Eczema Essay

The city grew very fast, and over the next ten years the population of West Tennessee expanded from 2500 to 100,000. The majority of families at the time took to farming, and with the help of the fertile land, Memphis soon became the center for growing and trading cotton in the Southeast. Indeed, cotton became king in Memphis, bringing in black men and women to work forced labor on plantations. According to the census of 1820, 20% of the populations of Memphis were slaves, and that number is said to have risen throughout the 19th century. Racial tension in Memphis began with its first and second mayor. Marcus B. Winchester was the irst mayor of Memphis and is said to have committed political suicide when he married a woman who was a quarter black. The second mayor of Memphis, Isaac Rawlings, had a common-law marriage with black woman, having multiple children with her. These encounters were socially unacceptable and looked down upon by white plantation owners, even though the majority of whites in Memphis at the time favored the gradual emancipation of slaves. As the civil war approached, the black population grew as more black slaves were brought to Memphis to work the cotton fields. During the Civil War, Memphis sided with the Confederates, but it was uickly captured by the Union who strategically wanted the town as the sight of a naval yard. The Union also made Memphis a freedman’s colony, and many neighboring black slaves came to Memphis once they were emancipated to partake in education and paid labor. Memphis fell so easily to the Union that most of its infrastructure remained in place. For a brief moment, Memphis became a city of opportunity to black Americans and a place where black communities thrived. Yet as white Memphians saw themselves competing with former slaves, tensions ran high on many of their newly arrived black neighbors in a riot that left forty-six dead, nearly wice that many injured, five women raped, approximately 100 blacks robbed, and ninety-one homes, four churches, and all twelve black schools destroyed. â€Å"[l] The riots did not stop until martial law was declared and troops from Nashville arrived in Memphis to force peace. For a good many years, the black community suffered and struggled to regain prominence, most of them being too poor to move away for new opportunities. With all the schools destroyed, the educational opportunities vanished and the ability of black to become literate and contribute to society became almost impossible. Yet this would not be the only disaster in Memphis. In the 1870s, a series of yellow fever epidemics plagued the city. In 1878, the worst of the yellow fever epidemics hit, and 25,000 people fled. 17,000 contracted the fever and over 5,000 died in the summer of 1878. Of the population that would remain in Memphis, 70% of those would be blacks who were too poor flee. The city of Memphis, which was already in debt, lost much of its tax base and went bankrupt. At the end of the 1800s and the early 1900s, Memphis would turn around again economically, yet many of the areas social problems would remain. Memphis fared reasonably well in addressing the problems of its white community, but the citys black citizens were shunted aside, their civil rights and their human dignity subordinated to a cult of white supremacy. â€Å"[2] Around this time, railroads connected Memphis to the Atlantic Ocean, allowing it to become one of the largest manufacturers and shippers of hardwood in the United States.

Research Paper on Fun Things Essay

For this assignment, you will compose a ten-page research paper on a great thinker chosen from the attached list. There are three components to this assignment. The first is that you will write about the major thought, idea, or system of belief for which the great thinker was known. Secondly, you will write about two or three ways in which the teachings of the great thinker influenced history. Thirdly, you will provide a thoughtful analysis of at least one idea touted by the great thinker. You will need to research and properly document eight to ten scholarly sources using the MLA standard for citation. In addition to writing the paper, you will be learning and implementing a way to organize your research and writing. You will receive two grades for this assignment: one for the final paper and one for the research and writing prep work. Research Component Research is an important skill for college students to learn and implement. When done well, research will help strengthen an argument, prove a point, verify facts, and explain with first-hand knowledge. At its most basic level, research is studying the works of experts in any given field. For instance, if I want to research heart surgery, I will locate experts in the field of cardiology and read what they say about the heart. In reading their works, which may consist of published studies, medical journal articles or medical books, or informative medical websites or blogs, I am presuming and appealing to their expert knowledge and skill in an area I know little or nothing about. In short, I’m reading and studying their work in order to learn. For your research paper, you will be appealing to the expert knowledge of a great thinker. You will research articles and/or books that were either written by the great thinker (we call this a primary source) or by someone who studied the great thinker (we call this a secondary source). For example, let’s say you are studying Jonathan Edwards (great puritan theologian). Were you to read and study his actual sermons, you would be researching a primary source. Were you to read and study what John Piper (present day pastor and avid Jonathan Edwards fan) has written about the theology of Jonathan Edwards, then you would be conducting secondary research. Both primary and secondary sources are very valuable when writing research papers. Usually primary research is considered to be the most reliable in that it is closest to the original (the idea originator, the eye-witness account, the photograph, etc†¦). A good research paper will be able to navigate both types of research and carefully integrate it appropriately. For the purposes of this assignment, you’ll be doing both because you’ll be concerned with what the great thinker actually taught and that will best be discovered by his or her actual writings, and you’ll be concerned with how his or her thoughts influenced history, which is most likely written by other scholars. In addition to conducting primary and secondary research, you’ll need to make sure that your sources are consider scholarly; that is, your sources come from people who have appropriate expertise in the field of you are studying. This will not be an issue with your primary sources because you will go straight to the source (i.e., you’ll study the writings or Marx or Lewis), but it will be more of an issue in secondary sources. How to tell if an article or book is considered scholarly or written by an authoritative expert in the field can be tricky, but here are a few rules: 1. Does the author have an educational background that would account for the knowledge in the article or book? 2. In the case of articles, is it published in a peer-reviewed journal? 3. Does the article or book offer analysis and critique of original sources or just give mere impressions or opinions? Another term I have mentioned in regards to research is MLA citation (Modern Language Association). MLA is a method of citation that is used primarily in the humanities that indicates the source of your research. Learning MLA citation rules takes time and lots of effort, but it is a necessary part of conducting and writing a well-researched paper. For this assignment, you’ll need at least eight scholarly sources that may include journal articles or books but may NOT include websites. There will be in-class lectures that will teach you where to locate scholarly resources and how to properly keep track and cite them. As I see it, there are two major avenues of research for this paper: the first is research involving the actual ideas of the great thinker. The second is research involving the analysis of the thinker’s ideas. Content of Research Paper As mentioned in the assignment overview, there are two components to this research paper. The first is that you will research the major idea or set of ideas for which the great thinker was known. This will be the first section of the research paper and will include a thorough explanation of the great thinker’s idea or ideas and he or she came to belief or think in such a way. The second section of the paper will include a discussion on two or three of the major ways the great thinker’s ideas influenced history. The third section is where you provide thoughtful analysis about at least one of the great thinker’s ideas. This is not, necessarily, an argument, but rather it is an opportunity for you to break down the idea and comment on it yourself. Although I’m more interested in your analysis, it may help to research how others have analyzed and critiqued your great thinker’s ideas. In this section you may 1. Critique the thinker’s idea 2. Break down the thinker’s ideas and provide additional clarification or implication 3. Show how different scholars have viewed or interpreted the great thinker’s ideas and whether agree or disagree 4. Show why the thinker’s ideas produced negative or positive effects on society, theology, history, etc. 5. Any combination of the above. Please note that I do not wanting you to write a biography; that is, I do not want detailed discussion on the great thinker’s family history, marriage, hobbies, physical appearance, personal health, friends, etc. unless is it absolutely pertinent to his or her intellectual contribution. Paper Organization Each of you will be required to create an outline for your paper and turn it in as part of the assignment. Additionally, you will be required to keep track of all research and turn in your research notes. Research Organization I will require you to keep track of your sources through Microsoft One Note or Word, whichever works best for you. For every source you look at, you’ll need to record the following information: 1. Source Title- title of the article or book 2. Source Location-book, ebook with web address, journal article with web address 3. Source Citation- MLA citation as it would appear in your works cited page 4. What-what information are you using from the source. If it is short, type it here. If it is a chapter or large section, write down the major idea or some key words and the necessary information to find it in your source easily (page or section numbers, headings, etc.). 5. Plan- how you plan on using the information You will have at least eight records with the above information, although it is much more likely you’ll have more and may not use all sources in the actual writing of the paper. That is okay. Note: Make sure you accurately record information for every source because you won’t remember. Grading Criteria: Each paper will be graded on the following: 1. Accurate, detailed, and thoughtful discussion and analysis of the great thinker’s ideas 2. Coherent, fluid prose 3. Accurate implementation of MLA citation 4. Integration of scholarly sources 5. Proper adherence to Standard English grammar and style 6. Proper adherence to page length and formatting guidelines

Advertising Statement Essay Example for Free

Advertising Statement Essay ? Rosser Reeves was the one who invented the term â€Å"USP†. The Unique selling proposition is sometimes referred to as â€Å"product difference.† In rare cases, some products or services have a unique and impressive proposition/benefit. A unique selling proposition is the ultimate proposition because its one that no other competitor can claim. It has to be something that you could also sell from. The concept of a unique selling proposition, or USP, is based on a benefit statement that is both unique to the product and important to the user. The heart of a USP is a proposition, which is a promise that states a specific and unique benefit you will get from using the product. If the product has a special formula, design, or feature, particularly if protected by a patent or copyright, then you are assured that it is truly unique. This is why a USP is frequently marked by the use of an â€Å"only† statement, either outright or implied. There are various methods that can be used to find a USP as it is demonstrated in advertising strategies and ideas. In advertising, â€Å"strategy† refers to the overall marketing or selling approach. It is the thinking behind the concept/idea. (The thinking behind the thinking, if you like.) Decisions about selling premises are central to the overall advertising strategy. The strategy (or strategic thought) can come from a proposition/benefit of the product, how it used, the market background, the choice of target audience, or any combination thereof. Every strategy should have an element of distinction ( small or large ) from the competition’s strategies, as should the proceeding concept and campaign. All strategies should be written in the form of a strategy statement, also known as â€Å"creative brief†. However, there are several questions we should seek to answer to cover the area of strategic analysis. Competitor, best prospects, and what buying appeals have the greatest leverage. At the corporate level what takes place in the advertising department would be seen as tac tical whilst in the advertising department this would be seen as strategic. * Corporate strategies are concerned with the major functions of the company, and cover finance, human resource management, production, administration, and marketing. * Marketing strategies are concerned with ANSOFF’s matrix and the marketing mix. * Promotional strategies are concerned with the promotional mix options (advertising, sales promotions, PR, publicity, selling, sponsorship, exhibitions). Advertising that is effective creates the message that best expresses the product-prospect relationship. In addition, the message has to be intrusive enough to battle through the clutter in the contemporary media marketplace. To reach the effectiveness in advertising, a creative strategy should be involved in the process. It has to sell the product effectively by promoting them through smart and well designed advertisement. When forming a strategy from which to create ideas, it needs to be written down in a black and white. This helps to focus and steer the formation of ideas from the onset. When people suggest that an idea is â€Å"off strategy,† they mean that it doesn’t relate back to the defined strategy, and will therefore be much harder to sell the idea to the client. By having a strategy statement at hand, you can keep referring back to it whilst generating ideas from that strategy. It is very hard, even if you are an experienced creative, to produce a great campaign idea (or even a single one shot) without a solid, tight strategy. In short, the better you are briefed, the easier your job will be. A poorly defined, vogue, â€Å"wooly† brief is no use to a creative person, nor is highly specific one that restricts the number of ideas. Below are the basic examples of the headings in a creative brief, * Client * Product/service * Product and market background (supposition) * Competition * Business/Advertising Objective (problem to solve) * Media * Target market/Group/Audience * Proposition/Promise/Benefit * Proposition Support Points * Tone of Voice * â€Å"Mandatories† (Inclusions/Exclusions) Media in Advertising The choice of media depends upon the type of product or service being advertised, the target market, and the client’s budget. Each campaign can be in one form of media, or multiple forms. Traditional advertising media includes print, TV, and radio. Non-traditional includes ambient and guerrilla concepts. In addition, there is direct media, and interactive media. In the UK, each type of media is defined by its relation to a hypothetical â€Å"line† that divides the two. Traditional media is â€Å"above the line,† whereas direct marketing and interactive advertising is â€Å"below the line.† Companies that produce work in all the forms are referred to as â€Å"through the line† agencies, or â€Å"full service.† Other divisions within advertising include sales promotion, and business-to-business. Whereas business-to-consumer advertising, once the skill is developed, the same creative process can easily be applied to these other forms of media an d advertising. All products and companies, as seen by the customer, occupy some kind of ‘position’ in the market whether they intend it or not. This might be, for example, high price-high value, low price-low value, high price-low value, good company-not-so-good company and so on, when compared with comparable competing products and competing organizations. Most modern organizations now attempt to actively influence this position in the market by matching product and corporate benefits with the needs of clearly identified segments. The managers have performed professionally, be integrated to match the identified needs of the target market. Positioning is how the marketer wants the consumer to view its product relative to the competition. Although product differentiation plays a role in creating a product position, product differences account for only part of a product’s position. A positioning strategy also includes the manner in which a product’s factors are combined, ho w they communicated, and who communicates them. The size (and value) of the advertising corporate and brand positioning is crucial to the well-being of any company and so should be left to the agency professional. Copywriting is an essential part of the design communication mix, and those of us who do it for a living will tell you that crafting massages and telling stories is a rewarding mental process, even in the business context. You’ll find that being able to generate a response from your audience is a valuable and highly sought-after skill. Copy (or text, or words) used in design is a very particular type of creative writing that requires the inspiration of an artist and the control of a craftsman or craftswoman. In comparison to the rails on which the copywriter runs, the novelist or poet has no limitations. Poetry and storytelling are flights of the imagination, with no client or news editor to bear in mind. Whether the personality of the writer shines through directly or indirectly, this is the purest creative writing – it can take off in any direction, be as fictional as it wants to be, and go wherever it pleases. Writing copy, however, is all about sticking to brief, while paying homage to the creativity and style of the poet and storyteller. Journalists and copywriters are commercial writers, but the essence of the role is completely different. In most cases journalist have to create the story from the scratch, usually by following leads. They will have to research the facts to get to the heart of the matter, discover the different viewpoints and opinions, and bring this material together accurately and coherently. Articles are often written to a tightly defined structure, while features can allow more room for individual expression and the interweaving of the writer’s viewpoint. The message has to be factually correct, balanced, and fair, but the writer is allowed to take a stance, which could reflect that of the newspaper or, in the case of regular column, the writer’s opinion. Copywriting borrows from all other fields of writing in its quest for creative expression, but ther e is no room for your personality in the copy that you write; you are simply a scribe, a hired mouthpiece for your client, and it is the brand’s voice that must come through, loudly and clearly. Advertising Statement. (2016, Oct 12).

Buddhism and Its Cultural Impact Essay Example | Topics and Well Written Essays - 1250 words

Buddhism and Its Cultural Impact - Essay Example The dissemination of such ideas and beliefs from one culture to another was facilitated through trade, migration, and warfare in the historic times. Throughout the post-classical periods, many groups of nomadic missionaries contributed to this cultural diffusion process. From such, groups, Buddhist missionaries more commonly referred to as priests or monks succeeded in inculcating the Buddhist way of life across much of the Asian region. Although Buddhism has its many forms across the Asian continent, the core belief system is reflected in all forms being practiced in countries as India, Sri Lanka, Thailand, Vietnam, Nepal, China and Japan. These core beliefs have contributed to the unique cultural aspects and practices that can be observed within these cultures. In order to discuss cultural manifestations resulting from religeious teachings, it is important to define what culture denotes. Culture according to Boas, â€Å"embrace all the manifestations of social habits of a community, the reactions of the individual as affected by the habits of the group in which he lives, and the products of human activities as determined by these habits† (Boas, 1930) Thus, it is important to understand the underpinning, belief systems and religious teachings which manifest in the form of distinct cultural characteristics, ideas and technologies. Considering the Asian countries, which influenced by, Buddhism, the cultures are rich with history, heritage, religious practices and philosophies. Buddhist way of life seeks to achieve morality and well being of life through harmony and self-moderation. Buddhists are motivated by the need of wanting to break the cycle of life referred to as â€Å"Samsara†, which is perceived as a sorrowful journey eve n at its most luxurious state of living.  

Healthcare and Law Essay Example | Topics and Well Written Essays - 1500 words

Healthcare and Law - Essay Example penses, once whatever coverage they would have runs out, or if the individual(s) in question would lack insurance, the greater question would become how to pay for the subsequent bill(s) in their entirety. There are countless health conditions that may require an individual to seek the assistance of a doctor at any point in their life. One such instance would be for those patients seeking to initiate the procedure of breast augmentation. While for some, the procedure of breast work would be for cosmetic reasons, for others, such work would come after the presence of a health condition, such as breast cancer that in turn, would require the reconstruction of the breast. With the introduction of the material added during such a procedure, there are risks that may result from the procedure being performed. Such as, the rupture of the material within the breast and as such, results in other health consequences as a result for the patient. For this very reason, there would exist laws state and nation wide, to serve as an education tool that would ideally, stave off any potential negative ramifications that may occur. From a legal standpoint, one such example of a case that would have been filed, due to issues surrounding breast implants, would involve a woman from San Diego, California. In this case, â€Å"A woman who claimed a design flaw in her silicone breast implants caused them to rupture and leak has lost her lawsuit against the manufacturer, Minnesota Mining and Manufacturing,† (â€Å"Woman†, 1999-2010, para. 1). By the action taken through the court system, the verdict rendered, would illustrate the apparent feeling on the part of the court that, in terms of rendering guilt, they would feel that the manufacturer of the implants in question, would not have been liable for what would have occurred to the patient in question. With this case, â€Å"Marva L Smith, 43, had sought damages against 3M for the cost of surgeries to remove the implants, to correct

The Valuation of Intellectual Property for Various Purposes Term Paper

The Valuation of Intellectual Property for Various Purposes - Term Paper Example Some approaches to valuation have evolved from others for evaluation, potentially confounding finance tools with those for measuring internal performance indicators; for example, where human resources may be regarded as the "knowledge assets" of a company (Andriessen, 2004, p.233). Other methods appear to have emerged primarily for external reporting of financial information. From the critical perspective, an evident reason for this accelerated activity is the growing importance of intangibles in the modern economy. Near the end of the 20th century, 79% of jobs and 76% of the American GNP were from weightless enterprise (Contractor, 2001, pp. XI-XII). There also is a greater call for valuations in modern business practice, including the purchase and sale of brands, international transfer pricing, mergers, and the formation of alliances. The proliferation of many methods and labels, all chasing many metrics, seems to be working against a common understanding of the fundamentals in the valuation of intellectual property. Even the word value has several meanings in English. For instance, Black's Law Dictionary details more than 40, all stemming from Black's view of value as a measure in a transaction: value is defined as the monetary worth or price of something; the amount of goods, services, or money that something will command in an exchange (Black's Law Dictionary, 1999). This equates value to the price achieved in a transaction. Different parties may have a different sense of the value of the item to be transacted; marketing professionals would have us believe value is determined by equation worked out, per situation, in each buyer's mind: "value is the combination of price, quality, convenience, service, ownership experience, and every other factor in the buying decision" (Calloway,2003, p.153). In many cases, this may produce results that differ in the mind of the buyer and the mind of the seller. Indeed, the likelihood for a transaction will increase when the perceived value of the item, to the buyer, exceeds the value suggested by the seller. To this extent, these varying ideas of market price, perception, and different parties may be resolved by thinking of value as the point at which the bid and ask prices converge. NYU Stern School of Business finance professor Aswath Damodaran, a leading authority on valuation, suggests value resides in the future: "The value of a firm is based upon its capacity to generate cash flows and the uncertainty associated with those cash flows...The value of a firm is still the present value of the expected cash flows from its assets" (Damodaran, 2001, p.11). From the critical perspective, this introduces a new set of variables, revolving around not the present circumstances of the market, but its possible future circumstances. It also creat es a distinction between assets that are (a) aggregated in the form of a go-forward business, producing income as a firm, (b) individually for sale, to the extent they may be separated from their present use, or (c) dormant in a business that has ceased operations, where no income is presently being generated. But what if item that is not able or meant to produce income in the future Gordon V. Smith, a leading author in the field of valuation, combines all variations into a single definition:

Forecasting Exchange Rates Case Study Example | Topics and Well Written Essays - 500 words

Forecasting Exchange Rates - Case Study Example The forecast, however, depends on probability which might cause uncertainty. Technical forecast will indicate the depreciation of the baht to help the company understand the how their money could be work for them understand these conditions (International Journal of Forecasting 2008). Technical forecast will however depends on fundamental forecast to get historical data of inflation and interest rate. The market-based analysis is based on the market and their implication of inflation and interest rate. The market analysis depends on demand-supply analysis. The market based analysis will help the company understand the consumption of their products in the Thailand market. It also assists management to know what quantity of export is needed. The analysis is also inclusive of interest rates and inflation rate and how it will affect the demand and supply of their products (International Journal of Forecasting 2008). If in the reality the value of the baht  90 days from now  is $0.22. Using technical forecast, baht will depreciates with six percent to give its vale at $ 0.22. Technical forecast shows that the expected results are equal to the real value. The fundamental forecast has three scenarios. The scenarios depend on the chances of depreciation and their percentage. If the chance for depreciation is 30% then baht will deprecates by 2% making the value of baht to be $0.18. 15 % chance of depreciates, baht will depreciates with by 5%, making the value of baht to be $ 0.21. 35 percent change, baht will depreciate with 10% making the value of baht to be $0.20. The market based analysis is involved in interpreting the calculation. In the calculations, it is true to say that fundamental forecast is accurate but tiresome. It offers a wide range of solution with different scenarios. The values from the fundamental are added

(the main differences between the NRA method of measurement , and Assignment

(the main differences between the NRA method of measurement , and CESMM3) Civil engineering measurement - Assignment Example It is not measured to construction joints, holes, ducts, sockets, mortices and the like that do not exceed 0.15 cubic meters in volume. The edge of blinding concrete should be less or equal to 75mm. An inclination angle of less than 15Â ° to horizontal plane is required. The pipes are measured in linear meters and made of clay, concrete, iron, steel, polyvinyl and glass reinforced plastic. The depth of excavation depends on the nominal pipe diameter and type of material for the pipe (CESSM3, 1991 p.48). NRA dwells puts a lot of emphasis on drainage and service ducts include drains, sewers, piped culverts and service ducts. It does not include filter drains, narrow filter drains and fin drains. The unit of measurement is linear meters. The measurement done from the center line between; For depth inverts that do not exceed 2m, average depth of invert to be stated the nearest 25mm. If the depths to invert exceed 2m but less than 4m and in steps of 2m, again the average depth must be stated to the nearest 25mm. From the CESSM3 point of view, unit of measurement for executed works is in sum form apart from scenarios where there is a specified unit of measurement. The divisions included are; contractual, specified requirements, method related charges, provisional sum and the rest. Contractual requirements are classified into performance bond, insurance of works and third party insurance. Specified requirements refer to accommodation, services, equipment, attendance for engineers’ staff and the like. The method-related charges focus on accommodation, services, temporary works, supervision and labour and plant. Provisional sums are measured in terms of day work and include; labour, materials, plant and their respective percentage adjustment. Method-related charges are inserted by the tenderer in the Bill of Quantities (CESSM3, 1991 p.16). NRA classifies this section under preliminaries and

Citizenship Curriculum Guidelines Changed the Methods of Teaching Essay

Citizenship Curriculum Guidelines Changed the Methods of Teaching Citizenship in English Secondary Schools - Essay Example As the report declares the active elements of teaching Citizenship usually engage pupils that are more difficult and if these elements were introduced into other mainstream lessons they could contribute to pupils engaging with learning, leading to less exclusion, less negative incidents improved attendance and improved classroom behaviour. The text of the current literature in Wright’s Teaching citizenship in the Secondary School supports student teachers, NQTs and practitioners in implementing the Citizenship Order in secondary schools - to be introduced in September 2002. This paper stresses that other changes in curriculum plans include providing further support for teachers. Recent efforts have been put forth to provide support for teachers in developing levels of religious and theological literacy, both of individual pupils and the society as a whole. It suggests tasks, activities and further reading designed to enhance the experiences of teachers. Resources are a factor that have been addressed in curriculum guidelines in the past and are a concern for the future. Tudor’s book is founded on the new curriculum for citizenship and the issues that arise from these changes. The book provides practitioners in secondary schools with the essential tools needed to teach citizenship. Tudor provides an overview of the citizenship order and gives guidance on how to fulfill its three main elements: social and moral responsibility, community involvement, and political literacy.

Questions (team decision) Assignment Example | Topics and Well Written Essays - 250 words

Questions (team decision) - Assignment Example The business operations should be aligned with the mayor’s goals in order stay in the business while addressing the mayor’s concerns. For instance, the hotels should create policies that will ensure that the business operate in environmental friendly way. The policies should focus on the social responsibilities of the firm. This means that in order stay in the business, the hotel should ensure that it restores the natural environment that has been destroyed by the business operations. The hotel should also agree to pay fines that are equivalent the destructions they cause on the natural surroundings. In this case, one would justify the decision to fulfill the responsibility and face certain loss by focusing on the benefits of the natural environment in the community. The case can be justified if the benefits of the natural environment outweigh the benefits of the hotels. For instance, it is worthy to eliminate the hotels as a long term solution since this will allows the beach plant to grow. This will attract more tourists, which will benefit the whole community. The hotels only benefit the hotel owners. One would also justify ignoring the responsibility by focusing on the benefits of the hotel. For instance, eliminating the hotels would lead to the loss of employment in the community. In addition, share holders will incur huge

Ri Paper For Internal Circulation Only Raffles Institution 2012 Essay Example for Free

Ri Paper For Internal Circulation Only Raffles Institution 2012 Essay Passage 1 1. What is the writer hoping to emphasise by italicising ‘does not’ and ‘cannot’ in line 6? [1] Lifted â€Å"Just because money does not buy happiness does not mean money cannot buy happiness,† says Elizabeth Dunn, a social psychologist and assistant professor at the University of British Columbia. Paraphrased The author hopes to emphasise †¦ (the contrast / difference between) what money often fails to do but actually has the potential to achieve. {MUST attempt to explain ‘does not’ and ‘cannot’. Merely saying that there is a difference/contrast is stating the obvious. E.g. ‘Does not’ is different from ‘cannot’. = 0m.} 1 2. Using your own words as far as possible, explain how we use money wrongly (line 8). [2] Lifted a. we tend to value GOODS over experiences, b. OURSELVES over others, Paraphrased a. People tend to think that commodities/things {must be tangible} are more important than experiences, b. People value themselves over others/self-centred†¦ OR spend on themselves rather than others, c. THINGS over people. c. People prefer spending on objects rather than people. d. [Inferred] We are spending on temporary pursuits and cravings that tend to disappear over time. {MUST capture the comparison (the reason it is wrong is because the preference is wrong). Need not follow the sequence in the answer scheme.} 3-4 points – 2 marks 1-2 points – 1 mark d. †¦ the spending that makes us happy, it turns out, is often spending where the money vanishes and leaves something INEFFABLE in its place. 1 For Internal Circulation Only 3. Using your own words as far as possible, explain three reasons why experiences are ‘inherently more social’ (line 28) [3] Lifted As experiences are inherently more social – when we vacation or eat out or go to the movies it is usually with other people, †¦we are liable also to RELIVE the experience when we see those people again. And past experiences can work as a sort of social adhesive even with people who did not PARTICIPATE with us, providing stories and conversational fodder in a way that a new watch or speedboat rarely can. In addition, †¦experiences do not usually trigger the same sort of PERNICIOUS comparisons that material possessions do†¦ (lines 2835) Paraphrased a. [Inferred] Experiences naturally involve interaction with people and 1 b. We are likely to recall / revisit / reminisce / recollect the experience when we meet those people again. c. People can also bond with people who were not present / strangers / others (through the sharing of anecdotes/conversational material). 1 1 d. Experiences do not cause hurtful comparisons†¦ OR experiences do not make people envious/jealous/ [inferred] insecure. 1 {MUST capture negative connotation.} 1m per point for any 3 of the above points. Maximum 3m. 4. Why does the writer use ‘seeming’ in the phrase ‘even for seeming essentials like cars, houses and, clothes’ (line 41)? Use your own words as far as possible. [1] Lifted Paraphrased Much of the impetus for a. Such needs are (could be) perceived to be crucial for discretionary spending everyday life even though they are not. OR†¦ even for seeming essentials like cars, b. Suggests that the author has doubts about whether houses, and clothes†¦ these objects are crucial. OR†¦ OR We might MISTAKE that c. The  writer feels that it is wrong to associate happiness with material/luxury goods. motivation for happiness, or for having a better life, but it is driven by something else, a human {Answer MUST include the idea of doubt and the need to compete or to fit explanation. in. Denied: sarcasm, cynicism, mockery Accepted: sceptical/scepticism (on its own), sceptical about this.} 1 2 For Internal Circulation Only 5. Explain what the writer means by ‘Talking about money and happiness in the same breath, it turns out, is not necessarily a surrender to crass materialism.’ (lines 47-48) [2] Lifted Talking about money and happiness in the SAME BREATH, it turns out, is not necessarily a SURRENDER to CRASS materialism†¦ Paraphrased a. Connecting / Linking money and happiness†¦ OR Talking about / discussing money and happiness together†¦ b. is in reality not always giving in to†¦ OR a submission to†¦ OR being a victim of†¦ c. a vulgar desire for material goods. OR excessive/shallow desire for luxury goods. 3 points – 2 marks 1-2 points – 1 mark. Passage 2 6. If you are a single male driving around in the Ferrari with nobody next to you, it is a glaring omission. (lines 14-15) a) Why does the writer refer to the Ferrari in line 15? [1] Pt Lift (not possible) Other trophies simply do not bring the payoff one expects. Says Loewenstein, If you are a single male driving around in the Ferrari with nobody next to you, it is a glaring omission. Inference a. A Ferrari is an example of a ‘trophy’. b. A Ferrari is a well-known example of a highly desired product. c. A Ferrari signifies high status / wealth in society. {Any of the above will be accepted.} 1 3 For Internal Circulation Only b) What does ‘glaring omission’ (line 15) imply here? Pt Lift (not possible) Other trophies simply do not bring the payoff one expects. Says Loewenstein, If you are a single male driving around in the Ferrari with nobody next to you, it is a glaring omission. [2] Inference [Inferred from ‘glaring’] a. It is strongly believed/popularly accepted that/very obvious {‘Emphasise’ and ‘highlight’ are not accepted because the question is not asking for the writer’s intention.} [Inferred from ‘omission’, reinforcing stereotypes.] b. that driving a Ferrari/luxury sports car will attract women / ladies / females / companions / partners. {Direct paraphrase of the quote is not acceptable.} 1 1 7. ‘A life that was constantly happy was not a good life’ (lines 30-31). Explain why this is a paradox. [2] Pt Lift Part of the meaning of life is to have highs and lows. A life that was constantly happy was not a good life. Inference [Must capture the clash.] a. We aim/wish/expect for maximum/optimum happiness in our lives all the time AND any of the following [Must capture what ‘good life’ entails.] b. Yet, a good life is one that has ups and downs, not just ups OR c. Yet, a good life is one with adequate/comparable negative experiences, not just positive experiences OR d. Yet, it is not just happiness that is ‘good’ but the ‘bad’ (of lows) is also seen to be ‘good’ {The key idea here is to question the value of ‘constant’ as a ‘good’ thing.} Answers will get either 2 marks or 0 mark. 1 1 4 For Internal Circulation Only 8. Summary Using material from paragraphs 2 to 5, summarise what Matthew Herper has to say about why wealth does not bring a constant sense of joy, why happiness is overrated, and the relationship between money and happiness. Wealth does not bring a constant sense of joy as Pt a. Lift Re-phrase Part of the reason is that people are not people do not know how to use their money very good at FIGURING out what to do to get happiness with the money, (lines 6-7) OR {Main idea of a lack of knowledge people are incompetent when it comes to understanding how to use money about how to manage money.} People generally OVERESTIMATE (line they have unrealistic expectations of the 8) {Students must capture the idea that it is off the mark.} the amount of long-term PLEASURE they will get from a given object. (lines 89) The way people spend money can make them less happy. (lines 9-10) Other trophies simply do not bring the (line 14) b. c. d. e. (Enduring) joy / happiness / positive outcomes that they can derive from something/object. The way people spend money can make them less happy. Other possessions also may not deliver f. g. {Do not accept ‘prizes’ / ’plaques’ / ’rewards’} PAYOFF one expects. (line 14) the rewards / utility / happiness we hope / wish to have. The central problem is that the human Next, our brain is accustomed to brain becomes CONDITIONED to pleasant/happy/joyful experiences as positive experiences. (lines 15-16) OR You can get used to anything, be it hanging by your toenails or making millions of dollars a day. (lines 19-20) Getting a chunk of unexpected money registers as a good thing, but as time passes, the response WEARS off. (lines 16-18) Mood may be set more by heredity†¦ (line 20) h. with time, their effect on us weakens/dies off/subsides. i. Lastly, our happiness level may depend on our genes 5 For Internal Circulation Only j. k. than by anything else (line 20) But this raises another question. How IMPORTANT is happiness anyway? (line 23) People with chronic illnesses describe themselves as happy, but they would still pay large sums for better health. And although healthy individuals are not much happier than quadriplegics, they would pay large sums of money to keep the use of their limbs. (lines 23-26) Some of lifes most SATISFYING experiences (line 26) do not bring happiness. (line 27) Part of the meaning of life is to have highs (line 30) and lows†¦Ã¢â‚¬  (line 30) A life that was CONSTANTLY happy (line 30) was not a GOOD life. (lines 30-31) more than other factors. OR predominantly / mostly [Inferred] However, the significance / necessity / crucial nature of happiness is still in doubt. [Inferred] Even supposedly happy people find happiness insufficient [Inferred] as they are willing to pay to ensure certain privileges†¦ OR health. l. m. n. o. p. q. r. s. Besides, some of the most gratifying/rewarding experiences do not make us happy. A meaningful life should also have its joyous and sad moments. A life that is always / perpetually happy is not fulfilling. t. u. {Do not award for point s in the absence of point r.} However, there may be at least one Yet, there remains a significant link IMPORTANT relationship between between money and happiness in that money and happiness (lines 31-32) †¦ happy people tend to have higher happy people earn more. incomes later on in their lives. (lines 3334) OR So, while money may not help make people happy, being happy may help them make money. (lines 34-35) Total: 21 points, 11 words that cannot be lifted Points ≠¥ 14 12 – 13 10 – 11 8–9 7 5–6 3–4 1–2 Marks 8 7 6 5 4 3 2 1 6 For Internal Circulation Only Wealth does not bring a constant sense of joy as†¦ people do not know how to use their money {a} and have unrealistic expectations {b} of the enduring joy that they can derive from something {c}. Other prized possessions {e} also may not deliver the rewards we wish to have {f}. Next, our brain is trained to be accustomed to pleasant experiences {g} as with time, their effect on us subsides {h}. However, the significance of happiness is still in doubt {k}. Even supposedly happy people find happiness insufficient {l} and are willing to pay to ensure their health {m}. Besides, some of the most gratifying experiences {n} do not make us happy {o}. A life that is always happy {r} is also not fulfilling {s}. Yet, there remains a significant link {t} between money and happiness – happy people earn more {u}. (16 points, 116 words) Vocabulary: 9. Give the meaning of the following words as they are used in the passage. You may write your answer in one word or a short phrase. Passage 1 Futility (line 3) noun Philosophers and gurus, holy books and self-help manuals have all warned of the futility of equating material gain with true well-being Answer should capture: ï‚ · Does not achieve purpose  OR lacks meaning ï‚ · Negative connotation Intriguing (line 8) adj The intriguing explanation for the poor wealth-to-happiness exchange rate is that the problem is not money, it is us. Answer should capture: ï‚ · Arousing intense curiosity OR very interesting ï‚ · Intensity 1 mark ï‚ · Hopelessness ï‚ · Uselessness ï‚ · Lack of purpose or meaning ï‚ · Efforts are in vain Accepted ï‚ · Ineffectiveness ï‚ · Meaningless ï‚ · Pointlessness 0 marks ï‚ · Wastage ï‚ · Worthless venture ï‚ · Failure ï‚ · Improbable ï‚ · Inability ï‚ · Foolishness ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · Ability to arouse intense curiosity Fascinating Deeply/very interesting Appeal strongly To captivate Very interesting and unexpected Very interesting and surprising ï‚ · ï‚ · ï‚ · ï‚ · Capture interest Arouse curiosity Extremely interesting Thoughtprovoking ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · Interesting {indication of intensity is not very apparent} Exciting Puzzling Ignorance Surprising Curious Intellectually stimulating 7 For Internal Circulation Only impetus (line 40) noun Much of the impetus for discretionary spending – even for seeming essentials like cars, houses, and clothes – comes from a desire to send certain signals about our buying power and our tastes. Answer should capture: ï‚ · The idea of a push or drive. Quell (line 45) verb And $5,000 worth of new stuff, or even $500,000 worth, is unlikely to permanently quell that need. Answer should capture: ï‚ · Suppressing. 1 mark ï‚ · (Sustained) drive ï‚ · Push ï‚ · Motivation ï‚ · A moving force ï‚ · Impulse ï‚ · Stimulus ï‚ · Boost Accepted ï‚ · Catalyst ï‚ · Thrust ï‚ · Spur ï‚ · Spark ï‚ · Trigger ï‚ · Incentive 0 marks ï‚ · Impulsive ï‚ · Penchant ï‚ · Impetuous ï‚ · Attack ï‚ · Assail ï‚ · Reason ï‚ · ï‚ · Subdue Suppress ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · Quiet Satiate / Quench Satisfy Appease Fulfil (incidental / part / whole) Allay Pacify Assuage ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · Vanquish / Defeat {wrong context} Stop / Halt / Cease Remove Overthrow / Overcome Repress Oppress Extinguish Destroy / Annihilate Kill Dispel Diminish Literally Figuratively Approximately {the idea of estimation, not accepted} Ostensibly Actually Effectually Basically Passage 2 Virtually (line 1) adverb Surveys have found virtually the same level of happiness between the very rich individuals on the Forbes 400 and the Maasai herdsmen of East Africa. Answer should capture: ï‚ · For the most part. ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · Nearly; Practically; Almost completely Almost wholly Almost entirely For the most part Just about ï‚ · ï‚ · ï‚ · Almost Effectively Essentially ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · ï‚ · 8 For Internal Circulation Only 10. Application Question: Drake Bennett talks about how pro-social spending can lead to happiness while Matthew Herper argues that money does not lead to happiness. Which writer’s views do you find more persuasive? Discuss the relevance of the arguments presented in both passages to you and your own society. [8] ï‚ · ï‚ · Answers that refer to only material from one passage would not be awarded more than 3 marks Explanation/evaluation should not just be a paraphrase of authors’ opinions Possible points From Passage 1 Bennett 1 Extract For deep-seated psychological reasons, when it comes to spending money, we tend to value goods over experiences, ourselves over others, things over people. (lines 9-11) Possible explanations S’pore context Relevant : In a society that is economically driven more than anything else, this is not a surprise. This drive, whether marked by early economic migration, post-war rebuilding, separation from Malaysia or simply Government push, remains sustained, if not stronger. Material success is celebrated and sheer narcissistic consumption is validated by commercial as well as social media. Examples : 2011 SMU study – S’porean women are significantly more materialistic than their American counterparts. Relevance of 5 ‘C’ [cash, credit card, car, condo, country club] notations despite more than a decade of use Not so relevant: There is a rising trend of S’poreans spending on people rather than objects; in 2007, Finance Minister Tharman had indicated clear tax concessions for charity spending and to focus Singapore as a ‘philanthropy hub’. More people, not just the rich, are donating generous sums to their alumni schools, religious charities and other organizations catering to  specific demographics such as the elderly, physically disabled, terminally ill etc. There is also increase in charity through CSR, whether by global MNCs or S’pore GLCs. Of course, the real intention behind the generosity is debatable but as corporations form a substantial percentage in a small workforce, the evidence is mathematically there that ‘we, the management/employees of company X, Y, Z†¦ are altruistic and caring’. There is a rising trend of social entrepreneurs opening restaurants to help the less fortunate and former prisoners. 9 For Internal Circulation Only 2 †¦ higher â€Å"prosocial spending† – gifts for others and donations to charity – was indeed correlated with higher selfreported happiness. (lines 18-20) Relevant : well-known philanthropists such as popiah king Sam Goi, Elim Chew of 77th Street, remisier Peter Lim, Chinese Chamber of Commerce ex-president Chua Thian Poh, Mustaq Ahmad of Mustafa Centre etc have gone on record to say so on newspaper interviews. Hong Kong billionaire loves to donate to Singapore, especially for education charity, for the same purported reason. The education system also introduces students to prosocial spending. The persistence of students in doing charitable acts also suggests happiness and satisfaction obtained. Not so relevant: This is not a technical disagreement. While the actual amount on ‘pro-social spending’ could be higher, it is usually by a small group of uber-rich AND older persons. The majority of Singaporeans, especially youth, have gone on record in many academic surveys that high levels of nuanced selfish material consumption is more rewarding than engaging in others’ welfare. The intense hours and anxiety spent on studying/work necessarily need to see material purchases as a direct reward. Perhaps this material reward can THEN lead to social happiness by the envy/admiration they evoke – e.g. for a Prada handbag, for the possession of a prized virtual weapon for a Diablo 3 PC online game character. This could be an unintended consequence of a meritocratic society. Most parents would also largely spend on their children and obtain great satisfaction. 3 money spent on experiences – vacations or theatre tickets or meals out – makes you happier than money spent on material goods. (lines 21-23) Relevant: Points to support this will  be relatively difficult based on available Singapore studies. Typically, Singaporeans vacationing will covet purchases from overseas; theatre ticket purchases locally may indicate a need to belong to a certain materially privileged crowd rather than the sheer love of the arts. Nevertheless, any qualified evidence is welcome to support agreement however the difficulty is in establishing the level of happiness obtained. Not so relevant: Typical youth population notes that playing World Of Warcraft or getting a new iPhone 4S is far more meaningful; middle-aged men and their toy figure collections; ladies of all ages on record saying that splurging on themselves, expensive cosmetic treatment more rewarding than anything else – refer to any Friday URBAN section of The Straits Times. As mentioned in earlier point, first the money has to be spent on material goods and THEN the experience related or shared can increase your happiness as a corollary. 4 As experiences are inherently more social †¦ we are liable also to relive the experience Relevant : the rise of alumni associations locally, aided by social media, have greatly increased networking; heritage and memorabilia gatherings are on the rise with 10 For Internal Circulation Only when we see those people again. And past experiences can work as a sort of social adhesive even with people who did not participate with us, providing stories and conversational fodder in a way that a new watch or speedboat rarely can 5 (lines 28-32) Talking about money and happiness in the same breath, it turns out, is not necessarily a surrender to crass materialism – it can also be a route to a new and more humane way to think about vitally important things like consumption, satisfaction, investment, and value. (lines 47-50) rich persons being key drivers behind these projects. Various hobby groups would also be relevant such as cycling clubs, dragon boating groups or cosplay groups. Not so relevant: There are hobby groups based on material goods such as car clubs or clubs for owners of toys. Material goods can also generate a sense of nostalgia. Relevant: SMU Board chairman Ho Kwon Ping and wife Claire Chiang regularly  promote pro-social spending and business; they are seen as new-age positive materialists and intellectuals. Christina Ong, boss of homegrown Club 21, is known for widespread charity outreach and growing her business, with equal pleasure. Not so relevant: For the majority of Singaporeans who lack such education or reflective practices, blatant monetary and property acquisition is the norm and only leads to a vicious cycle of debt, miscalculated risk-taking in investment etc From Passage 2 1 Extract People generally overestimate the amount of long-term pleasure they will get from a given object. Sometimes, Loewenstein notes, the way people spend their money can actually make them less happy. (lines 8-10) Possible explanations S’pore context Relevant: Just the consumer electronics industry here can attest to this; spending on upgrading of Apple smartphones from 3Gs to 4 to 4S; disposing of existing LCD for widescreen and 3D monitors, graphics cards in favour of new ones even though all are fully functional; rising number of secondhand computer parts shops is evidence of increasing technology shopping, which ultimately leaves one unhappy and insecure in the face of inability to keep up with the latest. While the tech spending may supposedly be mostly for males, the females display this ‘overestimation of long-term pleasure’ in copious fashion couture purchases and cosmetics. The increase in level of credit card debt is also an indication of less happiness as a result of overspending. Not to relevant: A basic level of income is necessary in Singapore to live a comfortable life. With a median household income of S$6000, anyone earning less would be hard pressed to live comfortably. In wired Singapore, having electronic goods is a necessity and can help attain more happiness in terms of education and interacting with peers. 2 The central problem is that the human brain becomes Relevant: Just looking at government financial decisions which affect the majority of the population after a while, 11 For Internal Circulation Only conditioned to positive experiences. Getting a chunk of unexpected money registers as a good thing, but as time passes, the response wears off. An expected paycheck does not bring any buzz at all  Ã¢â‚¬â€œ and does not contribute to overall happiness. (lines 15-19) Not so relevant: Since the Singapore population composition keeps changing with addition of foreigners, for the majority of the new citizens, this buzz is sustained. As Singaporeans also travel more, they might be more aware of societies which are not as fortunate as Singapore. Relevant: raising a family is universally seen as rewarding; the Government keeps promoting childbearing YET almost the first twenty years of a local child causes much stress for parents – working mothers post-maternity leave and the perennial maid/mother/mother-in-law management; young couples griping about childcare and choice primary schools; until the child is in a choice pri school and moving on to a premier sec sch, the parents put themselves under great duress. There is growing feeling that work-life balance is impossible and people are focusing more on their careers. Not so relevant: Hard to convince for majority local context but there is evidence that a positive change in attitudes to raising children is present, thanks to religious organizations, pro-family policies, national movements e.g. DadsForLife!. There are people who are giving up their careers for the sake of their children. There is also a push for women to return to the workforce after raising their children initially. 4 †¦ happy people tend to have higher incomes later on in their lives. So, while money may not help make people happy, being happy may help them make money. (lines 33-35) Relevant: There are professional who have given up their careers to take up their passions such as opening restaurants and bakeries, and they are doing well. There is an increasing number of entrepreneurs who pursue their passions. The main issue here is that happiness is independent of money and they do not have any links. Not so relevant In 2012, S’pore ranks high in benchmarks of physical comforts – healthiest country in the world, richest country in the world BUT ranks 90th in Happiness Index. Almost the entire local labour force can be on record for having technically higher incomes through their lives but the cost of living never lets up. Singaporeans are not happy or not happy enough but they keep making money anyway. the public becomes numb to the government’s welltimed monetary handouts and utility bill/town council fee reductions; the public expects more consultative engagement for sustained happiness. Recent feedback from the public seems to suggest that people overlook what is good in Singapore and focus on only the negative in terms of healthcare,  transportation, housing, and standards of living. Possible reasons could be a lack of sufficient information about life in other societies and countries. 3 Some of lifes most satisfying experiences do not bring happiness. For instance, having children actually makes people less happy over the short term (lines 26-28) 12

Today is a result of yesterday, tomorrow is a result of today Essay Example for Free

Today is a result of yesterday, tomorrow is a result of today Essay Today is a result of yesterday, tomorrow is a result of today. To what extent is this statement an apt representation of Buddhist ethics? Today is a result of yesterday, tomorrow is a result of today. This statement, as a representation of Buddhist ethics, would seem to suggest the realm of ethical action is effectively deterministic. This raises many questions about the nature of Buddhist ethics, most significantly: Is this a coherent position? If today is a result of yesterday how can one be said to be possessed of free will? And if one has no free will surely one cannot be considered a moral agent thus consequently, are responsible for ones actions. In this essay I will consider the evidence and arguments supporting this interpretation, together with counter-arguments in order to discuss the statement and reasoning behind this, resulting in my conclusion that will encompass all these arguments. Firstly I will look at morality in Buddhism as it is a Buddhists ethics which determine their behaviour which ultimately affects the results of tomorrow. Classifying Buddhist ethics can be considered an instance of virtue ethics, centred on the idea that the basis of morality is the development of good character traits which, in Aristotles system for example, include intelligence, wisdom (sila), and the ability to discern between good and bad. Peter Harvey contended that the Mahayana idea of skilful means (upaya) is similar to Christian situation ethics because it accepts ethical principles being overridden in certain situations in the name of wisdom and bodhichitta. Situation Ethics does not propose rules but rather suggests a guiding principle to decision making; Acting morally means acting in the most loving way in any situation. Unlike the approach of upaya in Mahayanan Buddhism, situation ethics do not ignore or reject traditional values but is bound by them. Both systems may allow for compassionate killing. There is a story in the suttas of how in one of his past lives the Buddha killed a robber to stop him from killing a number of bodhisattvas and thus prevented the robber from suffering in the hells for aeons. The difference though, is that in Buddhism only a very advanced bodhisattva is permitted to break with the traditional values, while situation ethics can be applied by anyone with a loving heart: This can thus be understood as in Buddhism, a Bodhisattva can act unethically and leave no imprint on the future, yet an unenlightened Buddhists actions will have a negative impact. Many religions emphasise the importance of the notion of good and bad actions, however in Buddhism, pre-determining actions to be good or bad would be a fruitless exercise Buddhists believe that reality of Dharma is beyond the concepts of good and bad; it contains both good and bad unseparated in a pre-conceptual state. Trying to remove half of reality, by definition, would be unachievable, thus trying to remove bad, would be unachievable, and pointless. More than that, the conscious effort to try to remove half of reality is also a kind of affirmation of the existence of just that part that you are proposing to remove. Buddhism does not say that there is no morality; it encourages the central importance of morals and ethical behavior in all areas of life. Although Buddhism believes in right action, it insists that right action is not the same as the Christian concept of right action; that moral action does not always match our conceived notions of morality. Buddhism believes that only this place and this moment are real and all else past and future are not real existence. It therefore follows that the only place where conduct can be right or wrong is here and now. So Buddhism emphasizes that right and wrong are concerned with the present moment, here and now. Acting morally means acting right at this very moment. Acting right at this moment is the only true morality. We can debate right and wrong as intangible concepts, but those abstractions are always detached from the real situation in front of us now, and so they are partial and can never be a complete guide to our action in the present; this notion would thus infer, in relation to the question I am researching, that past actions are not real existence, along with the future, therefore if the past is not real, how can it impact on tomorrow? It therefore follows that the only place where conduct can be right or wrong is here and now. So Buddhism emphasises that right and wrong are concerned with the present moment, here and now. Acting morally means acting right at this very moment. Acting right at this moment is the only true morality. It can be discussed right and wrong as abstract concepts, but those abstractions are always detached from the real situation in front of a Buddhist now, and so they are partial and can never be a complete guide to our action in the present. This therefore would conclude that if right and wrong actions are only connected with today, and not the past nor future No actions of yesterday impede on tomorrow. However this could be seen as a slight contradiction: Buddhism gives guidelines as to what good conduct is in the form of the Precepts these are not meant to be rigid and a broken rule will not result in committing sin, like the Christian Ten Commandments; However they are guidelines as to what right conduct is, but in actual situations conduct is decided by the state of the body/mind in the moment of acting, not by the precepts alone therefore if one is broken, Buddhism urges a buddhist to regain the balanced state and act in the present rather than be punished for past bad conduct, which has passed and can never be changed therefore if Buddhism states to live for today as the future and past are not real, yet the past bad conduct cant be changed therefore has left a mark in Buddhists life, how can past bad conduct even be conceived or considered to never be allowed to change if the past does not exist? To illustrate how a Buddhist might approach some of the ethical problems of today, you can look at the example of abortion. The early scriptures of Buddhism (The Pali Canon) are clear in seeing human life as starting with conception: when there is the union of the mother and father, and it is the mothers season, and the being to be reborn is present, through the union of these three things the conception of an embryo in a womb takes place. The word being, however, should not be thought of as a spirit or soul but consciousness being operated on by the force of (karma) that determines where the rebirth will be (according to previous deeds). When looking at such issue, we can see how it is an obvious moral problem as it has not been discussed at length in Buddhist literature; however there are references in the Pali canon that indicate the practice was regarded as wrong. Buddhist disapproval of abortion is related to the belief that in rebirth and teachings on embryology. It is widely held that conception marks the moment of rebirth, and that any intentional termination of pregnancy after that time constitutes a breach of the first of the Five Precepts (panca- sila), not to kill or injure living creatures, this could also be related the idea of euthanasia in Buddhism. This notion is an avowed view of most Buddhists however this position is not reflected in the abortion statistics in Buddhist countries: In more conservative countries such as South Asia, abortion is generally illegal, unless there is a threat to the mothers life, however illegal abortions are common with 300,000 per annum in Thailand, and in various east A sian countries abortions are even more numerous, such as one million per annum or greater is sometimes cited for countries such as Japan and South Korea. The fourth Noble Truth, the Noble Eightfold Path, sets out the main features of the Buddhist way of life addressing the nature of past behaviour affecting the future. The Buddha offered this path as the middle way, a way of life that does not fall into extreme views or extremes of behaviour but encourages balance and controlled moderation. Buddhist ethics are not based on the pursuit of sensual or other pleasures, and they also dont encourage extremes of deprivation, poverty or self-sacrifice. The various moral guidelines should be taken responsibly but with a light touch: Ethical conduct (sila) is built on the vast conception of universal love and compassion for all living beings, on which the Buddhas teaching is based (Walpola Rahula, What the Buddha Taught, London: Gordon Fraser, 1928). In relation to what is good or bad in Buddhism, the main principle is linked to intention and the determining factor is whether the act is out of selfish desire and craving, out of anger or hatred, or out of mindlessness and ignorance. Any actions that are motivated that way will result in caused suffering to us and to others. However, if the act is motivated by love and compassion they will bring well-being and happiness An action characterised by this moral quality (kusala-kamma) is bound to result (eventually) in happiness and a favourable outcome. Actions characterised by its opposite (akusala-kamma) lead to sorrow. This would thus infer that if actions result in happiness or sorrow , all actions of the past affect tomorrow. In Buddhism, ethical behaviour is ultimately dependent on the mind and not on the body. On the basis of the Buddhas advice, Buddhism has developed into many various types of ethical guidance that Buddhists do their best to follow. The ethical disciplines of Buddhism can be divided into the guidelines for lay people, and the guidelines for monastics. Within Theravada Buddhism, ethical action must always be motivated by ahimsa, the wish not to cause harm, and for Theravada monks, the main ethical guidelines are the 227 precepts of a monk. In Mahayana, ethical action must always be motivated by the bodhichitta, the wish to attain enlightenment for the benefit of others, in contrast to lay ethics where there is no need to attain enlightenment as there is no time to achieve such state. Ethical behaviour on the bodhisattva path consists of the six paramitas or transcendental actions. When one becomes a Buddhist one begins by taking the Three Refuges, refuge in the Buddha, Dharma, and Sangha. The main commitment is to refrain from harming others and to adopt the approach of non violence (ahimsa). Non violence is a prevalent principle of Buddhist ethics as stated in the Dhammapada 5: In this world hate never yet dispelled hate. Only love dispels hate (Dominique Side, Buddhism, Oxfordshire: Phillip Allan, 2005, p151). Lay people can also take one or several of the Five Precepts, which are five basic vows that strengthen the Buddhist way of life: I undertake to refrain from, 1) killing, 2) taking what is not freely given, 3) misusing sexuality, 4) harmful speech, and 5) taking intoxicants. As well as giving guidelines on what not to do, Buddhism encourages positive actions. In the Buddhas advice to a young man called Sigala, recorded in the Pali Canon, the Buddha explained the positive approach can be applied in daily life within the framework of the Six Relationships: 1) Take care of your family 2) Take care of your marriage 3) Keep good company 4) Develop good relationships between teachers and students 5) Develop good relationships between employers and employees, and 6) Develop a supportive and harmonious relationship with the monastic Sangha. The importance of love and compassion is also outlined in 118 of the Dhammapada, Set your heart on doing good. Do it over and over again and you will be filled with joy (ibid, p153). This statement could be interpreted as if you continuously do good the outcome tomorrow will leave you filled with joy, therefore suggesting the idea of free will in Buddhism. In the Noble Eightfold Path, ethical conduct includes three factors which overlap with the Five Precepts and Six Relationships: 1) Right Action, 2) Right Speech, and 3) Right Livelihood. Lay morality rests on the principle that lay people aim to minimise their bad actions and maximise their good so they have a better rebirth. Human actions (karma) in the Buddhist framework were to be determined based on both the intent or motive (chetanaa) and the consequences (vipaaka) of the action. In the Dharmaniyama (moral duty code), theories of causality in Buddhism were challenged in the view that human destiny was unaffected by the ethics or morality of human actions. It countered the doctrine of amoral causation (akriyavaada) whose supporters argued that there was no merit in doing good and no demerit for doing evil. In Buddhist teachings, individual karma is created by situations and moral predicaments, thus the outcome of a Buddhists future will be a result of yesterdays actions. Every action a Buddhist performs can leave an imprint and their karmic potential will allow for its own effect. Positive or virtuous actions give way to future happiness, and negative, non virtuous actions will result in future suffering. This connection between actions and their effects is known as the law of karma; this law is the foundations to Buddhist morality (sila). In the Buddhas teachings, Sila was spoken of vital importance as it allows for higher attainment of wisdom (panna) and concentration (Samadhi); this was mentioned in his Visuddhimagga: A wise man, after establishing well in virtue, develops consciousness and understanding (http://web.ukonline.co.uk/buddhism/panadpa8.htm). The law of karma is seen as a natural law just like physical laws like gravity. Karma is unlike morality in other religions as it is not operated by God or any other supreme being because Buddhism does not require supernatural intervention. The idea that karma works without any outside intervention means that the result of karma is not a reward, nor is it a punishment because they believe that this infers dependence upon a supreme power in judgement, whereas in Buddhism there is nobody to judge us, we determine our future by the way we act and the way we think, this idea is expressed clearly in the first verse of the Dhammapada, What we are today comes from our thoughts of yesterday, and our present thoughts build our life of tomorrow: our life is the creation of our mind. If a man speaks or acts with an impure mind, suffering follows him as the wheel of the cart follows the beast that draws the cart (Dominique Side, Buddhism, Oxfordshire: Phillip Allan, 2005, p130). This quote is an excellent example for evidence in supporting the question being asked in this essay, as it addresses the idea that not only our actions, but also our thoughts build our future and that impurities of the mind cause future suffering. A Common misinterpretation of karma is that it is a law of fate; however the idea of fate implies determinism, which would allow for no change or freedom, thus an idea seen in many theistic religions such as the notion that God determines life. This idea of determinism could be firstly viewed as a completely contrasting idea to that in Buddhism. Determinism is the philosophical proposition that everything is determined causally by an unbroken chain of prior events. However determinism has been expressed as the doctrine of Dependent origination in Buddhism and is an important factor in the evidence regarding my question. The doctrine of dependent origination is fundamental in connecting moral responsibility and causation, especially the ontological status of all matter. The doctrine teaches that all phenomena occur from dependence on causes and circumstances and lack intrinsic nature. The doctrine is expressed in its simplest form in phrase idam sati ayam bhavati, which means when this exists, that arises, which logically can be expressed as, when condition A arises, condition B arises, thus its reversal would be that when A doesnt exist, B wont. It is indicated in early sources that the Buddha became fully enlightened under the Bodhi tree when he fully realised the profound truth of Dependent Origination, thus that all matter is conditioned and arise and cease in a determinate chain of events. Therefore in relation to moral responsibility, it could be said that nothing happens out of its own volition, so there are no forces or metaphysical realties, such as a god or soul to act as a determiner, so it could be argued that moral responsibility is ones own, yet with a chain of causation, and no self, it could also be said one cannot be responsible if one doesnt exist. A way of classifying Buddhist ethics in Western terms is by relating it to soft determinism. Soft determinism is a midpoint between the hard determinism of philosophers such as Hobbes, Hume and John Stuart Mill, where the assertion of cause and effect is universal, therefore moral freedom is not possible; and libertarians who believe that uncaused, unconditional choices can be made and that free will exists, as Immanuel Kant stated free will was essential for morality, In morals, the proper and inestimable worth of an absolutely good will consists precisely in the freedom of the principle of action (Robert A Bowie, Ethical Studies, Nelson Thornes, 2001, p59). There is an important relationship between freedom and moral responsibility; it has been commonly held that we should be morally responsible for actions that we freely perform. If we can only blame or praise people for actions they freely and knowingly undertake, then it is vital that we have the freedom to act, morality depends on freedom. Immanuel Kant wrote ought implies can, therefore we cant blame someone for something they cannot do; If people are not free, the prospect of making moral decisions in denied, thus if an external influence causes human actions, people cannot be morally responsible. Hard determinists believe that we are not free and cannot be held morally responsible for our actions (Omar Khayyam R A Bowie, Ethical Studies, p91). This idea has some ethical similarity to the traditional Judeo-Christian idea of Predestination, it can be summarised as all our choices, decisions, intentions, other mental events, and all our actions are no more than effects of other equally necessitated events (Honedrich, ibid, p91). Hard determinism insists that all actions have a prior cause. However, this idea has a number of profound consequences; it puts doubt in our hopes for the future and how we consider the morality of others. If we praise a person for their good action, we are mistaken as it is not their action, it has been pre-determined. Additionally, if our actions are determined humans cannot deliberate rationally, we are, to an extent, illusive to the fact we cant decide for ourselves what we wish to do. Buddhism accepts the idea of determinism but rejects the idea of an agent and thus the idea that freedom is free will belonging to an agent. The Buddha said There is free action, there is retribution, but I see no agent that passes out from one set of momentary elements into another one, except the connection of those elements. Buddhism believes in neither absolute free will nor determinism, it preaches a middle doctrine, called pratitya-samutpada in Sanskrit, which is often translated as inter-dependent arising: When this exists, that exists; From the arising of this, that arises; When this does not exist, that does not exist; From the cessation of this, that ceases (Majjhima Nikaya, 1.262-64, D. Side, Buddhism, 2005, p97). It is part of the theory of karma. In Buddhism it is taught that the notion of complete freedom of choice is unwise, because it denies the reality of physical needs and circumstances. Similarly incorrect is the idea that we have no choice in life or that our lives are pre-determined. To deny freedom would be to undermine the efforts of Buddhists to make moral progress through our capacity to freely choose compassionate action. Pubbekatahetuvada, the belief that all happiness and suffering arise from previous actions, is considered an incorrect view according to Buddhist doctrines. Libertarianism however, rejects the idea of determinism. If we wish to retain the idea of moral responsibility and accept that a person can, when confronted with the choice between right and wrong, act as a free agent, one must accept the idea of libertarianism. Libertarians do not reject the idea of determinism completely, in general they agree that the inanimate world is mechanistic that all events are mechanically caused and therefore predictable and that all the mechanical chains of cause and effect may extend to the animate world. They deny the principle of universal causation applying to human action and that accordingly human behaviour is predictable. Libertarians distinguish between a persons formed character or personality and his or her moral self. David Hume described liberty in his An Enquiry Concerning Human Understanding: By liberty, then, we can only mean a power of acting or not acting, according to the determinations of the will; that is, if we choose to remain at rest, we may; if we choose to move, we also may (R A Bowie, Ethical Studies, 2001, p93). They believe we are not compelled to act morally by forces outside our moral consciousness. Moral actions are not chance or random events, but result from the values and character of the moral agent. An important argument for libertarianism is the human sense of decision making. While we have a sense of freedom, a sense of deliberating over our options, determinists maintain this is an illusion of freedom. Libertarianism does not explain human action, yet many would argue surely action needs a cause? Libertarianism attributes our moral judgement to an objective source, unmoved by environmental or upbringing, but this is questionable. Buddhists therefore do not agree with hard determinism in that Buddhists do not believe in an external cause, like libertarians, however they do not completely agree with the libertarian notion of complete free will. To resolve the debate on relating Buddhism to either Determinism or Libertarianism and thus create a type of determinism to which Buddhist causality can relate to, A.J. Ayer advocates a type of soft determinism which accepts that everything has a cause but which defines particular actions as free volitions. An action can be a free volition provided that 1) If you had had the volition not to do the action you would not have done it and 2) Nobody compelled you to do it. for Ayer we have responsibility for our volitional actions. In order to explain the difference between phenomena that are caused and free volitions, some soft determinists distinguish between the internal and external causes of an action, for example between its contingent and mental causes. It can be argued that while the external causes are determined, the psychological causes may not be; the Buddhist view is similar in that it states that physical objects and circumstances are always determined through causes and conditions. Physiological, physical and psychological causes are determined by mental decisions. Therefore a midway between the two contrasting theories is soft determinism and can most satisfactorily relate to Buddhist ethical principles. It would seem that we must choose between the belief in universal causation and, on the other hand, the belief in the existence of free will, it being accepted by both determinists and libertarians that these two beliefs are incompatible. The incongruity of the two however is rejected by soft determinists who say that human freedom and moral responsibility, is incomprehensible without determinism. Soft determinists state the assumption that determinism is inconsistent with free will is the result of considerable confusion about what is meant by free. It is true that freedom as incompatible with fatalism, the view that human beings are powerless to change the cause of events, but it is not conflicting with determinism. Therefore soft determinists have labelled the two ideas; when the cause of action is internal, therefore out of your own volition, you acted voluntarily and of your own free will; but when the cause is external, for example contrary to your wishes or desires, you acted involuntarily and under compulsion. According to the soft determinist, this distinction between internal and external causes explains why freedom and moral responsibility is not only compatible with determinism but actually requires it. Determinism is correct here in that for these actions to be uncaused would mean they would be completely unpredictable, impulsive and therefore irresponsible. Therefore when it is said a person acted freely it is not meant as his or her action was uncaused but rather they were not compelled to do it, that they were under no kind of external pressure, they themselves chose to act this way. We can relate the soft determinist notion of labelling the distinction between action and cause to Buddhist morality; in actual situations a Buddhists conduct is decided by the state of their body/mind in the moment of acting, not by the Precepts alone. They try honestly to follow the Precepts, but if they break one of the Precepts, Buddhism urges them to recover and regain the balanced state and act right in the present, rather than to forfeit for past bad conduct, which has passed and can never be changed as it does not exist anymore. Buddhism says that whether or not one can act morally or right in this moment does not depend on the concept or belief of what it right and what is wrong, but on the state of our body and mind at this present moment. The enlightenment (Bodhi) of the Buddha was both his liberation from suffering (dukka) and his insight into the nature of the universe; the Buddha was thus awakened to the truth of dependent origination. This is the idea that any phenomena only exists because of the existence of other phenomena in a complex web (Indras net) of cause and effect covering past, present and future. Everything is dependent on everything else: A human beings existence is dependent on the condition of everything else in the world (and universe) at that moment in time but equally the condition of everything in the world in that moment is dependent in an equally significant way on the character and condition of that human being. Indras Net is used in Buddhism as a metaphor for illustrating the concepts of emptiness of interconnectedness of all things. Everything in the universe is interconnected through this web of cause and effect thus all are interdependent. Therefore because all these things are transient (annicca) and conditioned it can be held that they do not exist, thus if they do not exist, can they be held morally responsible for their actions? The Heart Sutra disagrees with the idea of dependent origination and says that there is no such law as karma or cause and effect. This still however relates to the idea of emptiness (sunyata) as noted by Nagarjuna that dependent origination and emptiness are two sides of the same coin. For Nagarjuna, emptiness should not be interpreted ontologically, but rather in the way of the parable of the raft: The Buddhist teaching (especially shunyata), is like the raft one constructs for the crossing of a river. Once the river is crossed, the purpose of the raft has been served. Therefore the raft is not needed anymore. The same is true of emptiness: it should not be held on to; one who does hold on to it will have trouble functioning in life. Nagarjuna wrote extensively, and his teachings resulted in the formation of an Indian school called Madhyamika or the Middle Way School. Sunyata refers to the fact that no thing, including human existence, has ultimate substantiality, which in turn means that no thing is permanent and no thing is totally independent of everything else. In other words, everything in this world is interconnected and in constant flux. An appreciation of this idea of emptiness thus saves us from the suffering caused by our egos, our attachments, and our resistance to change and loss. Therefore all phenomena are dependent originations, which means that they dont exist, thus if they dont exist, they are empty which would thus mean that ethics itself is empty, thus it would be impossible to ever act ethically as there would not be such notion in existence. If everything is dependent causally, then it would seem impossible to act ethically: however a Buddhist would argue that the whole universe isnt dependent causally, there is only one determining factor, Karma. Thus one of the possible arguments against karmic determinism is that karma is only one influencing factor and the universe has a random aspect to it, which could account for a degree of freedom. Buddhist teachings explain how misleading states of mind keep you trapped in a recurring pattern of dissatisfaction by teaching about interdependent origination, the idea that things happen for a reason. Dependent Origination suggests that every event has a cause. For a Buddhist, life experiences, both good and bad, arent random, meaningless events, however, nor are they rewards, or punishments, as rewards and punishments require a controlling outside force sat in judgment such as a god, as there is no god in Buddhism blaming God or fate doesnt work, experiences come from a result of a series of causes and effects that begin in a buddhists mind. In teachings known as the Twelve links of dependent arising, Buddha described the mechanism that drives you from realm to realm within a cyclic existence and keeps you trapped in suffering and dissatisfaction in these links; Buddhas purpose for teaching these links, beginning with ignorance and ending with a corpse, was to make us aware of th e way our ignorantly motivated actions unavoidably lead to recurring suffering, this suffering motivates a Buddhist to seek a way out, thus nirvana. The Buddha taught that the way to achieve this freedom is by engaging in what he called the Three Trainings which form the foundations to which the entire structure of Buddhist practice rests. These three trainings were: Moral Discipline, the strength; Concentration, the sharp aim; And Wisdom, the tool. If a Buddhist practices the three trainings in combination with one another, the Buddha stated that cyclic existence would end and one would feel the inexpressible peace of liberation. With moral self-discipline as a base, concentration allowing for inherent focus, wisdom can break through ignorance and help you free yourself from the cycle of recurring misery. In relation to the asking if today is a result of yesterday, it would suggest it is, however a Buddhist can change tomorrows outcome by having moral self discipline, concentration and wisdom. If a Buddhist wants protection from suffering and from the danger of lower rebirth, they must try and not commit any more negative karma, however, this is also a problem in Buddhism as karma is generally negative, thus how can one create any more negativity if all karma is negative, thus everything really is Dukkha, therefore, in conclusion, it is apt to say that today is the result of yesterdays actions. However, a Buddhist can purify negative karma that has already been committed. There are ten principle non-virtuous actions that should be avoided; Three actions of the body, four of speech, and three of the mind. The three non virtuous bodily actions are killing, stealing and sexual misconduct; the four non virtuous verbal actions are lying, divisive speech, hurtful speech and idle chatter and the three non virtuous mental actions are covetousness, malice and holding wrong views. The best way for a buddhist to avoid negative actions is to practice consideration for others. All kind of non virtuous actions have three kinds of effects, the ripened effect, the effect similar to the cause and the environmental effect. The ripened effect of a negative effect is a rebirth in one of the three lower realms. A Buddhists previous actions will always have a definite outcome on tomorrow, however there are issues such as causality and free will which when taken into consideration make it hard to substantiate whether a Buddhist can be praised or blamed for such actions. The idea of Sunyata means that if everything in existence is empty, morality itself is empty which would mean acting ethically would be impossible, therefore actions are empty so the law of Karma is flawed, this is a contradiction in Buddhist philosophy as it would mean that there can be no actions of yesterday impacting upon the present, therefore I could reach a conclusion disagreeing with the statement Today is a result of yesterday, tomorrow is a result of today as it is impossible to be a result if there are no actions in existence allowing for a future; However, with many ideas in Buddhist philosophy contradicting each other it is hard to conclude on this question as there are too many conflictions between Karma being the universal law of cause and effect and dependent origination, as well as the idea that it is too simplistic a notion for today to be a result of yesterday as there are complicated ideas related to Buddhist ethics. Bibliography Damien Keown, Oxford Dictionary of Buddhism: Oxford University Press, 2003. Dominique Side, Buddhism, Oxfordshire: Phillip Allan, 2005. Geshe Kelshang Gyato, Introduction to Buddhism: Cumbria; Thorpe Publications, 1992. Michael Palmer, Moral Problems: The Lutterworth Press, 1991. Robert A Bowie, Ethical Studies: Nelson Thornes, 2001. Walpola Rahula, What the Buddha Taught: London: Gordon Fraser, 1926. www.buddhanet.co.uk www.wikipedia.org www.nibbana.com

Elucidating the p53 Signalling Network by Reverse Genetics

Elucidating the p53 Signalling Network by Reverse Genetics Reiyyan Tariq Nizami Aim In my research project I am working with four different strains Caenorhabditis elegans. I have been performing a double mutant screen using an RNAi knockout library. Cep-1 is a protein that causes apoptosis due to DNA damage in the worm. The cep-1 protein is an ortholog of the human tumor suppressor protein p53, which is found mutated in many cancerous cells. The aim of my experiment is to find genes that are regulators of cep-1 which cause lethality if mutated with cep-1. Alternatively mutants which are lethal as single mutants but survive as double mutants in combination with a cep-1 mutant are also potential genes of interest. These mutants have the potential to be regulated by drugs/proteins to change the levels of cep-1 and induce/prevent apoptosis in cells. Background Cancer is a disease of uncontrolled cell growth in our body. Under normal conditions a cell has regulatory proteins and check points that make sure that a cell is growing at the right pace and if for any reason the cell starts to grow irregularly then these proteins stop the cell growth and cause apoptosis. Apoptosis is the programmed death of a cell, as you can imagine it is tightly regulated and loss of regulation can result in catastrophe for the cell and the organism. When apoptosis is over active, healthy cells will die and this is often seen in diseases such as neurodegenerative diseases, hematologic diseases and many other tissue damage diseases. Loss of apoptosis in turn results in cancers, autoimmune diseases and inflammatory diseases. P53 is a tumor suppressor protein that was discovered in 1979. Since then a lot of research has been done into p53 and its functions. P53 is a tumor suppressor protein that is involved in defense of the cell. It is activated when a cell undergoes many various kinds of stress, such as DNA damage, hypoxia, metabolic stress and oncogene activation. It is one of the most important barriers to cancer in many mammals. P53 works in many various pathways, primarily p53 is involved in binding to transcription factors that then activate pathways involved in cellular defense, such as preventing angiogenesis and cellular growth. Malfunctioning p53 is one of the greatest hallmarks of cancer. The majority of mutations found in p53 are due to single nucleotide substitutions in the amino acids of the DNA binding domain of the protein. Similar cancerous phenotypes are observed when either p53 loses function due to a loss of function mutation or when negative regulators of p53 are overactive or positive regulators of p53 lose their function. However p53 is quite unique amongst tumor suppressor proteins because different missense mutations in the protein can cause different levels of p53 activity and hence have varying effects on the host. The effect of the mutation is further modified depending on the genetic background of the person with the mutation. Due to the different effects of various amino acid substitutions and the varying genetic background of patients it is quite difficult to study large populations. This is because high-throughput sequencing and genome wide single nucleotide polymorphism maps are expensive to obtain. The prices are going down as new technologies are becoming available however until now there have been no larger scale studies relating different mutations to varying levels of risks for different types of cancers. It is also hard to perform molecular studies on entire humans and so research is done on cell lines, mice and on cep-1 the Caenorhabditis elegans ortholog of p53. Caenorhabditis elegans is a worm in the nematodes phylum. They are a very well-studied organism due to their ease of study. C. Elegans are one of the simplest organisms that have a nervous system and that make them a very good model organism for neuronal studies. They are transparent creatures and so many molecular and cellular processes especially those involved in development have been thoroughly researched. They have a short generation time and are very cheap to maintain. They are an extremely good model organism for performing screens because any of their nonessential genes can be knocked out easily by using RNAi. The cep-1/p53 pathway is highly conserved throughout evolution as it is an extremely important pathway that is essential for cellular survival over time. The cep-1 protein works through the following pathway to cause apoptosis in cells suffering DNA damage. Cep-1 à ¯Ã†â€™Ã‚   Egl-1/Ced-13 –| Ced-9 –| Ced-4 à ¯Ã†â€™Ã‚   Ced-3 à ¯Ã†â€™Ã‚   Apoptosis A similar pathway is observed in humans p53 à ¯Ã†â€™Ã‚   BH3 –| Bcl2 –| Apaf1 à ¯Ã†â€™Ã‚   Caspase à ¯Ã†â€™Ã‚   Apoptosis A majority of these proteins are orthologs to one another. It was hoped that through my screen further proteins which interact with Cep-1 would be found and then their orthologs in humans could be found and then researched and targeted to regulate p53. Materials and Methods The experiment in itself was an extremely simple but time consuming experiment. Screens were performed with mutant worm strains which were then fed RNAi, through Escherichia coli (E. coli), to silence the gene of interest and create double mutants. To begin with a liquid screen is preferred over a solid screen. This is because with a liquid screen you can screen a larger sample of double mutants more easily than a solid screen. Liquid Screen The following strains of bacteria and worms were utilized, E. Coli – OP50, C. Elegans – N2, C. Elegans – GK138, C. Elegans – LG12501. E. Coli – OP50: Food source for C. Elegans C. Elegans – N2: Wild type worms C. Elegans – GK138: Cep-1 Mutant worms C. Elegans – LG12501: Cep-1 Mutant worms RNAi Knockout library for C. Elegans Chromosome 1 Day 1: Grow worms on Nematode Growth Medium (NGM) plates with OP50 as a food source for the worms. Day 3: Bleach worms that were plated on day 1 so as to only have eggs remaining on plates. Bend glass rod into L shape using a high temperature flame (Bunsen burner) Pipette bleach onto plates with worms on them and gently scrape the surface to mix worms and eggs with bleach Pipette fluid into Eppendorf tube and centrifuge at max speed for 3 minutes Aspirate most of the fluid and keep as much of the pellet as possible Suspend pellet with bleach Repeat steps 3 and 4 Suspend pellet using M9 solution Centrifuge at high speed for 1 minute Aspirate most of the fluid and keep as much of the pellet as possible Repeat 7 and 8 Suspend pellet using M9 solution and vortex Eppendorf tube at low speed to mix solution Place Eppendorf tubes in slow rocker in a 20 ° fridge overnight to allow eggs to hatch Day 4: Replicate bacteria containing RNAi from knockout library using a sterile 96 pin replication tool into a 96 well containing Liquid Broth (LB) with Ampicillin and allow the bacteria to replicate overnight at 37 °C in an incubator Induce transcription of RNAi using adding 0.1 Molar IPTG into wells and place in a shaker for 1 hour. Pellet bacteria by centrifuging in a cold centrifuge at 5 °C for 5 minutes at 2,500 g Remove the supernatant by flipping over the well quickly but carefully so as to keep pelleted bacteria in the wells Suspend bacterial pellet in wells using NGM Pipette worms into wells and place in 37 °C shaker Day 8: Remove worms from shaker gently making sure not to tilt the 96 well plates Observe and record phenotype of the worms Compare phenotype between the 3 different strains of worms Leave worms in 20 °C fridge overnight Day 9: Remove worms from fridge gently making sure not to tilt the 96 well plates Observe and record phenotype of the worms Compare phenotype from previous day Compare phenotype between the 3 different strains of worms Sequencing After potential hits were found in the liquid screen the RNAi from these bacteria were sequenced to ensure that the sequence of the RNAi was correct and hadn’t randomly mutated over time. RNAi was prepared by using a Qiagen miniprep spin kit. Suspend bacteria in 250  µl Buffer P1 and place in a microcentrifuge tube Mix 250  µl Buffer P2 and shake the mixture by flipping tube over a few times To the mixture add 350  µl N3 buffer, mix well quickly Centrifuge at 13000 rpm for ten minutes Aspirate supernatant into new tube Centrifuge again for a roughly 1 minute and discard the flow through Wash spin column with 0.5 ml PB buffer and centrifuge for 1 minute, discard flow through Wash spin column with 0.75 ml PE buffer and centrifuge for 1 minute Discard flow through and centrifuge at maximum speed for 1 minute Place prep column in a sterilized 1.5 ml microcentrifuge tube Add 50 ÃŽ ¼l of water to prep spin column and allow to rest for 1 minute after which centrifuge for 1 minute After the Qiagen miniprep is complete the tubes were sent for sequencing to The Centre for Applied Genomics where it was sequenced and results were obtained within a week. Solid Screen Solid screens were performed on genes which were found to have increased lethality with cep-1 deletion or increased survivability with cep-1 deletion. The following strains of bacteria and worms were utilized, E. coli – OP50, C. Elegans – N2, C. Elegans – TG12 E. Coli – OP50: Food source for C. Elegans C. Elegans – N2: Wild type worms C. Elegans – TG12: Cep-1 Fluorescent tagged (GFP) worms C. Elegans – Ned-8: Positive control C. Elegans – HT115: Negative control RNAi Knockout library for C. Elegans Chromosome 1 Day 1) Streak RNAi bacteria of interest from RNAi Library to obtain single colonies. Allow them to grow overnight in 37 °C incubator Day 2) Pick and grow a single colony in 5 ml of LB + Amp + Tet overnight in a 37 °C shaker Day 3) Add 0.1M IPTG for 4 hours to induce RNAi Plate 100  µl of induced bacteria on RNAi plates incubate at 37 °C incubator overnight Day 4) Pick 5 worms at the same stage and plate onto RNAi plates Allow to grow over 4 days at 20 °C Day 8) Score phenotypes and compared between different strains Day 9) Score phenotypes again on next day and compare between different strains and the previous day Fluorescent Microscopy We used a Differential interference contrast (DIC) microscopy to observe localization of cep-1 in TG12 worms. Worms were mounted using the following method Place a drop or two of hot liquid agarose onto microscope slide Immediately place a second slide perpendicularly on top of the agarose Allow agarose to settle for one to three minutes Gently slide off second slide so as to leave an agarose patch behind Add a drop of 1mM Levamisole to paralyze worms and prevent their movement Pick worms and place them on the slide Slowly place coverslip on top of worms, be very gentle Once prepared the slides were observed using a DIC microscope to find any irregularities in the localization or amount of cep-1 in the worms, especially in the germline and the eggs. Results The aim of this experiment was to find genes that cause lethality as double mutants with cep-1 mutants but not in wild type worms, these genes would be positive regulators of cep-1 and cause apoptosis through cep-1. The screen was also designed to find worms that cause lethality in wild type worms but not in cep-1 mutants, these would be negative regulators of cep-1 and cause apoptosis through cep-1. The results were gathered and tabulated to allow for an easier and better analysis of data. Note: There is a lot of data and so only data that is relevant is shown. Liquid Screen Results Legend L = Synthetic LethalE = Embryonic LethalG = Slow GrowthB = Egg laying abnormal R = No RNAi C = Contamination A = Larval Arrest S = Sterile V = Variable Morphology P = Lethal progeny W = No Worms Cells of interest are highlighted Chromosome 1 – Plate 1 N2 1 3 5 7 9 11 13 15 17 19 21 23 A R G G/L L C A/C A/C E R E/S G R R I S/L S R R R R K R R R R M R R R R S O R R S/E R cep-1 (gk) 1 3 5 7 9 11 13 15 17 19 21 23 A R L C/L C/L G/E C G/S G/L C/L E R E/S G R R I E/S E/S R R R R K R R R R M R G/E R R R /G/S O R R S R cep-1 (lg) 1 3 5 7 9 11 13 15 17 19 21 23 A R C/L L L E/P C G/L G L E R E/S G R R I S/B S/B R R R R K R R R R M R E/G R R R S O R R S/E R Chromosome 1 – Plate 5 N2 2 4 6 8 10 12 14 16 18 20 22 24 A C E G E I E K R M R R R O G R cep-1 (gk) 2 4 6 8 10 12 14 16 18 20 22 24 A C E G I E K R M R R R O R cep-1 (lg) 2 4 6 8 10 12 14 16 18 20 22 24 A C E G I E K R M R R R O R Key results of interest in liquid screen Lethality in gk and lg but not in N2 Well Gene N2 gk lg A23 F53G12.5 G/E E/P M03 Y95B8A_85.h G/E E/G Lethality in N2 but not in gk or lg Well Gene N2 gk lg G04 F25H2.10 E Solid Screen Results 3 hits of interest were obtained from all the plates that were screened. These 3 hits were then repeated multiple times on solid media to make sure the results were accurate. N2 lg gk A23 -1 A23 -2 A23 -3 N2 lg gk M03 -1 E M03 -2 E M03 -3 N2 lg gk G04 -1 L A G04 -2 E G04 -3 The M03 strain showed opposite results in the solid screen in comparison to the liquid screen. However since the results were consistent we decided to study both M03 and G04 under the microscope with fluorescent markers. Fluorescent microscopy results No major mislocalization of GFP tagged cep-1 was found and no extra or lack of GFP tagged cep-1 was observe