the world. The University's research partners and clients include Boeing, Rolls Royce, Unilever, Boots, AstraZeneca, GSK, ICI, Slazenger, and many more household names, as well as UK and overseas government agencies and charitable foundations.

The University has well-established partnerships with a number of universities and major corporations, both in the UK and abroad. Its partnership with Leeds and York Universities in the White Rose Consortium has a combined research power greater than that of either Oxford or Cambridge.

For further information please contact: Paul Mannion, Media Relations Officer, on 0114 2229851 or email P.F.Mannion@sheffield.ac.uk

To read other news releases about the University of Sheffield, visit: http://www.shef.ac.uk/news

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AAAS and EurekAlert! are not responsible for the accuracy of news releases posted to EurekAlert! by contributing institutions or for the use of any information through the EurekAlert! system.

Public release date: 14-Aug-2012
[ | E-mail | Share ]

Contact: Paul Mannion
p.f.mannion@sheffield.ac.uk
01-142-229-851
University of Sheffield

Decoded process could hold the key to future treatments for a wide range of chronic health problems including Motor Neuron Disease, myotonic dystrophy and a wide range of cancers, University of Sheffield scientists have revealed.

Experts from the University's Department of Molecular Biology and Biotechnology, collaborating with scientists from Harvard Medical School in the USA, have revealed how a complicated set of proteins called TREX act as a passport for the transfer of cell blueprints which create proteins that are essential for life.

The researchers believe their better understanding will mean they can ultimately fix problems in the process which cause fatal health conditions like Motor Neuron Disease, myotonic dystrophy and a wide range of cancers.

Professor Stuart Wilson, who led the groundbreaking project, said: "Protein production is an essential part of life for all organisms. This process involves reading the code in genes and converting this to a message which is ultimately decoded to make a protein.

"The message is made of a special molecule called mRNA. In all organisms from yeast through to man the mRNA is made in a compartment in the cell called the nucleus, but then has to be transported to a separate compartment, called the cytoplasm, where it is decoded to make a protein. This process of mRNA transport is essential for life and when it malfunctions in humans it can lead to diseases such as Motor Neuron Disease or cancer."

Researchers have shown how proteins called TREX provide a mark on the mRNA which acts as the passport which unlocks a transporter protein called Nxf1, allowing it to land on the mRNA and transport it to the cytoplasm to create proteins.

Professor Wilson added: "When a car is built in a factory, it goes through various stages of production with parts being added and, hopefully, quality control checks before it is driven out of the factory and into the salesroom.

"Similarly mRNA goes through various modifications in the nucleus, with bits added on and other sections removed. Only when the mRNA reaches the end of the production line in the nucleus and passes all the quality control checks is it given the passport which allows it to be transported to the cytoplasm and then used to make proteins.

"Until now, it has not been clear how the cell knows when the mRNA has reached the end of the production line in the nucleus and thus when it should be given a passport allowing passage to the cytoplasm. Now we have identified how the passport is issued, allowing the mRNA transport and the production of proteins in the cell which is essential for life."

###

Notes to editors:

The University of Sheffield

With nearly 25,000 students from 125 countries, the University of Sheffield is one of the UK's leading and largest universities. A member of the Russell Group, it has a reputation for world-class teaching and research excellence across a wide range of disciplines. The University of Sheffield has been named University of the Year in the Times Higher Education Awards for its exceptional performance in research, teaching, access and business performance. In addition, the University has won four Queen's Anniversary Prizes (1998, 2000, 2002, and 2007).

These prestigious awards recognise outstanding contributions by universities and colleges to the United Kingdom's intellectual, economic, cultural and social life. Sheffield also boasts five Nobel Prize winners among former staff and students and many of its alumni have gone on to hold positions of great responsibility and influence around the world. The University's research partners and clients include Boeing, Rolls Royce, Unilever, Boots, AstraZeneca, GSK, ICI, Slazenger, and many more household names, as well as UK and overseas government agencies and charitable foundations.

The University has well-established partnerships with a number of universities and major corporations, both in the UK and abroad. Its partnership with Leeds and York Universities in the White Rose Consortium has a combined research power greater than that of either Oxford or Cambridge.

For further information please contact: Paul Mannion, Media Relations Officer, on 0114 2229851 or email P.F.Mannion@sheffield.ac.uk

To read other news releases about the University of Sheffield, visit: http://www.shef.ac.uk/news

NR4049/ends

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AAAS and EurekAlert! are not responsible for the accuracy of news releases posted to EurekAlert! by contributing institutions or for the use of any information through the EurekAlert! system.

Source: http://www.eurekalert.org/pub_releases/2012-08/uos-sdt081012.php

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Success of engineered tissue depends on where it's grown

ScienceDaily (Aug. 14, 2012) ? Cells grown on different types of scaffolds vary in their ability to help repair damaged blood vessels. Tissue implants made of cells grown on a sponge-like scaffold have been shown in clinical trials to help heal arteries scarred by atherosclerosis and other vascular diseases. However, it has been unclear why some implants work better than others.

MIT researchers led by Elazer Edelman, the Thomas D. and Virginia W. Cabot Professor of Health Sciences and Technology, have now shown that implanted cells' therapeutic properties depend on their shape, which is determined by the type of scaffold on which they are grown. The work could allow scientists to develop even more effective implants and also target many other diseases, including cancer.

"The goal is to design a material that can engineer the cells to release whatever we think is most appropriate to fight a specific disease. Then we can implant the cells and use them as an incubator," says Laura Indolfi, a postdoc in Edelman's lab and lead author of a paper on the research recently published online in the journal Biomaterials.

Aaron Baker, a former postdoc in Edelman's lab and now an assistant professor at the University of Texas at Austin, is also an author of the paper.

Shape matters

For the past 20 years, Edelman has been working on using endothelial cells grown on scaffolds made of collagen as implantable devices to treat blood vessel damage. Endothelial cells line the blood vessels and regulate important process such as tissue repair and inflammation by releasing molecules such as chemokines, small proteins that carry messages between cells.

Several of the devices have been tested in clinical trials to treat blood vessel damage; in the new Biomaterials study, Edelman and Indolfi set out to determine what makes one such tissue scaffold more effective than another. In particular, they were interested in comparing endothelial cells grown on flat surfaces and those grown on more porous, three-dimensional scaffolds. The cells grown on 3-D structures tended to be more effective at repairing damage and suppressing inflammation.

The researchers found that cells grown on a flat surface take on a round shape in which the cells' structural components form a ring around the perimeter of the cell. However, when cells are grown on a scaffold with surfaces of contact whose dimensions are similar in size to the cells, they mold to the curved surfaces, assuming a more elongated shape. In those cells, the structural elements -- made of bundles of the protein actin -- run parallel to each other.

Those shapes determine what types of chemokines the cells secrete once implanted into the body. In this study, the researchers focused on a chemokine known as MCP1, which recruits inflammatory cells called monocytes.

They found that the architecture of the cytoskeleton appears to determine whether or not the cell turns on the inflammatory pathway that produces MCP1. The elongated cells grown on porous surfaces produced eight times less of this inflammatory chemokine than cells grown on a flat surface, and recruited five times fewer monocytes than cells grown on a flat surface. This helps the tissue implants to suppress inflammation in damaged blood vessels.

The researchers also identified biomarkers that correlate the cells' shape, chemokine secretion and behavior. One such parameter is the production of a focal adhesion protein, which helps cells to stick to surfaces. In cells grown on a flat surface, this adhesion protein, known as vinculin, accumulates around the edges of the cell. However, in cells grown on a 3-D surface, the protein is evenly distributed throughout the cell. These distribution patterns serve as molecular cues to inhibit or activate the pathway that recruits monocytes.

Precise control

The findings could help scientists manipulate their scaffolds to tailor cells to specific applications. One goal is using implanted cells to recruit other body cells that will do a particular task, such as inducing stem cells to differentiate into a certain type of cell. "By designing the matrix before we seed the cells, we can engineer which factors they are going to secrete," Indolfi says.

The work should also help researchers improve on existing tissue-engineered devices and test new ones, Edelman says. "Without this kind of understanding, we can't extend successful technologies to the next generation," he says.

The research was funded by the National Institutes of Health, the American Heart Association, and the David H. Koch Foundation.

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Story Source:

The above story is reprinted from materials provided by Massachusetts Institute of Technology, via EurekAlert!, a service of AAAS. The original article was written by Anne Trafton.

Note: Materials may be edited for content and length. For further information, please contact the source cited above.

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Journal Reference:

1. Laura Indolfi, Aaron B. Baker, Elazer R. Edelman. The role of scaffold microarchitecture in engineering endothelial cell immunomodulation. Biomaterials, 2012; 33 (29): 7019 DOI: 10.1016/j.biomaterials.2012.06.052

Note: If no author is given, the source is cited instead.

Disclaimer: This article is not intended to provide medical advice, diagnosis or treatment. Views expressed here do not necessarily reflect those of ScienceDaily or its staff.

Source: http://feeds.sciencedaily.com/~r/sciencedaily/most_popular/~3/HjBDE6H73pc/120814111001.htm

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Video: Saving Detroit?s dogs

White House feels heat to waive ethanol mandate

President Barack Obama announced emergency measures to ease the impact of the worst drought in half a century, but stopped short of waiving the government?s requirement that a large portion of the now-shriveled corn crop be diverted to make ethanol.

Source: http://www.msnbc.msn.com/id/40153870/vp/48653709#48653709

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New Afghan police attack on NATO forces; no deaths

KABUL, Afghanistan (AP) ? The new U.S. ambassador to Afghanistan called a sudden rash of attacks on international forces by their Afghan partners "troubling" Monday, after an Afghan policeman opened fire on NATO forces in the fifth such assault in a week.

No international service members were killed in the latest attack. A Taliban spokesman claimed responsibility for the shooting in the eastern province of Nangarhar, claiming the attacker was a police officer who had been in contact with insurgents before the assault.

A spike in so-called "green-on-blue" attacks, in which Afghan security forces or attackers wearing their uniforms turn their guns on coalition troops, has raised concerns as NATO aims to turn over control for security to Afghan forces in a little more than two years.

"It's obviously very troubling, not just to us, but it's also very troubling to our Afghan partners," U.S. Ambassador James Cunningham told reporters in the capital in his first public comments after taking over the post earlier in the day.

"There's a lot of work being done to understand why this is happening," Cunningham said. He said it was not clear if all of the attackers were Taliban infiltrators, but noted that the strikes still threaten the "confidence and trust" needed for the two military forces to work together.

"Obviously this undermines or attacks that confidence and trust," Cunningham said.

Taliban insurgents are eager to exploit any such rift.

The trend also raises renewed concern that insurgents may be infiltrating the Afghan army and police, despite intensified screening.

At least seven American service members have been killed in the past week by either their Afghan counterparts or attackers wearing their uniforms.

NATO spokesman Charlie Stadtlander said an initial investigation indicated that Monday's attacker was an Afghan police officer, though the man was wearing civilian clothes.

He said there were no NATO deaths but would not say if any international service members were wounded in the attack, citing coalition policy.

At least one Afghan intelligence agent was wounded in Monday's shooting, according to according to Ahmad Zia Abdulzai, spokesman for the Nangarhar provincial governor.

"The shooter has escaped, and Afghan security forces are looking for him," Abdulzai said.

Taliban spokesman Zabiullah Mujahid issued a statement Monday afternoon praising the shooting. He said the police attacker had been "waiting for such an opportunity to attack" international forces.

Mujahid said three Americans were killed, though the insurgents often exaggerate the results of their attacks and are quick to adopt any incident that allows them to claim support among Afghans.

The coalition on Monday sought to dispel any idea that the rising number of turncoat attacks signals any shift in public sympathy toward insurgents, increased Taliban infiltration or growing resentment toward the mostly American coalition forces.

Insurgents were behind only about 10 percent of this year's reported green-on-blue shootings, a NATO spokesman said, citing investigations into attacks before those of the past week.

"Obviously, the Taliban want to take credit for these things, but the fact of the matter is the majority of these attacks result from individual grievances, cultural misunderstandings or personal or battle stress," said James Graybeal, deputy political affairs officer for the coalition.

He insisted the deadly violence is relatively small scale.

"We're talking about 31 individual bad actors out of a total of almost 340,000 (Afghan security forces)," he said.

Coalition officials say the attacks have not impeded plans to hand over security to Afghan forces ? which are supposed to reach a strength of 352,000 in a few months ? by the end of 2014.

Green-on-blue attacks are on the rise. So far this year, 34 coalition troops have been killed in 27 attacks, compared to 11 attacks and 20 deaths in 2011, according to an Associated Press count, and five attacks in each of the previous two years.

Six died in two separate attacks on Friday in different areas of the volatile southern province of Helmand.

NATO has said both attackers have been detained, though it has released little information about the shootings, and accounts from other officials differ.

Also Monday, Afghan President Hamid Karzai appointed his former defense minister, Abdul Rahim Wardak, as one of his senior advisers. The move came after parliament voted earlier in the month to dismiss Wardak over allegations of corruption in his ministry, as well as failure to prevent cross-border shelling from Pakistan.

___

Associated Press writer Rahim Faiez contributed to this report.

Source: http://news.yahoo.com/afghan-police-attack-nato-forces-no-deaths-134743662.html

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Rebel video claims to show captured Syrian pilot

BEIRUT (AP) ? Syrian rebels circulated dramatic video Monday of what they claimed was the downing of a warplane and armed men later holding the captured pilot who ejected as the MiG fighter was engulfed by flames. Syria acknowledged a pilot bailed out of a disabled plane but blamed the crash on a technical malfunction.

The authenticity of the images or the claims could not be independently verified. If the rebels did bring down their first aircraft, that could signal a significant jump in their firepower and give opposition forces their most high-profile military captive.

But wider questions remain even if the rebel reports are confirmed, including whether this could be just a one-time blow against expanding air offensives by the forces of Bashar Assad's regime. Just days ago, protesters across Syria pleaded for the rebels' main backers ? including Turkey and Gulf states ? to send anti-aircraft weapons for outgunned fighters.

Assad's military has significantly stepped up aerial attacks in recent weeks. Strafing from warplanes and close-range missile strikes from helicopter gunships have pushed back rebels in key fronts such as Aleppo, the country's largest city and the scene of fierce attacks to dislodge rebel positions.

As the sun was setting on Monday, an Associated Press reporter saw two fighter jets over the village of Marea, 30 kilometers (20 miles) north of Aleppo.

Terrified residents collected on street corners and near the doors to their houses to watch and point as the jets dived low, dropping bombs that sent up clouds of smoke and firing machine guns that crackled over the village.

On one crowded market street, a handful of rebels with rifles ran toward the site of the bombings.

"What are you going to do, bring down a jet with a rifle?" a man screamed.

After the jets left, young men on motorcycles rushed to the bombing site on the edge of the village to find two craters the size of cars in a dirt field next to a swimming pool.

A man working at the privately owned pool said only three people were there at the time and that none was injured. He didn't give his name and tried to chase away journalists and residents seeking to film the pool.

It is unclear why the area was targeted. Residents said there was no rebel base nearby.

"In the summer, it's hot so the guys gather here to swim," said Abdullah Najjar, 21, adding that some of them could have been rebels.

"This is the only place we have in town to come for entertainment."

Nationwide, the relentless bloodshed ? including alleged massacres by pro-regime mobs and retaliation killings by rebels ? has already claimed more than 20,000 lives, activists say, and will be further examined in a report expected Wednesday by the U.N. Human Rights Council's independent commission probing abuses in Syria.

In another crack in Assad's diplomatic corps, a Syrian diplomat who worked with the U.N. rights council in Geneva said he left his post to join the opposition. A spokesman for the council, Rolando Gomez, identified the Syrian as Danny al-Baaj and described him as a junior member of his country's U.N. mission. Syria is not a member of the 47-nation council, but al-Baaj worked with it as part of his duties.

The claims of bringing down the warplane and capturing the pilot, meanwhile, are likely to become a key propaganda tool to rally rebel fighters.

Activists released a video which they say showed a government Soviet-made MiG warplane catching fire after it was hit by ground fire over Deir el-Zour province, an area near the Iraqi border where the opposition has strongholds. Hours later, another video shown on the pan-Arab network Al-Arabiya purported to show the captured pilot surrounded by armed rebels. "Introduce yourself," says another speaker with his back to the camera.

The alleged captive identified himself as Col. Rafik Mohammed Suleiman and says he was on a mission to attack a rebel-held area.

"What do you tell the officers of the Assad army?" the speaker asks the man, whose beard and hair are flecked with gray. The man who identified himself as the pilot urges them to defect.

The speaker ? whose face remained hidden ? said the hostage will be treated according to tenets of Islam and the Geneva Convention. The later reference could be an indirect reply to recent international outrage over videos posted on the Internet claiming to show summary executions and torture by rebels, including bloodied prisoners being gunned down against a wall as people cried: "Free Syrian Army Forever!"

Syria's state-run SANA news agency said the pilot ejected from a warplane after a technical failure during a "training mission." It added that a search was under way to find the pilot.

The Britain-based activist group Syrian Observatory for Human Rights said the plane was hit as it was conducting air raids on the town of Muhassan, which is close to a military airfield. The group quoted activists in the area as saying the plane was hit with fire from a heavy machine gun used by rebels in the area.

Syria has such anti-aircraft weapons in its arsenal and it's possible that some could have fallen into rebel hands. In June, Syria said it used anti-aircraft machine gun fire to bring down a Turkish F-4 Phantom fighter jet it claimed crossed into Syrian aircraft. Turkey said the plane was in international flight zones.

Theodore Karasik, a regional security expert at the Dubai-based Institute for Near East and Gulf Military Analysis, said the rebel claim ? if true ? would suggest a stepped-up flow of outside military assistance. Older anti-aircraft weapons, possibly looted from Syrian arsenals such as variations on the Soviet-era SA-7, are considered overmatched by later model MiGs in the Syrian air force.

"If this is true, the conjecture would be that covert aid to the rebels is expanding with higher-grade anti-aircraft capabilities," Karasik said.

Gulf states such as Saudi Arabia and Qatar have pledged to help supply the Syrian rebels, but the extent of the aid is unclear. Rebel commanders have consistently complained about lack of ammunition, including during recent battles in Aleppo. Last week, anti-regime protesters across Syria staged rallies calling for greater anti-aircraft firepower.

The possibility of a high-ranking military captive also could raise pressure on Assad's regime after a series of abductions, including 48 Iranians taken earlier this month and 11 Lebanese Shiites seized in May.

Rebels claim the Iranians include members of Tehran's Revolutionary Guard and were on a "reconnaissance mission" in Damascus. Iran insists the men were on a religious pilgrimage. The Lebanese are apparently held to try to pressure the government in Beirut to show greater support for the rebels ? which is unlikely because of the strong influence of the Lebanese militant group Hezbollah, which backs Assad.

On Sunday, the head of Syria's main opposition group in exile renewed a plea for international powers to impose a no-fly zone in border areas to protect civilians who are coming under increasingly intense attacks by regime warplanes and helicopters.

The site of the plane incident ? the eastern, oil-rich region of Deir el-Zour ? has been the site of heavy clashes between government troops and rebels over the past week. Syrian Prime Minister Riad Hijab, who defected to Jordan earlier this month, comes from the area.

Besides the rising death tolls in clashes ? more than 50 on Monday according to the activist Observatory group ? at least three Syria-based journalists have been killed in recent days. Al-Ikhbariya TV also said two other journalists and their driver are being held by rebels near the capital Damascus.

Source: http://news.yahoo.com/rebel-video-claims-show-captured-syrian-pilot-161712920.html

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What Makes E-Commerce Website Development Crucial for Online ...

Article by Joseph Martin

What Makes E-Commerce Website Development Crucial for Online Business ? Web Development

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The burgeoning popularity of online trading has completely changed the business scenario of the era. With millions of service providers vying for prospects across the web, catching the attention of online visitors and holding them for long and turning them into buyers have become a challenging task for every e-business owner. Thanks to the e-commerce website development that has simplified the problem to a large extent. A website needs to be highly functional and professionally developed to hold a successful business promotion and visitor satisfaction. E-commerce development has made it easier for the webmasters to keep balance between their online business and target audience.

Two websites meant for the same online business may not achieve the same success. If you analyze the reasons, you can feel that website design has a major role in the success of an e-business. An ordinarily designed website can never solve the demands of modern marketing needs. It needs a professionally and strategically designed website to penetrate the world of online business and create a strong platform for the company to enlarge its customer base. The online shoppers have millions of options to choose from across the web, but it is the design of the website that can hold their attention and influence their buying decision. E-commerce website development plans serve the basic purpose of the e-business and make a lot of difference in the success of the business.

Strategic Design for Target Audience

Relevant visitors are crucial for any online business. However, it is the design on a website that attracts or diverts online traffic to it. If a website or e-business has right kind of e-commerce website development, it can get faster user attention than its competitors. Different types of online businesses have different requirements considering their audience. If a site is designed in a user-friendly way, it will provide a hassle-free shopping experience for the online buyers. Detail information regarding services and shopping, easy navigation structure, handy payment modes, etc helps a lot in the shopping decision of the customers. These factors are considered carefully in the e-commerce website development to give a boost to the number of visitors to a site.

Design to Fit the SEO Plans for the Business

Unless a website is properly promoted across the web, it?s difficult to get visibility as well as visitors to the site. However, it needs a carefully designed site so that it can fit into the SEO plans to market the online business. These days, the search engines have changed their algorithms and giving preference to sites that are useful in all respect for the visitors. In short, a website designed in a well-planned and SEO-friendly way can receive better attention.

It is difficult to meet the demands of online marketing without having all round approach for the e-business. That?s why E-commerce website development has increased with it popularity as well as functionality. There are various aspects of e-commerce development and webmasters can select the site design based on the kind of business they have or their requirements from the site.

About the Author

Joseph Martin is a web developer who writes useful articles on Web design, Web development, CMS website design, E-commerce website development and Iphone applications development. For more information on CMS website design and E-commerce development solutions, you can also visit http://www.ukwebsitesdesigner.com.

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Source: http://www.citizensmith2010.com/2012/08/what-makes-e-commerce-website-development-crucial-for-online-business/

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