Genetic Transformation in E Coli

Genetic Transformation in E Coli Genetic Transformation is the act of changing of DNA in an organism by adding new genes, which may be done in multiple ways. The addition of new genes to DNA could have an almost infinite amount of advantages, ranging from studying the cultures of bacteria that become immune to modern medicine, to making artificial animal proteins. In a CNN article written by Matt Ford, scientists are using genetic transformation to do research on the use of growing animal proteins that the scientists claim will be healthier for the plant and mean less animal cruelty. However, the idea of artificially grow animal protein is still very controversial.ÂÂ   In the experiment performed by our lab, we used the idea of heat shock to genetically transform E coli. Heat shock is the process of exposing the cells to a temporary yet extreme increase in temperature, which temporarily opens the membranes of the cells. The purpose of opening these membranes is that the genes that are placed in the surrounding area will slip into the cell and become part of the DNA of that cell. In this experiment, we were testing whether or not the heat therapy opened the membranes of the cells, and therefore attempting to complete genetic transformation.ÂÂ   In the paper Nonchromosomal Antibiotic Resistance in Bacteria: Genetic Transformation of Escherichia coli by R-Factor DNA by Stanley N. Cohen, Annie C.Y. Chang, and Leslie Hsu is also an example of this kind of genetic transformation on E Coli. After the E. Coli was exposed to CaCl2, the E coli did not fully become resistant to antibiotics. The reason was that the E coli also needed the right temperature and conditions for the genes to fully become effective in the E coli. After the E coli was introduced to a heated environment for a short time, and then allowed to change and grow in an incubated environment, the resistance for the antibiotics increased in the E coli. While there have been cases where it was found that heat shock therapy was no t necessary to engage the genetic transformation cycle, as shown in the article One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution, we still used the idea of heat shock therapy for our experiment. To begin the experiment, we took two microcentrifuge tubes and labeled them +pGLO and -pGLO. Next, taking a micropipette with a clean tip we put 250 microliters of transformation solution into each of our microcentrifuge tubes. We then put ice into a beaker large enough for ice and our two tubes, and put these materials into the beaker. After, a sterile loop was used to take a single colony of bacteria into each of our tubes, using separate loops to keep them sterile and avoid contamination. After obtaining another new sterile loop, we put the loop into a tube marked pGLO plasmid DNA. This loop was then put into the tube labeled +pGLO and mixed. After this, we left both tubes in the ice beaker for at least ten minutes to get them and their contents to a lower temperature. While these are on ice, we obtained 4 Luria Broth (LB) nutrient agar plates from our lab provider; one LB plate, two LB/ampicillin plates, and one LB/ampicillin /arabinose plates were given to us. After the ten minu tes were over, both tubes were place in water that was 42 degrees Celsius for 50 seconds. After this warm water treatment, we immediately place the tubes back into the ice beaker. After two minutes in the ice beaker, we removed the tubes from the ice beaker. Using a clean tip for each tube on the micropipette, we added 250 microliters of LB nutrient broth to the +pGLO tube and the -pGLO tube and let mixtures sit for ten minutes. After the ten minutes, we gently flicked the tubes to mix the contents of the tubes. Then, we added 100 microliters of +pGLO to the LB/amp nutrient agar plate, 100 microliters of +pGLO to the LB/amp/are plate, 100 microliters of -pGLO to the LB/amp plate, and 100 microliters of -pGLO to the LB plate. Using a new clean and sterile loop for each plate, spread the mixtures of each plate so that they are mixed up well, while being sure not to press hard into the plate. We then closed the plates with their lids and stacked them on top of each other, putting tape around them to keep them in order. We then placed the plates into an incubator for one week. In this experiment, we introduced the pGLO plasmid to E. coli bacteria so that the cells were genetically transform a resistance to ampicilin as well as the ability to produce the protein that causes a glow. We used heat shock therapy in order to introduce the pGLO plasmid stored in an incubation unit the bacteria in agar plate containing ampicilin, arabinose and nutrient broth. As a result, the agar plate containing nutrient broth with the bacteria that had not been given the pGLO plasmid had bacteria grow in the plate. The plate containing nutrient broth and ampicilin with the bacteria, which was not given any pGLO, did not have any bacterial growth in the plate. The plate with nutrient broth and ampicilin that had the bacteria that had been given pGLO did grow new bacteria, but it did not glow. The final plate containing nutrient broth, ampicilin and arabinose and the bacteria that had been given pGLO both grew new bacteria and also glowed under the light. I stated that I believed that the E. coli bacteria that had been given pGLO would not only grow in the presence of ampicilin, but would also glow in the light when there was also arabinose. The results of the experiment did not disprove my hypothesis since the bacteria that had been given pGLO grew in both of the plates with ampicilin present, and glowed in the plate with arabinose present as well. The results of this experiment were consistent with other similar experiments with the same use of heat therapy on genetic transformation. A prime example is the experiment conducted by Cohen, Chang and Hsu in which the method of heat shock was used to introduce antibiotic resistance to E. coli bacteria (Cohen, Chang, Hsu, 1972). The results of the experiment showed that the introduction of R-factor DNA could genetically transform E. coli bacteria to have certain resistances. This experiment helps support our findings since their procedure and outcomes were very similar to our experiment. A few possible errors that occurred in our experiment could include the fact that the bacteria sat for a week after the first part of the experiment instead of being examined after 24 hours, which may have altered the amount of bacteria that was cultured. Also, it was almost impossible to get two halves of the same colony so it is possible that the two samples of E. coli were not genetically identical. However, we do not believe that our experiment had been sufficiently flawed to cause significant error Citations: 1. Meat is murder? Well, perhaps not for much longer. By Matt Ford. http://www.cnn.com/2009/TECH/science/08/07/eco.invitro.meat/index.html Accessed 11-11-2009 2. Nonchromosomal Antibiotic Resistance in Bacteria: Genetic Transformation of Escherichia coli by R-Factor DNA by Stanley N. Cohen, Annie C.Y. Chang, and Leslie Hsu. http://www.pnas.org/content/69/8/2110.abstract.ÂÂ   Accessed 11-10-2009 3. One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution by C T Chung, S L Niemela,, and R H Miller. http://www.pnas.org/content/86/7/2172.abstract accessed 11-10-2009 Donna Weedman, 2009 Life 102 Attributes of Living Systems, Cache House Inc. Eden Prairie, MN

What Is A Hero? Essay -- essays research papers

When one thinks of heroes, names such as Ghandi, Martin Luther King, and Mother Theresa often come to mind. These individuals have all helped society tremendously through their determination and unselfish deeds. However, we may not notice some of society’s other heroes. One such heroic person that comes to my mind is Arnold Schwarzenegger. Many people perceive Arnold as only a blockbuster movie star and not as the hero he truly is. People see the glittery and glamorous fascade of the actor but not the truly heroic acts of the humanitarian. Through his charity work, Arnold has truly touched the lives of those who are less fortunate.   Ã‚  Ã‚  Ã‚  Ã‚  There is a growing concern in the United Sta  Ã‚  Ã‚  Ã‚  Ã‚  tes, where health problems are increasing at an alarming rate. The diets of many men, women and children are unhealthy, and lead to high blood pressure strokes, diabetes, osteoporosis, hypertension, and mental health problems. The poor state of America’s health is a serious concern for Arnold, and he is constantly striving for ways to improve it. Arnold is an advocate of health and fitness as displayed by his own lifestyle. He uses his celebrity status to showcase himself as a living example of health, which encourages a healthy lifestyle in the general population. The fact that â€Å"a quarter of a million deaths each year can be attributed to physical inactivity† (Modern Maturity) is utterly shocking. I firmly believe that Arnold’s relentless motivation to educate and teach people the benefits of a healthy lifestyle can reduce these horrific numbers dramatically. Arnold was the chairman of the President’s Council on Physical Fitness and is the author of many books that promote health and fitness. Arnold began a fifty-state tour to encourage health and fitness by going to schools, meeting with governors, legislators and various community leaders. This tour took a tremendous amount of his own time and money. Nevertheless, he undertook this task to improve the health of people he did not even know. Arnold’s three goals for his tour were to â€Å"improve fitness of American children by returning a regimen of daily physical activity to the school, to increase parental involvement in their children’s fitness, and to promote community based fitness programs.† (Schwarzenegger) His extraordinary effort shows his genuine concern for the greate... ... a program of sports training and athletic competition for over one million kids and adults with physical and mental disadvantages. Arnold played a key role in developing the weightlifting program for the Special Olympics; he has worked with countless special Olympians and inspired them to participate to the best of their abilities. He said, â€Å"when you train with Special Olympics athletes, you go in thinking that you are there to inspire them. But the fact is, you feel inspired by what they can do with all those obstacles.† (Schwarzenegger ) If a hero is defined by his or her selfless contributions to society, and through their positive influences on other people, then Arnold Schwarzenneger is truly an unsung hero. Through Arnold, the general population of America is more informed on health issues, inner city kids have been given a chance to succeed, and the mentally or physically handicapped are truly made to feel good about themselves. A real hero doesn’t care if they are noticed for their work, as the knowledge that they have helped somebody is reward enough. Arnold has unselfishly given his money, but more importantly, his time and dedication to improve the lives of others.

Minimising risk in fashion

Explain how Buyers and Merchants minimise hazard when planning ranges for their manner company. Gabrielle ( Coco ) Chanel stated ‘Fashion does non merely exist in frocks, manner is in the air, it is brought in by the air current, one feels it coming, breathes it in the sky and on the paving, it depends on thoughts clients and happenings’ ( Charles-Roux, 2005 p.11 ) . The undertaking of construing how the changing universe is likely to impact the consumer’s desire for manner merchandises is hard, and this is why manner purchasers and merchants turn to command tools.There are many control tools which a manner company will utilize throughout its concern construction in order to accomplish maximal net income borders and to assist understate hazard and to give themselves a strong competitory advantage over rivals. Control tools are a critical portion of a manner company in order for it to turn successfully and to stay competitory. Despite supplying a figure of advantages to a manner company, control tools besides present a figure of disadvantages and restrictions whereby they are merely utile to a certain point, and this is when other control tools need to be put into the equation. A cardinal control tool used in the purchasing procedure is Range Planning. Range Planning involves roll uping a commercially acceptable aggregation of garments within fiscal and design parametric quantities, prior to production and bringing ( Goworek 2007 ) .The development of a scope of merchandises for a new season raises of import strategic issues for manner retail merchants about new merchandises and markets. In any season there is ever a balance struck between updated rudimentss that will be repeated in some signifier, and wholly new manner merchandises. A utile tool to assist entree the scope of merchandises for a season and to assist understate hazard is the Ansoff product-market matrix. This planning tool helps separate the basic options for increasing gross revenues by pull stringsing merchandise and market chances. It works good for the manner industry where merchandises are of all time altering and new markets continually developing, hence this will assist to understate h azard when planning ranges. Buyers and merchants should recognize the strategic options indicated by the Ansoff Matrrix, and the differing deduction of each to developing merchandise ranges in a season ( Watts, 1998 ) . Basically these options represent different methods of increasing gross revenues which will basically assist to maximize net incomes. The trouble manner retail merchants face is where to strike the balance between the breadth of the scope they offer and the deepness of pick available in their scope of merchandises. ‘Range width’ refers to how broad a pick of merchandises is to be offered to clients. Some manner retail merchants e.g. Miss Selfridge, began selling vesture and accoutrements but have widened their merchandise ranges to include jewelry, make-up and a whole host of other merchandises reflecting the lifestyle demands of the consumer. The deepness of a scope refers to the picks of manners, colorss, sizes and monetary values available to clients in important Numberss of units within production classs. When purchasers and merchants are be aftering the scope, they need to look at their mark consumer to place what merchandise categorization should be included and how many of each categorization, this is to assist understate hazard. Product Mix is an of import component of Range planning. If assess ed right, harmonizing to consumer demands and wants, the right merchandise mix will assist to maximize net incomes. The advantage of holding a merchandise mix is that it can potentially increase the volume of gross revenues, widening the scope on offer helps to distribute the fiscal hazards which are potentially involved. In add-on adding new scopes to the trade name will assist to diversify the manner company and convey approximately new concern chances and aid to increase possible gross revenues and net incomes. Having a wide merchandise mix will besides assist to aim new clients every bit good as making and prolonging the involvement of current consumers. Furthermore Product Mix is a cardinal driver in the scope planning procedure. The disadvantage of Product Mix as a control tool is an increased hazard of stock markdowns if Product Mix on offer is excessively broad ( Shaw, 2005 ) . When be aftering a scope program construction many retail merchants and purchasers struggle to make up one's mind whether to go on with merchandises which have sold good in the yesteryear but which are going ‘old fashioned’ such as the Laura Ashley jumper daisy print frock ( Shaw, 2005 ) . A clear and up to day of the month statement of the retailers’ market place and mark client, based on uninterrupted research, will give the purchaser assurance to be ruthlessly matter-of-fact over what to include and what non to maintain in a scope. This will assist to understate hazard within the manner company. When be aftering a merchandise scope, the Product Mix and categorization program are of critical importance ; each merchandise will hold its ain single merchandise life rhythm ( Doha, 2013 ) . The merchandise lifecycle of a merchandise has four really clearly defined phases, each with its ain features, that average different things for concerns that are seeking to pull off the life rhythms of their peculiar merchandises. These four distinct phases consist of: Introduction, Growth, Maturity and Decline. These four phases will change well harmonizing to the type of merchandise which is being sold, and whether it’s a authoritative, craze or manner merchandise. Product lifecycle direction is an of import control tool within calculating gross revenues and seasons. It makes it possible to command the whole lifetime of a merchandise and the information connected with it. Efficient merchandise lifecycle direction enables companies to vie successfully in international planetary markets ( Stark, 2006 ) . Product life rhythm direction is the activity of pull offing a company’s merchandises all the manner across its lifecycles in the most effectual manner. In so making, it enables the company to take control of its merchandises. If a company loses control of its merchandises this can hold black effects. Therefore merchandise life rhythm direction is a cardinal control tool, for merchants and purchasers. During the development of a merchandise, it doesn’t physically exist, so during that stage of its life it’s hard to command. PML helps to convey better merchandises to market faster, and enable better support of customersâ€℠¢ usage of merchandises. It is of import to convey a merchandise to market rapidly, otherwise the client will take a competitor’s merchandise before yours get to market. An advantage for purchasers and merchants of utilizing the merchandise lifecycle as a control tool is that it helps cut down the cost of a merchandise. The merchandise life rhythm can be used by purchasers and merchants to assist understate hazard for their manner companies by reexamining old products’ lifecycles, where they peaked and where they started to worsen. This can be used to an advantage to assist be after the merchandise life rhythms of similar futuristic merchandises which will assist to command and gauge the life rhythm of a merchandise. It’s of import to cut down merchandise costs ; otherwise consumers may halt utilizing your merchandise and utilize a competitor’s alternatively. PLM enables the value of a merchandise to be maximised over its lifecycle. With accurate consolid ated information about mature merchandises available, low-priced ways can be found to widen their revenue-generating life-times. PLM gives transparence about what is go oning over the merchandise lifecycle ; it offers directors visibleness about what is truly go oning with products.. However the disadvantage of utilizing the PLC is that the merchandise lifecycle is merely a theoretical theoretical account and the merchandise lifecycle is non needfully reflective of all manner merchandises. Forecasting gross revenues and seasons is a critical control tool which purchasers and merchants have to implement in order to understate hazard with planning scopes for their manner company. Manner prediction looks at the fiscal parts and the comparing of the seasons’ gross revenues last twelvemonth. Additionally manner companies have to equilibrate the customer’s pick with the demand to accomplish the best return of stock investing. The get downing point for many manner retail merchants is gross revenues history, as it identifies what clients expect to purchase from the concern based on their current and past demands. Gross saless history may mention to last week’s gross revenues, and as such besides reflects customers’ current seasonal demands ( Douglas 1997 ) . There is much valuable information available to the purchaser from the gross revenues history of old seasons. Forms have likely been established of the type of manner ware which the retailers’ clients have purchased in the yesteryear. Best sellers from old seasons need to be replaced with new yet every bit profitable ware, The purchaser needs to through ally analyse which fact ors contributed towards an points bestselling position ( Goworek 2007 ) . Manner season is an of import component of the control tool ‘Forecasting gross revenues and seasons’ . Historically there have been two clearly defined and traditional manner seasons, which are Autumn/Winter and SS. Easy ( 1995 ) explains that retail merchants have organised themselves around consumer demand that has traditionally been influenced by conditions forms. In the yesteryear there have been two clearly defined and traditional seasons and these are still steadfastly ingrained in our civilization, even though they have progressively less relevancy to consumers and manner concerns. With a greater accent on fulfilling the manner demands of customers’ altering life manners, manner retail merchants have to purchase more rapidly and maintain scopes focused on what clients want at peculiar times of the old ages, as opposed to fulfilling two big period of demand, i.e. summer and winter. Another factor which has influenced client demand is greater travel, as clien ts require vesture for a assortment of climes and occasions out of season, e.g. swimsuit in winter. In order to understate hazard manner retail merchants have to purchase more rapidly and maintain scopes focused on what clients want at a peculiar clip of the twelvemonth, as opposed to purchasing to fulfill two big periods of demand. In order for purchasers and merchants to understate hazard is it critical that they besides ‘buy’ in order to fulfill discreet ‘user occasions’ . Increasing consumer demand is coercing retail merchants to retink merchandise ranges about changed client behavior. Product categorization is a cardinal component of Range Planning. It is critical for the merchant and purchaser to calculate the merchandise lifecycle of merchandises with different categorizations such as Fashion, Fad and Classic. The merchant plays a cardinal function in working with the purchaser to accomplish the right balance of merchandises within the seasons range. This is done by utilizing a combination of gross revenues history combined with forward gross revenues and tendency anticipations. The sum of manner and fad merchandise in the Product Mix will find the sum of flexibleness needed. It is noted that the manner manner tendency reviews begin many months in progress of a season. Normally at the terminal of the old season the purchasing and selling squad will hold undertaken what is known as a ‘seasons station mortem’ . The advantages of set abouting a seasons station mortem is that it is a utile papers for be aftering the future manner way of a manner concern and is helpful during the planning of a new season. It is of import to command the degree of stock and type of stock that is in the concern at any given clip ; this is carried out through manner prediction, and the scrutiny of merchandise classs to utilize in a season, such as Authoritative trench coats manner or crazes. The ware program is an of import control tool in order to understate the hazard of holding excessively much ‘cost’ that hazards being unsold ( Shaw 2005 ) . This relates to be aftering and profitableness. It is critical to observe that one of the most of import ends in successful selling is to help the procedure of accomplishing the planned degree of purchasing profitableness. ‘Buying profit’ can be defined as the difference between the cost monetary value paid for the garment and the retail merchandising monetary value in the store. The grade up or net income made between the two monetary values can be explained by the undermentioned merchandising monetary value equation. Retail merchandisin g monetary value ( RSP ) -Cost Price ( CP ) =Gross Margin ( or gross net income ) . WSSI is an of import control tool which is used every twenty-four hours in the life of a purchaser and merchant. Throughout the twelvemonth, it is critical for the merchant and purchaser to construct stock degrees in expectancy of trading extremum periods such as Christmas and high summer. The WISSI is really good at assisting the purchaser and merchant program frontward stock consumption into the DC. Every Monday gross revenues figures are awaited in order that purchasers and merchants can place fast selling lines that need reiterate bargains to guarantee that they are kept in stock. On the other manus it is besides the Monday figures which reveal the worst Sellerss that finally will necessitate holding their monetary values marked down. Thus the WISSI helps to pull off and command hazard. The WISSI is one of the most valuable studies used by the purchaser. It includes gross revenues and stock list degrees by manner for the old hebdomad. The WISSI acts as an overall control mechanis m to guarantee that the flow of stock and money in the concern is go oning expeditiously. WSSI is an of import control tool for purchasers and merchants because it significantly works alongside the bringing agenda. It lists the hebdomad and the twenty-four hours when the providers have been requested to present the garments to the DC. Being cognizant of precisely what is traveling to be delivered is besides of cardinal involvement to shop and promotional direction maps of the administrations ; this can assist pull off hazard and supply a competitory advantage for the manner company. One of the cardinal selling activities controlled by utilizing the WISSI is the facet of line degree monetary value change that regularly occurs within all manner concerns. Ideally at the start of the season, the purchaser and merchant purpose to make a monetary value construction that relates to the type of clients to whom they are selling. Monetary values must besides be competitory to guarantee they a re in line with major competition. Overall WISSI acts as the cardinal selling direction papers within manner retailing ; it is lone portion of a wider scope of be aftering paperss. It is shamelessly an internal fiscal control papers which is a sum-up of all the trading activity within the section and finally the full concern. There are disadvantages of WISSI as a control tool. Taken out of context, it can be a nonmeaningful page of figures. It requires careful reading, utilizing experience to pull strings both it and the purchasing, if the section is to win. It is of import to retrieve that manner is non formulaic ; merchants try to utilize it in a formulaic manner, bring forthing it without understanding. Therefore it is of import that merchants and purchasers do non respond excessively rapidly to sudden good and bad gross revenues forms ; reaction to the WSSI must be both considered and careful. Overall the of import of understanding the workings of WSSI should non be underestimated. Using this control document the merchant has considerable advantages and is able to maintain hebdomadal updates of yesteryear, nowadays and likely future public presentations of the section and direction to on a regular basis look into their public presentations against the original program. More significantly it acts as a decision-making tool to assist the concern make up one's mind what action to take on an on-going regular footing. This will more significantly assist to understate hazard and aid to maximize net incomes. To reason there are a figure of control tools which purchasers and merchants can utilize in order to understate hazard and maximize net incomes when planning ranges for their manner companies. There are considerable advantages of utilizing a figure of control tools in order to derive competitory advantages over similar manner companies and rivals and to assist increase net income borders. Manner companies will strategically utilize a combination of control tools in order program in front to give themselves the greatest opportunity of success. For a manner company to win it has to â€Å"reflect the demands and the desires of a coevals at a peculiar point in clip. It has to be of the minute, but with ageless values that will ever be recognised† ( Menkes 2001 ) Mentions: Ahmed Doha, Ajay Das, Mark Pagell, ( 2013 ) â€Å" The influence of merchandise life rhythm on the efficaciousness of buying patterns † , International Journal of Operations & A ; Production Management, Vol. 33 Iodine: 4, pp.470 – 498 Charles-Roux, E. ( 2004_ .The universe of Coco Chanel. Friends Fashion Fame. London: Thames and Hudson, p.11 Douglas C. West, ( 1997 ) â€Å" Pull offing Gross saless Forecasting † , Management Research News, Vol. 20 Iodine: 4, pp.1 – 10 Easy, M. ( 1995 )Manner Selling.Oxford: Blackwell Science. Goworek, H ( 2007 ) .Manner Buying. London: Blackwell. p44-45. Hines.T, et Al. ( 2007 )Manner Selling: modern-day issues.Routledge ; 2 edition ( 2006 ) Jackson.T, et Al. ( 2000 )Manner Buying and trading direction #Gerald Watts, Jason Cope, Michael Hulme, ( 1998 ) â€Å" Ansoff’s Matrix, hurting and addition: Growth schemes and adaptative acquisition among little nutrient manufacturers † , International Journal of Entrepreneurial Behaviour & A ; Research, Vol. 4 Iodine: 2, pp.101 – 111 Stark, J ( 2006 ) .Product Lifecycle Mangament 21stCenturyParadigm for Product Realisation. London: Dordrecht: Springer 2006. P 4-5

Smoking A Therapeutic Approach to Quitting Essay - 981 Words

Smoking A Therapeutic Approach to Quitting While smoking is a problem that affects millions of people in The United States, several different approaches are available to assist in breaking the habit; specifically, therapeutic approaches, when utilizing group sessions, one-on-one interaction, or self direction, do offer the ability to modify personal views, behaviors or other challenging issues to achieve predetermined results. Since each of the aforementioned options provide varying amounts of contact with trained therapists and other people, it is necessary to make a remedial selection based upon the comfortability and willingness of a person to openly talk about their dilemma with others. However, in order to successfully establish†¦show more content†¦14), I would personally prefer using covert sensitization. I choose this option because of its exclusivity, and only I can determine what reasons of quitting smoking will be effective in my situation; as case and point, there are different reasons to stop smoking, such as: inflated cost of cigarettes, severe health risks, pregnancy, and vanity. Many people decide to use one or all of these choices, but I believe that the most advisable way to utilize these powerful motivators is through self-reflection and action. Why Would This Address the Problem? With my chosen approach, the importance is in the choice of control, which comes from within me; for, â€Å"you have to change your mind, if you want to change your life†¦ You have to change your attitude towards smoking to get rid of it. Until you†¦begin to feel good about quitting, you will get nowhere† (unknown author, 2006, para. 3). 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