Summary of marketing article Essay Example | Topics and Well Written Essays - 500 words

Synopsis of showcasing article - Essay Example Of these specialists, Converse was one of the main to underline on clients which has now become the center of the idea of advertising blend. The creator has given a reasonable course of events which traces the different creators and their commitments to the idea since 1911 until 1960. The creator has given a reasonable and brief clarification of the different contributions from a few creators in an unmistakable way. The creator further proceeds onward to talk about the ‘blossoming of the advertising mix’, where he has brought up how Culliton (1948) proposed the name for the idea. He proceeds onward to give a clarification of how the name created and how the 1950s saw a development as far as the idea and the related speculations. Again the creator has given in a compelling and effective way the commitment of a few creators throughout the years and how the promoting blend idea developed and framed shape. The creator additionally features how the 4Ps of promoting had been presented and created in 1960. Besides, the creator has proceeded onward to talk about a fundamental part of the idea where the future contemplations for the showcasing blend have been made. Here the creator considers and talks about the commitments of Park, MacInnis and Silverman and their recommendations of the 4Ps setup. The idea appeared dependent on the few monetary speculations that were available in the prior years. Anyway there have been inquiries regarding the motivation behind the 4Ps and the general convenience as far as animating interest. The creator has addressed some significant parts of the current day thought of the field of showcasing and has drawn out some appropriately new considerations where the promoting has been perceived to be the authoritative culture and theory. The creator has additionally brought out fundamental focuses which feature the requirement for a fresher view on the idea which can be found in the light of some useful dynamic procedures of the current

The Portrayal of Women in Art 1962-2002 Free Essays

We have, pretty much, as a group of people become used to the romanticized portrayal of ladies. Regularly, especially in old style styles, they were depicted as leaning back nudes who were there for the viewer’s delight. With deflected eyes, they contacted themselves exotically, normally guiltless and careless that there is somebody painting her for all to see. We will compose a custom paper test on The Portrayal of Women in Art: 1962-2002 or on the other hand any comparable theme just for you Request Now At the point when they weren’t sexual-dream grub, they were servile and obedientâ€particularly in the 1940?s and 1950?s after the finish of the tough ladies period of World War II. They wore their hair in flawless twists, with their ideal dresses and worked cheerfully away in their ideal kitchens. In Jack Levine’s Girl with Red Hair there is a move away from the ideal, lighthearted lady that preceded. Or maybe, bareness is held onto as a part of the woman’s power as opposed to the viewer’s object. The subject stands up to the watcher with her look. This picture isn't a representation of an exposed young lady, yet rather, a young lady who happens to be stripped. There is no hint of provocativeness or sensualityâ€we are attracted to her face with the goal that we may endeavor to observe what this young lady is thinking. In spite of the fact that her bosoms are there, they are ineffectively rendered contrasted with the profundity of her face and don't trap the eye like the conveniently delineated tissue of the leaning back nudes. Hotline for Troubled Teens, 1970. Joe DeMers (1910-1984). Acrylic ready, 22 ? x 18 ? in. New Britain Museum of American Art, Gift of Walt Reed, 2000. 45. Through both this aesthetic strengthening of ladies and the then stimulated Feminist Movement, ladies turned out to be less items for a viewer’s delight and rather autonomous characters. In Joe DeMers’ Hotline for Troubled Teens, the sex is almost expelled from the young lady. She is seen wearing an over-shirt that shrouds her bosoms and her other ladylike highlights are limited. The young lady is gone into a narrativeâ€no longer is there a dislodged naked body simply lying around. Rather, we are demonstrated a young lady in her not really perfect life. Her face is concerned and the phone string is wrapped about her shoulders and wrist. She gives off an impression of being totally pretentious of her viewersâ€be they out in the city around her, or somewhere else. She is self-serving and worried about just her current circumstance. The title even proposes that this young lady is connecting (at that point, even that would have been untouchable) so as to help herselfâ€a rule that started to enable ladies during the Feminist Movement. Laneisha II, 1996. Dawoud Bey (b. 1953). Polacolor ER prints, 90 x 45 3/4 in. New Britain Museum of American Art, Members Purchase Fund, 2000. 34. This is one of my preferred bits of the assortment for some reasons. Transcendently, the portrayal of ladies has revolved around the â€Å"ideal woman†Ã¢â‚¬which, on the off chance that you haven’t got a magazine recently, is ordinarily white, appealing, youthful, meager and energetic. The lady here, in any case, is the absolute opposite. Despite the fact that she is alluring, she doesn't have the â€Å"elegant† highlights that a painter may have searched for in the main portion of the century. She is divided into six pieces and keeping in mind that they for the most part coordinate upâ€in that there are no tremendous holes of informationâ€there is a noteworthy distortion of her figure. Her face is extra wide and left arm appears to be strangely long. An unmistakable distinction between the superbly kept and rendered ladies of the past, this cutting edge lady permits her imperfections and her dissension to be reflected in the middle of each casing. She is a lady, not an item to view. Untitled, 2000. Cindy Sherman (b. 1954). Shading photo, release 1/6, 32 1/2 x 22 in. Individuals Purchase Fund, 2000. 88. I especially appreciate this piece for a few reasons: like the piece above, she isn't regularly â€Å"beautiful†Ã¢â‚¬particularly for the time wherein it was taken. Or maybe, her appearance is outdatedâ€thick, dim eyebrows, slicked back hair and that horrendous blue shirt she is wearing. Rather than excusing the lady as terrible, we can see past her rawness. She bears a face that nearly says â€Å"Yeah, so what? ’ to her crowd. She isn’t hip, nor is she youthful and excellent (as directed by the measures of society) any more. There is an obtuseness to this photo that debilitates the sitter; it nearly appears as though she’s the one judging and not the other way around. Magnificence I, 2002. Imprint Catalina (b. 1965). Acrylic on canvas, 72 x 72 in. New Britain Museum of American Art, Gift of the Artist, 2003. 01 Lastly, this piece appears to me to be the most powerful out of the bundle. We are not conscious of the â€Å"real† picture, however just its negative. In structure, we may perceive the individual as a female. They have bosoms, long streaming hair, jewelry†¦ a portion of the key markers of what we may connect with being a lady. Be that as it may, with the modified hues, we are indicated somebody with masculine highlights and in this manner, the lines of sexual orientation are obscured. Obviously, the cosmetics the subject is wearing is exaggeratedâ€dark lips and feline like eyeshadowâ€and further veils the individual’s sexual orientation. This piece is so relentlessly attached to the manner by which sex and sex are isolated and characterized. In this, the craftsman is reclassifying the presence of ladies, in that ladies may not be â€Å"feminine† by any means. This piece introduces the topic of womanliness and womanhood in a totally new manner, and is completely proper in the developing setting of ladies in workmanship. What's your opinion about the depiction of ladies in craftsmanship? How has it changed over the most recent 500 years? 50 years? 5 years? By what method would women be able to pick up power through portrayal in craftsmanship? How does this contrast with men in craftsmanship? Instructions to refer to The Portrayal of Women in Art: 1962-2002, Papers

Blog Archive Mission Admission Give Short Answers Adequate Attention

Blog Archive Mission Admission Give Short Answers Adequate Attention Mission Admission is a series of MBA admission tips; a new one is posted each Tuesday. Many candidates will work painstakingly on their essays, prepare rigorously for their interviews and endlessly contemplate their choice of recommenders. However, applicants tend to leave the short answer section of their applications to the last moment, and some will simply paste in information from their resumes or from their applications to other schools. However, the short answer sectionsâ€"meaning the actual questions within the application about your work and community experiences, hobbies, etc.â€"should definitely  not  be ignored, and should be completed with the same spirit of diligence that you would bring to all the other aspects of your application(s). We advise candidates to refrain from just pasting bullet points from their resumes into the short answer section, and to instead take time to truly contemplate the questions that are asked and write out the answers in full sentence form. (Further, bullets from a resume often lack the necessary context, so they rarely make effective answers for this portion of the application.) Although the information conveyed in the short answer section is important, we believe that what is really crucial is that candidates give care and thought to  all  aspects of their applications. Share ThisTweet Mission Admission

The Raven By Edgar Allen Poe - 1203 Words

Edgar Allen Poe was an American writer stretching his life span from 1809 to 1849. His style of writing, while gloomy and erratic, becomes thrilling through the metaphorical resemblance of life. While covering his stories in a morbid tone there is a bright sense of hope and reason at the end of his poems or short stories many people relate to. After all nobody is happy without a sense of overcoming a hardship or obstacle in their life. â€Å"The Raven† is one of Edgar Allen Poe’s most famous poems. In the poem the character experiences a loss and travels through the grief process. While doing so he becomes ill at ease with his emotions. Expressing wrath, despair, and sorrows he may soon find the answer to his emotional crises. â€Å"Once upon a midnight dreary, while I pondered, weak and weary, Over many a quaint and curious volume of forgotten lore, While I nodded, Nearly napping Suddenly there came a tapping,† Edgar Allen Poe, â€Å"The Raven†, (1845). The classic poem â€Å"The Raven† consists of a dark gloomy setting. The character heard somebody at his door. Assuming that the knock is from a visitor he remains sitting in a depressed state of mind. In the first stage of grief you refuse to accept the loss of a loved one. You ignore and avoid the loss. A disturbance comes from outside the charter’s window. â€Å"Open here I flung the shutter, when, with many a flirt and flutter, In there stepped a stately Raven of the saintly days of yore,† (â€Å"The Raven†). When he opens the shutters a ravenShow MoreRelatedThe Raven By Edgar Allen Poe1233 Words   |  5 Pagesproper use of it a poem may not be as strong as others. Poetry is a form of art, and a poet uses language as a painter would use color to portray their art. Edgar Allen Poe is one of the greatest poets in history because his mastery of literary elements such as allusion, allegory, symbolism, and figurative language. â€Å"The Raven† by Edgar Allen Poe is a fascinating poem in which he displays excellent use of language. There are many symbols in this poem which help the reader understand the change in theRead MoreThe Raven By Edgar Allen Poe1494 Words   |  6 PagesIn 1844, Edgar Allen Poe published one of his most famous poems â€Å"The Raven.† â€Å"The Raven† is about a man reading books trying to keep his mind off a woman named Lenore he loved that died. Later, while he is napping, he wakes up from a knocking on the door. He gets up and opens the door of his chamber, but no one is there. He goes to his window then opens the shutters. A raven flies right in and sits on a bust of Pallas, goddess of wisdom, on top of his chamber door. The narrator sits there askingRead MoreThe Raven By Edgar Allen Poe1049 Words   |  5 Pages Poetry has long been an art form that has entertained readers for many years. Edgar Allen Poe, a poet from the 1900’s, is known for his deep dark poems. He wrote many poems that are now considered classics. One of his greatest works, â€Å"The Raven†, was written in 1845; just two years before his wife Virginia Clemm died of Tuberculosis. The entirety of â€Å"The Raven† is confined in a man s chamber. It is a December winter and it is midnight. The narrator, the person who resides in the chamber, is readingRead MoreEdgar Allen Poe s The Raven1398 Words   |  6 Pagesthat are difficult to understand. However, Edgar Allen Poe’s poem, The Raven presents the concept of insanity in an enchanting way. In this paper, I am going to argue that Poe’s use of the image of the raven represents the narrators budding madness and could possibly be viewed as a symbol for his subconscious mind in the poem. Poe seems to structure the raven in this way through his abstract language, form, tone, symbolism and imagery. Although The Raven is rich in symbolism, the plot is actuallyRead MoreSymbolism Of The Raven By Edgar Allen Poe1045 Words   |  5 Pagesmeaning within the narrative itself. As a poet Edgar Allen Poe occupied his narratives with literary elements. Symbols become one of the most predominant elements in his works of literature. Consequently, in his concise poem â€Å"The Raven,† the literary element most noted by the reader is symbolism. Characters, places, and objects all have a deeper meaning within this poem. In â€Å"The Raven† the most noted of the symbols are the bust, Lenore, the time, and the raven. Before discussing the topic proper, theRead MoreThe Raven, by Edgar Allen Poe Essay896 Words   |  4 Pages The poem, â€Å"The Raven,† written by Edgar Allen Poe shows the deep depression and confusion that the narrator is experiencing since the death of his beloved wife. The gloomy setting of the poem predicts the visit of the Raven, whom is a sign of misfortune, darkness, and death. Throughout the poem, the narrator is continually mourning his wife, Lenore. He secretly hopes that the Raven will bring good news regarding his wife and his future; however, the Raven informs him that he will forever remainRead MoreThe Raven, By Edgar Allen Poe1164 Words   |  5 Pagesn the discussions of Edgar Allen Poe’s â€Å"The Raven,† one issue has been the explanation of the bird and it’s presence in the poem. Majority agrees that the feathered creature does have a symbolic meaning of the conscious mind of the speaker in the poem. The agreement usually ends with how we communicate with our selves. Some focus more on who or what a person is talking to with their conscio us while others direct their attention to how we can better understand our conscious conversations with ourRead MoreEssay about Edgar Allen Poe: The Raven1151 Words   |  5 PagesEdgar Allen Poe: The Raven Creating the Melancholic Tone in â€Å"The Raven† Edgar Allan Poe’s The Raven, representing Poe’s own introverted crisis of hell, is unusually moving and attractive to the reader. In his essay entitled The Philosophy of Composition, Poe reveals his purpose in writing â€Å"The Raven† and also describes the work of composing the poem as being carefully calculated in all aspects. Of all melancholy topics, Poe wished to use the one that was universally understood, death; specificallyRead MoreAnalysis Of The Poem The Raven By Edgar Allen Poe1376 Words   |  6 Pagesproper use of it a poem may not be as strong as others. Poetry is a form of art, and a poet uses language as a painter would use color to portray their art. Edgar Allen Poe is one of the greatest poets in history because his mastery of literary elements such as allusion, allegory, symbolism, and figurative language. â€Å"The Raven† by Edgar Allen Poe is a fascinating poem in which he displays excellent use of language. There are many symbols in this poem which help the reader understand the change in theRead MoreAnalysis Of Edgar Allen Poe s The Raven984 Words   |  4 Pageslive in a world where money can only get you so far. The one thing that we ultimately strive and mourn the loss for is love. After reading Edgar Allen Poe’s â€Å"The Raven,† you begin to realize the downside of experiencing love is loss because at that point your loneliness creates a veil of uncertainty in everything you do. Through imagery, repetition, and allegory, Poe conveys to the reader that the loss of a loved one results in lonesomeness and alienation not just from one’s self-but from the world.

How Libertarianism Is The Best Of The Three Views

The aim of this essay is to prove that Libertarianism is the best of the three views, which refers to the idea of human free will being true, that one is not determined, and therefore, they are morally responsible. This essay will be strengthened with the views and arguments on C.A. Campbell. Campbell demonstrates that one could have done otherwise and one is the sole author of the choices they make. I will present a few arguments in the essay in support of Libertarianism and attempt to rebut the views of others in this process. . As a result, this essay will prove that you are held morally responsible for any act that you perform. The Libertarian view consists of one’s actions not being determined, which means that a person would have†¦show more content†¦A prime example of these inner acts can be seen in a decision for a teen to decide to smoke a cigarette. Two teens are given the same opportunity in smoking their first cigarette. The first teen comes from a bad upb ringing with questionable role models in their life, the second teen comes from a good home and from parents who have instilled strong moral values. After reviewing both teens environments one would most likely come to the conclusion that the first teen with the poor upbringing would smoke and the teen with the good home life would resist the urge to smoke. This conclusion may well be correct but from the aspect of a Libertarian the end result of whether these two teens choose to smoke has no correlation on whether they have free will. According to Campbell, When we decide to exert moral effort to resist a temptation, we feel quite certain that we could withhold the effort; just as, if we decide to withhold the effort and yield to our desires, we feel quite certain that we could exert it- otherwise we should not blame ourselves afterwards for having succumbed (Campbell). When we apply Campbell s views to the teens issue of smoking we can see why the final choice these teens make o n whether to smoke doesn t matter. It is indeed the inner struggle to resist smoking that matters. If both teens struggle with the decision to smoke that means that both have the opportunity

Designing A Design Services For Sprinkler Systems

You asked the following questions: â€Å"We have a storage arrangement of large wooden boxes stored on top of each other to approximately 20 ft. The boxes are filled with household items of Class III commodities. Section 5.6 does not specifically address this commodity. Would Class III be a proper description of this commodity? Would Chapter 14 be appropriate to determine density? Specifically Figure 14.2.4.2 with high temperature sprinklers?† In response to your questions we have reviewed NFPA 13, 2010 edition that you indicated as the applicable standard. Our informal interpretation is that there are few questions that we are unable to answer; unfortunately, yours is one of them. The protection criteria will have to be discussed with†¦show more content†¦I hope you understand our dilemma regarding basic design for facilities. To date, we ve answered over 2000 written requests for interpretations and countless numbers of verbal requests. I hope we can assist you in the future. The following is some guidance to help get you started. First, A.3.9.1 indicates that a six-sided portable storage container can be a combustible or noncombustible enclosed storage unit with varying dimensions ranging from small portable rental storage units to large freight containers. The Technical Committee on Sprinkler System Discharge is unaware of fire tests, and no design guidance is furnished. A table was also added as Table A.5.6, Examples of Commodities Not Addressed by Classifications where one item listed is Storage Container - Large container storage of household goods. That applies to your scenario. To get you started, check with the client on the commoditi es that will be stored inside the portable moving containers. Typically, household items have a range of commodity classifications from Class I through expanded Group A plastics in the form of foam cushions or mattresses. Since the commodities may vary, the worse case scenario of expanded Group A plastics may be a starting point. The container storage would not qualify as rack storage and would match closer to solid pile storage arrangement. The decision tree in Figure 15.2.2 provides guidance on selecting a density in Table 15.2.6(a) for Control Mode

Accounting Issues in XYZ Company-Free-Samples-Myassignmenthelp.com

Question: Analyze the Financial and Accounting Issues in XYZ Company. Answer: Introduction: This report has been prepared to analyze the financial and accounting issues in a company. It is quite normal for a company o face accounting and financial issues in its routine life. There are various ways and techniques which is not accepted by the accounting board and used by the companies for convenience creates an issue for the company. It is required for every company to manage the accounts and finance according to the set rules and regulations to offer an exact result to the user of the reports and internal and external stakeholders of the company. Accounting and financial rules have been set by the boards according to the international accounting and finance rules and regulations to make a common set of rules and regulations for every state and the companies. Singapore culture is diverse in nature. It has been found through analyzing the Singapores accounting board that the rules and regulations in Singapore are similar to the international rules and regulations. More, in this report corporate governance of Singapore has also been studied to analyze the rules of corporate governance in the country. Company act of Singapore has briefed about the corporate governance code of the country which must be followed by every company in the state. Further, it has been analyzed that how the concerned company is applying corporate governance code in its business and what are the issues which could be faced by the company due to corporate governance code. This report briefs the user about many issues which could be faced by a company due to many internal and external factors. Further, many other factors have also been analyzed on a Singaporean company to understand the factors of the company briefly. Further, this report briefs the user about the entire internal and external factor of the company. Company overview: For this report, XYZ plc has been taken into consideration. This company is operating its business functions in Singapore from last many decades. This company has been established in 1980. This company is operating in retail industry. Many stores and warehouses are operated by the company to manage and enhance the sales of the company. Further, this company also offers various services and products to its customers. This company is enhancing its productivity rapidly. Annual report of this company has also been analyzed and it has been found that the liquidity, profitability, capital structure, efficiency etc condition of the company is in the favor of the company or vice versa. Further, many other aspects related to finance and accounts has been analyzed over the company to make a better decision[1]. Following are the reports of the company: XYZ Plc INCOME STATEMENT Fiscal year ends in June. MVR In millions except per share data. 2016-06 2015-06 2014-06 2013-06 2012-06 Revenue 3954 3652 3484 3308 3128 Cost of revenue 3089 2854 2745 2610 2480 Gross profit 865 798 739 699 648 Operating expenses Sales, General and administrative 1006 931 884 839 771 Other operating expenses -361 -334 -336 -318 -284 Total operating expenses 644 597 548 521 486 Operating income 221 201 191 178 161 Interest Expense 4 6 9 10 14 Other income (expense) 1 1 0 1 1 Income before income taxes 218 196 183 168 148 Provision for income taxes 66 59 54 51 44 Minority interest 0 0 Other income 0 0 Net income from continuing operations 152 137 128 117 105 Other 0 0 Net income 152 137 128 116 105 Net income available to common shareholders 152 137 128 116 105 Earnings per share Basic 1.51 1.36 1.27 1.16 1.05 Diluted 1.5 1.35 1.25 1.16 1.05 Weighted average shares outstanding Basic 100 100 101 100 100 Diluted 101 101 102 101 100 EBITDA 263 241 227 211 193 XYZ Plc BALANCE SHEET Fiscal year ends in June. MVR In millions except per share data. 2016-06 2015-06 2014-06 2013-06 2012-06 Assets Current assets Cash Cash and cash equivalents 52 49 43 67 40 Total cash 52 49 43 67 40 Receivables 98 81 71 64 58 Inventories 546 479 459 426 428 Prepaid expenses 4 4 4 5 Other current assets 6 3 2 2 2 Total current assets 703 617 578 564 534 Non-current assets Property, plant and equipment Other properties 419 385 366 333 301 Property and equipment, at cost 419 385 366 333 301 Accumulated Depreciation -236 -208 -185 -152 -119 Property, plant and equipment, net 184 176 182 181 182 Equity and other investments 0 0 0 0 Goodwill 37 36 36 35 30 Intangible assets 49 49 49 49 49 Deferred income taxes 21 17 15 15 16 Total non-current assets 290 278 282 280 277 Total assets 992 895 860 843 811 Liabilities and stockholders' equity Liabilities Current liabilities Accounts payable 302 254 244 335 365 Deferred income taxes 11 9 8 15 4 Deferred revenues 29 Other current liabilities 134 117 70 93 70 Total current liabilities 447 380 352 442 439 Non-current liabilities Long-term debt 110 139 180 124 150 Other long-term liabilities 31 32 33 33 37 Total non-current liabilities 141 171 213 158 187 Total liabilities 588 552 565 600 627 Stockholders' equity Common stock 49 57 58 63 62 Retained earnings 328 269 220 169 118 Accumulated other comprehensive income 27 18 16 12 5 Total stockholders' equity 405 343 295 243 185 Total liabilities and stockholders' equity 992 895 860 843 811 XYZ Plc Statement of CASH FLOW Fiscal year ends in June. MVR In millions except per share data. 2016-06 2015-06 2014-06 2013-06 2012-06 Cash Flows From Operating Activities Other non-cash items 185 180 41 156 215 Net cash provided by operating activities 185 180 41 156 215 Cash Flows From Investing Activities Investments in property, plant, and equipment -52 -42 -36 -35 -46 Property, plant, and equipment reductions 0 0 1 1 1 Acquisitions, net -2 -3 -4 Net cash used for investing activities -52 -44 -38 -38 -45 Cash Flows From Financing Activities Long-term debt issued 54 Long-term debt repayment -30 -40 -26 -84 Common stock issued 6 3 22 1 4 Repurchases of treasury stock -13 -5 -26 Cash dividends paid -93 -87 -77 -65 -77 Other financing activities 0 -1 0 -1 0 Net cash provided by (used for) financing activities -131 -130 -28 -91 -158 Effect of exchange rate changes 0 0 1 1 0 Net change in cash 3 6 -24 28 12 Cash at beginning of period 49 43 67 40 27 Cash at end of period 52 49 43 67 40 Free Cash Flow Operating cash flow 185 180 41 156 215 Capital expenditure -52 -42 -36 -35 -46 Free cash flow 133 137 5 121 169 Supplemental schedule of cash flow data Cash paid for income taxes -66 -60 -61 -40 -49 Cash paid for interest -4 -6 -7 -9 -13 Impact on stakeholders: Stakeholders of the company are those people or parties who are related to company through any monetary or non monetary relationship. Stakeholders are the main parties of the company. Mostly, shareholders are the main stakeholder of an organization. Shareholders are the investors who have invested their money into the company to raise the invested amount. Shareholders interests get affected with every change in the operations and business functioning of the company[3]. It is required for every shareholder to analyze the financial and non financial data of the company and then make an investment into the business of an organization. Through analyzing over XYZ plc it has been found that the performance of the company is enhancing rapidly. Minor changes have been faced by the company from last 5 years due to many internal and external issues. It has been found that from last 2 years, economy of the state is getting fluctuate due to international market and it is also impacting over the financial condition of the company. Following are the details over the financial data of the company from last 5 years which will depict the user about the liquidity condition, profitability situation, solvency analysis, efficiency situation etc. Computation of ratio analysis Liquidity ratio 2016 2015 2014 2013 2012 Current ratio 1.572706935 1.623684211 1.642045455 1.2760181 1.216400911 Quick ratio 0.351230425 0.363157895 0.338068182 0.312217195 0.241457859 Working capital 256.0 237.0 226.0 122.0 95.0 8% Profitability Ratios 2016 2015 2014 2013 2012 Operating Profit Margin 9.578907436 10.37102957 10.87112514 11.44951632 12.10837596 0.792122137 Net Profit Margin 0.038442084 0.037513691 0.03673938 0.035066505 0.033567775 -0.000928393 Return on Capital Employed 69.5 73.5 74.6 94.5 101.8 4.048276476 Return on Equity 0.375308642 0.39941691 0.433898305 0.477366255 0.567567568 0.024108268 Return on Total assets 0.153225806 0.153072626 0.148837209 0.137603796 0.12946979 -0.000153181 Solvency Ratios [4] Capital structure ratio 2016 2015 2014 2013 2012 Debt- equity 1.455445545 1.609329446 1.915254237 2.469135802 3.407608696 Interest coverage ratio 9468.75 6312.5 4208.333333 3787.5 2705.357143 Activity ratio Efficiency ratio 2016 2015 2014 2013 2012 Receivable turnover ratio 44.17877095 48.05263158 51.61481481 54.2295082 Creditor turnover ratio 11.11151079 20.5323741 9.481865285 21.0483871 Inventory turnover ratio 6.027317073 35.2345679 6.203389831 40.78125 Assets turnover ratio 4.190779014 4.161823362 4.091603053 4 Computation of return on capital invested Particular 2016 2015 2014 2013 2012 ROCI -13.03214596 -10.29929577 -10.29281278 -19.62365591 -23.52941176 Through, above are the details of the company which has been faced by the company from last 5 year. These details would help the entire stakeholders of the company to make decisions about the company. Ratio analysis assists every stakeholder of a company to investigate the relevant information and make a better decision about the companys performance and their investment into the company. Here, liquidity position of the company depict about the ability of the company which is useful for the company to met its short term obligation, this information is required by the debtors, creditors and short term investors of the company to analyze the short term financial situation of the company. Here, it has been found that the current ratio of the company has been decreased from last year but has been enhanced from 2012. Currently, the current ratio is 1.57 which depict that company is utilizing the minimum resources at maximum level. Further, it has been found that the quick ratio of the company has also been improved from last 5 years. Company is managing a 0.35 of quick ratio in 2016 which depict that 35% of total liabilities could be paid by the company quickly which a standard rate is according to the industry. Further, working capital management of the company depict about the improvement in the working capital from last year as the operations of the company has been enh anced so it is also required for the company to enhance the working capital to run the business smoothly[5]. More, profitability position of the company depict about the ability of the company to earn the profits in its short term period as well as long term period, this information is required by the debtors, creditors, banks, financial institutes, suppliers, short term and long term investors of the company to analyze the short and long term financial situation of the company. Here, it has been found that the operating profit margin ratio of the company has been decreased from last year and it has also decreased from 2012. Currently, the ratio is 9.57 which depict that company is earning 9.57% of total revenue as a profit. Further, it has been found that the net margin ratio of the company has also been decreased from last 5 years. Company is managing a 0.038 of quick ratio in 2016 which depict that 3.8% of total sales could be retained by the company as net profit of the company. Further, return on capital employed of the company has been analyzed which depict about the total return to t he stakeholders over the capital employed of the company. It has been found that the return on capital employed ratio of the company has also been decreased from last 5 years due to less profit[6]. Return on equity and return on total assets have also been analyzed to offer the information to the stakeholders of the company. The return on equity depicts about the total return to the equity holders of the company. It has been analyzed that the return amount has been lower due to less profit and return on total assets depict about the relationship about the revenue and the assets of the company and it has been found that the return on total assets ratio has been enhanced from last year. Here, solvency ratio of the company depict about the ability of the company to manage the capital structure of the company in long term period, this information is required by the debtors, creditors, banks, financial institutes, suppliers, short term and long term investors of the company to analyze the short and long term financial situation of the company. Here, it has been found that the debt equity ratio of the company has been decreased from last year and has also been decreased from 2012. Currently, the debt equity ratio is 1.45 which depict that company is managing the 1:1.45 ratio of debt and equity. Further, it has been found that the interest coverage ratio of the company has also been decreased from last 5 years. Company is managing a 9468.75 of interest coverage ratio in 2016[7]. More, activity position of the company depict about the ability of the company to efficient the activities and business functioning of the company, this information is required by the debtors, creditors, banks, financial institutes, suppliers, short term and long term investors of the company to analyze the short and long term financial situation of the company. Here, it has been found that the receivable turnover ratio of the company has been decreased from last year and it has also decreased from 2012. Currently, the ratio is 44.17. Further, it has been found that the creditor turnover ratio of the company has also been decreased from last 5 years. Company is managing an 11.11 of creditor turnover ratio in 2016. Further, inventory turnover ratio of the company has been analyzed which depict about the total time period in which the inventory is ordered by the company for further production. It has been found that the return on inventory turnover ratio of the company has also been de creased from last 5 years due to less profit and strategy changes of the company. Asset turnover ratio of the company has also been analyzed to analyze the asset condition in the company[8]. The asset turnover ratio has been analyzed and found that the 4.19 times the revenue of the company is higher than the total average assets of the company. Lastly, return on capital invested of the company depict about the ability of the company to pay the returns to the investors in its short term period as well as long term period, this information is required by the debtors, creditors, banks, financial institutes, suppliers, short term and long term investors of the company to analyze the short and long term financial situation of the company. Here, it has been found that the ROCI of the company has been enhanced from last year and it has also enhanced from 2012. Currently, the ratio is -13.03 which depict that company is paying -13.03% of total revenue as a dividend to the shareholders. Accounting and finance problems: Further analysis has been done over the companys financial and accounting data to analyze the process of preparing the financial statement of the company. It has been analyzed through study over the accounting and financing books, it has been found that there are many issues which could be faced by an organization due to the final statements of the company and the process through which these statements are created by the company. It has been found through this study that it is required for every company to understand the accounting concept deeply and prepare the data accordingly. For it, companies could hire a professional accountant, who has knowledge about the accounting rules and regulations so that the financial report and accounting analysis could be done easily in the company. In concern of XYZ plc, it has been found that there are various issues in the process of preparing the final financial data of the company which is depicting wrong information to the user. It has been found that company is not following entire rules and regulations perfectly. It has also been found that company is required to look over the international regulations too while preparing the final data. It has been evaluated that company is making changes into many techniques every year just to depict wrong information to the stakeholders so that they could be impressed through the company and make an investment in the company. In the reports of the company, it has been found that company is not focusing over the conceptual framework totally. Company is still using some traditional approach to prepare the financial reports while new policies have been implemented by the country for the listed companies to prepare the final financial reports accordingly. It has been analyzed that the depreciation methods used by the companies to depreciate the machineries and other fixed assets of the company are not same in each year. Company makes the changes in it according to the convenience. Further, it has also been analyzed that company is not focusing over the conceptual framework totally[9]. Company is still using some traditional approach to prepare the financial reports while new policies have been implemented by the country for the listed companies to prepare the final financial reports accordingly. Further, GAAP rules have been studied and found that there are many rules of GAAP have not been accepted by the com pany and still it is working like before. Lastly, the assumptions which have been taken by the company while making financial and accounting decision analyzed. The issues of the company are as follows: Depreciation techniques: Depreciation is a tool which is used by the companies to figure out the exact value of fixed assets of the company. Depreciation over the assets includes a significant part of the assets in many organizations. Depreciation could have an important effect in presenting and determining the results of operation of this organization. The depreciation objective is to stipulate the depreciation accounting treatment[10]. Depreciation accounting rules and regulation depict that property, equipments and plants of the company must be depreciated, and it does not consider the natural resources of the company. Goodwill of the company must not be depreciated. Depreciation is the distribution of depreciable amount over an asset in estimated useful life. It is charged in a financial year through the net profit of the company either directly or indirectly[11]. In this case of XYZ Plc, it has been found that depreciation techniques used by the company are quite similar according to the depreciation accounting standard but the consistency of depreciation methods have not been followed by the company[12]. It has been found that in each year, company make changes in the depreciation % as well as depreciation techniques. Such as in 2013, company has used straight line method to depreciate the equipments but in 2015, company has changed the method and applied diminishing method to depreciate the equipments of the company. Further, it has been found that various changes have been made by the accountant of the company to make the attractive financial data. It has been found through analyzing over the depreciation techniques of the company, it has been found that company wanted to reduce the tax charges and enhance the net profit of the company thus it had made changes into the depreciation techniques. Depreciation also creates an issue for the company n terms of selecting the total estimated useful life and take assumption about the scrap value of the company. MASB also depict that it is required for every company to disclose the depreciation technique in its annual report while preparing the financial reports[13]. Company is required to depict entire information related to depreciation such as the estimated life of an asset, residual value of the asset, scrap value of the assets; depreciation allowed in a period, gross depreciable amount in a financial year etc must be disclosed by the company in its annual reports. Through the annual report of the company, it has been analyzed that there are various points which have not been mentioned by the company in its annual reports. It has been found through this report that the annual report of the company is not depicting the entire information about the depreciation of the company[14]. It has been found through the reports that the accountant of the company is either have less information about the accounting standards or he is pressurize by the company to make the annual reports more attractive and thus he ignored many accounting regulations[15]. Conceptual framework: Basically, it is not some rules and regulations or accounting standards which makes the principles and concepts to assist the accountant or the top level manager in preparing the final statements and annual reports of the company. Consequently, conceptual framework is a set of regulations and rules which should be concerned in a firm while making the financial statements and annual reports of the company[16]. It makes sure about the financial statements reliability to the external stakeholders of the company. MASB has launched a framework to help out the accounting professionals to prepare and present the financial statements in 1989. This framework has been known as conceptual framework. This conceptual framework locates the conception which positions the preparation and presentation of statements of final financial reports for the external as well as internal stakeholders[17]. It also assists a firm to prepare and present the financial statements with the assistance of many international rules and regulations; it offers depth information about the preparation and presentation of final statements as well as it also considers the accounting standards. Conceptual framework assists the auditors in shaping their own opinion about financial statement of a firm. It assists financial statements user to understand the figures according to their requirement. Internal users of final financial statement and annual reports of a firm are top level managers like production manager, finance director, marketing manager etc. These financial concepts are used by the firm to make decision about future strategy. Every stakeholder considers the financial statements for different objectives. Like a manager could look for decision making, entrepreneur for the objective of growth and development of business, personnel for the reason of take an idea about financial performance and health of the firm. Creditors of the company analyze the risk; politics and government look to examine the tax return[18]. It has been explained about 4 characteristics, which are as below: Understandability Relevance Reliability Comparability[19] In the case of XYZ plc, it has been found that the company is not following the conceptual framework while presenting and preparing the final reports of the company. It has been found through an analysis over the annual report of the company that company is taking many assumptions while preparing the reports rather than it must consider the conceptual framework to maintain the financial detail of the company[20]. Further, it has been found that this framework is not followed by the company due to the accountant of the company. Either the accountants have less information about the accounting standards or they are pressurized by the company to make the annual reports more attractive and thus he ignored many accounting regulations. GAAP rules: GAAP rules are also known as generally accepted accounting principles. GAAP is a compilation of commonly-followed standards and rules of accountants for financial and accounting reporting[21]. GAAP specifications contain explanation of principles and concepts, as well as some rules related to industry. The main purpose of GAAP is to make sure that the reporting of financial and accounting figures are consistent and transparent from one firm to another[22]. There is no general GAAP customary and the particulars vary from one location (geographical) to another. In the Singapore, the MASB permissions that financial reports hold on to GAAP necessities. The IFRS, IASB, FASB, MASB etc boards stipulates GAAP general and the GASB (Governmental Accounting Standards Board) describes GAAP for local and state government[23]. Companies which are publicly traded must observe with both GAAP and SEC requirements. Many states around the globe have espoused the IFRS (International Financial Reporting Standards)[24]. It has been designed to offer a worldwide framework to public limited companies to prepare and reveal their financial statements and reports. Accepting a single set of accounting global standards simplifies the procedures of accounting for international countries and offers auditors and investors with a unified view of investments. In the case of XYZ plc, it has been found that the company is not following the global standards while presenting and preparing the final reports of the company. It has been found through an analysis over the annual report of the company that company is taking many assumptions while preparing the reports rather than it must consider the conceptual framework to maintain the financial detail of the company[25]. Further, it has been found that these GAAP standards have not been followed by the company due to the accountant of the company[26]. Either the accountants have less information about the accounting standards or they are pressurized by the company to make the annual reports more attractive and thus he ignored many accounting regulations. Assumptions: Fundamental accounting assumptions or basic assumptions mean the accounting conceptual which have been concerned and chased while recording and presenting the financial information. As required informations are different from stakeholder to stakeholder, firm to firm and condition to condition so it could be said that assumption varies. Basically, there are 3 assumptions which are considered by the companies while preparing and presenting their financial data. Following are there assumptions: Going concern concept Accrual concept Consistency Further, some accountants depict that there are 5 assumptions which must be used by the companies, which are as follows: Business entity concept Going concern concept Accruals concept[27] Money Measurement concept Consistency Though, through the MASB, it has been analyzed that following assumptions could be used by the company for preparing and presenting the financial information of the company: Going concern this assumption depict that the business will always run. Accrual basis of accounting this statement depict that entire financial statements apart from the cash flow statement must be made according to the accrual basis[28]. Consistency this statement depict that consistency in classification and presentation must be there while preparing the financial statement. Materiality this statement depict that material class of same items which are presented individually[29] Offsetting this statement depict that assets alongside incomes and liabilities against the expenses which could not be offset except required or permitted by IAS. Among the above five, beginning 3 are of more importance as they has been least affected by requirements of IAS and situation of the business. Singapore corporate governance and issues regarding the Singapore corporate governance to the company: Singapore corporate governance has been studied to analyze the corporate governance. It is a procedure which is apprehensive about how firms could be managed, how managers are administered, what queries could be faced by BOD and the responsibility a firm corporation has to shareholders. Corporate governance is an important aspect which is followed by the companies to manage the effectiveness of the company and meet its social responsibilities[30]. It has been found through the reports that it controls over entire matters of the company. It assists the company to manage the contribution and development of the firm. Singapore corporate governance offers the good relations and policies to the companies to adopt and enhance their business functioning. In this case, it could be investigated that the XYZ Plc has been able to execute efficient corporate governance that directs the firm to become more spirited in the market. According to the investigation over the retailing industry of Singapore and XYZ Plc the issues has been concerned that the board must include the skill level and care which is expected from the directors and insufficient financial necessities and requirements. It has been analyzed that board of directors and managers are working on the same phase to resolve these issues[31]. In positive aspects terms, the board of director, managers and other employees of the company has been capable to manage entire stuffs of the company and also manage the competitive performance of XYZ Plc. The BOD (board of directors) of this firm makes sure that entire actions of the company are legal and stick to business ethics. Further, they also make sure that the social responsibility of the company is also included with their responsibilities. Hence, the firm has some positive aspects as well as some negative aspects. One of which remains to the concept that, due to the inconsistencies and mistakes of the entity who is involved, it could not be assured always that there would be no errors or losses which would occur[32]. In capability to grip the differences and diversities also makes a negative aspect which could be attached with corporate governance practice of the company. Lastly, incapacity to make sure shareholder worth is one more negative aspect of the firm. It could be said that shareholders value must be enhanced. Lastly, it could be concluded that these corporate governance practices help the company and individual to maintain the competitive advantages of the company[33]. Conclusion: Thus through this report, it could be concluded that Accounting and financial rules have been set by the MASB boards according to the international accounting and finance rules and regulations to make a common set of rules and regulations for every state and the companies. Singapore culture is diverse in nature. It has been found through analyzing the Singapores accounting board that the rules and regulations in Singapore are similar to the international rules and regulations[34]. More, in this report corporate governance of Singapore has also been studied to analyze the rules of corporate governance in the country. Company act of Singapore has briefed about the corporate governance code of the country which must be followed by every company in the state. Further, it has been analyzed that how the concerned company is applying corporate governance code in its business and what are the issues which could be faced by the company due to corporate governance code[35]. This report briefs the user about many issues which could be faced by a company due to many internal and external factors[36]. Further, many other factors have also been analyzed on a Singaporean company to understand the factors of the company briefly. Further, this report briefs the user about the entire internal and external factor of the XYZ plc References: Edwards, D. Wolfenzon, B. Yeung, Corporate governance, economic entrenchment, and growth, Journal of economic Literature, 2005 Sep 1;43(3):655-720. Epstein, AA. Mirza, Wiley IFRS 2006: interpretation and application of international financial reporting standards, Wiley; 2006 Feb 3. Epstein, EK. Jermakowicz, WILEY Interpretation and Application of International Financial Reporting Standards 2010, John Wiley Sons; 2010 Feb 5. Doidge, GA. Karolyi, RM. Stulz, Why do countries matter so much for corporate governance?, Journal of financial economics, 2007 Oct 31;86(1):1-39. Nobes, RH. Parker, Comparative international accounting, Pearson Education; 2008. Hay, WR. Knechel, N. Wong, Audit fees: A Meta?analysis of the effect of supply and demand attributes, Contemporary accounting research, 2006 Mar 1;23(1):141-91. Helleiner, S. Pagliari, The end of an era in international financial regulation? A postcrisis research agenda, International Organization, 2011 Jan;65(1):169-200. Choi, GK. Meek, International accounting, Pearson Higher Ed; 2011 Nov 21. White, AC. Sondh, D. Fried, Analysis of Financial Statement, Analysis, 2005. Chen, Q. Tang, Y. Jiang, Z. Lin, The role of international financial reporting standards in accounting quality: Evidence from the European Union, Journal of International Financial Management Accounting, 2010 Sep 1;21(3):220-78. Davies, D. Green, Global Financial Regulation: The Essential Guide (Now with a Revised Introduction), John Wiley Sons; 2013 May 8. Jackson, Variation in the intensity of financial regulation: Preliminary evidence and potential implications, Yale J, on Reg,, 2007;24:253. Filatotchev, R. Strange, J. Piesse, YC. Lien, FDI by firms from newly industrialised economies in emerging markets: corporate governance, entry mode and location, Journal of International Business Studies, 2007 Jul 1;38(4):556-72. Gray, J. Hamilton, Implementing financial regulation: Theory and practice, John Wiley Sons; 2006 May 1. Solomon, Corporate governance and accountability, John Wiley Sons; 2007 Mar 16, Kim, JS. Tsui, HY. Cheong, The voluntary adoption of International Financial Reporting Standards and loan contracting around the world, Review of Accounting Studies, 2011 Dec 1;16(4):779-811. Revsine, DW. Collins, WB. Johnson, Mittelstaedt HF, Financial reporting and analysis, New York, NY: Pearson/Prentice Hall; 2005. Radebaugh, SJ. Gray, EL. Black, International accounting and multinational enterprises, New York, NY: Wiley; 2006 Jan 4. Iqbal Khadaroo, Business reporting on the internet in Malaysia and Singapore: A comparative study, Corporate communications: An international journal, 2005 Mar 1;10(1):58-68. Barth, WR. Landsman, MH. Lang, International accounting standards and accounting quality, Journal of accounting research, 2008 Jun 1;46(3):467-98. Barth, WR. Landsman, MH. Lang, International accounting standards and accounting quality, Journal of accounting research, 2008 Jun 1;46(3):467-98. Young, MW. Peng, D. Ahlstrom, GD. Bruton, Y. Jiang, Corporate governance in emerging economies: A review of the principalprincipal perspective, Journal of management studies, 2008 Jan 1;45(1):196-220. Brown, International Financial Reporting Standards: what are the benefits?, Accounting and business research, 2011 Aug 1;41(3):269-85. Brown, International Financial Reporting Standards: what are the benefits?, Accounting and business research, 2011 Aug 1;41(3):269-85, Gourevitch, J.Shinn, Political power and corporate control: The new global politics of corporate governance, Princeton University Press; 2005. Ball, International Financial Reporting Standards (IFRS): pros and cons for investors, Accounting and business research, 2006 Dec 1;36(sup1):5-27. Ball, Robin A, Sadka G, Is financial reporting shaped by equity markets or by debt markets? An international study of timeliness and conservatism, Review of accounting studies, 2008 Sep 1;13(2-3):168-205. Ball,International Financial Reporting Standards (IFRS): pros and cons for investors, Accounting and business research, 2006 Dec 1;36(sup1):5-27. Tricker, RI. Tricker, Corporate governance: Principles, policies, and practices, Oxford University Press, USA; 2015. Weber, DW. Arner, Toward a new design for international financial regulation. Gilson, CJ. Milhaupt, Sovereign wealth funds and corporate governance: A minimalist response to the new mercantilism, Stan, L, Rev,, 2007;60:1345. Holthausen, Accounting standards, financial reporting outcomes, and enforcement, Journal of Accounting Research, 2009 May 1;47(2):447-58. Holthausen, Accounting standards, financial reporting outcomes, and enforcement, Journal of Accounting Research, 2009 May 1;47(2):447-58 Li, Does mandatory adoption of International Financial Reporting Standards in the European Union reduce the cost of equity capital?, The accounting review, 2010 Mar;85(2):607-36. Jeanjean, H. Stolowy, Do accounting standards matter? An exploratory analysis of earnings management before and after IFRS adoption, Journal of accounting and public policy, 2008 Dec 31;27(6):480-94. Wu, Corporate governance and corruption: A cross?country analysis, Governance, 2005 Apr 1;18(2):151-70 Ball, R,, International Financial Reporting Standards (IFRS): pros and cons for investors, Accounting and business research, 2006 Dec 1;36(sup1):5-27. Epstein BJ, Mirza AA, Wiley IFRS 2006: interpretation and application of international financial reporting standards, Wiley; 2006 Feb 3. Barth ME, Landsman WR, Lang MH, International accounting standards and accounting quality, Journal of accounting research, 2008 Jun 1;46(3):467-98 Chen H, Tang Q, Jiang Y, Lin Z, The role of international financial reporting standards in accounting quality: Evidence from the European Union, Journal of International Financial Management Accounting, 2010 Sep 1;21(3):220-78. Holthausen RW, Accounting standards, financial reporting outcomes, and enforcement, Journal of Accounting Research, 2009 May 1;47(2):447-58 Epstein BJ, Jermakowicz EK, WILEY Interpretation and Application of International Financial Reporting Standards 2010, John Wiley Sons; 2010 Feb 5. Iqbal Khadaroo M, Business reporting on the internet in Malaysia and Singapore: A comparative study, Corporate communications: An international journal, 2005 Mar 1;10(1):58-68. Brown P, International Financial Reporting Standards: what are the benefits?, Accounting and business research, 2011 Aug 1;41(3):269-85. Brown P, International Financial Reporting Standards: what are the benefits?, Accounting and business research, 2011 Aug 1;41(3):269-85 Choi FD, Meek GK, International accounting, Pearson Higher Ed; 2011 Nov 21. Holthausen RW, Accounting standards, financial reporting outcomes, and enforcement, Journal of Accounting Research, 2009 May 1;47(2):447-58 Kim JB, Tsui JS, Cheong HY, The voluntary adoption of International Financial Reporting Standards and loan contracting around the world, Review of Accounting Studies, 2011 Dec 1;16(4):779-811. Revsine L, Collins DW, Johnson WB, Mittelstaedt HF, Financial reporting and analysis, New York, NY: Pearson/Prentice Hall; 2005. Gray J, Hamilton J, Implementing financial regulation: Theory and practice, John Wiley Sons; 2006 May 1 Radebaugh LH, Gray SJ, Black EL, International accounting and multinational enterprises, New York, NY: Wiley; 2006 Jan 4. Ball R, International Financial Reporting Standards (IFRS): pros and cons for investors, Accounting and business research, 2006 Dec 1;36(sup1):5-27, White GL, Sondh AC, Fried D, Analysis of Financial Statement, Analysis, 2005. Li S, Does mandatory adoption of International Financial Reporting Standards in the European Union reduce the cost of equity capital?, The accounting review, 2010 Mar;85(2):607-36. Barth ME, Landsman WR, Lang MH, International accounting standards and accounting quality, Journal of accounting research, 2008 Jun 1;46(3):467-98 [20] Li S, Does mandatory adoption of International Financial Reporting Standards in the European Union reduce the cost of equity capital?, The accounting review, 2010 Mar;85(2):607-36. Weber RH, Arner DW, Toward a new design for international financial regulation Hay DC, Knechel WR, Wong N, Audit fees: A Meta?analysis of the effect of supply and demand attributes, Contemporary accounting research, 2006 Mar 1;23(1):141-91, Davies H, Green D, Global Financial Regulation: The Essential Guide (Now with a Revised Introduction), John Wiley Sons; 2013 May 8 Nobes C, Parker RH, Comparative international accounting, Pearson Education; 2008. Helleiner E, Pagliari S, The end of an era in international financial regulation? A postcrisis research agenda, International Organization, 2011 Jan;65(1):169-200 Ball R, Robin A, Sadka G, Is financial reporting shaped by equity markets or by debt markets? An international study of timeliness and conservatism, Review of accounting studies, 2008 Sep 1;13(2-3):168-205 Jackson HE, Variation in the intensity of financial regulation: Preliminary evidence and potential implications, Yale J, on Reg,, 2007;24:253. Solomon J, Corporate governance and accountability, John Wiley Sons; 2007 Mar 16 Wu X, Corporate governance and corruption: A cross?country analysis, Governance, 2005 Apr 1;18(2):151-70 Edwards B, Wolfenzon D, Yeung B, Corporate governance, economic entrenchment, and growth, Journal of economic Literature, 2005 Sep 1;43(3):655-720 Tricker RB, Tricker RI, Corporate governance: Principles, policies, and practices, Oxford University Press, USA; 2015Gourevitch PA, Shinn J, Political power and corporate control: The new global politics of corporate governance, Princeton University Press; 2005 perspective, Journal of management studies, 2008 Jan 1;45(1):196-220 Doidge C, Karolyi GA, Stulz RM, Why do countries matter so much for corporate governance?, Journal of financial economics, 2007 Oct 31;86(1):1-39. Filatotchev I, Strange R, Piesse J, Lien YC, FDI by firms from newly industrialised economies in emerging markets: corporate governance, entry mode Gilson RJ, Milhaupt CJ, Sovereign wealth funds and corporate governance: A minimalist response to the new mercantilism, Stan, L, Rev,, 2007;60:1345

Harmful Health Effects of Nuclear Energy

Introduction The world has experienced an exponential growth in its energy demands over the course of the last century. This demand is expected to increase as the world’s population raises and the standards of living for people in the developing nations increase.Advertising We will write a custom research paper sample on Harmful Health Effects of Nuclear Energy specifically for you for only $16.05 $11/page Learn More The demand has put a strain on traditional sources of energy such as fossil fuel. Nuclear energy has emerged as a feasible alternative to renewable energy and most of its advocates argue that it is the only means through which sustainable development can be achieved in an energy intensive world. However, nuclear energy poses significant danger to the society especially if an accident in a power plant occurs. Khripunov notes that accidents at nuclear power facilities pose numerous risks since they affect systems that society depends on such as healthcare, transportation, and environment (19). This dangers posed by nuclear energy plants were best exemplified following the devastating Chernobyl accident of 1986. The accident led to the release of radioactive material to the population with various health implications. This paper will engage in a detailed discussion on the health effects of the Chernobyl nuclear accident so as to demonstrate that nuclear energy poses a significant threat to human existence. The Chernobyl Accident The Chernobyl nuclear power station accident is the most devastating nuclear accident in history. The accident took place during a planned systems text when an unexpected power surge led to the damage of a reactor leading to an explosion. This incident resulted in severe contamination of vast territories of the former USSR and other parts of Europe as radionuclide releases were spread by winds for over one week (Chudley 221). The nuclear accident in Chernobyl in 1986 demonstrated the risk t hat nuclear power generation poses to mankind. Reports indicate that the amount of radiation fallout from the Chernobyl accident was over 400 times that caused by the atomic bomb detonation at Hiroshima (Chudley 221). The massive explosion that destroyed the react led to six million people from Ukraine, Belarus, and Russia being exposed to radioactive contamination (Chudley 221).Advertising Looking for research paper on environmental studies? Let's see if we can help you! Get your first paper with 15% OFF Learn More Health Impacts of Nuclear Energy Direct Health Impacts Exposure to radiation from nuclear energy sources increases the risk of cancer. The risk of developing thyroid cancer following exposure to nuclear radiations increased with a decrease in the age of the subject. Younger individuals are therefore more likely to develop thyroid carcinogenesis after X-ray exposure than more mature people are. After the Chernobyl accident, an increase in thyroid cancer was found in children exposed to fallout from the accident (Baverstock and Dillwyn 1312). As of the year 2000, the number of individuals who suffered from thyroid Cancer due to exposure as children had reached 2,000. This number increased to 4,000 in the years 2005 (Baverstock and Dillwyn 1312). This observation is supported by the OECD/Nuclear Energy Agency, which notes that there has been an increase in incidents of thyroid cancers following the Chernobyl accident (81). Exposure to high doses of radiation results in radiation sickness; a condition that can lead to the death of an individual. Acute radiation sickness results in future health consequences such as cancer and premature aging of the patient. The natural background levels of radiation in Europe average at 3mSv/year. However, people in the highly contaminated regions experienced radiation doses that were 30-70 times greater than these average natural levels (Chudley 222). The OECD/Nuclear Energy Agency confirms that in the C hernobyl accident, all cute deterministic health effects occurred among the personnel of the plant (78). Fire workers and clean-up workers were exposed to high doses of radiation and over 200 emergency workers suffered from acute radiation sickness with 28 dying because of this. The early symptoms of radiation sickness included vomiting, diarrhoea, and fever. Chudley asserts that the effects of exposure to high dose or radiation can range from† death following acute radiation exposure, to cancer or genetic re-arrangements if doses are small and exposure is spread over time (222).Advertising We will write a custom research paper sample on Harmful Health Effects of Nuclear Energy specifically for you for only $16.05 $11/page Learn More Nuclear accidents also result in a suppression of the body’s immune system. The OECD/Nuclear Energy Agency states that the immunity system is suppressed due to a depression of bone-marrow function following exp osure to radiation (80). Patients who suffer from this immunity suppression have to receive doses of anti-fungal agents and antibiotics in order to help the body fight off common ailments. There is a high probability of genetic changes occurring due to exposure to nuclear radiation. As of 2001, no radiation-induced genetic diseases had been observed in the population affected by the Chernobyl disaster. However, Chudley asserts that this is not an indication that genetic disorders were caused by the accident since such changes are difficult to identify in the human population (224). An observation of irradiated pine trees in the Chernobyl region demonstrates that these plants have undergone major genetic changes due to radiation exposure. The trees exhibit a marked change in the shape and size of needles and their seed quality had degraded over the years. In addition to this, there are some pine trees that suffer from impaired fertility due to radiation exposure (Chudley 224). It can therefore be deduced that genetic changes occurred in human beings as well and the only reason why they are not visible is because of the differences in life and reproduction cycles. Exposure to radioactive material also increases the risk of leukaemia. Baverstock and Dillwyn state that leukaemia is strongly associated with radiation (1313). As such, intensive efforts have been made to detect this condition in the population affected by the Chernobyl incident. While at the moment there have been no significant increases in the levels of leukaemia among the population, scientists predict that the level will increase in the coming years. Baverstock and Dillwyn demonstrate that leukaemia is a disease of older age and as the victims of the Chernobyl grow older, there will be increases in chronic lymphatic leukaemia (1313). In addition to the already mentioned health impacts, nuclear accidents result in a rise in all types of disease. Report by the Russian National Medical Dosimetric Re gistry indicate that malignant diseases have been on the increase between 1989 and 1992 and this can be attributed to the radiation exposure (The OECD/Nuclear Energy Agency 89).Advertising Looking for research paper on environmental studies? Let's see if we can help you! Get your first paper with 15% OFF Learn More The mortality rate within the Russian Federation exhibited a 2% increase following the accident. The death rate from respiratory cancer was especially elevated and while this could be blamed on smoking, the findings suggested that the nuclear disaster contributed to it (OECD/Nuclear Energy Agency 89). Indirect Health Impacts The people who live in the contaminated zone face additional health risks. Abbott, Wallace and Matthias note that the diet of the people living in the Chernobyl area was negatively affected with most people not being able to have a good diet in the zone (116). This is consistent with research by Baverstock and Dillwyn, which indicates that dietary changes to avoid perceived contamination may be an indirect consequence of a nuclear accident (1313). These forced dietary changes negatively affect the health of the individual. Nuclear accidents have psychological consequences on the community surrounding the nuclear site. Such accidents are followed by forced evacua tion from home and land and people are left unsure of what the future holds. Baverstock and Dillwyn state that following the Chernobyl accident, there was an increase in the number of psychological illnesses (1313). There was an increase in the consumption of alcohol and cigarettes as people tried to deal with the stress they experienced because of this incident. The accident therefore had an indirect consequence of causing some deaths from suicide, cirrhosis, or lung cancer. In addition to the health effects caused by radiation, nuclear accidents have the ability to degrade the social fabric in the affected territories. The Chernobyl accident best exemplifies this considering the various physical and psychological effects that have persisted since the accident. The Chernobyl accident increased public mistrust of the administration and the government. The OECD/Nuclear Energy Agency notes that public scepticism towards authority was heightened due to the poor governmental response in the early days following the accident and the lack of public education on the polluting hazards of the disaster (92). Discussion The paper has observed that the effects of a nuclear disaster affect the population in an indiscriminate manner. In addition to the immediate negative health effects caused by exposure to radiation, long-term health effects continue to be felt by the community decades after the nuclear accident happened. Abbott et al. document that the radioactivity levels that resulted from the accident at the Chernobyl Nuclear Power Complex are likely to remain at high levels for around 300 years (105). The Chernobyl incident forced the world to reconsider the use of nuclear power to provide for the energy needs of the society. The disaster continues to cost the affected regions millions of dollars in health care costs almost three decades after the accident occurred. For this reason, the Russian society and the rest of the international community remains deeply divided about the nuclear power infrastructure because of the accident in Chernobyl, which resulted in immense damages to the citizens of Ukraine. Conclusion This paper set out to discuss the health impacts of nuclear energy with particular focus on the Chernobyl accident of 1986. The paper began by providing an overview of the accident to underscore the fact that the accident could not have been foreseen. It then proceeded to discuss the many negative health implications that people exposed to the radiation suffered from and they include; radiation poisoning, cancer, and genetic disorders. The paper has also observed that immune suppression is one of the inevitable consequences of exposure to high doses of radiation. In addition to the direct health implications or radiation, there are indirect implications, which include stress, suicidal ideation, dietary problems, and degradation of the social fabric. From the discussions presented in this paper, it can be authoritatively stated that nu clear facilities are a threat to man’s survival since accidents at nuclear facilities have a large-scale social impact. Works Cited Abbott, Pamela, Wallace Claire and Matthias Beck. â€Å"Chernobyl: Living with risk and uncertainty†. Health, Risk Society 8.2 (2006): 105 – 121. Web. Baverstock, Keith and Dillwyn Williams. â€Å"The Chernobyl Accident 20 Years On: An Assessment of the Health Consequences and the International Response†. Environmental Health Perspectives 114.9 (2006): 1312-1317. Web. Chudley, Albert. â€Å"Genetic implications and health consequences following the Chernobyl nuclear accident†. Clin Genet 77.1 (2010): 221–226. Web. Khripunov, Igor. â€Å"How Safe Is Russia? Public Risk Perception and Nuclear Security†. Problems of Post-Communism 54.5 (2007): 19–29. Web. OECD/Nuclear Energy Agency. Chernobyl: Assessment of Radiological and Health Effects. Paris: OECD, 2002. Print. This research paper on Harmful Health Effects of Nuclear Energy was written and submitted by user Nancy Lindsay to help you with your own studies. You are free to use it for research and reference purposes in order to write your own paper; however, you must cite it accordingly. You can donate your paper here.

Tumor-Stroma Interaction †Biology Research Paper

Tumor-Stroma Interaction – Biology Research Paper Free Online Research Papers Tumor-Stroma Interaction Biology Research Paper Abstract Matrix metalloproteinases (MMPs) are metal-dependent endopeptidases that play pivotal roles in tumor disease progression. In many solid tumors, MMPs are indeed produced by tumor stromal cells, rather than by tumor cells. This expression pattern is, at least in part, regulated by tumor-stroma interaction via tumor cell-associated extracellular matrix metalloproteinase inducer (EMMPRIN). In vitro, recombinant EMMPRIN dose-dependently stimulated MMP-1 production by primary human fibroblast cells. Interestingly, in addition to stimulating MMP expression, EMMPRIN also induced its own gene expression. To further explore this potential positive feedback regulatory mechanism, we generated human breast cancer cells expressing different levels of EMMPRIN. Coculture of EMMPRIN-positive tumor cells with fibroblast cells resulted in a concomitant stimulation of MMP-2, MMP-9, and EMMPRIN production. This induction was EMMPRIN dependent, was further enhanced by overexpression, and was reduced by antisense suppression of EMMPRIN expression in tumor cells. Increased expression of membrane-associated EMMPRIN was accompanied by an MMP-dependent generation of a soluble form of EMMPRIN representing a proteolytic cleavage product lacking the carboxyl terminus. On the basis of these findings, we propose a model in which tumor cell-associated EMMPRIN stimulates MMPs, as well as EMMPRIN expression in tumor stroma. Increased MMP activity in tumor local environment results in proteolytic cleavage of membrane-associated EMMPRIN, releasing soluble EMMPRIN. Soluble EMMPRIN in turn acts in a paracrine fashion on stroma cells that are both adjacent and distant to tumor sites to further stimulate the production of MMPs and additional EMMPRIN, which consequently contributes to tumor angiogenesis, tumor growth, and metastasis. Introduction Degradation of basement membrane by matrix metalloproteinases (MMPs) is one of the most critical steps in various stages of tumor disease progression, including tumor angiogenesis, tumor growth, as well as local invasion and subsequent distant metastasis (1–3). MMPs are a family of more than 25 endopeptidases that are metal dependent and share a common modular domain structure. MMPs are overproduced in tumor local environment. Collectively, these enzymes are capable of cleaving all of the extracellular matrix components of the parenchymal and vascular basement membranes, normally mechanical barriers to tumor cell migration and invasion. High levels of MMPs have been correlated with increased tumor invasion capacity both in vitro and in vivo (4, 5). Clinically, elevated levels of MMPs detected in the body fluids of cancer patients correlate with adverse prognosis in various cancers (6, 7). Imbalances in the production of MMPs and their endogenous inhibitors, tissue inhibitors o f metalloproteinases (TIMPs), result in tumor angiogenesis and metastasis (2, 3, 8–12). Interestingly, recent studies of clinical tumor specimens provided evidence that most MMPs are generated by peritumoral fibroblasts in the stromal compartment, instead of by tumor cells themselves. Peritumor stromal cells are indeed responsible for the production of tumor-associated interstitial collagenase (MMP-1), stromelysin-1 (MMP-3), stromelysin-3 (MMP-11), and gelatinase A (MMP-2), in breast, colon, lung, skin, and head and neck cancers (13–16). The induction of MMP production is, at least in part, mediated by tumor-stromal cell interaction via a tumor cell surface glycoprotein, extracellular matrix metalloproteinase inducer (EMMPRIN; 17, 18). EMMPRIN was originally purified from the plasma membrane of cancer cells as a glycoprotein of Mr 58,000, and was designated tumor collagenase stimulating factor (TCSF) because of its ability to stimulate fibroblast synthesis of collagenase-1 (MMP-1; 19, 20). Subsequent studies further demonstrated that EMMPRIN also induced fibroblast synthesis of MMP-2, MMP-3, as well as the membrane-type 1 MMP (MT1-MMP) and MT2-MMP that function as endogenous activators of MMP-2 (17, 21, 22). Therefore, EMMPRIN functions as an upstream modulator of MMP production in tumor local environment. EMMPRIN-positive tumor cells stimulate neighboring fibroblast cells to express MMPs and therefore facilitate tumor invasion and metastasis. This rationale for the function of EMMPRIN has been supported by results from several clinical studies that demonstrated high levels of EMMPRIN expression in tumor compartments as compared to peritumoral stromal tissues. These tumors include the lung (16), breast (16), bladder (23, 24), and glioma (25). Examination of EMMPRIN expression in these clinical samples by a variety of means, including Northern blot, in situ hybridization, and immunostaining, revealed that in most cases, EMMPRIN is expressed by tumor cells, but not by the neighboring stromal cells. In contrast, MMPs are expressed by peritumoral stromal cells. The biological significance of increased expression of EMMPRIN in tumor cells was investigated by in vitro studies using recombinant EMMPRIN or native EMMPRIN purified from tumor cells. EMMPRIN has been shown to stimulate the expression of various MMPs produced by fibroblasts (17, 18). The induction occurred at the transcription level and is at least in part mediated by a mitogen-activated protein kinase (MAPK) p38 kinase signaling pathway (18). The role of EMMPRIN in tumor growth and metastasis was directly illustrated using EMMPRIN-overexpressing human breast cancer cells. MDA MB 463 cells are normally slow-growing cells when they are implanted into nude mice. However, when these cells were transfected with EMMPRIN, they adopted a more aggressive growth pattern, with both accelerated growth rate and metastatic phenotypes (26). To further study the interplay between tumor cell-associated EMMPRIN and fibroblast cells, we used recombinant engineering to generate MDA MB 231 human breast cancer cells expressing different levels of EMMPRIN. We discovered a novel positive feedback regulatory mechanism controlling the interaction between tumor and fibroblast cells. Our results demonstrate that EMMPRIN positively regulates its own gene expression, in addition to stimulating MMP expression. Furthermore, we also detected generation of soluble EMMPRIN in the coculture of tumor and fibroblast cells that occurs via an MMP-dependent cleavage. On the basis of these new findings, we propose a new model in which the interplay between tumor and stromal compartments is regulated by both tumor cell surface EMMPRIN, as well as by soluble EMMPRIN, generated from MMP-dependent cleavage. Results Induction of MMP-1 Expression by Soluble Recombinant Human EMMPRIN in Primary Fibroblast Cells The stimulatory effects of EMMPRIN on cultured fibroblast cells were studied using a soluble recombinant EMMPRIN protein (srEMMPRIN) corresponding to the extracellular portion of the molecule. srEMMPRIN was added at different concentrations to primary normal human fibroblast cells in culture. Serum-free media conditioned by these cells were collected at 1 or 3 days after EMMPRIN stimulation. MMP-1 production in response to EMMPRIN stimulation was determined by assaying MMP-1 activity secreted into the conditioned medium. EMMPRIN dose-dependently stimulated MMP-1 production by normal human lung fibroblast (NHLF) cells (Fig. 1 ). The stimulation of MMP-1 production by srEMMPRIN also occurred on the transcription level as determined by TaqMan analysis (Fig. 2A ). FIGURE 1. Induction of MMP-1 expression by soluble recombinant human EMMPRIN in human primary lung fibroblast cells. srEMMPRIN dose-dependently stimulated MMP-1 production by NHLF cells. Ten thousand NHLF cells (passage 1) were plated in each well of a 96-well cell culture plate. Cells were challenged with different concentrations of EMMPRIN. After 1 day and 3 days, conditioned medium was collected from each well and MMP-1 activity was determined. Points, mean values of duplicates from three independent experiments; error bars, SDs. FIGURE 2. Induction of EMMPRIN expression by soluble recombinant human EMMPRIN in primary fibroblast cells. Primary normal human fibroblast cells of the lung were challenged with 5 Â µg/ml of srEMMPRIN. A. Total RNA was collected from untreated cells or at 4, 24, and 48 h after EMMPRIN treatment. EMMPRIN and MMP-1 gene expression level was determined by quantitative TaqMan analysis and was expressed as percentage to that in untreated cells, which was designated as 100%. B. Western blot analysis of EMMPRIN expression in fibroblast cells in response to stimulation with recombinant EMMPRIN. Cell lysate containing 50 Â µg of total protein was loaded into each lane, G361 high EMMPRIN-expressing melanoma cells (lane 1), fibroblast cells treated with EMMPRIN (lane 2), and unstimulated fibroblast cells (lane 3). Positive Feedback Regulation of EMMPRIN Gene Expression in Primary Fibroblast Cells Interestingly, when we examined the expression level of EMMPRIN in fibroblast cells after stimulation with 5 Â µg/ml EMMPRIN, we observed a substantial increase in EMMPRIN gene expression in these cells. As determined by quantitative TaqMan analysis, a 2.2-fold increase in EMMPRIN gene expression was induced at 4 h after cells were stimulated with EMMPRIN (Fig. 2A). EMMPRIN gene expression continuously increased to 4.6-fold at 24 h and 3.7-fold at 48 h after treatment. On EMMPRIN stimulation, MMP-1 gene expression level also increased (Fig. 2A). This induction occurred more rapidly and transiently, with a peak expression of a 3.3-fold increase at 4 h, and dropped to near pretreatment level at 48 h. When EMMPRIN protein expression in fibroblast cells treated with EMMPRIN was examined with Western blot analysis, we observed a strong increase in EMMPRIN protein expression (Fig. 2). In contrast, only minimal levels of EMMPRIN protein were detectable in unstimulated fibroblast cells. Tak en together, these results suggest that in addition to increased expression of MMP-1, fibroblast cells also respond to EMMPRIN stimulation by up-regulating EMMPRIN gene expression. Generation of MDA MB 231 Tumor Cells Expressing Different Levels of EMMPRIN To further explore the potential regulatory mechanism of EMMPRIN gene expression in fibroblast cells, especially in the context of tumor-fibroblast cell-cell interaction, we used recombinant DNA engineering technology to generate cancer cells that express different levels of EMMPRIN. As described in Materials and Methods, MDA MB 231 human breast cancer cells were transfected with expression vectors harboring the full-length coding region of human EMMPRIN in either sense or antisense orientations. Stably transfected cells were cloned and EMMPRIN expression level in these cells was determined by Western blot analysis. As shown in Fig. 3 , EMMPRIN protein expression in one of sense transfected cell clones S1-3 was approximately 2-fold higher as compared to that of wild-type control cells. In contrast, EMMPRIN protein expression was substantially inhibited in two antisense transfected cell clones, AS1-5 and AS2-5, to only 47% and 62% of that in wild-type cells. Changes in EMMPRIN protein expression in these engineered cells were also independently confirmed by ELISA analysis (Fig. 4 ). In addition, changes in EMMPRIN protein expression also occurred on cell surface in sense and antisense transfected cells, respectively, as determined by fluorescence-activated cell (FAC) analysis (data not shown). FIGURE 3. Generation of MDA MB 231 tumor cells expressing different levels of EMMPRIN. A. Western blot analysis of EMMPRIN protein in cell lysates. Samples containing 20 Â µg of total protein from each cell type were loaded in duplicates, MDA MB 231 wild-type cells (lanes 1 and 2), S1-3 cells (lanes 3 and 4), AS1-5 cells (lanes 5 and 6), and AS2-5 cells (lanes 7 and 8). A single EMMPRIN protein band of approximately Mr 52,000 was detected in all of the samples. B. Quantitative determination of EMMPRIN expression by scanning densitometry. EMMPRIN Western blot was scanned and the pixel density of each band was determined by scanning densitometry. EMMPRIN expression level in each cell type was expressed as the percentage of density of that in wild-type cells (lane 1), which was designated as 100% error bars, SDs. FIGURE 4. Stimulation of both cell-associated and soluble EMMPRIN expression in coculture of tumor cells and fibroblast cells. EMMPRIN protein concentration was determined by ELISA analysis in total cell lysate (A) or in conditioned medium (B) of breast cancer cells expressing different levels of EMMPRIN either in monoculture or in coculture with NHDF cells. All concentrations were normalized to total protein amounts. WT, wild-type MDA MB 231 human breast cancer cells; S1-3, sense transfected EMMPRIN-overexpressing cells; AS1-5 and AS2-5, antisense transfected cells. Columns, representative of two independent experiments; error bars, SDs. We next used these cells with different levels of EMMPRIN protein expression to address the role of tumor cell-derived EMMPRIN in regulating its own expression during tumor-fibroblast cell interaction. Stimulation of Both Cell-Associated and Soluble EMMPRIN Expression in Coculture of Tumor Cells and Fibroblast Cells In monoculture of tumor cells or fibroblast cells, only low levels of EMMPRIN protein were detected in cell pellets or conditioned medium by ELISA analysis. Transfection with sense or antisense expression constructs resulted in substantial changes in cell-associated EMMPRIN expression, with an increase from 3.32 ng/ml in wild-type to 7.2 ng/ml in sense transfected S1-3 cells, and a decrease to 0.85 and 1.43 ng/ml in antisense transfected AS1-5 and AS2-5 cells, respectively (Fig. 4A). Only low levels of soluble EMMPRIN were detected in conditioned medium of these cells in monoculture. Transfection of tumor cells with sense and antisense expression constructs resulted in only minor changes in soluble EMMPRIN protein levels (Fig. 4B). The expression of cell-associated EMMPRIN was readily induced when tumor cells were cultured together with fibroblast cells. In coculture, EMMPRIN was detected at 10 ng/ml as compared to 0.8 ng/ml in fibroblast cells and 3.32 ng/ml in wild-type tumor cells in monoculture (Fig. 4A). This increase in EMMPRIN expression in coculture was mediated by tumor cell-associated EMMPRIN. When EMMPRIN expression in tumor cells was inhibited by antisense transfection in AS1-5 and AS2-5 cells, the increase in EMMPRIN expression stimulated by coculture was concomitantly reduced to minimal levels (Fig. 4A). It is known that coculture of EMMPRIN-positive tumor cells with fibroblast cells stimulates MMP expression (22, 27). These MMPs can in turn cleave cell surface proteins to generate soluble proteins. We therefore postulated that there may be an increase in soluble EMMPRIN protein level in conditioned medium of these cells in coculture. In monoculture, the levels of soluble EMMPRIN were extremely low in conditioned medium of fibroblast cells (0.31 ng/ml) and wild-type tumor cells (0.53 ng/ml; Fig. 4B). However, coculture of tumor cells with fibroblast cells induced the release of substantial amounts of soluble EMMPRIN, 7.48 ng/ml by wild-type and fibroblast coculture and 10.39 ng/ml by S1-3 and fibroblast coculture (Fig. 4B). Similar to cell-associated EMMPRIN, the increase in soluble EMMPRIN expression was also correlated with the degree of EMMPRIN expression in tumor cells. Inhibition of EMMPRIN expression by antisense transfection virtually abolished the generation of soluble EMMPRI N in coculture (Fig. 4B). Taken together, these results demonstrate a positive feedback regulatory mechanism of EMMPRIN gene expression during tumor-fibroblast cell-cell interaction. In addition to stimulating EMMPRIN expression and MMP expression, tumor cell-associated EMMPRIN also mediates the release of soluble EMMPRIN during tumor-fibroblast cell interactions. MMP-Dependent Generation of Soluble EMMPRIN in Coculture of EMMPRIN-Positive Tumor Cells and Fibroblast Cells Western blot analyses using antibodies against different regions of EMMPRIN were performed to further characterize soluble EMMPRIN generated in the coculture of tumor cells and fibroblast cells. Antibodies against the extracellular portion of EMMPRIN molecule detected a single band of approximately Mr 52,000, consistent with the size of intact protein, in total cell lysates. Meanwhile, this same antibody detected a slightly smaller molecule in the conditioned medium from coculture, indicating a loss of a portion of EMMPRIN protein (Fig. 5A ). Interestingly, antibodies specific to the carboxyl terminus of EMMPRIN only detected EMMPRIN in total cell lysates, but failed to detect any EMMPRIN protein in conditioned medium (Fig. 5A). Taken together, these data suggest that soluble EMMPRIN secreted into coculture medium of tumor and fibroblast cells is likely a cleaved product of the full-length transpanning form of membrane EMMPRIN. FIGURE 5. Soluble EMMPRIN in coculture of EMMPRIN-positive tumor cells and fibroblast cells- proteolytic cleavage products generated in an MMP-dependent fashion. A. Western blot analyses of cell lysates or conditioned medium of coculture of wild-type MDA MB 231 human breast cancer cells and fibroblast cells. Membranes with 50 ng recombinant EMMPRIN extracellular domain (lane A), cell lysates of 50 Â µg total protein (lanes B and D), and conditioned medium of 50 Â µg total protein (lanes C and E) were blotted with antibodies that recognize extracellular domain (lanes A–C) or intracellular carboxyl terminus (lanes D and E). B. Soluble EMMPRIN concentration in conditioned medium from coculture of tumor cells and fibroblast cells was determined using ELISA analysis. 1,10 phenanthroline was included in the study from 0.1, 0.5 to 1.0 Â µM to inhibit MMP enzymatic activity. WT, wild-type MDA MB 231 human breast cancer cells. Columns, representative of two independent experiments; error bars, SDs. , NHDF; , WT; , WT + NHDF; , WT + NHDF + 0.1 Â µM 1,10 PA; , WT + NHDF + 0.5 Â µM 1,10 PA; , WT + NHDF + 1.0 Â µM 1,10 PA; *, P < .05 compared to WT + NHDF. Because the generation of soluble EMMPRIN occurred concomitantly with increased production of MMP in coculture of tumor and fibroblast cells, we speculated that the release of soluble EMMPRIN might be mediated by MMP-dependent proteolytic cleavage. To further explore the mechanism of soluble EMMPRIN generation, 1,10 phenanthroline (1,10 PA) a general metalloproteinase inhibitor, was used at noncytotoxic concentrations to inhibit the enzymatic activity of MMPs. As shown in Fig. 5B, inhibition of MMP activity by 1,10 PA resulted in significant decreases in the level of soluble EMMPRIN in a dose-dependent fashion. These results demonstrate that the generation of soluble EMMPRIN stimulated by tumor fibroblast cell-cell interactions is the result of proteolytic cleavage of membrane-associated EMMPRIN by increased MMP activities in coculture. Discussion It is now known that the microenvironment of the tumor-host interface plays a proactive role during malignant disease progression, including the transition from carcinoma in situ to invasive cancer, tumor cell proliferation, and tumor cell dissemination and metastasis (28). On the host side, the stromal compartment is comprised of a variety of cell types, including immune cells, inflammatory cells, muscle and myofibroblast cells, and vascular cells. Tumor cells express molecules, either secreted or presented on cell surface, to interact with surrounding stromal cells. In the current study, we have demonstrated that EMMPRIN exists in both soluble and membrane-bound forms. Generation of soluble EMMPRIN resulted from tumor-fibroblast cell interaction in an EMMPRIN-dependent fashion, and was mediated by MMP proteolytic activities. These novel findings suggest a positive feedback regulatory mechanism of EMMPRIN expression that occurs at tumor-host interface. EMMPRIN is normally expressed as a glycoprotein on tumor cell surface that migrates at an apparent molecular size of Mr 58,000 (19). The domain structure of EMMPRIN consists of an extracellular portion which contains two immunoglobulin-like domains, a transmembrane domain, and a short cytoplasmic domain (19). It has been suggested that tumor cell-associated EMMPRIN exerts its MMP stimulatory effects on fibroblast cells via close cell-to-cell contact. However, the production of soluble EMMPRIN by tumor cells has recently been indicated in several studies (29, 30). In these studies, tumor cells affect fibroblast cells in the absence of physical contact in an EMMPRIN-dependent fashion, resulting in increased MMP-2 expression and activation (29), or enhanced cell migration or invasion (30). In addition, soluble full-length EMMPRIN has been purified from concentrated culturing medium of breast cancer cells, presumably secreted in shed vesicles (31). However, secreted soluble EMMPRIN in ca ncer cell monoculture is estimated to account for only 2–3% of EMMPRIN produced by these cells (31). In the present study, we have demonstrated the release of a substantial amount of soluble EMMPRIN into cell culture medium of cocultured tumor and fibroblast cells. Because fibroblast cells respond to EMMPRIN stimulation in producing increased levels of EMMPRIN, it is likely that the soluble EMMPRIN detected in coculture is derived from both tumor and fibroblast cells. In addition, the soluble EMMPRIN generated in coculture lacks the intracellular carboxyl terminus as determined using an antibody against the intracellular region of the molecule. In contrast, this carboxyl terminal region was preserved in cell-associated EMMPRIN. More interestingly, generation of soluble EMMPRIN in coculture could be efficiently blocked via inhibiting MMP activity. Therefore, this soluble EMMPRIN likely represents the proteolytic product of membrane-associated EMMPRIN. The generation of soluble EMMPRIN at the interface of tumor-host interaction may have profound biological significance. The soluble EMMPRIN could diffuse away from local tumor invasion site and stimulate stromal cells at distant sites. We and others have shown that soluble EMMPRIN consisting of only the extracellular portion of the molecular is functional in stimulating MMP expression in fibroblast and endothelial cells (17, 32). Increased MMP activity in the stromal compartment in turn promotes tumor invasion and angiogenesis. The receptor on fibroblast cells that is responsible for EMMPRIN-mediated stimulation of MMP production has remained elusive (17, 21). It has been suggested that EMMPRIN may serve as its own counter-receptor in cancer cells, thus stimulating MMP via a homophilic interaction (33). One could speculate that EMMPRIN on fibroblast cells could also serve as counter-receptors for tumor cell-associated EMMPRIN in stimulating MMP expression. However, this hypothesis is not supported by the fact that only extremely low levels of EMMPRIN expression could be detected in normal fibroblast cells. Our finding of a novel positive feedback regulatory mechanism of EMMPRIN expression provides an alternative explanation for the potential role of EMMPRIN as its own counter-receptor in fibroblast cells. When fibroblast cells were exposed to EMMPRIN stimulus, EMMPRIN expression is up-regulated in these cells. Newly synthesized EMMPRIN could then be presented on the cell surface and serves as the counter-re ceptor for EMMPRIN-dependent signaling between tumor and fibroblast cells. This novel regulatory mechanism is supported by recent findings of EMMPRIN mRNA expression in peritumoral fibroblasts in ovarian carcinoma (34). However, this homophilic interaction likely only accounts for EMMPRIN-mediated tumor-host interaction in some tumors because EMMPRIN expression has not been detected in the stroma of many other tumors including melanoma (35), or lung and breast cancer (16). In these samples, despite of a lack of EMMPRIN expression in stromal tissue, MMP expression is stimulated in stromal tissues adjacent to EMMPRIN-positive tumor cells, suggesting that there might be an unknown receptor(s) on stromal cell surface mediating these interactions. In conclusion, we have identified a novel positive feedback regulatory mechanism of EMMPRIN expression in fibroblast cells and discovered a soluble form of EMMPRIN. These findings suggest an amplification of EMMPRIN-mediated, MMP-dependent signaling events at the tumor and host interface, as well as at distant sites (Fig. 6 ). FIGURE 6. Proposed model for signal loops between EMMPRIN and MMP and tumor cells and host cells. Tumor cells use cell surface EMMPRIN to initiate contact with surrounding fibroblast cells (1), signaling fibroblast cells to synthesize MMPs (2). MMPs secreted by fibroblast cells subsequently cleave cell surface EMMPRIN and generate soluble EMMPRIN (sEMMPRIN; 3). These sEMMPRIN molecules, in turn, act on cells either in local tumor environment or diffuse away to act on distant cells (4) to further stimulate MMP and EMMPRIN expression (5) and augment the migration and invasion potential of tumor cells (6). Materials and Methods Cell Culture MDA MB 231 human breast carcinoma cells were purchased from American Type Culture Collection (Manassas, VA) and were cultured under recommended conditions. Briefly, cells were cultured in DMEM containing 10% fetal bovine serum (Life Technologies, Inc., Grand Island, NY). NHLF or normal human dermal fibroblast (NHDF) cells were obtained from Clonetics (Walkersville, MD) and were cultured in fibroblast growth medium containing 1 Â µg/ml human recombinant fibroblast growth factor (hFGF), 5 mg/ml insulin, 50 mg/ml gentamicin, and 50 mg/ml amphotericin B, at 37Â °C, 5% CO2. For coculture experiments, 100,000 cancer cells (MDA MB 231, MDA MB 231 S1-3, MDA MB 231 AS1-5, or AS2-5) were cultured together with 200,000 NHDF cells in a six-well cell culture plate in complete DMEM culturing medium. After 24 h, culturing medium was replaced with fresh serum-free DMEM and cells were cultured for 2 days. Culture medium was then replaced with 1.0 ml of fresh serum-free DMEM. Conditioned medium was collected 3 days later and secreted EMMPRIN protein concentration was determined using ELISA. Cells were lysed with 200 Â µl lysis buffer [50 mM Tris (pH 7.8), 150 mM NaCl, and 1% NP40] to determine cell-associated EMMPRIN concentrations. Cell Treatment and MMP-1 Activity Assay Recombinant EMMPRIN corresponding to the extracellular domain of human EMMPRIN protein was produced in NSO cells (RD Systems, Minneapolis, MN). MMP-1 activity in serum-free medium conditioned by fibroblast cells treated with different amounts of recombinant EMMPRIN protein was quantitatively determined using an MMP-1 Activity Assay Kit according to product manual (RD Systems). Briefly, MMP-1 contained in 150 Â µl of standards or samples was captured by anti-MMP-1 antibodies immobilized on the bottom of assay wells. Captured MMP-1 was subsequently activated by 4-aminophenylmercuric acetate (APMA). MMP substrate added into each well was cleaved by activated MMP-1 and the resulting fluorescence was determined using SpectraFluor Plus Plate Reader (TECAN, Research Triangle Park, NC) with the following parameters: excitation wavelength at 320 nm and emission wavelength at 405 nm. Generation of Sense and Antisense Expression Constructs and Establishment of Stable Transfectants To study the function of EMMPRIN in mediating tumor and stromal fibroblast cell interaction, the cDNA sequence corresponding to human EMMPRIN open reading frame (ORF) was PCR amplified and subcloned into pcDNA3.1TOPO vectors (Invitrogen, Carlsbad, CA). The EMMPRIN open reading frame was cloned in both sense and antisense orientations for cytomegalovirus promoter-driven expression in mammalian cells. Transfection of MDA MB 231 human breast carcinoma cells was performed using Effectene transfection reagents according to manufacturer's recommendations (Qiagen, Valencia, CA). Stably transfected cells were selected using Genicitin at 1000 Â µg/ml (Life Technologies). Individual transfectant colonies were established by cloning antibiotic-resistant cell colonies. Effects of sense and antisense transfection on EMMPRIN protein expression were determined using Western blot analysis and ELISA. RNA Sample Preparation and TaqMan Quantitative Reverse Transcription-PCR Total RNA was extracted from cells using RNeasy Kit (Qiagen) following manufacturer's instructions. Five micrograms of total RNA were used in the reverse transcription (RT) reactions which were carried out in a 50-Â µl total volume using Superscript First-Strand Synthesis System for RT-PCR kit following manufacturer's instructions (Invitrogen). Briefly, RNA samples were mixed with 0.5 Â µg/Â µl Oligo(dT), 0.4 mM dNTP mix in a total volume of 25 Â µl and were incubated at 65Â °C for 5 min, followed by quick cooling on ice for 2 min. The tubes were spun briefly and added with 5 Â µl 10x RT buffer, 10 Â µl 25 mM MgCl2, 2 Â µl 0.1 M DTT, 1 Â µl RNaseOUT, 1 Â µl (50 units) of Superscript II RT, and 6 Â µl nuclease-free water. The tubes were gently mixed and incubated at 42Â °C for 50 min. The reaction was terminated at 70Â °C for 15 min followed by quick cooling on ice for 2 min. The tubes were spun briefly and 1 Â µl of RNase H was added and incubated for 20 min at 37Â °C before proceeding to amplification of the target gene. Real-time PCR analyses were performed in 50 Â µl volume containing 2x TaqMan Universal PCR Master mix (Perkin-Elmer, Norwalk, CT), appropriate amounts of cDNA or genomic DNA, and 200 nM of TaqMan primers and 400 nM of TaqMan probes. Primer pairs and probes used in the study were designed using Primer Express software (Perkin-Elmer), with sequences listed in Table 1. Probes were labeled at the 5'-end with fluorescent reporter dye Fam and at the 3'-end with fluorescent quencher dye Tamra by Synthegen (Houston, TX) to allow direct detection of PCR products. Real-time PCR amplification and detection were performed in 96-well optical plates using ABI 7900HT sequence detector (PE Biosystems, Norwalk, CT). PCR conditions included thermal cycles of 30 s of 95Â °C for denaturing, 30 s of 60Â °C for annealing, and 60 s of 68Â °C for elongation. Relative gene copy numbers were calculated using a standard curve generated using PCR standards, serially diluted human genomic DNA (Clontech, Palo Alto, CA). Protein Electrophoresis and Western Blot Analysis Stably transfected cells were lysed in cell lysis buffer containing 50 mM Tris (pH 7.8), 150 mM NaCl, and 1% NP40. Protein concentration of cell lysate or conditioned medium was determined using the MicroBCA method (Pierce, Rockford, IL). Equal amount of proteins was loaded onto 4–15% gradient gels and was separated by SDS-PAGE under reducing conditions. Resolved proteins were electrophoretically transferred to nitrocellulose membranes (Invitrogen). The membranes were blocked with 5% low fat dry milk in TBS-T [10 mM Tris (pH 7.2), 50 mM NaCl, 0.5% Tween 20] for 1 h at room temperature, followed by incubating with primary antibody at 4Â °C overnight. Anti-EMMPRIN monoclonal antibody purchased from Research Diagnostics Inc. (Flanders, NJ) recognizes the extracellular portion of the protein. C-19 polyclonal antibody from Santa Cruz Biotechnology (Santa Cruz, CA) was raised against peptides of the carboxyl terminus of EMMPRIN protein. Blots were extensively washed with TBS-T and incubated with 1:5000 dilution of horseradish peroxidase (HRP) conjugated secondary antibody (Vector Laboratories, Burlingame, CA) diluted in TBS-T for 1 h at room temperature. Labeled proteins were visualized with Western Blotting Luminol Reagent (Santa Cruz Biotechnology). Pixel density of protein bands on Western blot was determined using Un-Scan-It software (Silk Scientific Corporation, Orem, UT). EMMPRIN ELISA EMMPRIN protein concentration in conditioned medium was determined using EMMPRIN ELISA. Briefly, 96-well Nunc Immuno Plates (Nunc, Denmark) were coated overnight at 4Â °C with 50 Â µl of anti-EMMPRIN antibody (RD Systems) at 5 Â µg/ml in buffer A (pH 7.4, 20 mM Tris-HCl, 150 mM NaCl, 1 mM CaCl2, 1 mM MgCl2, and 10 Â µM MnCl2). After one wash in buffer A, nonspecific binding sites were blocked with 150 Â µl of blocking buffer [3% bovine serum albumin (BSA) in buffer A] for 1 h at 37Â °C. Wells were then rinsed two times with binding buffer (buffer A with 0.3% BSA and 0.2% Tween 20). Either recombinant human EMMPRIN or conditioned medium was added into each well and was incubated at 37Â °C for 2 h. After three washes with binding buffer at RT, plates were blocked with 150 Â µl of blocking buffer (3% BSA in buffer A) at 37Â °C for 1 h. Fifty microliters of 1:5000 diluted biotin-conjugated anti-EMMPRIN antibodies (RDI-147, Research Diagnostics) were added into each well and incu bated at 37Â °C for 2 h. After three washes with binding buffer, 50 Â µl of horseradish peroxidase-conjugated streptavidin 1:10,000 diluted in binding buffer were added into each well, and were incubated at 37Â °C for 1 h. After three washes with binding buffer at RT, 50 Â µl of developing buffer (1 mg/ml O-phenylenediamine, 0.1 M citric acid, 0.2 M sodium phosphate, 0.01% H2O2) were added to each well and incubated at RT until color develops. Colorimetric reactions were stopped by adding 25 Â µl of 4 N H2SO4. ELISA data acquisition was performed using VersaMax Tunable MicroPlate Reader (Molecular Devices, Sunnyvale, CA) at 490 nm. Data were analyzed using Softmax Pro 3.1 software (Molecular Devices). 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