Enron Scandal Essay

Accounting scandals are political or business scandals which arise with the disclosure of financial misdeeds by trusted executives of corporations or governments. These days, not too often, these scandals are splashed as headlines across media. Why? Because there are complex groups of stakeholders who might be seriously affected by the scandals. Enron scam was the most remarkable scandal in 20 centuries by their institutionalized, systematic, and creatively planned accounting fraud. The scandal also brought into question the accounting practices and activities of many corporations in the United States. The scandal also affected the greater business world by causing the dissolution of the Arthur Andersen accounting company. This report will reveal the whole story of Enron scam and auditors role from the situation in more depth. It will also include the current situation of those responsible for the fraud. 2. Background of U. S in 1990s The 1990s of U. S were a time of prosperity and this prosperity period is originated from growth in IT corporates and in accordance with increase in productivity as technology develops with IT. In the mid- to late-90s, society’s expectations of what the Internet could offer were unrealistic. Many investors foolishly ignored the fundamental rules of investing in the stock market and instead, investors and entrepreneurs became preoccupied with new ideas that were not yet proven to have market potential. Furthermore, they ignored the blatant signs that the bubble was about to burst. (IT Bubble in 90s) There are two main reasons for the bubble; with investors` optimistic expectation, there were a lot of ‘Window dressing’ in accounting. As Enron collapsed, the growth of the tech sector proved to be illusory, and bubble started to sober. 3. Enron- who are they? Enron Corporation is an energy trading, communications company which was formed in 1985 based in Houston, Texas. Enron marketed electricity and natural gas, delivered energy and other physical commodities. Company branched into many non-energy-related fields as well, including such areas as high-speed Internet bandwidth, and financial and risk management with 21,000 employees at mid-2001. The company reported revenues of $101 billion in 2000. It has stakes in nearly 30,000 miles of gas pipeline, owns or has access to a 15,000mile fiber optic network and has a stake in electricity generating operations around the world. 4. Enron- what they did? The Enron fraud case is extremely complex. There are several main characters who are spearheading the Enron scam. One of them was Jeff Skillng. Jeff Skillng, who was a consultant in Mckinsey, took charge in consulting Enron. He found problems and also potentiality of Enron and he proposed the idea of â€Å"gas bank† to Enron, which is a system that is combining financial system and gas supply and demand system, and taking the margin from the two system(as bank does). This was an ideal idea in theory and Enron asked him to take charge for this business, and later he became the president of Enron’s trading operations. Also, he convinced federal regulators to permit Enron to use an accounting method known as â€Å"mark to market. † Using this method allowed Enron to count projected earnings from long-term energy contracts as current income. This was money that might not be collected for many years. It is thought that this technique was used to inflate revenue numbers by manipulating projections for future revenue. The problem is that it doesn`t match realised profit and real cash flow. especially, Enron`s main trading was long term future contract which is hard to make valuation for the future. Use of these techniques made it difficult to see how Enron was really making money. The numbers were on the books so the stock prices remained high, but Enron wasn’t paying high taxes. When the telecom industry suffered its first downturn, Enron suffered as well. Eventually, the house of cards began falling. When Enron’s stock began to decline, the Raptors began to decline as well. On August 14, 2001, Enron’s CEO, Jeff Skilling, resigned due to â€Å"family issues. † Enron chairman Ken Lay stepped in as CEO. 5. One example of fraud Enron`s â€Å"too much Off-Balance Sheet Transactions† Enron used â€Å"off-balance-sheet†technique for anytime, for many purposes, because it would enable Enron to present itself more attractively as measured by the ratios favored by analysts and investors. Skillng used securitisation to supply more liquidity and also to clean up the assets that is hardly generating income from it. He also hided most of the debt by securitisation. So, Enron needed Special purpose entities(SPE) for the securitization purpose. JEDI was one of the SPE. California PERS and Enron invested by $25000m each. As soon as the JEDI established, Enron started to sell energy related stock to JEDI and it grew JEDI by 23% per year on average. It made Skillng`s ECT business bigger and bigger. 6. Consequences for the stakeholders The key stakeholders affected by the collapse of Enron were its employees and retirees. Stakeholders and mutual funds investors lost $ 70billion market value. Banks were also affected by the meltdown of the company. Not only the stakeholder and bondholder lose out, the confidence in the company also fell. This was the major setback for the company. The actions of Enron management left a deep scare for its 4000 employees which lost out their jobs and also impacted others around them. Some blamed Arthur Andersen; Enron’s accounting firm and some blame the board of directors for insufficient oversights. The damage was so big that it was likely to take years for the court to sort the wreckage. The company did not think of its future and took many bad steps just to earn money. The CEO should have looked into the company matters long time ago and took action so that hundreds of jobs could have been saved. The companies who were associated with the big firm were affected on a very large scale. This was the biggest bankruptcy of a firm with $63. 4 billion in assets. 7. Auditors in this scandal, and their role The external auditing body of Enron company was Arthur Anderson LLP, formerly one of the ‘big 5’ accounting firms, providing auditing, tax, and consulting services to large corporations. Andersen definitely Knew Enron Was in Trouble but they overlooked at it and even conspired with Enron to manipulate the financial statements. They knew Enron was in trouble as early as Feb.  2001, a company memo showed, and Andersen debated dropping the collapsed energy firm all together, Reuters reported. Additionally, Andersen knew in mid-August of a senior Enron employee’s concerns about improprieties in the energy company’s accounting practices. Andersen confirmed that a memo dated Feb. 6 recounted a meeting between Andersen executives about whether Andersen should retain the now-bankrupt Enron as a client. Auditors are responsible directly under the law especially the international standards to report directly to the shareholders on the status of the company’s or a bank’s account at a particular point in time. They heavily misconducted as auditors as they received money and hided about Enron`s truth. 8. Ramifications It is not easy to implement rigorous standards without changing Incentives. This situation can be seen in South East Asian countries like: Singapore, Thailand, Hong Kong, China. Each country can implement its own accounting standards, but did not implement the substantial institutional changes required to make these standards effective. According to various studies conducted in this area, new standards did not result in better-quality financial reporting.9. Conclusion: In search of better standards and ethics The ENRON Scandal is considered to be one of the most notorious within American history; an unofficial blueprint for a case study on White Collar crime. Enron’s behaviour has confirmed that the treatment of off-balance-sheet dodge, American accounting standards are too lax. It is time for another effort to realign the system to function more in shareholders’ interests. Companies need stronger non-executive directors, paid enough to devote proper attention to the job.

Marooned in Iraq Essay

Marooned in Iraq is a film by Bahman Ghobadi set on the border of Iran and Iraq. Wherein people who lives in Kurdistan experiences a precarious life. The Kurds experiences how it feels to live in a cold, sleazy camp for refugees near Iran’s border. The cruelty of Saddam Hussein was greatly felt by the loved ones who remained alive. They were the ones who are left facing the daily deluge brought about by Iraq’s bomber jets that are streaking across the sky. However this film is made   more meaningful by Ghobadi by having a mixture of laughter and tears. This made the film more appealing to the viewers and more enjoyable to watch.   Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚   The movie revolves on the life of a Kurdish family who are musically inclined. It is lead by the elder Kurdish whose name is Mirza who is in quest for the woman who was gone them two decades before. He was left by his wife and goes with another man to another place. Mirza then decided to take courage and travel beyond the borders of Iran together with his two sons Barat and Audeh.   They experienced great squabbling among themselves but their interaction is really humorous. The humor added to lessen the tension of the scenes makes up a light and enjoyed viewers. Some example of this is when they are caught in a troubled situation like when their motorcycles are stolen by some bandits.   Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚   There are metaphorical and symbolisms used by Ghobadi on the film Marooned in Iraq. He carefully unveils the situation of the Kurdish people under the regime of Saddam Hussein. The film was set during 189 and 1991 where Kurdish are in great depression under Hussein. Many lives were killed and families whoa re left behind greatly suffered. The Iranian revolution is the time when Barat was left by his wife. In the film there is no clear image given about the Kurdish wife. This implies how women during that revolution have been robbed of their humanity. Women singers like Hannareh (Mirza’s wife) are silenced and are not capable of speaking of themselves. The title itself of thefilm symbolizes how the Kurdish people were isolated during that time. Chaotic, traumatic and harsh experienced under the claws of Hussein. On the other hand, there are some exemptions on being Marooned in Iraq this is because some of the characters in the film do find their ways. A ray of hope is still visible in the film which gives the Kurdish people a situation where they can think and do ways to reach lberty.   Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚   As the pursuit of Mirza and his two sons Barat and Audeh continues for Hannareh, they came to learn and understand more the life of being a Kurdish. Through this characters Ghobadi tries to expose the situation during that time. The three lead characters witnessed the frightful devastation received by Iraq from both U.S bombs and Saddam Hussein’s bloody troops. To lighten up the situation Ghobadi uses an approach of adding up humor and beauty along the journey of the three characters. This made the viewers understand more about Kurdish identities. How strength and determination allows the characters to stand amidst the adversities in their way.   Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚   Ghobadi made the film to make a distinction about the Kurdish people. The diversity of the characters in dealing with the situation and how the essence of family floats amidst conflicts and depression. Mirza portaryed his father image by commanding his two sons to go with him for his search of his wife.   Barat shows willingness to cooperate with his father but but Audeh made some complaints because of leaving behind his 7 wives and 11 daughters. This shows the lifestyle of some Kurdish male who settles for many families. But sooner in the film Audeh decided to go withhis father and brother because it is an opportunity for him to find another wife who will bear him a son. This depicts the fact how Kurdish values having a male born offspring in their families. The director used the idea of making the characters in the film portray being musicians to lighten the atmosphere in exposing the situation of the country in that time. While the three are set to travel and does performances along the way, they came across victims of chemical warfare. They also experienced being abducted by thieves who buried them alive upto their necks as their motorcycle and warm clothes are stolen. Some practices were also exposed by the director in the film like the scene when the lead characters came to attend an Iranian wedding. Later on the two sons finds their solace when Barat met a woman he constantly sees in the film but this grieving woman is always being stopped by the police and forced to do some military services. Audeh on the other hand no longer finds his supposedly 8th wife but he was able to find some orphans who will make nice sons. Mirza was also able to find his own surprise and his two sons made their happy return to their home.   Ã‚   The film is set in a rhythmic and dramatic mode that made it more interesting. This is a story of reality with some moments of humor to break the harsh reality being experienced that time. Although villages have been scourged by Saddam Hussein during his reign of terror, the people are resilient enough to find their ways of surviving.

Solar Energy Research Paper Example | Topics and Well Written Essays - 500 words - 1

Solar Energy - Research Paper Example This means that there is no limit to the amount of solar energy that can be captures since the energy itself is renewable. The maintenance costs of solar power are also quite low compared to other forms of energy (Thomas 11). It is also quite easy to install solar panels which are available at relatively low costs. Solar energy can also be used in any setting, be it at home or an industrial setting. Another main pro of solar energy is its reliability. This is an energy source that will never be affected by geographical, economic or political upheavals (13). One of the major disadvantages of solar power is the fact that it is only available in places which have lots of sunshine. People who come from countries that are usually cold throughout cannot get as much solar energy as they would need for their daily use. Another disadvantage of solar energy is that it can only be captured during the day when there is enough sunlight. This means that one needs to have other forms of energy storage devices with which to store solar energy that has been captured during the day (Benduhn 34 and Thomas 22). Today, solar power is being used for many different purposes. It is used for domestic purposes that include heating, cooking, lighting, communications, operating spacecraft many others. People are turning to solar power as an alternative source of energy for use both at home and in industries. Solar panels are the most commonly used solar energy capturing equipment. In many cases, solar energy is being used as a source of backup energy in case of power failure. Healthcare facilities are embracing the use of solar energy in emergency situations. Production industries are also slowly opening up to the idea of using solar power for their everyday operations. The future does look brilliant for solar energy. More and more people are becoming aware of the effects that fossil fuels have on the environment and they are turning to the safe

Global Communications Gap Analysis Research Paper

Global Communications Gap Analysis - Research Paper Example That is, a particular organization will not be successful due to failed organizational strategies, decision-making process, etc, as well as the inept functioning of the employees. In order to correct these mistakes and to put the organization on the successful path, effective at the same time strong decisions have to be taken, without any impassiveness. One of the strong and impassive decisions should layoff of the employees, who are expendable. Although this decision will be a difficult one to take, both from the organizational and ethical point of view, it has to be taken for organizational well being. From employees’ perspective, they can be taken care, by finding other alternate options. â€Å"They got the right people on the bus, moved the wrong people off, ushered the right people to the right seats - and then they figured out where to drive it† (Collins cited in Grissom, 2006). So, this paper will discuss how Global Communication can reach its two important targe ts of achieving optimum growth as well as the tag of being the global player, through layoff of expendable workers, with the help of more innovations and plus outsourcing route. Situation Analysis Issue and Opportunity Identification Global Communications (or GC for short) is in a tight situation because of financial downslides and also due to tough competitions. To further add to its woes, the entire telecommunications sector was also not showing good growth. The fact that validates the GC tough scenario is the 50 percent depreciation it suffered in its stock value, leading to a struggling financial situation. With many competitors fighting for the same local, national as well as international markets, offering slew of new services like complete solutions for computers, televisions and plain old telephone service (POT), Global is facing very tough competition. To overcome this tough situation, beat off the competitors and increase the profits, the GC’s senior leadership team has come up with a two-pronged aggressive approach. The first plan is to, come up with their own new services, which can compete with the local telephone and cable companies. In that direction, GC got associated with a satellite provider, and has plans to offer value added services like video services and satellite version of broadband. This partnership with a wireless satellite provider can actualize a scenario where the small business owners can have anytime Internet access, just by using their wireless telephones or PC cards. The second strategy as part of the two-pronged approach is cost-cutting through the removal of employees. For that, they have identified number of job designations, which can be removed, while absorbing a minor portion of employees in other departments. The laid off jobs will be moved to India and Ireland as part of outsourcing strategy. With GC’s analysis showing that outsourcing could reduce unit costs for handling calls by nearly 40%, and with GC wanting to become a global corporation, this plan appears feasible. However, when viewed from another perspective, it raises some organizational and ethical dilemmas. Stakeholder Perspectives/Ethical Dilemmas GC GC to overcome its tough financial situation has come up with a twin strategy. Although GC’s first plan to introduce new services including value added servi

Finance report Essay Example | Topics and Well Written Essays - 4750 words

Finance report - Essay Example We divided its capital into four equal portions of QR 250,000 each to invest in each of the four investment platforms available. As such, we invest QR 250,000 in the Qatar Stocks Exchange, which is the principle stock market of Qatar. It invests another QR 250,000 in Qatar National Banks as bank deposits. It then invests the third portion of their capital, another QR 250,000 in gold and other metals. Some of these gold businesses are international and not available within the country. Furthermore, it invests the last portion of their quarter million in Forex markets, mainly dealing with currencies exchange with the Qatar Riyal. We has an investment philosophy that is of a risk taking capacity, which means that we will easily go for the high risk investments because of the high returns associated with these investments. Furthermore, we is aware that these high-risk investments may either lead to the loss of all the invested capital, or lead to a significant profit way beyond their imagination. We practices on the â€Å"risk-return tradeoff† investment strategy. This investment strategy operates on the principle that the potential of returns raises with an increase I the risk of investment. As such, lower levels of uncertainty, such as loss risk investments, have a high likelihood of bringing in low potential returns on the other hand, high levels of uncertainty, such as in high-risk investments, there is a high likelihood that the investments will yield high potential returns. Consequently, in accordance with the concept of â€Å"risk-return trade off† in business investments, money invested by an investor can render higher profits only if it is subject of a high possibility of losses. This model of business investment strategy requires traders to be always aware of their personal risk tolerance whenever they make choices of their investment portfolios owing to the

Gender Segregation in Education Essay Example | Topics and Well Written Essays - 2000 words

Gender Segregation in Education - Essay Example Studies have shown teachers spend more time with the boys in the classroom than with the girls. (MacNaughton, 2004, 82)This may be due to disruptions by the boys, although the teacher is often unaware of the amount of time spent with boys over girls. In the personality domain, there are various notable traits with gender differences. These include aggression, conformity, emotional adjustment, values and life goals and achievement orientation. Numerous studies have shown that whilst adolescent boys are more aggressive than girls in terms of both physical and verbal aggression, during adolescence the differences between males and females are negligible at primary school level (Vialle et al, 2000, p.154). This is not saying that girls become physically aggressive but perhaps resort to verbal aggression or the use of covert forms of aggression such as social alienation (malicious gossip and rumors, exclusion) (Vialle et al, 2000, p.154). Statistically, girls are more conformant than boys and are more likely to seek help from adults than are boys. (Grieshaber, 2004, 22) This difference is attributed to the observation ...that girls tend to participate in adult-structured activities on the other side boys are more likely to participate in activities with little adult involvement in school. (Vialle et al, 2000, p.154). Emotional behavior by boys is expressed differently, depending on gender. While females are more likely to freely display their emotions, males are more likely to suppress their feelings, with the exception of anger. (Walkerdine, 1999, 172) Most researchers attribute these differences to societal expectations rather than biological differences. This can be supported by studies of boys under five years of age. (Vialle et al, 2000, p.154).

News Essay Example | Topics and Well Written Essays - 750 words - 1

News - Essay Example Allegedly several secrets were established and the worst trouble ensued when the public came to know the fact that she possible had known about the attack of US embassy in Libya without sharing the delicate information with the relevant authorities, this amounted to the Benghazi case on an attack on the U.S. embassy (Collinson, 2015). Fox Network news also aired another controversial story on Clinton. The broadcast network placed the story as a second airing. It entailed the claims that a gentleman named George Stephanopoulos had donated to the benevolent Clinton foundation. The donations, however, were undisclosed to the public. That instigated controversial questions surrounding the probable intents of the whole act. The airing was brief as compared to the CNN’s case (it took six minutes). From this observation, it is notable that both the broadcasts had stories in their headlines about Hillary Clinton, which were interestingly controversial and detrimental to her reputation. George Stephanopoulos had served in the Bill Clinton presidential campaign in 1992 and even held some top positions in the administration including being the spokesperson and senior advisor of Bill Clinton. This story was viewed as more credible as compared to the previous case where the public had claimed that the emails had nothin g incriminating against the former first lady. In a second CNN coverage, it aired a planned nationwide memorial day despite the death of 17 bikers who died in the supposedly biker shoot out in Waco, Texas between rival groups (Karimi, 2015). The news lasted for about 8 minutes. The news portrayed the bikers as a violent and unlawful lot that threatened public peace. On the dreadful day set to honor the fallen soldiers of the Second World War, the bikers threatened to honor the17 lost lives of their fellows (Karimi, 2015). The bikers had promised to hold motorcycles’ rallies in a manner to honor their fallen mates.

Revolutionizing the Retail Sector Case Study Example | Topics and Well Written Essays - 1000 words - 1

Revolutionizing the Retail Sector - Case Study Example This system enabled merchants to changeover only parts that needed upgrade unlike the whole system in previous system. Motherboard and hard disk are removable for maintenance and upgrading to match with the latest and high performance blade. This makes retailers easily acquire high performance technologies while keeping conformations and fixings. This design decreased downtime and minimized cost and maintenance time. Blades can power and connect a variety of devices that enabled retailers to try new technologies while in store (Parry, 2011). These solutions offered by DigiPoS, are advantageous to the retailers compared to the later system that did shut down for a complete changeover. Changing the only parts of a system which needs upgrade is a great achievement as it allows the retailers continue serving their customer and no losses due to change over or loss of reputation. Since DigiPoS produced both hardware and software, an alternative of implementing the new technology could be, have a robust type of hardware that most software upgrades could not affect the performance of the whole system (Parry, 2011). DigiPoS could have made the hardware energy efficient with the required high technology capabilities and customize their software to offer solutions to the upgrade of their system unlike the hardware upgrades which caused closure of retailers businesses due to changeover of systems. This can be a good solution to the issue of upgrade. Make software upgrades match with the latest technology and only upgrade hardware at rare conditions with backups to enable retailers continue with their businesses unlike the later where they had to shut down for each upgrade. Most a times, upgrade of software does not affect much the performance of a system though change of configurations and compatibility might change. Quantum blade system is an upgrade of retail blade system with increased power efficiency and increased performance. Quantum blade runs on SSD made by Intel c ompared to retail blade that used rotating hard disk drives. These SSD used in quantum blade systems has no spinning parts and this increases the reliability and productivity of the systems. Quantum blade has a 60% reduction of power consumption compared to retail blade that increases the return on investment. Unlike retail blade systems, quantum blade have reduced carbon footprint, which saves a lot of cost. With better heat dissipation, there is a further decrease of the cost of ownership with the quantum blade systems. Quantum has a 30 percent increase in performance compared to the retail blade due to the introduction of the new processor. The storage of data in quantum has increased volumes due to flash drives added compared to retails blade (Digipos, 2013). Both quantum and retail blade systems have low consumption of power and decreased downtime. They also offer little time for upgrade or replacement as hard disk, power supply, and motherboard in both technologies. Furthermor e, both have a higher reliability due to the fan less power supply and capability of connecting a variety of devices powered by the Blade. Data storage and diagnostics due to the use of diskette on component flash drives. In addition, both systems have the capability of mounting on different locations depending on retailer’s wish (DigiPoS Store Solutions, 2013). Several methods said to help reduce the cost of owner

Discuss and analyse Essay Example | Topics and Well Written Essays - 1500 words

Discuss and analyse - Essay Example In her writing, the past is no longer a prologue but something hardly different from the current, (John and Smith, 2001, p.56). Smith’s novel is a whirlwind multiculturalism and immigration investigation and their impacts on individuals and the ways such cultural events get into London life. The tale begins with the Second World War and ends before the start of the millennium, taking about fifty years, a period that witnesses the change of all aspects of life. In exploring story about the reformation of the contemporary London, the novel joins Archie and Samad lifestyles, a click of colleagues whose friendships is established on common experiences in the last days of the struggles. Additionally, addressing issues to do with race, culture, and generation gaps with irony and brevity, White Teeth focuses on the relationship between Archie, Samad and their families. This bond was formed during the last days of the WW11 when they met at the main tank mishap survivors, (Kavc?ic? and Maver, 2007, p. 45). After the incident the two parted their ways with Samad going back to his country to attend college and Archie pursing his daily struggles in England. Almost 30 years later, the blue-collar Briton and the educated Muslim meets again. This happened when Samad immigrated to England with his family and tried to look for his ancient day’s friend, Archie. ... The story begins with Archie in his vehicle, with sealed shut windows and a running vacuum hose from the exhaust pipe of the car. Depressed because of a divorce, and very confused, he rests on an unremarkable street side, selecting a location across butchery - halal. As the smoke mixes up his thoughts, a reader gets a figure of a person who will serve as their protagonist. This depressing moment has occurred to Archie about 20 years back when he saw Daria’s smile that made him shed tears of joy. Archie has seen Daira in thoughts – a pretty girl in a doorway very attractive – and he discovered that he regretted not leaving her. If he had another opportunity of having such a sight again, Archie wished for a second chance and an extra time, (Smith and Sterlin, 2001, p. 63). Therefore, a burcher man saves his life and the story begins. Mostly, Zadie Smith’s story is just an investigation of how Archie uses his leisure time. He falls for a pretty, black woman, Clara Bowden, who has a London origin and a backslidden Jehovah’s Witness believer with roots from Caribean. Archie and Samad, who had recently a girl less young than him, feel the generation gap among their wives and themselves, and this gap broadens even when their kids are born. The two men do not understand each other, but all through the narrative, their bond keeps them together. Similarly, the two men understand not their partners, children, or their universe, but they clearly comprehend each other, even though their behavior and actions show a parting of fervent insanity to the outsiders. Although, Archie is the protagonist of the story, the author orients the audience between characters

History Of Oslo Essay Example for Free

History Of Oslo Essay Oslo is Norway’s capital city. Oslo came from the Old Norse word â€Å"†¦os meaning mountain and lo meaning plain or river â€Å". In other words, Oslo can be described as a place beneath the mountains (â€Å"Oslo† 2006). People of German origin may have settled in the settlements along the Oslo Fjord who moved up to the north from mainland Europe (Zelko 63). The first evident settlement in Oslo was in the 8th century. (â€Å"Oslo† 2006). The first people of who inhabited Oslo lived in â€Å"humble wooden houses with turf roofs† that was surrounded with pens or sheds for goats, sheep and cows (â€Å"Oslo: City† 2006). Like all the other settlers of Norway, the people were grouped together in the form of a tribe and those who were landowners met with his other landed neighbors in a public assembly called Thing. Legislative assemblies were called Lagtings (Zelko 63). During the Viking age, AD800-1000 Oslo settlers grew rapidly as it had become the center for trade and shipbuilding activities and later it earned the name the† Viking Capital† (â€Å"Oslo† 2006). Viking means â€Å"a man from â€Å"Vik†, a huge bay between the Cape Lindesnes in Norway and the mouth of Gota River in Sweden (Britannica 1033). Norwegian Vikings, like other Vikings of Denmark and Sweden, were feared all over Europe as they had â€Å"superior ships and weapons as well developed military organization†. They were basically endowed with extraordinary hunger for adventure (Britannica 1034). Norwegian Vikings were known to be ruthless and brave fighters who killed their victims and greedily loots their conquered settlements and when they were done plundering destroyed the place with fire. They changed the face of Europe as they engaged in raiding and trade and then finally attracted by their conquered lands settled to live there. Their target places were mostly western European countries such as England, Scotland, and Iceland. Their active participation in trading had renewed the neglected European commerce of the middle Ages. Their contact with Western Europe was instrumental for their Christianization and eventually unification of Norway (Britannica 1033). In 885 the first Viking Monarch, Harold the Fair-haired united Norway (Halsey 239). During his reign the settlers of Norway, including Oslo, were commonly engaged in blood feuds, fightings that involve almost, if not all, disagreeing families. Even minor disputes can trigger the killing and maiming of both members of the quarreling families and if not settled can start an endless round of retaliation. The violence can be finally stopped when offending families pay the wronged families which they called â€Å"bot†. Sometimes family feuds were brought to the attention of the Lagtings who will decide the outcome of the disputes and impose penalties. The worst penalty they can give at that time was known as outlawry—a sentence that regarded the offender as if he died already. All his goods were taken away from him and he cannot exercise or claim any legal rights. In fact, anyone can kill him without facing the risk of penalty. These condemned people were left with no choice but to leave their homeland or live in the forest as an outlaw (Tseng 24). In 1050 Harald Hardrade or King Harald III officially founded Oslo and made it the center of southern Norway. This site was located at the eastern side of the harbor on the left bank of the small Akers River (Halsey 239). Harald died in 1066 and his death ended the Viking period as raidings stopped (â€Å"Norway† 2007). Oslo experienced a great era during the reign of Haakon V who was crowned in 1299. He decided to build the Akershus fortress in Olso to serve as his home with his wife Euphemia of Rugens, a Northern German princess (â€Å"Oslo† 2006). He made Oslo the capital of Norway replacing Bergen. This time Oslo and the rest of Norway enjoyed relative peace (â€Å"Norway† 2007). In 1318 Norway was united with the kingdom of Sweden when Duke Erik of Sweden married Princess Ingebjorg, daughter of Queen Euphemia and Hakon V Magnusson. The unification of both countries was officially signed in the Bishops castle; now presently know as Oslo Ladergard (â€Å"Oslo: City† 2006). The saddest event that occurred in the early history of Oslo was the Black Death that terrorized the world in 1349. It was estimated that half of Oslo’s population died during the time of the Black Death or â€Å"bubonic† plague. Bubonic plague was carried by fleas in rats from England (â€Å"Oslo† 2006). Because of the loss of its inhabitants and damage to agriculture Norway united together with Denmark and Sweden via the Union of Kalmar 1397(â€Å"Norway† 2007). During this time Copenhagen, instead of Oslo, was selected as the actual capital of Norway. Consequently Oslo lost it political importance and was only regarded as a provincial administrative center while the kings lived in Copenhagen and Stockholm from 1400-1500 (â€Å"Oslo: City† 2006). In 1523, however, Sweden dropped out of the union, and weaker Norway was left in the care of Danish Kings(â€Å"Norway† 2007). Part 2 Modern Era Reformation Period Like the rest of mainland Europe, Oslo was greatly affected by the Lutheran Protestant Reformation of 1537 when the German Monk Martin Luther questioned the laws of Roman Catholic Church and sought to reform the religious beliefs of the Europeans. Oslo citizens were engaged in religious conflicts(Thodock 2003). The catholic bishop of Oslo, Hans Rev was converted to Protestantism despite the reluctance of the citizens( â€Å"Oslo:City† 2006). Since Oslo had slightly lost its political and economic importance as a city at this time as it was being ruled by Denmark most of the buildings constructed there was only made of wood so that the city was easily destroyed by fire in 1624. King Christian IV of Denmark and Norway decided to move the town to the right bank of the Akers River and rebuild it near the Akershus Fortress. The city buildings were constructed in a Renaissance city style with rectangular blocks and renamed it Christiania (Halsey). Scientific Revolution The scientific revolution changed the world’s beliefs entirely, and Norway is in the heart of it Religious beliefs were proven to be wrong and questions were answered. Copernicus first with his opinion on the heliocentric theory, Galileo second with proof of Copernicus theory, and Newton third with the laws of gravity to explain how and why the planets revolved around the sun. Enlightenment During the 1700’s the Age of Reason or the Enlightenment flourished in Europe particularly to the Norwegian trading partners of Great Britain and Holland. In these places people were trying to improve their lives with the use of reason instead of following traditional religious or accepted beliefs. Contact with the â€Å"enlightened† British and Dutch affected the daily life of the inhabitants of Christiania. Norwegian traders brought home with them not only the prized cargoes of tobacco, coffee, tea and spices but also enlightened ideas. They constructed luxurious houses with magnificent gardens in accordance with the style western Europe (â€Å"Oslo: City† 2006). Napoleonic wars At first, Denmark and Norway attempted to remain neutral in the Napoleonic wars between France and England and their respective allies in 1805. However, England attacked the entire Danish fleet in 1807 as a result Denmark joined with France against Britain (Britannica 1034). Britain cut-off trading with Norway and set up a continental blockade against Denmark and Norway wherein British navy prevented the goods from both Norway and Denmark in reaching its trading partners . This action greatly affected the economy of Norway. The export of fish and timber from Christiania as well as the import of grain from Denmark were blocked. As a result, citizens of Christiania faced an economic crisis and suffered hunger. To end their trouble, Britain loosened up its blockade against Norway in 1810-13(â€Å"Oslo† 1991). In 1814 the king of Denmark was forced to give up Norway to Sweden in a Treaty of Kiel when Napoleon I was defeated (â€Å"Oslo† 2006). Eager to rebuild their government the Norwegian created its first constitution on May 17 the same year and Christiania became the capital city. In time, Christiania replaced its rival Bergen as Norway’s largest city. The people celebrated with joy as Christiania was expected to reclaim its old glory without giving much thought at first that they were still under the control of a foreigner, Sweden. As a capital city, Christiania once again becomes important politically and economically. For its newly acquired role, new monumental buildings were erected as a venue for important functions. . They were the Royal Palace, Bank of Norway and the stock exchange Oslo Bors. Later, most Christiania citizens joined with the rest of Norway in demanding for a complete control of their own affairs. They did not want to recognize the provisions of the Treaty of Kiel and instead preferred a Danish King to rule them. Because of the political unrest , Sweden was forced to make the Act of Union of 1815,that gave Norway the privilege of having its own army, navy and parliament(Storting) and was permitted to control their own internal affairs in exchange for their strict compliance to the Treaty of Kiel(â€Å"Oslo† 2006). Renaissance Period Because of their internal independence, Christiania’ economic and political power rose. By mid 1800’s, Christiania grew into a major administrative, economic and military center. The economic success brought about by the shipping and agricultural industry made Christiania a dominant economic city of Norway (World Book 1981). The site of rapid industrial revolution in Oslo started at the Aker River around 1840. The development of Industries relied in the electricity provided by this River. Old buildings along the River were demolished and replaced with concrete large textile industrial buildings . Because of the increased jobs offered by industries the population of Christiania increased as peasants and rural populace moved to the cities hoping find work. In the beginning of the Industrial Revolution Christiania’s industrial workers, like most of European industrial cities, lived in poverty since income was low. They mostly lived in crowded houses meant for workers in which sanitation was bad. As a result, sickness was common and spread easily. To augment family income, the children aged below ten were forced to work in the industries, an environment that was not conducive for their well-balanced growth. In fact, industries were noisy and dusty. They caused water and air pollution. Because of this, the health of the children suffered and their resistance to sickness decreased as they lacked the time to play and sleep. In fact, schoolwork was unavoidably neglected (â€Å"Off â€Å"2007). Revolutions of 1848 The political revolutions in Europe in 1848 had increased the desire of Norway to demand for independence from Sweden. In the late 1800’s popular Norwegian playwrights Henrik Ibsen and Bjornstjerne Bjorjornson staged a play in Christiania’s theater with a theme about political hypocrisy, rights of women and social problems. These plays helped Norwegians develop a sense of nationalism. In 1901 the city of Christiania was selected by the notable Swedish inventor, Alfred Nobel to be the site for the awarding of the most prestigious Noble Peace Prize (Lundestad 2001). By 1905 Sweden granted independence to Norway and Christiania at last was freed from foreign interference that began 1300’s (Halsey 240). Industrial Revolution World War One was inevitable due to industrialization and the desire each country had to become supreme. Firstly, relationships between the nations of the world were dramatically altered by the Industrial Revolution of the nineteenth century. Tensions between the industrial powers began to rise. The intensity between the industrial powers grew more and more as each country sought to expand its sphere of influence at the expense of others. Secondly, there was a growing desire by ethnic populations within the larger countries to become independent nations. Norway separated from Sweden and revolts occurred in colonies such as China, Zimbabwe, Namibia, and Ireland. War broke out in the Balkans in 1912 and again in 1913, as Bulgaria, Serbia, Macedonia, and Greece fought first the Ottoman Empire and then Austria-Hungary. Lastly, Colonies around the world were a valuable asset for supporting the host countrys economy. The great European powers needed raw materials and outlets for their goods. They wanted to expand into new colonies while still protecting the ones they already had. World War I The economy of Christiania steadily grew until before World War I. When the First World War broke out in 1914 Norway declared its neutrality. However, Christiania benefited economically from the war by carrying British cargoes inspite the loss of Norwegian lives and ships that were caught in the war (Compton 610). After the war Christiania settled back to its normal life. In 1924, Christiania was renamed back to Oslo (â€Å"Oslo: City† 2006). World War II. On April 9, 1940 Oslo was defeated by the Germans almost with only a slight resistance due the betrayal of Norwegian Officer and Fascist politician Vidkun Quisling who collaborated with the Nazis. As a result, the city was only slightly damaged during the war (Halsey 239). In 1942 Germany installed a puppet government in Norway under National Union leader Vidkun Quisling. However, the Norwegians resisted the Germans and its puppet regime. Oslo became a place of opposition strikes. Other citizens were secretly involved in the large-scale industrial sabotage and espionage on behalf of the allied powers. (â€Å"Oslo† 2006). During their occupation of the city the Nazis used Akershus as a prison and place of execution for those convicted of allied conspiracy and today its the site of Norways Resistance Museum, which provided a detailed account of German takeover and the Norwegian struggle against it (Thodock 2003). The German forces that overrun Norway surrendered in May 8, 1945 enabling the exiled King Hakon VII to return to Norway from London in June. Oslo then began to rebuild its ruin buildings while at the same time prosecuted â€Å"about 90,000 alleged cases of treason and defection†. Convicted traitors faced execution foremost of which was Vidkun Quisling (â€Å"Oslo† 2006). War had been going on for a very long time when it came to The Cold War, the postwar to World War II. After World War II, as the Cold War began two major alliances were created. Although, in World War I a systems of alliances did not work, the systems of alliances that formed after World War II were bigger and they werent buddy-buddy alliances. These two alliances were formed to bring peace throughout the world, to end The Cold War, and to prevent anymore future high scale wars. Toward the end of the war, hostility between Norwegians and the German occupying forces grew considerably stronger. A prime cause was the German withdrawal from all fronts. Retreating German units from the fronts in the north, the USSR and Finland, withdrew to Norwegian territory. In the autumn of 1944, the Red Army followed the Germans into Finnmark, where it liberated Kirkenes and the northeastern areas. As the occupying troops retreated, they ordered an evacuation of the entire Norwegian population in Finnmark and in Troms south to Lyngen. In Lyngen, three German army corps dug in. Then the entire region north of Lyngen was burned and destroyed. The destruction included 10,400 homes, bridges, power stations, factories, fishing vessels, telephone facilities and other types of infrastructure. The scorched earth area was much larger than all of Denmark. After the Soviet Red Army crossed into North Norway, a Norwegian military mission and a small number of Norwegian soldiers from Great Britain and Sweden followed. The Russians withdrew from Norway in September 1945, a few months after the German capitulation.

Polymerase Chain Reaction (PCR) Steps

Polymerase Chain Reaction (PCR) Steps We owe the discovery of the polymerase chain reaction to Kary B Mullis in the year 1983. He was the actual proponent of PCR. Few people are aware that in 1971, Kleppe and the Nobel laureate Gobind Khorana published studies including a description of techniques that are now known to be the basis for nucleic acid replication. However, it is unfortunate that Kleppe and Khorana were ahead of their times. Oligonucleotide synthesis wasnt as simple as it is today; genes had not been sequenced and the idea of thermostable DNA polymerases had not been described. Hence, the credit for discovering the PCR remains with Kary Mullis. The Polymerase Chain Reaction is essentially a cell-free method of DNA and RNA cloning. The DNA or RNA is isolated from the cell and replicated upto a million times. At the end, what you get is a greatly amplified fragment of DNA. The PCR is quick, reliable and sensitive and its variations have made it the basis of genetic testing. WHAT KARY B MULLIS SAYS ABOUT HOW HE DISCOVERED THE POLYMERASE CHAIN REACTION I was just driving and thinking about ideas and suddenly I saw it. I saw the polymerase chain reaction as clear as if it were up on a blackboard inside my head, so I pulled over and started scribbling. A chemist friend of his was asleep in the car. Mullis says that Jennifer objected groggily to the delay and the light, but I exclaimed I had discovered something fantastic. Unimpressed, she went back to sleep. Mullis kept scribbling calculations, right there in the car. He convinced the small California biotech company, Cetus, he was working for at that time, that he was up to something big. They finally listened. They sold the patent of PCR to Hoffman-LaRoche for a staggering $300 million the maximum amount of money ever paid for a patent. Mullis meanwhile received a $10,000 bonus. BASIS OF THE METHOD The purpose of PCR is to generate a huge number of copies of a segment of DNA, which could be a gene, a portion of a gene, or an intronic region. There are three major steps in a PCR, which are repeated for 30 or 40 cycles. This is done on an automated cycler, which can either heat or cool the tubes containing the reaction mixture, as required, in a very short period of time. There are three major steps in a PCR, which are repeated for 30 or 40 cycles. DenaturationDuring this process, the double stranded DNA melts and opens to form single stranded DNA. All enzymatic reactions, such as those carried over from a previous cycle, stop. This will be explained in the next paragraph. The temperature for denaturation is not fixed but it usually occurs at about 95 °C. It is important to realize that the denaturation temperature is largely dependent on G:C (guanine:cytosine) content of the DNA fragment to be analyzed. This is reasonable when one considers that the G:C bond is a triple hydrogen bond and the AT bond is a double bond. Logic dictates that a triple bond should be 1.5 times harder to break than a double bond. Therefore, when the segment of DNA to be analyzed has a very high G:C content, the denaturation temperature can reach even upto 99 °C. AnnealingThis requires temperatures lower than those required for denaturation. In this process, the primers anneal to that very specific segment of DNA that is to be amplified. The primers are jiggling around, caused by the Brownian motion. Ionic bonds are constantly formed and broken between the single stranded primer and the single stranded template. The more stable bonds last a little bit longer (primers that fit exactly) and on that little piece of what is now double stranded DNA (template and primer); the polymerase can attach and starts copying the template. Once there are a few bases built in, the ionic bond is so strong between the template and the primer, that it does not break anymore. ExtensionThis is done at 72 °C. This is the ideal temperature for working with polymerase. The primers, which are complementary to the template, already have a strong ionic attraction to the template. This force is stronger than the forces breaking these attractions i.e. the high temperature. Primers that are on positions with no exact match (non complementary) get loose again (because of the higher temperature) and dont give an extension of the fragment. The nucleotide bases are added from the 5 end to the 3 end. The phosphate group of the dNTPs is coupled with the hydroxyl group of the extending DNA strand. The extension time depends on two factors; the type of polymerase used and the length of the DNA fragment to be amplified. Usually, Taq polymerase adds dNTPs at the rate of about 1000 bases per minute. It is important to realize that each component of the PCR including the input DNA, the oligonucleotide primers, the thermostable polymerase, the buffer and the cycling parameters has a profound impact on the sensitivity, specificity and fidelity of the reaction. The three steps of the first cycle are shown, that is, denaturation, annealing and extension. At the end of the first cycle, two strands have been synthesized. At the end of the second cycle, four strands have been synthesized (the three steps of the cycle have not been shown). At the end of the third cycle, eight strands have been synthesized. The number of strands increases exponentially with each cycle. Nuggets The Polymerase Chain Reaction is essentially a cell-free method of cloning DNA and RNA. There are three steps involved in every cycle; these are denaturation, annealing and extension. At the end of each cycle, the DNA doubles. Therefore, theoretically, if there are n cycles in a reaction, the number of DNA fragments at the end of the reaction will be 2n. COMPONENTS OF THE POLYMERASE CHAIN REACTION The components that are essential for a successful PCR are elaborated here. TEMPLATE DNA This is that portion of the DNA/gene that is to be amplified. Usually the concentration is  100 ng genomic DNA per PCR reaction. However, this can vary depending on the target gene concentration and the source of DNA. The PCR reaction is inherently sensitive. It is not necessary for the template DNA to be abundant or highly purified. Higher amounts of template DNA can increase the yield of nonspecific PCR products, but if the fidelity of the reaction is crucial, one should limit both template DNA quantities as well as the number of PCR cycles. DNA in solution may contain a large number of contaminants. These contaminants may inhibit the PCR. Some of these reagents are phenol, EDTA, and proteinase K, which can inhibit Taq DNA polymerase. However, isopropanol precipitation of DNA and washing of DNA pellets with 70% ethanol is usually effective in removing traces of contaminants from the DNA sample. Effects of Fixation This is of particular interest to the pathologist since he has to deal with formalin fixed tissue. DNA extracted from fresh tissue or cell suspensions forms an optimal template for PCR. The tissue is best stored at -70 °C at which the nucleic acids can be stored indefinitely. A temperature of -20 °C is sufficient to preserve the DNA for several months and at 4 °C, the DNA can be stored for several weeks. At room temperature, the DNA has been successfully stored for hours to days; however, mitochondrial DNA is very sensitive to temperature and may degrade in thawed tissues. DNA extracted from fixed tissue has been used successfully for PCR. The type of fixative and the duration of fixation are of critical importance. Non crosslinking fixatives like ethanol provide the best DNA. Formaldehyde is variable in its DNA yield. Carnoys, Zenkers and Bouins are poor fixatives as far as DNA preservation is concerned. Not surprisingly, formaldehyde is the fixative which has been evaluated the most, because it is more commonly used worldwide. The studies have demonstrated that a successful PCR depends on the protocol to extract the DNA and the length of fixation. Formaldehyde reacts with DNA and proteins to form labile hydroxymethyl intermediates which give rise to a mixture of end products which include DNA-DNA and DNA-protein adducts. Purification of DNA from formalin fixed tissue, therefore, includes heating to reverse the hydroxymethyl additions and treatment with a proteinase to hydrolyze the covalently linked proteins. However, there is no way to reverse the DNA-DNA links and these links inhibit the DNA polymerases. This accounts for the low PCR yield which is seen with formalin fixed tissue. Usually, the PCR reaction with formalin fixed DNA as a template yields products which are not more than 600 bp in size. Nuggets Template DNA is required in a concentration of 100ng for each PCR reaction. Contaminants in DNA may inhibit the reaction. Fixation of tissues provides DNA which is not as good as DNA obtained from fresh/ frozen tissues. Different fixatives give different DNA yields. Alcohol is the best fixative and Carnoys, Zenkers and Bouins are poor fixatives as far as DNA preservation is concerned. Formalin is intermediate in DNA yield. Purification of DNA from formalin fixed tissue involves heating to reverse the attachment of hydroxymethyl intermediates and treatment with a proteinase to hydrolyze the covalently linked proteins. The DNA obtained after fixation can be used for reactions in which the PCR product is not more than 600 bp. PCR BUFFER The purpose of using buffers in PCR is to provide optimum pH and potassium ion concentration for the DNA polymerase enzyme (usually obtained from bacteria Thermus aquaticus) to function. Most buffers are available in a 10X concentration and require dilution before use. Although most protocols recommend the final buffer concentration of 1X, a concentration of 1.5X might result in increased PCR product yield. The PCR buffer contains many components. Some important ones are discussed here: Divalent and monovalent cations These are required by all thermostable DNA polymerases. Mg2+ is the divalent cation that is usually present in most of the PCR buffers. Some polymerases also work with buffers containing Mn2+. Calcium containing buffers are ineffective and therefore, rarely used. Buffers can be divided into first and second generation buffers on the basis of their ionic component. The second generation buffers, as opposed to first generation buffers, also contain (NH4)2SO4 and permit consistent PCR product yield and specificity over a wide range of magnesium concentration (1.0 to 4.0 mM MgCl2). The overall specificity and yield of PCR products is better with second generation buffers, as compared with first generation PCR buffers. Buffers also contain KCl. Salts like KCl and NaCl may help to facilitate primer annealing, but concentration of 50 mM will inhibit Taq polymerase activity. Interactions between K+ and NH4+ allow specific primer hybridization over a broad ran ge of temperatures. Magnesium is one of the most important components of the buffer. Mg2+ ions form a soluble complex with dNTPs which is essential for dNTP incorporation; they also stimulate polymerase activity and influence the annealing efficiency of primer to template DNA. The concentration of MgCl2 can have a dramatic effect on the specificity and yield of PCR products. Optimal concentration of MgCl2 is between 1.0 to 1.5 mM for most reactions. Low MgCl2 concentration helps to eliminate non-specific priming and formation of background PCR products. This is desirable when fidelity of DNA synthesis is critical. At the same time, however, too few Mg2+ ions can result in low yield of PCR products. High MgCl2 concentration helps to stabilize interaction of the primers with their intended template, but can also result in nonspecific binding and formation of non specific PCR products. It is important to be aware that many PCR buffers (often sold in 10X stocks) already contain some amo unt of MgCl2. Therefore, the addition of further amounts must be carefully monitored. In the best possible scenario, the PCR would work well with the amount of Mg2+ already present in the buffer solution. However, if this does not occur, it is necessary to standardize the amount of Mg2+ in the reaction mix. This can be difficult because the dNTPs and the oligonucleotide primers bind to Mg2+. Therefore, the molar concentration of Mg2+ must exceed the molar concentration of the phosphate groups contributed by dNTPs and the primers. As a rule of thumb, the magnesium concentration in the reaction mixture is generally 0.5 to 2.5 mM greater than the concentration of dNTPs. The optimal concentration of Mg2+ should, therefore, be standardized for each reaction. Tris-Cl The concentration of tris-Cl is adjusted so that the pH of the reaction mixture is maintained between 8.3 and 8.8 at room temperature. In standard PCR reactions, it is usually present in a concentration of 10mM. When incubated at 72 °C which is the temperature for extension, the pH of the reaction mixture falls by more than a full unit, producing a buffer whose pH is 7.2. Other components Some buffers also contain components like BSA (Bovine serum albumin) and DMSO (dimethyl sulphoxide). BSA reduces the amount of template sticking to the side of the tube, making it available for amplification and reducing the risk of primer dimer. Primer dimers are products obtained when the primers anneal to each other instead to to the template DNA. DMSO has been shown to facilitate DNA strand separation (in GC rich difficult secondary structures) because it disrupts base pairing and has been shown to improve PCR efficiency. In effect, it is wise not to tamper with the buffer provided with the Taq polymerase. The buffer is usually standardized for the vial of Taq and there is no need to add additional MgCl2 or stabilizers like DMSO and BSA. However, some Taq buffers come with the buffer in one vial and MgCl2 in a separate vial. Under such circumstances, it is advisable to start with 1 µL of MgCl2 and increase its concentration in aliquots of 0.5  µL, if the initial reaction fails. Nuggets The PCR buffer contains divalent and monovalent cations, Tris Cl and other components. The PCR buffer is used to give the correct pH and potassium concentration for the DNA polymerase to function. The most common divalent ion used is magnesium in the form of MgCl2. MgCl2 concentration is vital for PCR. Tris Cl is used to maintain the pH between 8.3 and 8.8 at room temperature. Salts like NaCl and KCl may facilitate primer annealing Other components like BSA and DMSO help to increase the sensitivity and specificity of the reaction. OLIGONUCLEOTIDE PRIMERS What are Oligonucleotide Primers? PCR primers are short fragments of single stranded DNA (17-30 nucleotides in length) that are complementary to DNA sequences that flank the target region of interest. The purpose of PCR primers is to provide a free 3-OH group to which the DNA polymerase can add dNTPs. There are two primers used in the reaction. The forward primer anneals to the DNA minus strand and directs synthesis in a 5 to 3 direction. The sequence of primers is always represented in a 5 to a 3 direction. The reverse primer anneals to the other strand of the DNA. How to design a primer? The predominant goal kept in mind while designing a primer is specificity. Each member of the primer must anneal in a stable fashion to its target sequence in the template DNA. The longer the primer, the higher is its specificity. Unfortunately, the longer the primer, the less likely it is to anneal to a particular sequence in the template DNA. Conversely, if the primer length is small, it is likely to anneal, but its specificity will be poor. A compromise is reached by designing primers between 20 and 25 nucleotides long. Inclusion of less than 17 nucleotides often leads to non specific annealing, while presence of more than 25 nucleotides may not allow annealing to occur at all. Remember that the DNA sequence in the human genome appears to be a random sequence of nucleotides. When designing primers, it is important to calculate the probability that a sequence exactly complementary to a string of nucleotides in the human genome will occur by chance. Several formulae are designed to calculate such probabilities. However, mathematical expressions are not necessarily correct and in this case, the predictions maybe wildly wrong. The distribution of codons is non random with repetitive DNA sequences and gene families. It is advisable to use primers longer than the statistically indicated minimum. It is also advisable to scan DNA databases to check if the proposed sequence occurs only in the desired gene. For a practicing pathologist, it is best not to attempt designing of primers. What a pathologist requires is the primer sequence for an established test. If, for example, a pathologist requires primer sequence for the diagnosis of sickle cell anemia, all he has to do is search the web for papers related to molecular testing of sickle cell anemia. The primer sequences will be provided in the paper. Custom made primers can be commercially synthesized. Several biotechnology companies provide this facility. Before the primers are ordered, it is essential to check that the sequence is correct and that there are no missing nucleotides in the sequence. That is where, BLAST is invaluable. Before the intricacies of the BLAST search are elaborated upon, it is necessary to mention that designing a primer does not depend only on the sequence of nucleotides. Other factors like the GC content and melting point are also important considerations. They will be dealt with later in the chapter. BLAST and its uses BLAST is an acronym for Basic Local Alignment Search Tool. It is an algorithm comparing information about primary biological sequences with a library or database of sequences. A BLAST can be performed for different organisms, but in this book, we will concern ourselves with nucleotide BLAST in humans only. BLAST searches the database for sequences similar to the sequence of interest (the query sequence) by using a 2-step approach. The basic concept is that the higher the number of similar segments between two sequences, and the longer the length of similar segments, the less divergent the sequences are, and therefore, likely to be more genetically related (homologous). Before perfoming a BLAST search the oligonucleotide sequence is first identified. The sequence is fed into the programme. BLAST first searches for short regions of a given length called words (W). It then searches for substrings which are compared to the query sequence. The program then aligns with sequences in the database (target sequences), using a substitution matrix. For every pair of sequences (query and target) that have a word or words in common, BLAST extends the search in both directions to find alignments that score greater (are more similar) than a certain score threshold (S). These alignments are called high scoring pairs or HSPs; the maximal scoring HSPs are called maximum segment pairs (MSPs). The BLAST search as outlined in fig 7.2 shows the results of the search. If we scroll down further, we can see the sequences producing significant alignments. Note that in this BLAST search, there are 49 BLAST hits in the query sequence. In the list shown in figure 7.2, there is a list of hits starting with the best (most similar). To the right of the screen is the E-value. This is the expected number of chance alignments; the lower the E value, the more significant the score. First in the list is the sequence finding itself, which obviously has the best score. To the left is the accession number. This refers to a unique code that identifies a sequence in a database. It is important to know that there is no set cut-off that determines whether a match is significant or similar enough. This must be determined according to the goals of the project. The sequences provided in the figure 7.2 show a significant alignment with Pseudomonas japonica. It shows a high score (bits) and a low E-value. Note that the lower the E value, the greater the likelihood that the sequence is a good match. BLAST output can be delivered in a variety of formats. These formats include HTML, plain text and XML formatting. For the NCBIs web-page, the default format for output is HTML. When performing a BLAST on NCBI (National Centre for Biotechnology Information), the results are displayed in a graphical format showing the following: The hits found A tabular form showing sequence identifiers for the hits with scoring related data Alignments for the sequence of interest and the hits received with corresponding BLAST scores for these. The easiest to read and most informative of these is probably the table. The main idea of BLAST is that there are often high-scoring segment pairs (HSP) in a statistically significant alignment. BLAST searches for these high scoring sequence alignments between the query sequence and the sequences in the database. The speed and relatively good accuracy of BLAST are among the key technical innovations of the BLAST programs. Sequence of events to be followed when performing a BLAST search.: Go to PUBMED (http://www.ncbi.nlm.nih.gov/pubmed/) Scroll down to reach a heading called POPULAR Under POPULAR click on BLAST Click on nucleotide blast Under the heading, enter accession number(s), gi(s), or FASTA sequence(s), type or paste the sequence that you want matched. Click BLAST Wait for the results. Analyse the nucleotide sequence as it appears. Calculation of Melting Temperature The melting temperature or Tm is a measure of stability of the duplex formed by the primer and the complementary target DNA sequence and is an important consideration in primer design. Tm corresponds to the midpoint in transition of DNA from the double stranded to its single stranded form. A higher Tm permits an increased annealing temperature that makes sure that the annealing between the target DNA and the primer is specific. The Tm is dependent on the length of the oligonucleotides and the G+C content of the primer. The formula for calculation of Tm is given in table 7.1. Table 7.1: Formula for calculation of the melting temperature. Length of Primer Tm ( °C) Less than 20 nucleotides long 2(effective length*) 20 to 35 nucleotides long 22 + 1.46(effective length) *Effective length = 2(number of G+C) + number of (A + T) Primers are usually designed to avoid matching repetitive DNA sequences. This includes repeats of a single nucleotide.. The two primers in a PCR reaction are not homologous to each other and their complementarity can lead to formation of spurious amplification artifacts called primer dimers. The 3 end of a primer is most critical for initiating polymerization. The rules for selecting primers in addition to those already mentioned are as follows: The C and G nucleotides should be distributed uniformly throughout the primer and comprise approximately 40% of the bases. More than three G or C nucleotides at the 3-end of the primer should be avoided, as nonspecific priming may occur. The primer should be neither self-complementary nor complementary to any other primer in the reaction mixture, in order to avoid formation of primer-dimer or hairpin-like structure. All possible sites of complementarity between the primer and the template DNA should be noted. The melting temperature of flanking primers should not differ by more than 5 °C. Therefore, the G+C content and length must be chosen accordingly (a higher G+C content means a higher melting temperature). The PCR annealing temperature (TA) should be approximately 5 °C lower than the primer melting temperature. G+C content in each primer should not be more than 60% to avoid formation of internal secondary structures and long stretches of any one base. Primer extension will occur during the annealing step. Primers are always present in an excess concentration in conventional (symmetric) PCR amplification and, typically, are within the range of 0.1M to 1M. It is generally advisable to use purified oligomers of the highest chemical integrity. Primer Dimers A Primer Dimer (PD) consists of primer molecules that have attached or hybridized to each other because of strings of complementary bases in the primers. As a result, the DNA polymerase amplifies the PD, leading to competition for PCR reagents, thus potentially inhibiting amplification of the DNA sequence targeted for PCR amplification. In the first step of primer dimer formation, two primers anneal at their respective 3 ends. The DNA polymerase will bind and extend the primers. In the third step, a single strand of the product of step II is used as a template to which fresh primers anneal leading to synthesis of more PD product. Primer dimers may be visible after gel electrophoresis of the PCR product. In ethidium bromide stained gels, they are typically seen as 30-50 base-pair (bp) bands or smears of moderate to high intensity. They can be easily distinguished from the band of the target sequence, which is typically longer than 50 bp. One approach to prevent PD formation consists of physical-chemical optimization of the PCR system, i.e., changing the concentration of primers, MgCl2, nucleotides, ionic strength and temperature of the reaction. Reducing PD formation may also result in reduced PCR efficiency. To overcome this limitation, other methods aim to reduce the formation of PDs only. These include primer design, and use of different PCR enzyme systems or reagents. Nuggets Oligonucleotide primers are short fragments of single stranded DNA (17-30 nucleotides in length) that are complementary to DNA sequences that flank the target region of interest. They dictate which region of DNA in the PCR will be amplified. Primer sequences can be obtained by reviewing previously published literature. A confirmation of the sequence can be done by using BLAST (Basic Local Alignment Search Tool). The melting temperature is the midpoint in the observed transition from a double stranded to a single stranded form. A higher annealing temperature ensures that the annealing between the target DNA and the primer is specific. A primer dimer consists of primer molecules that have attached or hybridized to each other because of strings of complementary bases in the primers. Taq polymerase amplifies the primer dimer leading to competition for the PCR products. Several methods are used to reduce primer dimer formation including changing the concentrations of primers, MgCl2, nucleotides, ionic strength and temperature of the reaction. TAQ DNA POLYMERASE The initial PCR reaction used the Klenow fragment of Escherichia coli DNA polymerase. However, this was unstable at high temperatures and it was necessary to add a fresh aliquot of enzyme after every denaturation step. The annealing and extension temperatures had to be kept low and as a result, there was formation of non specific products in abundance. The discovery of the thermostable Taq DNA polymerases ensured that the PCR did not remain a laboratory curiosity. The extension and annealing temperatures could now be kept high and the formation of non specific products was greatly reduced. Taq became famous for its use in the polymerase chain reaction and was called the Molecule of the Year by the journal Science. Why Taq? Taq is the enzyme of choice in PCR because of the following reasons: Taq works best at 75 °C80 °C, allowing the elongation step to occur at temperatures which make non-Watson-Crick base pairing a rare event. It can add upto 1,000 nucleoside triphosphates to a growing DNA strand. Taq has a half-life of 40 minutes at 95 °C and 9 minutes at 97.5 °C, and can replicate a 1000 base pair strand of DNA in less than 10 seconds at 72 °C. Because of all these properties, Taq is the enzyme of choice in the PCR. How does Taq polymerase act? The first requirement is a primer. The primer is annealed to the template strand having free hydroxyl group at its 3 end. During the extension phase, the Taq synthesizes a new DNA strand complementary to the template by adding dNTPs in a 5 to 3 direction condensing the 5 phosphate group of the dNTPs with the 3 hydroxyl group of the end of the extending DNA strand. Since Taq works best between 70 °C- 80 °C, a temperature of 72 °C is usually chosen as the optimum annealing temperature. Where does Taq come from? In Thermus aquaticus, Taq polymerase is expressed at very low levels and commercial production is not economically viable. However, the enzyme can now be produced from different versions of the engineered Taq gene so as to obtain high levels of expression in E coli. What other polymerases are available for use in PCR? Taq is not the only polymerase; other polymerases are available but Taq is the one that is generally used in a PCR. A few other polymerases with their uses are as follows: PFU DNA polymerase -Found in Pyrococcus furiosus, it functions in vivo to replicate the organisms DNA. The main difference between Pfu and alternative enzymes is the Pfus superior thermostability and proofreading properties compared to other thermostable polymerases. Unlike Taq DNA polymerase, Pfu DNA polymerase possesses 3 to 5 exonuclease proofreading activity, meaning that it works its way along the DNA from the 3 end to the 5 end and corrects nucleotide-misincorporation errors. This means that Pfu DNA polymerase-generated PCR fragments will have fewer errors than Taq-generated PCR inserts. As a result, Pfu is more commonly used for molecular cloning of PCR fragments than the historically popular Taq. However, Pfu is slower and typically requires 1-2 minutes to amplify 1kb of DNA at 72 ° C. Pfu can also be used in conjunction with Taq polymerase to obtain the fidelity of Pfu with the speed of Taq polymerase activity. TFL DNA polymerase Obtained from Thermus flavus, it is useful for the amplification of large segments of DNA. WHAT IS FIDELITY? All DNA polymerases have an intrinsic error rate that is highly dependant on the buffer composition, pH of the buffer, dNTP concentration and the sequence of the template itself. The types of errors that are introduced are frameshift mutations, single base pair substitutions, and spontaneous rearrangements. Therefore, the PCR reaction generates a product that is very similar, but in many cases, not identical to the original sequence. The quantity of dissimilar product obtained is obviously related to the cycle in which the mismatch took place. Under normal circumstances, this does not make any difference; however, these errors may become significant during sequencing when the role of fidelity comes into play. Fidelity is the ability of the polymerases to avoid the incorporation of wrong nucleotides during the reaction. Under normal circumstances, it really does not make a difference if a wrong nucleotide is incorporated because the size of the PCR product remains the same and that is what we have to look for. However, there are some polymerases like Pfu which have a high fidelity. In addition to reading from the 5 to the 3 direction, they can also read from the 3 to the 5 direction and correct the wrong nucleotides wh

Eve Kosofsky Sedgwicks Tendencies: Queerness and Oppression Essay

Eve Kosofsky Sedgwick's Tendencies: Queerness and Oppression Over the last two decades or so, the idea of queerness is one that has been utilized and considered by individuals and communities of marginalized sexualities and genders. The concept is one that has attempted to broaden and deconstruct traditional notions of gender and sexuality in order to include all of their incarnations as valid experiences and identities. Queerness endeavors to include all of those who feel they are a part of it yet, seemingly, not everyone can be queer without changing the very nature of queerness. Or can they? Queerness is a concept which resists borders and structure yet it seems as though there must be certain commonalities among all queer identities and behaviors. In her book, Tendencies, Eve Kosofsky Sedgwick constructs queerness as a seemingly all-inclusive and individually determined space, writing that: queer can refer to: the open mesh of possibilities, gaps, overlaps, dissonances, resonances, lapses and excesses of meaning when the constituent element's of anyone's gender, of anyone's sexuality aren't made (or can't be made) to signify monolithically. (8) She expands queer beyond the bounds of "same-sex sexual object choice" making queerness about performative behavior rather than sexual mechanics (Sedgwick 8). For example, Sedgwick's idea of queer includes "feminists... masturbators... lesbian-identified men...[and] people able to relish, learn from, or identify with such" among others(8). She posits that the fundamental precondition, "to make the description 'queer' a true one is the impulsion to use it in the first person" (Sedgwick 9). Yet is this self-determined queerness valid? Can actual queerness be claimed s... ...cepting one's status as incongruent allows one to claim a singular identity without inviting oppression because one's incongruence prohibits a singular identity claim to completely describe the self. The manner in which Foucault and Sedgwick construct queerness allows for oppression because they assume that the self is a singular cohesive body. Warner supposes that by realizing the fragmentation the self, one can claim an identity and escape oppression. Works Cited Foucault, Michel. The History of Sexuality Volume 1: An Introduction. Trans. Robert Hurley. New York: Vintage Books, 1980. Sedgwick, Eve Kosofsky. Tendencies. Durham, NC: Duke University Press, 1993. Warner, Michael. "Tongues United: Memoirs of a Pentecostal Boyhood." The Material Queer: A LesBiGay Cultural Studies Reader. Ed. Donald Morton. Boulder, CO: Westview Press, 1996.

Packaging :: essays research papers

Successful and Unsuccessful Packages   Ã‚  Ã‚  Ã‚  Ã‚  Budweiser package and Foster package does more than they contain and protect products. They also tell their great deal about the people from different countries that buy the products, as well as the people from different countries who sell the products. In addition, both packages are made in different countries, and both packages have differences and similarities.   Ã‚  Ã‚  Ã‚  Ã‚  Budweiser package is manufacture in America and contains various bottle sizes and various can sizes. Budweiser bottle is small and easy to hold and the cap is easy to open. In addition, it is made in thick glass so when it drops from the knee down it does not crack, and it ¡Ã‚ ¯s appearance is attractive to people. Budweiser can is made in different beer quantity from 6 ounces to 24 ounces. Moreover, it encloses different item quantity from case of 6 to case of 24.   Ã‚  Ã‚  Ã‚  Ã‚  On the other hand, Foster package is manufacture in Australia and contains one bottle size and one can size. Foster bottle is large and hard to hold and it is hard to open. In addition, it is made in thin glass so it ¡Ã‚ ¯s easy to break when it drops. Foster can contains only one beer quantity, which is 6 ounces and doesn ¡Ã‚ ¯t enclose various options to meet people preferences. In addition, Foster only has one item quantity, which is case of 6 that most people prefer case of 24. Budweiser package is a successful selling product in America because the price of the products is affordable to buy and it on advertisement (billboard, radio, and TV). In addition, the company and the product are popular to US citizen. Besides, Budweiser alcohol level is low around .02, which most American prefers that level of alcohol to be. For example, in New York Time Magazine say most American like to drink Budweiser because they say it tastes better and the alcohol level is no t dangerous for driving. On the other side, Budweiser package is a failure in Australia because it considers a foreign product and Australian prefers to have higher alcohol level.   Ã‚  Ã‚  Ã‚  Ã‚  Foster package is a failure in the United State for the reason that it ¡Ã‚ ¯s a foreign product and doesn ¡Ã‚ ¯t meet US citizen preferences. In addition, it contains lack in advertising and contains high alcohol level. Besides, Foster company and products are unpopular in the US, so the percentage of selling the product really low.

My Role As The School Librarian Essay -- eduaction

Everyday I walk into my school library with the hope of influencing a student or a teacher to read a new book, use a new search strategy, or to collaborate on a unit of study. My goals for students range from getting to know them and their interests and then guiding the student to these new sources or literature. I look upon the library as a learning lab or, as some in my profession have dubbed it, the Learning Commons. Every morning I greet anywhere from 50 to 80 students who are waiting for the first bell of the day. These students are in the library because they have made a decision that this is their place; a hangout for those students who are drawn to a vibrant environment that is teeming with learning and discussion. The Library Commons at Olathe Northwest is a natural extension of the secondary library’s traditional mission in a digital world. My library offers a wide range of elements to foster student learning in new and creative ways. My goal is that this space fits the need of every student. Students who need to complete an assigned task that calls for collaboration can meet in the library and plan, discuss and complete their assignment. At the same time other students can be searching for a great book or looking online for sports scores or prom dresses. The forty computers in the library offer access for any student who walks in the door. And just in case more computers are needed, a wireless laptop can be called into service. My school library is a vibrant place that welcomes all students, there is something for everyone and as the teacher-librarian I am proud to say that I listen to my patrons and work to achieve the goal of providing a learning space that fills the needs of all who enter. The course to ac... ... time together outside of school hours. In my role as the school librarian I have access to every student and teacher in the building. I try to reach all of these people through creative and relevant library-based programs. I want the library to invite all users through its doors. I entice the I don’t like to read with great technology and computer resources. For the students who can’t get enough of the latest Manga book I am willing to stop at the bookstore and pick up the newest entry in that series. Regarding teachers, I am all about perfecting the collaborative effort to meet student learning. I have established a climate of respect. Respect for the learner, the teacher and the content material. Students genuinely like the library and my hope is that they will continue to use libraries as adults and they will be strong advocates for schools and libraries.

White Settlement

â€Å"What was the impact on White Settlement on Indigenous life between 1788- 1861? Describe the meeting of two cultures that we see in Bennelong and William Buckley. † 1. Early years in Sydney- Arthur Phillip and Bennelong In 1788, led by Captain Arthur Phillip (Australia’s first governor), the Europeans had arrived in Australia with the first fleet. They saw no sign of life and had no idea that the Indigenous people had been living in Australia for thousands even tens of thousands of years. But the Indigenous people were very aware that they had come and were deeply fascinated by them. At first they thought they were ghosts or evil spirits but they soon realized that these white-skinned and strangely-dressed people were planning to stay, and that they were. These â€Å"white-skinned’ people were in fact convicts who was shipped to do hard laborious work for their committed crime. Life before Europeans had settled in Australia was a very different place. There was either clear land and/or lots of bush. There were no boats, building or roads, in fact the Indigenous people probably had no idea there was such thing. This was probably why Arthur Phillip presumed the land could be theirs because no-one held sovereign right. But the Indigenous people were living there. The Indigenous people lived happily and peacefully with guidance from their elders who knew local dream time stories and customs. They comfortable living together, helping and sharing with one another and stayed alive by eating animals and berries from the bush. They didn’t need fancy clothes, building or roads unlike the Europeans to live a good life. On the 25 November 1789, Captain Arthur Phillip, captured elder Bennelong and used him as a way to learn about the language and customs of the indigenous people. Bennelong willingly liaised between the cultures, and adopted European dress and other ways. 2. Victoria- Buckley and batman After convict William Buckley’s escape from the Victorian settlement he was discovered by the Wathaurang people who thought this pale, 198cm giant carrying a spear was the ghost of one of their leaders. Buckley had arrived at Port Phillip Bay from England in 1803 with about 300 soldiers, settlers and convicts after being sentenced to transportation for life. Before the European settlement was abandoned, Buckley escaped. He wandered alone for weeks before he was befriended by the Wathaurang people. Over the next 32 years Buckley lived with the Wathaurang, learnt their language and customs, married and had a daughter. In 1835 he finally emerged to meet Batman’s colonising party and tried to work as an intermediary between settlers and aborigines, but felt he wasn’t trusted by either. 3. Overview of their decline and dispossession of land Victoria was originally the home to many Indigenous tribes that had been living there for a long period of time. Although when the Europeans settled in Australia Batman planned to negotiate his way into ‘buying’ the land of the Indigenous people. Once Batman arrived in Victoria he approached the local Indigenous leaders with a contract, to ‘buy' their land. His negotiations were successful, and he walked away with 240,000 hectares of prime farming terrain – almost all of the Kulin nation’s ancestral land even though they were technically not eligible to sell the land as it wasn’t really their land. However the transaction went ahead, and it was agreed that batman was to on a annual basis, give them 40 blankets, 30 axes, 100 knives, 50 scissors, 30 mirrors, 200 handkerchiefs, 100 pounds of flour and 6 shirts in exchange for the land. Because of this transaction between the Indigenous People and Batman the majority of Victoria’s population is, as the Indigenous people originally called us, â€Å"white-skinned†.

Small Scale Industries Problems

Article: Information technology: a growth navigator for small scale industries in India Small scale industries (SSI) have a significant role in the Indian economy in view of its contribution to production, employment, and export. However, since 1991 small scale industries in India find themselves in an intensely competitive environment due to globalization, domestic economic liberalization, and dilution of sector specific protective measures. The formation of World Trade Organization in 1995 also forced the member countries (including India) to drastically scale down the restrictions on import. This article analyzes the various factors influencing the growth of small scale industries in India. Information Technology (IT) is one of the significant factors, which is, in general, not used by the small scale industries in India for their production and business process, in the growth of small scale industries in India. Further on the basis of Nolan's ‘Stages of growth model' the stages of use of information technology in small scale industries are studied. Small Scale Industry in India: An Analysis in the Context of Liberalization Small and Micro producers are crucial in developing economies, and their role is even greater in the largely rural economies of South Asia. In India as well, the sector is the second largest employer, after agriculture, and accounts for nearly 6 percent of the country's GDP. India was an exception in that it gave the small-scale sector large incentives, and protection, in the period 1948-1991, going to the extent of reserving certain production lines solely for the sector. In the historical context of this, this paper shall attempt to analyze the issues peculiar to a ‘smalls scale of production' in India in an increasingly globalised scenario. It shall also look at some of the other issues plaguing the sector such as credit availability and maintaining quality standards. Movement of toxic metals from small-scale industrial areas: a case study from Delhi, India In India, an estimated 70% of the total industrial pollution load is attributed to Small and Medium Enterprises (SMEs). Among SMEs, small-scale industries continue to use obsolete technologies with no stringent rules for pollution control. Consequently, waste generated from them is generally dumped without treatment. These facts are strengthened from the analysis conducted on solid waste and wastewater samples from five industrial areas, water samples of river Yamuna, the tributary of river Ganges and solid waste samples of landfill areas in Delhi. Toxic metal concentrations were analysed and found in high concentration in collected samples. Besides that, fractionation study was also done. The data obtained from this study identify that the wastes with high concentrations of toxic metals, calling for the introduction of point source control of waste generation.

Wisdom, Morality, and Meditation

The Fourth Noble Truth is the Noble Eightfold Path, which is also referred to as â€Å"Magga. † The Noble Eightfold Path essentially has three main parts: Wisdom, Morality, and Meditation. These three sections represent the eight sections of the Noble Eightfold Path. Wisdom is broken down into â€Å"Right View† and â€Å"Right Intention. † Next, morality consists of â€Å"Right Speech,† â€Å"Right Action,† and â€Å"Right Livelihood. † Finally, meditation consists of â€Å"Right Effort,† â€Å"Right Mindfulness,† and â€Å"Right Concentration. † One may think that these eight parts must be followed in a specific order, however, all eight parts work mutually dependent of each other. Right View is a part of Wisdom and, according to our class lectures, is the â€Å"Middle Way between eternalism and nihilism; the emptiness of all things. † Right View distinguishes wholesome (beneficial) things from unwholesome (harmful) things. A few examples of unwholesome things from our class notes are: onslaught of living beings, taking what is not given, sensual misconduct, lying speech, divisive speech (idol speech), harsh speech, covetousness, and wrong view. The roots of unwholesomeness can be narrowed down to three things: greed and desire, hatred and anger, and ignorance and confusion. Thich Nhat Hanh describes the importance of Right View and what it is within chapter 9. Right View is known as samyag drishti. TNH talks about how seeds are planted within our bodies, and everyone has them. I thought it was the coolest analogy when TNH taught of these seeds within our bodies. It seems like everyone has each kind of seed of all different traits, but it depends on whether or not those seeds are watered within our bodies. He says: If you live in an environment where your seed of loyalty is watered, you will be a loyal person. But if your seed of betrayal is watered, you may betray even those you love. You’ll feel guilty about it, but if the seed of betrayal in you becomes strong, you may do it. (TNH, 51) This is such an amazing statement because I am a fairly strong believer that you are the product of your environment. Most people do whatever the â€Å"status quo† is in their neighborhood and rarely does anyone make a big jump to do something drastically different. I feel like all people are created the same, at least mentally, and it is up to the upbringing to form how someone acts in life. The reading of TNH’s chapter 9 discussed how it is up to the individual to decide which seed grows more than others within one’s body. In class we discussed how one can try to keep the seed of anger, for example, from growing. It is up to the individual to essentially stunt the seed of anger’s growth when one feels any possibility of anger coming in. While pushing the feeling of anger away, one should try to grow the seed of loving-kindness instead. Within my own life, I try to live by the idea of â€Å"killing people with kindness. † This is my third year as RA here on St.  Bonaventure and when I confront a situation, I try to always be as nice as possible. There’s nothing better than when we are documenting a room for a violation, usually alcohol related, and being overly nice to them. They have no idea how to handle the niceness in the situation. It just makes the situation so much better in the long run. Most people act very mad and rude to us when they are being documented and they don’t expect us, the RA’s, to be nice to them, but when we are nice to them and don’t let their obscene â€Å"hate words† affect us, they don’t know what to do. I feel like this could be a small example of growing my seed of kindness because I could get very angry about the students calling me hateful names for simply doing my job. Instead, I try to do what TNH said in Chapter 10, â€Å"†¦replace an unwholesome thought with a wholesome one by ‘changing the peg,’ just as a carpenter replaces a rotten peg by hammering in a new one. † (TNH, 62) In this example of my RA duties, I replace the unwholesome thought of harsh speech with loving-kindness, compassion, and clarity (education) to why the students are being documented. Discussing â€Å"Morality†, I read a part in which Kornfield was talking about his teacher, Maha Ghosananda (the Gandhi of Cambodia). Kornfield was telling how his teacher would teach the survivors of the 1975-88 genocide in Cambodia practices of compassion and loving-kindness for their own loss and that of others. He said, â€Å"You have lost so much. Now you know how precious everything is in this world. You must love again and let new things grow. † (Kornfield, 81) This quote can be related to â€Å"Right View,† but more importantly the concept of compassion which is within â€Å"Right Conduct† or â€Å"Morality. I absolutely love this quote because I feel like way too many people take their great lives for granted. I am sad to say that I am sometimes right in that category of people. I am always humbled so much when I meet someone who has endured a great amount of pain, or those who have already had cancer and are the same age as me. Right now, while at college I have two close friends who have already battled cancer and are now back at college. It makes me feel like I should be so incredibly thankful for the life that I have been blessed with. Many times one can become attached to something that is not all that important. Meditation can resolve this. In chapter 12 of TNH, it talks about how we have become so efficient and able to talk to places on the other side of the planet, however, he also explains that people have a harder time with one-on-one interactions and speech nowadays. This is an example of becoming attached to technology instead of listening and speaking with people in person. When reading through Kornfield’s 24th chapter, I noticed the stories about Dipama Barua, one of the greatest meditators of the Theravada lineage. They told of how she lost two out of three of her young children to illness and lost her husband due to a heart attack soon after. Most people would feel like there is no longer a reason to live after something like that, and she was one of those people. However, after a year of lying in bed full of grief, she started doing meditation and then eventually became a master of meditation. (Kornfield 382-384) Kornfield had gone to see Dipama and had such an encounter! When he was leaving from seeing her, she touched him and said a 10 minute prayer in which he started to have a realization and see everything in a positive light. After this, he could not stop smiling at everything. (Kornfield 382-384) This encounter between Kornfield and Dipama reminds me of times that I feel like nothing can go right, but all it takes is seeing and talking to someone who you really like and respect. Then, after talking to this one person, you have a totally new positive outlook on life. This short story tells me that how you go through life is all about perspective. This â€Å"halt† in life represents a meditation. Sometimes one has to take a break from their busy lives and just reflect on their life and spirituality. When I have done this in the past, it feels so incredibly rewarding to just take a break from things and reflect on how great life is. When one is thinking about the Noble Eight-Fold Path, one has to remember that all of the â€Å"Rights† link into each other. We need to be compassionate for others, practice loving-kindness, and embrace wisdom, morality, and meditation within our lives to better understand everything.