Okefenokee Swamp Essay

Swamps can be seen in various perspectives and can convey different atmospheres associated with the respective swamp. The two passages on the Okefenokee Swamp both convey two different atmospheres and tones for the swamp, almost as if it was two different swamps. The author use of diction, detail and figurative language conveys how the swamp in the first passage is more inviting and safe versus the swamp in the second passage which is displayed in a more malicious tone. The author’s use of diction in the first passage and the second passage regarding the swamp is extremely different from one another. Throughout the first passage the author uses diction such as â€Å"primitive†, â€Å"festooned† and â€Å"exotic† to describe the swamp. The author’s formal tone and word choice helps convey that the swamp is a peaceful and safe environment that is appealing to others as well as to the author. The diction helps support as if the author is trying to persuade others to come to this swamp and enjoy its beauty Unlike the first passage, in the second passage the author uses diction such as â€Å"unfathomable†, â€Å"unconquerable† and â€Å"misery of life† to convey the atmosphere of this swamp. The author sees the swamp in the second passage in a more malicious and dark light that almost seems to be repelling the reader. The tone the author holds is almost as if he is trying to repel the reader from coming and visiting the swamp. It is negative and does not display any of the attributes that are associated with the swamp; instead it is very critical and focuses on all the peculiarities of the swamp. The word choice that is used in both the passages varies from one another and conveys different atmospheres for the swamp. Detail is a major factor throughout the two passages that helps distinguish the swamps in the passages from one another. In the first passage the author uses detail that have positive context such as, â€Å"Saucer-shaped depression of approximately 25 miles wide and 40 miles long that covers an area of more than 600 square miles.† This detail holds a revealing and formal tone that relays a pleasant atmosphere. It also has an alluring tone that persuades the reader to come visit the swamp and explore its vastness. In the second  passage the author uses details such as, â€Å"Four hundred and thirty thousand acres of stinging, biting and boring insects.† Although the second passage is also describing the enormous land size it is presented in a much more malicious and unappealing way. The author uses various literary devices throughout the two passages to show that he appreciates one swamp more than the other. In the first passage the author uses personification to expand and emphasize the beauty of the swamp. It states, â€Å"Exotic flowers. Among them floating hearts. Lilies. And rare orchards abound†, the floating hearts help convey an atmosphere of peace and playfulness that attracts the reader’s attention as well as persuades them to come visit the swamp. In the second passage the author uses a simile to show the distaste and lack of appreciation they held for the second swamp. The author describes the swamp â€Å"like some hellish zoo† almost conveying as if the swamp was every readers nightmare. The figurative language the author uses in the second passage is primarily focused on repelling the reader and persuading them not to come to the swamp and stay in the comforts of their own home. Unlike in the first passage where the figurate language was solely focused on inviting the reader to visit the swamp and enjoy the beauty and serenity the swamp had to offer. The author holds two very different tones when discussing the different swamps. The author uses different devices throughout the passages to create and convey a certain tone for each swamp. In the first passage the swamp is seen as something that holds beauty and is inviting unlike in the second passage where the swamp seems more malicious and repelling. The author uses various levels of diction, detail and figurative language to convey the tone and attitude they had toward each of the respective swamps, almost making it seem as if it were two different swamps entirely.

Conscious Capitalism

In the introduction of conscious capitalism, John Mackey gives us a brief history on how he established Safer Way which is now known as Whole Foods Market with no prior business education background. His intention was to create a business that was ethical and purpose driven. His insight on free enterprise capitalism is much focused throughout the introduction and first chapter, its importance and how it has changed the world. I agree with the findings because, it free enterprise capitalism has led to innovations and social cooperation that any other system.It has freed millions of individuals from extreme poverty and enabled businesses to open up around the world that have contributed to the welfare of humanity. Raj Sisodia, the co-author of the book together with Mackey, have pursued to change the way business is thought about, taught and practiced through their movement. This is both noble and I believe it’s our human responsibility to find solutions to the world and innovat ions that can take us to the next phase of how business is done.Capitalism has been misunderstood and has been tagged with bad titles such as crony capitalism, profit maximization for investors as top priority and unethical business activities as inferred by products from its activity. Crony capitalism is one of the most tagged names of big corporations because they have the financial resources to corrupt and bribe the government and inturn use this power for their own enrichment.Although the above examples are true and real, conscious capitalism is able to resolve this issue by ensuring that businesses not only focus on profit maximization but in creating value for all its stakeholders. Examples of how it does this, is by strongly opposing greed, exploitation, manipulation and by building environments based on trust and love that positively transform the world. Business are good because they create value, they are ethical because they are based on voluntary exchange.The book provid es turning points in history that were made possible by free capitalism, examples include the fall of the berlin wall on November 9, 1989, which brought about the end of communist regimes and the birth of the web by Tim Berners in 1989 which has brought about access to unlimited information and has connected almost everyone in the world with social media such as Facebook and Twitter. No known business is in isolation; all exist and have interaction with people in different environments in their pursuit of profit maximization. In  order to do this and create more value for all its stakeholders we are introduced to the four tenets of conscious capitalism, they include higher purpose, stakeholder integration, conscious leadership and conscious culture.First and foremost the reason I agree with Conscious capitalism is that they are subjected to the discipline of the market and they need to deliver strong financial results just as any other business however, they do this by doing what is right because it is right Secondly, I agree with John Mackey is his description of a business based on conscious capitalism by using imagination.Imagine a business based on love and caring whose team member are passionate and committed to their work; a business that cares profoundly about wellbeing of its customers and above all a business exists in a virtuous cycle of multifaceted value creation, generating social, intellectual, emotional, spiritual, cultural and physical wealth for everyone. John Mackey ends this description with giving examples of businesses that exist with such a description, from Google to Starbucks to the container store; this proves that conscious capitalism does exist in reality and therefore it can be achieved.Traditional businesses tend to use corporate social responsibility as a defensive mechanism and to build its reputation with external stakeholders. I solely agree that CSR mindset should be built in to the core of the business because it integrates the interests of all stakeholders and incorporates higher purpose and caring culture. An example is Google, with its Google grant donation program awarding free Ad Words advertising to select charitable organizations; this is centrally aligned with its business.Another example is Johnson and Johnson, its purpose is to alleviate pain and suffering however remains focused in making profits to ensure sustainability and growth. Thirdly, I also agree with the statement that happiness cannot be pursued unless for short term gratification. True happiness comes from living a life of meaning and purpose, which I believe goes the same for businesses. If a business pursues achieving its purpose then happiness will be achieved through satisfaction of all stakeholders.Matching individual purpose with business purpose is one of the ways to enhance passion for work and increase performance. This also makes it easier for team members to overcome challenges with ease. In conclusion, although some o f his view on some issues such as reduction in violence levels and majority becoming aware of consuming healthier foods is brought about by conscious capitalism is contentious generally I agree with his thought. By aligning business in a set of four categories the good, the true, the beautiful, the heroic.Conscious businesses should strive to provide service to its stakeholders that improve the quality of life such as Wholefoods in its endeavor of providing food products that are healthy to consume; they should be in the forefront of furthering human knowledge by being innovative and researching on better products or services for their customers, an example is Google; they have been able to create a way to access any information in the internet by a click of a button and last but not least a business should be heroic, the courage to do what is right to change and improve the world an example given is Grameen bank in India that has been able to empower poor people in the eradication of extreme poverty. Finally, as businesses pursue profit maximization they should also strive to discover and grow in their purpose; this is the only way they can be able to wholesomely create value for all stakeholders and adapt to an ever changing and developing world.

Representation of Women in Bollywood

Cinema is meant and believed to entertain. It is a popular media of mass consumption which plays a key role in molding opinions, constructing images and reinforcing dominant cultural values. The representation of women characters in the main stream bollywood movies. Hindi cinema has been a major point of reference for Indian culture in this century. It has shaped and expressed the changing scenario of modern India to an extent. Bollywood cinema has influenced the way in which people perceive various aspects of their own life. Bollywood’s journey is nearly hundred years.Bollywood has been essentially male-centric, leaving a little space for the female counterparts to evolve and grow as performers. In earlier films, they were based on mythology or historic kind, where male were given importance and female were showcased here and there. Now, it’s different. As women in India redefined the status and role in the society, it is seemed to be reflected in the films as well. Bu t up to what extent is it true it should be a healthy practice. There is no reason to think women as sensitive creatures and huddled behind a veil.The way of living of real life women is the inspiration for reel life act. But is it an exact portrayal of what we see in real life? The changes found in the characterization of women in the last decade are vast. WOMEN CENTRIC FILMS During 2002 – 2012, there is a lot of notable women centric films. Women being protagonist is either portrayed as a positive or negative character. But there are also movies like â€Å"JODHA-AKBAR† which is not under this category but the characterization of Jodha, makes us feel that movie is being pulled by her.Likewise, â€Å"ROCKSTAR† the entire movie is scripted on the heroin; but the character of her is negative (i. e. against the social ethics). Women are either portrayed as modern life â€Å"Sati-Savithri’s† or entirely the negative version. But women are hardly portra yed as they are in real life. The movies are made for the audience. Therefore, they portray in such a way , how they could grab the attention of their audience. There is a blind mindset always makes the filmmakers to insists their heroin to dress up in more of glamour kind.The teenagers are also found to be in modern outfits. But, not as they portray in cinema. WOMEN IN REALISTIC FILMS In real life depicted films like â€Å"RAAJNEETHI† , â€Å"SARKAR† , â€Å"SARKAR-RAJ† women are showcased boldly and courageously. In a movie like â€Å"GURU†, the women’s role is soft and subtle. This type of characterizations inspires and develops the respect for women in society. Like wise, movies like â€Å"RAAGINI MMS† should not be encouraged. They should be a ban. As this will definitely have affect on the social dignity of women.

Define TQM Essay Example | Topics and Well Written Essays - 250 words

Define TQM - Essay Example ic upon seeking to manage the organization as a whole and develop all of the processes and levels of integration that exist within it in such a manner that a level of synergy can be realized between all of these factors (Alfalla-Luque et al., 2012). Obviously, as can quickly be noted, such an approach is difficult to achieve on any macro scale and ultimately breaks down into a situation in which micro processes will dictate whether or not total quality management, or some lesser achievement, will ultimately be realized. Whereas many different approaches to management had existed over the past several decades, total quality management, although still employed within a litany of different fields, has not experienced the same widespread level of success that it did during the late 1980s and early 1990s (Zatzick et al., 2012). Instead, approaches such as ISO 9000 and Six Sigma have permeated the management field and slightly turned the focus away from total quality management; at least a s compared to what it was two decades ago. Alfalla-Luque, R., Marà ­n-Garcà ­a, J. A., & Medina-Là ³pez, C. (2012). Is worker commitment necessary for achieving competitive advantage and customer satisfaction when companies use HRM and TQM practices?. Universia Business Review, (36), 64-88. Zatzick, C. D., Moliterno, T. P., & Fang, T. (2012). Strategic (MIS)FIT: The Implementation of TQM in Manufacturing Organizations.Strategic Management Journal,  33(11), 1321-1330.

Management of hypoglycemia Essay Example | Topics and Well Written Essays - 3000 words

Management of hypoglycemia - Essay Example Diabetes is increasingly common throughout the world.Its prevalence is well over 5% in many developed countries and is rising in the United Kingdom.While talking about its prevalence,it is to be remembered that diabetes is caused by our genes and our personal environment, which is created by our lifestyles. For people with diabetes, whether type 1 or type 2, the disorder can have a profound influence on all aspects of life and can affect most organs of the body. There is no cure for diabetes, and once occurs, it is present for life. The increasing appreciation throughout the healthcare industry for clinical decisions to be driven by sound scientific evidence represents an opportunity for the patients, practitioners, and healthcare planners alike in that it shapes the delivery of care on the basis of evidence of effectiveness. The past two decades have seen considerable progress in producing evidence to support treatments aimed at reducing the risk of diabetes and its complications. T his indicates a shift of focus from treatment of effects to treatment of cause, where prophylactic interventions are more important than just therapeutic measures. This means caregiving now does not await the disease to happen; rather, the care tends to identify population or individual who would develop diabetes in a later life (Foster, 1998, p 2069-2070).Definitions: Diabetes mellitus occurs either because of lack of insulin or because of the presence of factors that oppose the action of insulin. Therefore, it can be defined as a chronic disease caused by inherited and/or acquired deficiency in production of insulin by the pancreas or by the ineffectiveness of the insulin that is produced. The result of insufficient action of insulin is an increase in blood glucose concentration or hyperglycaemia. This increased glucose would damage many of the body systems, most significantly blood vessels and nerves. Data from genetic, epidemiological, and aetiologic studies have led to the impr ovement of the understanding of the aetiology and pathogenesis of diabetes. From my experience in the clinical placement, I have come across patients during clinical practice with varied presentations. Whatever may be the mode of diagnosis, persistently elevated plasma glucose in the fasting state would lead to the diagnosis of diabetes mellitus even if the patient is asymptomatic. In the ward while working with the patients of diabetes, frequent monitoring of plasma glucose and adjusting the diet or medication is a normal routine nursing activity. I have see a venous plasma glucose concentration of 140 mg/dL in at least two separate occasions or plasma glucose concentration of 200 mg/dL 2 hours following ingestion of 75 g of glucose would constitute a diagnosis of diabetes mellitus(Foster, 1998, p 2080). Insulin: Diabetes is treated with insulin in the inpatients with or without oral hypoglycaemic agents. Since the disease at least partly results from insulin deficit, insulin is required for many patients with diabetes mellitus type 2. If no oral agents are used for treatment, all diet-responsive type 2 diabetics must be treated with insulin. My observation suggests that it is fairly easy to control the symptoms of diabetes with insulin, and as a nurse working in the ward, I know it is difficult to maintain a normal blood sugar throughout the day even with the use of multiple injections. It is also to be mentioned that the patient treated with insulin by injection cannot reproduce the physiologic changes in a normal patient in response to a diet load. Thus, if enough insulin is given to keep the postprandial glucose normal, too much insulin will be present during the postabsorptive phase, and hypoglycaemia will result. As a result, insulin therapy is guided mainly by adjustments

Working out Alternatives Case Study Example | Topics and Well Written Essays - 1250 words

Working out Alternatives - Case Study Example As a safety precaution, the teams are provided with emergency radios when there is need to report cases such as loss or being hurt among the team members. Information such as the starting point of the race is usually given a day before the event (Kumar, Petersen and Robert70). The team members therefore, have to find means to get through and survive as fast as possible. Adventure racing has its roots in a race set in New Zealand, in the year 1980. The wilderness endurance racing was started when one person accidentally overheard a conversation between Americans to create a wilderness challenge. FAR was founded by Dave Ziestma in the year 1998. In a span of three years, FAR’s ‘raid the north’ series served as an introduction to adventure racing to several Canadians. It also became included in the list of the top race series in North America (Lewis 536). FAR’s races were termed as difficult (Holland and Jackie 248). Of the teams sent out, only about 25% would make it to the finishing line intact and in time. The founder of the race, Zietsma, would usually take part in the race. As a result, many got assured that the race would be safe. His involvement in the race as a top competitor also aided in making the name of the company, making it be at the forefront. Frontier Adventure Racing (FAR) is a popular firm in the North Racing series. The organization has been popular among lovers of adventure racing. Adventure racing may take the definition of a race of individuals using vehicles over some terrain. The event is multi-sport, non-stop and with a mixture of teams. It is therefore essential for team members to ensure that they cross the finish line first and in time. The event borrows from kayaking, mountain biking and running. Its popularity grew in New Zealand, Australia and in Europe. Therefore, in the last decades, the event has attracted media attention as well as many players. In retrospect, FAR one of the players of Adventure

Collective Behavior and Social Change Essay Example | Topics and Well Written Essays - 750 words

Collective Behavior and Social Change - Essay Example So, if a business company decides to modernize, interested community of people who are apt to get a job in the same company must gain the formal education as a credential fit for the jobs available. Conversely, some ideological link may hamper a society to adapt fully to innovations. For example, the use of internet has limitations to some sectors because of religious convictions in terms of moral standards. But, most schools today encourage students to utilize information technology to help them facilitate learning. As in the past, educational institutions were exclusive, either for whites only or for blacks only. Today, most schools no longer recognize race and color. (479-481) 3. Switch over of ideas, barter of technologies, and exchange of people, allows the flow of knowledge, technology, and people into many varied directions bringing about awareness, education and eventual social change after people realize the benefits of what had just been learned. (482) 4. Crowds are swarm of people gathered in a milieu as commonplace bystanders, usual spectators, but, can also be active partakers as in theatrical presentations, and expressive participants as in religious rites. This throng of people does not necessarily interact for deeper interpersonal relationships. Conversely, all these kinds of crowds taken together compose the aggregate group. (Scribd chapter7) 5. Bell-bottom blue-jeans were actually in for sometimes, replacing tight-fitting cowboy look. Living in one's own home also became a fad because in economics reality, it is not categorized as consumption, but, an investment which most people did not understand, but went with the crowd because it was the style. 6. Universally, people panic upon hearing news of an upcoming danger to human properties and lives like storm, fire, volcanic eruptions, war, foreclosures, bankruptcies, outbreak of infectious diseases etc. which are all beyond normal living conditions, and this is the usual reaction of an active kind of crowd. (484) 7. Leaders in social movements are the polarizing factors. They are usually persuasive and fluent at crafting a united stand, and clever at convincing people to supply time, money, place, etc, for the cause. (Chapter 18 491) 8. Relative deprivation theory means that the deficiency in life of people within a society is calculated from end to end evaluation of similarities and differences of lives and living conditions within the same society (Chapter 18 490). But, apparently, not every member of social movements is motivated by the same mental attitude because of individual differences which may be contributed by family backgrounds, education, and social exposures. 9. Reactionary social movements cling to what went before the present generation. They may be called the traditionalists who intend to keep white supremacy over everything considering the applicability of old cultural beliefs and idiosyncrasy. Conversely, the conservative social movements intend to preserve the current practices and strive to resists growth and developments (Chapter 18 491). 10. Arrays of civil rights movements come and go. Usually, they start conglomerating because of a prevailing problem which does not have a solution in sight. Then they band together and elect a polarizing individual to solidify their stand on the perceived

Evaluate an Argument Essay Example | Topics and Well Written Essays - 500 words - 1

Evaluate an Argument - Essay Example The author further states that the information may be true but, but since the experts do not review the site’s access; there is a substantial peril in relying on this resource for your academic works (Isites.harvard.edu, n.d.). As with the conclusion of the argument, the author clamps the whole argument against the use of Wikipedia and suggests that Wikipedia is good for use for some functions. For instance, they can be used for becoming accustomed or background information of what one is researching (Isites.harvard.edu, n.d.). For the validation of the argument to hold, it means that all of their premises that support the argument are all true and the conclusion must ultimately be true. The argument in this article is valid because the premises are well linked to the conclusion. In terms of soundness, the argument is not sound even if it is deductive and logically valid; it has some mythical arguments that habitually make it unsound. When viewed from an inductive perspective, which holds on the basis of probability, that if the premises given are true then conclusion is most likely to be true. The inductive argument employed here is strong- it holds on the assumption that the conclusion of X is probably true if the premises of X are true. The deductive is valid as per the author’s premises for a number of reasons, which include; the contributors are not expertise and are anonymous and, therefore, their credibility is questionable; the information or articles on the website are out-of-date and may be posted by someone who wants to just mislead the audience; and because the administrators do not review the site’s access. Based on these validations, it clear that the deductive argument is true and valid since its conclusion sustains all the premises stated therein. However, the deductive argument in the article is not sound because of the following. Several

Human Resource Information Systems Essay Example | Topics and Well Written Essays - 500 words

Human Resource Information Systems - Essay Example In this industry, there is need for the employees to work in a friendly and comfortable environment. Their working environment can only be comfortable if they have things to do with their health looked into; if they get their salaries effectively and other things related to work made easier for them. This will definitely boost the morale of the employees and they do their work effectively (Gupta & Chhabra, 2004). When Mr. Morgan approached me regarding all this for advice as a HR Consultant, I decided to introduce him to the Human Resources Information System (HRIS). Since he would like to travel less due to the cost of fuel, the HRIS should be able to help him execute most of his work effectively and at cheaper costs. This will save him time, energy, and money. IDENTIFIED PROBLEMS: The HR problems Mr. Morgan is facing will be no more as soon as the use of HRIS is put in place. The main problem was travelling every week, which wasted so much time and was expensive in terms of fuel. The manner in which he made the payroll checks and the use of Excel spread sheet to prepare them would greatly be improved through HRIS. Some of the HR functions, which will be efficient, in this case are: Salary determination is one of the most important functions in this business. The work of HRIS in this field acts as the software for clarifying for data entry, data information, and data tracking. It helps the Human Resources to manage accounting functions and payrolls within the company or business. It is the most efficient and effective way of transacting accounting calculations. It will provide data, which is very accurate. It can also keep and retrieve big amounts of data making work much easier (Gupta & Chhabra, 2004). HRIS NEEDS ASSESSMENT: The computerization used in the HRIS might create more effective process in the functions above, due to the fact that it has the ability to perform very heavy tasks within a short time. For instance, if the HR

Social media and its usage for marketing and advertisements Research Paper

Social media and its usage for marketing and advertisements - Research Paper Example This research will begin with the statement that in this information technology-based era every business is trying to enhance its services and products over its competitors. In this case, online support is considered to be a crucial factor in any advertising policy and arrangement. Additionally, companies and industries initiated promotion and publicity operations focused on people with the help of written sources and audio/video channels. However, this policy is expensive and its efficacy is hard to determine. On the other hand, it is generally not obvious that how broadcast marketing will play a major role in the latest information financial condition that allows for exceptionally cooperative and focused interactions. Thus, in this customer-oriented financial system, companies are trying to discover additional money-making methods to promote their business services and products-line, access existing clients, and make new users/clients across the business boundaries. Business people sometimes talk about the Return on Investment (ROI) in Social Networking. In fact, it sounds great, but what does it mean to the business people? How does it actually help them grow their business? The simple answer to these questions is that businesses could be successful through Social Networking. However, normally, social networking is a process that helps organizations establish the relationship and then continue and maintain them by means of customary practices such as telephone, e-mail, and face-to-face meetings.... Thus, if a company builds identity on the social networking site, it will allow drawing people who share their viewpoints and want to work with you (Tubb, 2011). At the present, the social networking websites have become a family name. Despite the fact that, it is someone’s confidential or professional life, they are in fact wonderful process for communicating with friends, colleagues, and making new relationships online (Sedycias, 2009). Since, a social networking web site is a web site where people are able to develop an online profile; and they mention their interests in profile, as well as they can insert links to other profiles. Typically, the users of social networking sites are able to dispense personal information, encompassing photographs, blog entries and videos. MySpace is an example of social networks, which is very popular in the same way with individuals and with music groups, who are able to market their tour dates, and transfer their songs on their profile page s (Hawkins, 2009). A social networking site is a very helpful and functional way to keep in contact with a large number of people (friends, colleagues, relatives, etc). Since, if someone has some news which he/she needs or wishes to distribute to others, then he/she can insert it on his/her personal bulletin board and it will be transferred or distributed to the people to whom he wants to share. Thus, the social networking sites can play a significant function for meeting with new people, whether on a business or friendship (Hawkins, 2009). Phillips & Phillips (1998) define the social network as a set of organizations, businesses, individuals, or other social entities

The Need for Variance Analysis Term Paper Example | Topics and Well Written Essays - 500 words

The Need for Variance Analysis - Term Paper Example Baker and Baker (2014) define variance as "The difference between standard and actual prices and quantities" (p. 201), and variance analysis as "Analysis of these differences" (p. 201). Variance analysis underscores or brings attention to deviations from what was initially intended. It is only after deviations have been identified that adequate and effective decisions can be made by the individuals in management positions (Baker and Baker, 2014). According to Berger (2011), lack of conducting a variance analysis may prompt leaders to overlook areas causing these deviations and hence fail to make relevant decisions. For variance analysis to be performed, a prior formulation of standards ought to be conducted, for instance, budgetary targets (Baker and Baker, 2014). This shows that variance analysis drives managers and other leaders to set targets or standards in advance. Decision making goes hand-in-hand with accountability in that individuals overseeing areas proved by the variance a nalysis to be causing the variations are answerable. Therefore, departmental heads are forced to make effective decisions and implement relevant strategies that forces them to operate within the budget (Berger, 2011). In the case study, new procedures for discharging patients have been implemented. A variance analysis conducted after six months showed that costs were running 25 percent higher than was expected. After conducting the variance and proving that there is actually a deviation from the budget by 25 percent, the next step would be to identify the causes of the deviation (Berger, 2011). There are many areas that may cause the deviation, for instance, extended working hours that increase the cost of labor. After identifying the cause of the deviation, then my next step would be to evaluate action plans under implementation to determine their effectiveness and investigate how they can be improved (Berger, 2011). If the action plans

Is Wal Mart Good For America Essay Example for Free

Is Wal Mart Good For America Essay The growth and dominance of Wal-Mart over the years has indeed turned it into an economy all its own. There is no single corporation in the world that has as much impact as Wal-Mart has, save perhaps for oil corporations, on any single domestic or even international economy. The volume of sales which Wal-Mart generates is enough to keep the rest of the world happily employed and is also enough to keep the consumerist population of the United States happy in their spending habits. Every single commodity manufacturer who is interested in surviving must be able to gain the good graces of Wal-Mart. The impact of Wal-Mart is such that it â€Å"has life-or-death decision over [almost] all the consumer goods industries that exist in the United States (Gereffi 2006). † This virtually unbridled power of the life or death of consumer goods industries, however, presents the question of whether or not Wal-Mart is indeed good for America. While it certainly does create jobs and sustains the consumer goods industries of America, it also has the same power of taking such benefits away and perhaps leaving the American economy in a far worse off situation than it is at the present. Wal-Mart has become so powerful that any shift in its purchasing and production policies will certainly result in some imbalance on a certain economy in the world. An example of the power of Wal-Mart is the fact that because of the immense volume of sales that Wal-Mart generates it is able to dictate where goods are to be manufactured in the world despite the fact that it is not a producer but simply a retail-chain. For a retail chain to be able to dictate to suppliers where they are to produce their items in order to be able to sell to Wal-Mart at a lower cost means that the retail chain has either a huge stake in the ownership of the supplier or buys so much from the supplier that it is able to dictate the price that it is willing to buy at and by doing so dictate where such goods are to be produced. In determining whether or not Wal-Mart is good for America, the basic economic principles of any market must be analyzed. Every market is governed by two basic forces, supply and demand. Wal-Mart is able to control both these forces because of the immense size that it has. It controls supply by deciding what items it chooses to retail. It also decides the demand for the item by pricing competitively. In its early years, Wal-Mart provided jobs for most Americans because a majority of the goods supplied were produced in the United States. With globalization and the theory of a flat world, other countries have become more competitive industrially and have now taken those production jobs that were previously held by Americans. The outsourcing of consumer goods industries to other countries can be theorized to have been created by the Wal-Mart demand for cheaper products from their suppliers. The problem with this scenario is that it creates unemployment for Americans who are the main buyers from Wal-Mart. In order to counteract this scenario, Wal-Mart must then reduce prices lower to meet the increasing lower income bracket of unemployed Americans who have lost their jobs because of the global production tilt to other countries. As this trend continues, it may be theorized that Wal-Mart will eventually strangle itself by driving the prices of goods down too much without protecting its major market which is the United States. The status of Wal-Mart, therefore, as either a boon or a bane for the American economy solely depends on whether or not it is willing to protect its major market which is the United States. Wal-Mart not only determines which consumer goods industries are to survive but also which economies are to benefit.

Analysis of the Term Victorian

Analysis of the Term Victorian How useful is the term ‘Victorian’? The era of Queen Victoria’s reign witnessed the passing of milestones in social, economic, and personal progress. It was the age of industrialisation, a time of travel, a battleground for the conflict between science and religion. Yet further to these great markers by which many of us recognise the nineteenth century, and indeed because of them, Victoria’s reign inspired change within the individual; a revaluation of what it meant to be a human being. The literary artists gave new form to the questions on the lips of the society around them: questions that were no longer so easily answered by Christianity. This essay will explore how the term ‘Victorian’ does or doesn’t fit into the context from which it supposedly arises. I will look at trends such as the development of literary criticism, pioneering scientific discoveries, the exploration into psychic phenomenon, the increasing independence of women, the mapping of the world, all of which contribu te to what we know and understand as ‘Victorian’, and have in some way shaped the work of authors such as Eliot, Conan Doyle, and H.G Wells. Using some close textual analysis I hope to identify the nature of the inspiration behind the literature of the time and whether or not such work transcends the limits of the term ‘Victorian.’ Many great literary minds of the time such as Arnold, Dickens, and Ruskin helped define the era in their critical attitudes towards it. (Davis 2002, p.10). Criticism appears to have become a form of exploration in an attempt to turn what concerned and worried the artist into something that questioned and reassured. Arnold, in his Essays in Criticism (Arnold, 1865, p.V) explains how he perceives the difference between logical and artistic thought The truth is I have never been able to ht it off happily with the logicians, and it would be mere affectation in me to give myself the airs of doing so. They imagine truth something to be proved, I something to be seen; they something to be manufactured, I as something to be found. It is this growing awareness of difference that was to become a defining feature of Victorian literature. Differences appeared in the very perception of things, which led to feelings of isolation, despair, alienation all prominent themes in nineteenth century work. In Arnold’s A Summer Night (Arnold 1913, p.167) we see the poetic mind struggling to find meaning on a moonlit street where the windows, like hostile faces, are ‘silent and white, unopening down’: And the calm moonlight seems to say Hast thou then still the old unquiet breast That neither deadens into rest Nor ever feels the fiery glow That whirls the spirit front itself away, 30 But fluctuates to and fro Never by passion quite possessd And never quite benumbd by the worlds sway? And I, I know not if to pray Still to be what I am, or yield, and be Like all the other men I see. Arnold recognises that the society around him is unfulfilled, that men are giving ‘their lives to some unmeaning taskwork’ and he questions whether he should be questioning at all. He is aware of a gap between the reality of working life and life outside of work; a difference that he strives to find explanation for. Arnold appears to be lost amidst the streets of his own mind afraid of not being able to define who he is, what he is. These feelings in part express what it meant to be a Victorian struggling to place thoughts and feelings which appear to no longer fit into society. The Victorian era contained much of what had past and much of what was still to come it cannot be seen as an isolated time, nor as an isolated term. It contained aspects of the Romantic period for instance in Arnold’s poem, The Buried Life, we see vestiges of Wordsworth’s legacy of Ode to Immortality. In both poems there is a sense of something lost an old passion or instinct that has gone with the passing of time yet Arnold, unlike Wordsworth, finds it more difficult to come to terms with this: ‘A longing to inquire / Into the mystery of this heart that beats / So wild, so deep in us, to know / Whence our thoughts come and where they go.’ (Arnold 1913, p.170). The language is more passionately discontent than the resolute tone of Wordsworth’s visionary acceptance: ‘We will grieve not, rather find/Strength in what remains behind.’ (Wordsworth 1928, p.136). The styles are obviously connected, but the trouble with defining the era usi ng literary terminology is that it is clearly neither a quirky extension of the Romantic’s vision, nor is it a straightforward path to the modernists. The 1870’s saw the maturation of authors such as Anthony Trollope who brought out his later novels, yet only twenty years later in 1896 these publications are sitting beside the considerably different form and subject matter of work such as H.G. Wellls’ The Time Machine and The Island of Dr. Moreau, with literary experiments with the modern such as Richard Jefferies’ The Story of My Heart occurring between in 1883. A growing concern in nineteenth century life was the potential loss of the Romantic link between human nature and the natural world, and the gap which sudden industrial progress highlighted between nature and mechanisation. As technology developed so did the notion of artificiality. It is worth noting J.S.Mill’s essay on Nature (Mill 1874, p.65) where he says that it is man’s nature to be artificial, to remedy nature by artificial pruning and intervention. Further to this, a contemporary of Mill’s Richard Jennings also drew a line between the ‘province of human nature’ and the ‘external world.’ (Lightman 1997, p.80). In the countryside more efficient methods of farming were employed (see the contrast between Henchard’s methods and Farfrae’s ‘ciphering and mensuration’ in Hardy’s Mayor of Casterbridge, (Hardy 1886, p.122)), and new machines introduced which no longer required the labour force to run the m, encouraging people to migrate to towns and cities. The urban reality was harsh in 1851 roughly four million people were employed in trade and manufacture and mining, leaving only one and a half million in agriculture. (Davis 2002, p.13). City life, as portrayed by Dickens, was a cruel, unhealthy and unwholesome existence for many (see Bleak House and Little Dorritt). However, much of his work was set in the period of his youth and childhood which was pre-Victorian. (Lawton (ed) 1995, p.xvi). Working conditions in cities were often cramped, unhygienic and poorly ventilated, and living conditions could be even worse. Mrs. Gaskell, living in Manchester, witnessed the appalling pressures that these conditions forced upon family life, and in North and South depicts the difficulties of urban living, offering that salvation for the working classes lay with themselves and their employers, working together. (see North and South 1855) However, city life was not all desolate based in cit ies, the development of the detective novel brought the city back to human scale (Lehan, p.84). Detectives pieced together and reconstructed past events through clues for example, the murder of Bartholomew Sholto in The Sign of Four by Conan Doyle: As far as we can learn, no actual traces of violence were found upon Mr Sholto’s person, but a valuable collection of Indian gems which the deceased gentleman had inherited from his father had been carried off. The discovery was first made by Mr Sherlock Holmes and Dr Watson () Mr. Athelney Jones, the well-known member of the detective police force, happened to be at the Norwood police station () Mr Jones’ well-known technical knowledge and his powers of minute observation have enabled him to prove conclusively that the miscreants could not have entered by the door or by the window but must have made their way across the roof of the building, and so through a trapdoor into a room which communicated with that in which the body was found. (p.66) The city provided an exciting backdrop to crime scenes its labyrinthine streets similar to the mapping of the pathways of the human mind so that the two became inextricably linked. As Joseph McLaughlin says in Writing the Urban Jungle, ‘the urban jungle is a space that calls forth a pleasurable acquiescence to something greater, more powerful, and, indeed, sublime () also an imaginative domain that calls forth heroic action: exploring, conquering, enlightening, purifying, taming, besting.’ (McLaughlin 2000, p.3). Further to what McLaughlin suggests, the Victorians’ perception of time and space in the city and the countryside was changing radically from the medieval perceptions that still existed in the Romantic period. People saw the finished products in both manufacturing and farming no longer involving the long, drawn-out means to an end, instead the end result was being achieved faster and with more control. Here developed the root of modern industry which continues today in intensive farming and factory lines. Yet here too the beginnings of waste and excess. Richard Jefferies, a nineteenth century naturalist and mystic, known for his essays on nature, remarks on the abundance of food in the natural world in his essay Meadow Thoughts: The surface of the earth offers to us far more than we can consume the grains, the seeds, the fruits, the animals, the abounding products are beyond the power of all the human race to devour. They can, too, be multiplied a thousandfold. There is no natural lack. Whenever there is lack among us it is from artificial causes, which intelligence should remove. (Jefferies, 1994, p.26). Unfortunately there was plenty for those who could afford it but not enough to spare for the poorer lower classes. (Ritvo 1997, p.194). Trends of over production and wastage which became a worry in Victorian times are reflected in the literary concerns of Jefferies’ spiritual autobiography The Story of My Heart, and in his children’s story, Bevis, where words, despite their abundance, are in danger of becoming an insufficient medium of expression and not filling the metaphysical space on the page. In describing a sunrise and the thoughts and feelings associated with watching it, Jefferies struggles to articulate the beauty before him: The sun had not yet stood out from the orient, but his precedent light shone through the translucent blue. Yet it was not blue, nor is there any word, nor is a word possible to convey the feeling. (Jefferies 1881, p.391) We see too in James Thomson’s City of Dreadful Night (Thomson 1892, p.2) the desperateness of trying to articulate thoughts and feelings: Because a cold rage seizes one at whiles To show the bitter old and wrinkled truth Stripped naked of all vesture that beguiles, False dreams, false hopes, false masks and modes of youth; Because it gives some sense of power and passion In helpless impotence to try to fashion Our woe in living words howeer uncouth. In both passages there is a sense of trying to convey so much more than the words will allow. And that is the essence of the problem of defining the era with a word which the era itself selected ‘Victorian’ like the authors of its time struggles to convey the enormity and the condensed nature of its changing environment. Victorian literature is thus perhaps best studied between the lines of its texts rather than for what it offers at face value. Thomson’s words ‘to try to fashion our woe in living words’ although appearing dismal could actually withhold a more positive message: it deals with the notion of perseverance that by creating words, however difficult, the author is refusing to give in to despair by trying to transform it into creative energy. There is a sense of crisis in the work of Thomson, just as there is to be found in Jefferies futuristic After London where the lone explorer Felix discovers the land after humanity has overreached itself to sociological disaster and has lost the harmonious relationship between mankind and nature. London becomes no more than a crystallised ruin in a ground oozing with poison ‘unctuous and slimy, like a thick oil.’ (Jefferies 1885, p.205). Through work like this we see that ‘Victorian’ was an era of possibility where visions of the future suddenly became tangible concerns and possible realities, and where contemporary conceptions of language and life might no longer hold up to the pressures of the time. In H.G. Wells’ the Time Machine, the time traveller discovers a land in the year 802,701: The air was free from gnats, the earth from weeds or fungi; everywhere were fruits and sweet and delightful flowers; brilliant butterflies flew hither and thither. The ideal of preventative medicine was attained. diseases had been stamped out. I saw no evidence of any contagious diseases during all my stay. And i shall have to tell you later that even the processes of putrefaction and decay had been profoundly affected by these changes. (Wells 1995, p.28) In this description of a futuristic age the Victorian imagination still retains the idea of a paradise a place full of ‘butterflies’ and ‘flowers.‘ This Christian concept is a literary hangover from Milton’s Paradise Lost, and remains an important theme for the moderns such as D.H. Lawrence. (see The Rainbow). The Victorian age suffered from a dualistic split between a bright future on the one hand promised by leaps in technology, education and economical success and an increasingly alienated, confused society on the other. There were those writers like Huxley who believed that by human intervention within a political and economic framework humans could evolve out of their condition seeing ‘no limit to the extent to which intelligence and will () may modify the conditions of existence’ (Huxley 1911, p.149), and there were those like Hardy whose characters were destined to fail because they were not emotionally fitted into the cosmos ou t of which they evolved. (see Tess and Jude the Obscure). It was the nineteenth century spiritual crisis which precipitated the literary shift into the new genre of the realist novel. By the mid-nineteenth century, society had begun to grow away from the idea of atonement for sin within an omnipotent religion, where judgement would come solely in heaven, and towards the more humanistic idea of God as in-dwelling, so that salvation could be achieved on earth: We have now come to regard the world not as a machine, but as an organism, a system in which, while the parts contribute to the growth of the whole, the whole also reacts upon the development of the parts; and whose primary purpose is its own perfection, something that is contained within and not outside itself, an internal end: while in their turn the myriad parts of this universal organism are also lesser organisms, ends in and for themselves, pursuing each its lonely ideal of individual completeness. (Gore (ed) 1890, p.211) A spiritual lack created a need to define, order and categorise a world that suddenly appeared chaotic. When Darwin published On the Origin of Species in 1859 he raised issues of public concern as to the truth of the bible and the essence of Christianity. However, it’s content and its methodology were seriously criticised (Appleman 2001, p.200). It was a difficult work to accept as it caused the public to rethink and redefine their history that they were a product of evolution and not a tailor made being came as a shock. The future of thought and literature was suddenly changed as people tried to sew together the threads of the past. Natural Science became a national obsession exotic flora and fauna from across the world were brought into London daily, to be displayed in the British Museum or Kew Gardens (Lightman, 1997 p.1). In literature, we see the author begin to play the part of evolutionist: Eliot’s Middlemarch although concerned with the evolving character of D orothea Brooke follows the threads of sub-plots and the successes and failures of other characters which form a pattern of development. As Gillian Beer says: There is not one primitive tissue, just as there is not one key to all mythologies () emphasis upon plurality, rather than upon singleness, is crucial to the developing argument of Middlemarch. (Beer 2000, p.143). Gone is the tradition of the valiant hero or heroine singularly conquering their environment (a trend set by classics such as Homer’s The Odyssey) and in its place a landscape upon which the author grafts and nurtures developing shoots of life. It is this sort of growth that is in danger of remaining unseen to the contemporary historian or critic as it can become shrouded by generalising concepts which are so often prescribed to the term ‘Victorian’ concepts such as ‘repression‘, ‘old-fashioned’ and ‘prudish.’ (http://www.victorianweb.org/vn/victor4.html). These sort of terms restrict the individual’s perception of the era when it was a time when growth was encouraged rather than restricted. Authors used the metaphor of pruning and nurturing plant life to symbolise the development of the self for example in North and South Gaskell discusses the problem of the working individual who struggles to reach his or her potent ial when the manufacturers are ‘unsparingly cutting away all off-shoots in the direction of literature or high mental cultivation, in hopes of throwing the whole strength and vigour of the plant into commerce.’ (Gaskell 1865, p.69). For Gaskell, it is through the everyday interaction between people that such difficulties are given the chance to be overcome. And this was the essence of the realist novel set amidst a world which had witnessed such alteration to transform the lostness felt by society into a seeing of the smaller things in life which could withhold qualities of greater spiritual value. As Philip Davis says, ‘the realist novel was the holding ground, the meeting point, for the overlapping of common life.’ (Davis 2002, p.144). And it was within this common life that a more calm acceptance of the new state could be achieved. Gillian Beer suggests that through her novel’s organisation Eliot creates order and understanding of the evolving process of novel-writing. In Middlemarch, the naming of Casaubon‘s books ‘Waiting for Death, Two Temptations, Three Love Problems draws attention to the book’s organisation by emphasising categorisation: But the process of reading leads into divergence and variability. Even while we are observing how closely human beings conform in the taxonomy of events we learn how differently they feel and think. For Dorothea and Casaubon waiting for death means something very different from what it means for Mary Garth and Featherstone. The relations are different. The distances between people are different. Lydgate, here at one with the project of the book, longed to demonstrate the more intimate relations of living structure (1:15:225). In this double emphasis on conformity and variability George Eliot intensifies older literary organisations by means of recent scientific theory. In Darwinian theory, variability is the creative principle, but the type makes it possible for us to track common ancestry and common kinship. (Beer 2000, pp.143-4) Writing itself was becoming an almost divine representation, an inner order of a chaotic external world. The idea that humans had evolved from primates meant that the boundaries between what was one thing and what was another were no longer so clearly defined. There developed a fear of the animate and a fear of the inanimate, and efforts were sought to understand them. As Harriet Ritvo says in The Platypus and the Mermaid: Depending on the beholder, an anomaly might be viewed as embodying a challenge to the established order, whether social, natural, or divine; the containment of that challenge; the incomprehensibility of the creation by human intelligence; or simply the endless and diverting variety of the world. And beholders who agreed on the content of the representation could still disagree strongly about its moral valence whether it was good or bad, entrancing or disgusting. (Ritvo 1997, p.148). In a world where categorisation was important but not so easily achievable, the novel too became neither one thing nor another; realism became a melting pot for ideas, a sort of hybrid of styles. In Eliot’s The Lifted Veil realism is used as a vehicle for the exploration of her ideas into psychology and psychic phenomena. Latimers clairvoyance forces him to endure a painful insight into the minds of the people around him: I began to be aware of a phase in my abnormal sensibility, to which, from the languid and slight nature of my intercourse with others since my illness, I had not been alive before. This was the obtrusion on my mind of the mental process going forward in first one person, and then another, with whom I happened to be in contact: the vagrant, frivolous ideas and emotions of some uninteresting acquaintanceMrs Filmore, for examplewould force themselves on my consciousness like an importunate, ill-played musical instrument, or the loud activity of an imprisoned insect. But this superadded consciousness, wearying and annoying enough when it urged on me the trivial experience of indifferent people, became an intense pain and grief when it seemed to be opening to me the souls of those who were in a close relation to me when the rational talk, the graceful attentions, the wittily-turned phrases, and the kindly deeds, which used to make the web of their characters, were seen as if thrust asund er by a microscopic vision, that showed all the intermediate frivolities, all the suppressed egoism, all the struggling chaos of puerilities, meanness, vague capricious memories, and indolent make-shift thoughts, from which human words and deeds emerge like leaflets covering a fermenting heap.(Eliot 1859, pp.13-14) Latimer is no longer caught up in the ‘web’ of people’s characters. Eliot plays with the idea that his consciousness has the ability to transcend the mundane the ’rational talk’, the ’kindly deeds’ in order to gain insight into an alternative and not so rosy vision of the mechanics of the human mind where thoughts are ‘make-shift’ and ‘chaotic.’ The nineteenth century saw the acceptance of the concept of otherworldly phenomena into the working classes. Robert Owen, a social reformer, who influenced the British Labor movement (Oppenheim 1985, p.40) encouraged many working class ‘Owenites’ to ‘follow him into the spiritualist fold, where they enthusiastically continued their ongoing search for the ‘new moral world.’’ Interests such as spiritualism and psychology which had previously been more underground pursuits, were brought out into the open. The concept of telepathy, a term coined by Frederic Myers in 1882 (Luckhurst 2002, p.1) even helped to ‘theorize the uneasy cross-cultural encounters at the colonial frontier.’ (Luckhurst 2002, p.3) These developments suggest that the Victorians felt imbued with the power of their age they felt confident of their ability to communicate on different planes of consciousness. So it could be argued that ‘Victorian’ was not simply a time devoted to the discovery of the self and the workings of the inner mind, but a time that also focused on the projection of ideas and thoughts outside of the self; ideas which themselves stand outside of the category ‘Victorian.’ In 1869 the Spiritualist Newspaper began selling first as a fortnightly, then as a weekly publication. (Oppenheim 1985, p.45). This draws the discussion to the point of representation the social nature of Victorians seems to suggest that they enjoyed the focus being on themselves. Self-obsession is an aspect of the time which the term ‘Victorian’ usefully represents: by specifically referring to the rule of the Queen the term draws attention to the importance of the individual. The era saw the development of many different styles of fashion and the use of photography. As part of the Freudian influence great importance was placed on childhood and it was during the nineteenth century that the first laws concerning child welfare were passed. (Mavor quoted from Brown (ed) 2001, p.i) The focus on the central, the ego, was paramount. As Mavor says, it was as if the camera had to be invented in order to document what would soon be lost, childhood itself; and childhood had to be invented in order for the camera to document childhood (a fantasy of innocence) as real. (Brown (ed) 2001, p.27). Perhaps because of society’s awareness of change there seems to have been a necessity to record and keep track of the world around. Discovery took place on a much grander scale in the exploration of the world. The British Empire was global, yet as Patrick Brantlinger suggests in Rule of Darkness, (Brantlinger 1988, p.4) imperialism was not generally reflected in the literature of the time. What we do see evidence of however is the mapping of new worlds and territories (see Robert Louis Stevenson‘s Treasure Island and Richard Jefferies’ Bevis). The development of the adventure story suggests that Victorians desired to explore what lay outside of what they knew and in this respect the term ‘Victorian’ which people can think of as representing a society closed within in itself is misleading. The rise of imperialism began to shape the ideological dimensions of subjects studied in school (Bristow 1991, p.20) and so through literature the Victorian child was offered an exciting world of sophisticated representation and ideas with the knowledge that the world was theirs to explore. Does the term then encourage us to think of the society as a class of people set apart from the rest of the world? In The Island of Dr. Moreau it is not just the future of science that is explored but the concept of a new territory and its effects on the mind. For example, when the protagonist first sees the beast-servant on board the ship he is immediately frightened: I did not know then that a reddish luminosity, at least, is not uncommon in human eyes. The figure, with its eyes of fire, struck down through all my adult thoughts and feelings, and for a moment the forgotten horrors of childhood came back to my mind. Then the effect passed as it had come. An uncouth black figure of a man, a figure of no particular import, hung over the taffrail, against the starlight. (Wells 1997, p.31). The circumstances of being at sea is disorientating and causes the imagination to play tricks so that the man is first one thing ‘a figure with its eyes of fire’ and then suddenly becomes an ‘uncouth black figure of a man.’ The effect is that the protagonist suddenly regresses to the ’forgotten horrors of childhood.’ This sudden fluctuation is important as it represents the fluidity of the era and how change and discovery on a global scale, although empowering, also caused instability within the individual. Therefore, when considering the age in the context of its name we can understand that the term was perhaps created out of both the desire to represent achievement but also out of a need to belong. This desire to belong which manifested itself during an age ruled by one woman placed great importance on the role of the female in society. It was a time when women began to travel and write without the necessity of using a pseudonym (see Cheryl McEwan on Kingsley in West Africa, (2000, p.73)). In books such as Hardy’s Tess the idea of the fallen woman is tested: Tess attempts to rediscover paradise at Talbothays but ultimately a life with her lover is denied. The nineteenth century began to be more explicit concerning issues of gender: for example, the relationship between Arthur Munby and Hannah Cullwick (see McClintock 1995, pp.132-138) where Cullwick is photographed cross-dressed as a farm worker. A ‘Victorian’ man however appears to have had more stigma attached to him and in this context the term is commonly associated with heroism and English valour (Ridley/Dawson 1994, p.110). There is less flexibility surrounding the notion of Victorian men -as if the te rm somehow threatened their masculinity. However, this did not seem to affect the male authors of the time. Lewis Carroll captured the public imagination through Alice’s Adventures in Wonderland, which although following the story of a little girl, depicts many male characters. (see Carroll 2000). In conclusion, the term ‘Victorian’ although useful to refer to a specific time period in history, does however encourage us to make sweeping generalisations without investigating how diverse the era was. In terms of the subject matter of Victorian Literature there is no clear cut distinction between early, middle and late Victorian for example, Bulwer-Lytton (see Zanoni and A Strange Story) attempts at the beginning of the century what Richard Jefferies does at the end the difference is in style and form. Within that time frame there was condensed an incredible diversity of styles, tastes and attitudes, yet the term suffers from being associated with prejudices and assumptions about Victorians. However, it is worth bearing in mind that prejudices were indeed a part of Victorian society. When the Victorians explored the rest of the world they made generalisations and assumptions based on what they found (eg: The Island of Dr. Moreau) where experience and the nature of what is discovered defines behaviour. As a critic in 1858 wrote ‘we are living in an age of transition’ (quoted from Houghton 1957, p.1); therefore when considering the Victorian age we should remember that values and trends were evolving it was not a static time governed by repression or old fashioned values. From the research carried out for this essay it appears that through the gaining of knowledge, Victorians also realised how little they knew and how much more there was to discover. As Arnold says in The Buried Life: How fair a lot to fill / Is left to each man still. (Arnold 1913, p.168). In this context the term ‘Victorian’ can be dualistically representative: discoveries of the time, although revolutionary, were often rudimentary in nature, and it was humbling for the individual to consider how much further knowledge and discovery had yet to go. On the other hand, the term suffers too from being inadequate: a single word is too smaller term for the vast wealth and diversity of discovery, and it could be argued that the era is better realised if seen as a second revolution. Like the Victorian authors themselves we are left with no suitable words to convey the entirety of an era as John Lawton says in his introduction to The Time Machine (1995, p.xxvi) ‘the term ‘Victorian’ is used too loosely to encompass a sequence of eras, the diverse reign of a woman who lent her name to objects as diverse as a railway terminus and a plum.’ When studying Victorian Literature it is worth bearing in mind the fluidity of the time and the changeability which arose out of living on the cusp between the passing away of old values and the unknown territory of the new. Realism recognised the gaps which were forming in society such as the distancing of the self from religion and offered to paper the cracks through its vision of bringing people together on a mundane level. It’s territory stretched to include the darkest recesses of the mind to the smallest of everyday events, celebrating the grey area between extremes as we now know as ‘Victorian.’ Bibliography Arnold, M., 1913, The Poems of Matthew Arnold, 1840-1867. Oxford: Oxford University Press Appleman, P, 2001, Darwin. London: Norton Beer, G., 2000, Darwin’s Plots. Cambridge: Cambridge University Pr

Pseudomonas Aeruginosa Gene Expression

Pseudomonas Aeruginosa Gene Expression Pseudomonas aeruginosa is the gram negative rod shaped bacteria is found in all natural and man made environments. the organism is so stubburn so that it can grow in any environment and also at high temperatures such as 420C. The unique ability of the organism is forming biofilm. The biofilms are formed by the organism with the association with a surface. The surface may be anything such as cloth, paper, glass etc and any artificial surface. It is the opportunistic pathogen which behaves neutral to the normal tissues but it infects once the defences of them are compromised. The development of the biofilm starts with the attachment of the freely moving planktonic cell to a surface. Generally the twitching motility of the P.aeruginosa helps to use its unipolar flagellum to adhere a surface. The bacterium develops colonies known as microcolonies and adhere to the surface strongly which are known as pili structures (Baron S et al.,1996). During the biofilm development several factors pla ys a key role mainly the exopolymeric matrix. The EPS matrix is not visualised directly, but it can be done by using some fluorescent dyes. The functioning of the EPS matrix in holding cells of Pseudomonas aeruginosa together and forming the mature biofilms. The biofilm develops with cell division and recruitment. The cells interacts via quorum sensing and these interactions are studied under scanning electron microscopy. The biofilm formation takes place in stages and many changes are observed in its size and shape during the development. The EPS matrix is made up of polysaccharides, proteins and nucleic acids. The exopolysaccharide matrix is the important component of the biofilm. The exopolysaccharide genes such as pilB, pslA, lecA, ureB and alginate genes are responsible for the formation of pseudomonas biofilms. The locus of the psl polysacchide genes encode the cells to adhere to a surface and thus maintaining the biofilm structure. In this research project we try to quantify the gene expression of the above genes pilB, pslA, lecA, ureB using rpoD as the house keeping gene. The biofilms are grow invitro on microcarriers. The microcarriers made up of dextrons in the range of 60-90 µm in size are used as a surface for Pseudomonas aeruginosa to form biofilms. The microcarriers are spherical in size where the bacteria can easily adhere on surface. In this present work we try to grow planktonic cultures and biofilms cultures on the microcarriers in a fermentor separately. The genes responsible for the biofilm formation are quantified by comparing the planktonic and biofilm samples. The planktonic cells grow very rapidly whereas the bacterial adhering takes some time to form biofilm in other case. The samples are collected at regular intervals and mRNA is extracted. The gene expression is quantified using real time PCR. The relative quantification method of qPCR or real time PCR method does not require any standard curve for the analysis where the samples are normalised by using an internal control. The threshold CT values obtained from the curve are used to calculate relative quantification by comparative à ¢Ã‹â€ Ã¢â‚¬  Ãƒ ¢Ã‹â€ Ã¢â‚¬  CT method. The results from the planktonic and biofilm gene expression results are compared. On the other hand we also assay the nutrients such as glucose, ammonia, phosphorus and protease during the fermentation. The concentration levels of the nutrients are also compared for both biofilm and planktonic fermentation. Review of literature: Pseudomonas aeruginosa is a member of gamma proteobacteria class of bacteria. The bacteria is gram negative, rod shaped and aerobic belongs to the bacterial family of pseudomonadanceae. The recent developments in the taxonomy based on the 16s ribosomal RNA or conserved macromolecules, the family members are divided into eight groups. Pseudomonas aeruginosa is a species present in this group sharing the with other twelve members. Pseudomonas aeruginosa is a free-living bacterium, commonly found in soil, water, skin flora and in all man made environments. The organism regularly occurs on the surfaces of the plants and animals. Pseudomonas aeruginosa is an occasional pathogen to plants. It is also an increasingly emerging pathogen of clinical relevance. The organism not thrives in the normal atmosphere but also in the places with little oxygen and thus forms the colonies in natural and artificial environments. Pseudomonas aeruginosa is an opportunistic pathogen for humans, which means that it exploits some break in the host defences to initiate an infection. The bacteria never infects the uncompromised tissues, but it can infect any if the tissue defences are comprised. It can cause infections such as urinary tract, respiratory system infection, soft tissue infection dermatitis, gastro intestinal infections. It causes a variety of systemic infections in patients particularly with severe burns and immune suppressed in cases of cancer, AIDS, cystic fibrosis in lungs. Characteristics Pseudomonas aeruginosa is rod shaped gram negative bacteria. The size of it ranges between 0.5 to 0.8  µm by 1.5 to 3.0  µm. almost all the strains of Pseudomonas are motile with a unipolar flagellum. It is ubiquitous in soil ,water and on the surfaces in contact with water or soil. It has respiratory metabolism and can grow in the absence of oxygen if NO3 is available as respiratory acceptor for electron. Generally the bacterium is found in nature in the form of biofilm attached to a surface or substrate, or in the planktonic form as a unicellular organism swimming actively with its unipolar flagellum. It is one of the most vigorous and fast swimming organism found in pond waters. It can grow at temperatures at 42OC or high, but the optimum temperature is 30 to 37OC. it is resistible to a wide variety of physical conditions such as temperature and can even resist the salts, dyes, antiseptics and most commonly used antibiotics. Pseudomonas aeruginosa produces different types of colonies from the source they have isolated. They appear like small and rough colonies. Moreover the samples collected from the clinical sources look like a fried egg like structure which is smooth, large with elevated appearance. The other type of colonies obtained from the respiratory and urinary tract secretions looks like mucoid.this mucoid appearance is attributes for the production of alginate slime. These smooth, mucous colonies presumably plays an important part in the colonization and virulence. Pseudomonas aeruginosa produces two types of soluble pigments. Pyocyanin which is a blue-green and a yellow-green and fluorescent pigment called as pyoverdin. Pyocyanin is produced abundantly in the media with low levels of iron and also functions for the iron metabolism of the organism. Many strains of Pseudomonas aeruginosa produces pyocyanin but not all the strains does it. Although the organism is classified as the aerobic, sometimes the organism is considered as facultative anaerobe where it can adapt and proliferate the conditions with less or total O2 depletion. The organism can grow in the absence of oxygen if NO3 is available as respiratory acceptor for electron. It can ferment arginine by substrate level phosphorylation where the layers of alginate present surrounding the mucoid cells of the bacteria controls the diffusion of oxygen(Collins FM, 1955), this is witnessed in the cystic fibrosis of the lung infection. Pseudomonas aeruginosa which is found in the biofilm form is responsible for a variety of activities involved which are natural and artificial. Resistance to antibiotics Pseudomonas aeruginosa is opportunistic pathogen, has a notorious resistance to antibiotics. The organism is very resistance towards many antibiotics due to the potential permeability barrier provided by the gram negative membrane. Pseudomonas aeruginosa in the biofilm form makes the cells impervious to the concentrations of the therapeutic antibiotic. The organism gets associated with others like actinomycytes, moulds and bacillus because of the natural environment they live. Moreover the organism have antibiotic resistance plasmids R factors and RTFs, the genes are able transferred by conjugation and transduction following the mechanism horizontal gene transfer(HGT) (Todar.K,2008). Biofilms Biofilms are defined as a matrix enclosed bacterial populations which are adherent to each surfaces or interfaces. Microbial aggregates and floccules and also adherent populations within the pore spaces of the media are spoken in the definition (Costerton J.W, et al 1995).bacterial biofilms came into existence and became significant from their first recognition in their ubiquity. The past researches show that profoundly the growth phase of the biofilms is distinct from the planktonic. In response to the surfaces proximity, bacterial cells alter their phenotypes (Costerton J.W, et al, 1995). The early stages of biofilm formation the bacteria is in stable juxta position with those of same and other species. The micro environmental conditions, cellular juxtapositions and the extra cellular polysaccharide matrix constitute to the development of the bacterial biofilm. Different bacteria responds to the specific environmental conditions with certain growth patterns, and a structurally comp lex and mature biofilm develops(Hamilton W.A et al, 1987). Physiological cooperation is the major criteria in shaping structure and juxta positions establishment to make biofilms adherent for the surfaces. It has been proposed that large suites of genes are expressed differentially regulated. Formation The formation of the biofilm initially starts with the attachment of the freely floating bacterial cells to a surface. The bacterial colonies adhere to the surface with weak vander wall forces. These colonies can lead themselves to more adhesion called as pili structures (Baron S et al.,1996). The organism adheres more to the available surface by binary fission, exopolymeric matrix is produced to develop mature biofilms. The matrix produced holds the biofilm together. Biofilm develops with cell division and recruitment, the cell in the biofilm communicate via quorum sensing. We can observe changes in shape and size in the final stages of biofilm formation. Distribution and ubiquity of biofilms The general phenotypic strategy of organism is to change in the mode of growth that to adopt to the altering growth conditions. The cells of Pseudomonas aeruginosa which is a ubiquitous bacterial species respond to all favourable nutrient conditions. The organism adheres to the available surface by binary fission, exopolymeric matrix is produced to develop mature biofilms. The rod shaped bacteria grows in matrix which is enclosed in a sessile growth mode. The cells are protected from the biological, antibacterial, chemical agents and adverse environmental conditions. The periodically shed planktonic cells from the biofilm provide and sacrifice the protection to the new fresh habitats can colonize with the biofilm. The adherent biofilm populations has a significant metabolic activities( Fletcher.M, 1986). The extent of biofilm formation is predicted in the particular system is based on the following principles: Metabolically active bacteria show tendency towards the surfaces for adhesion in all natural environments. In all environmental conditions the extent of biofilm formation is controlled by the available nutrient for cell respiration and exopolysaccharide production. The organic nutrients sometimes tend to associate with the surfaces, in such cases the local biofilm is difficult to trigger. This is more in extremely oligotrophical environments, but bacteria normally does not adhere to surfaces in nutrient less environments. Pseudomonas aeruginosa Biofilms The communities of microbial bacteria(Pseudomonas aeruginosa) forms a definite structures with the association with surfaces are abundantly available environment. These structures are often called as biofilms which are less susceptible to antimicrobial agents. These biofilms causes chronic infections and very difficult to remove. The extracellular polymeric matrix(EPS) are known to holds the biofilm structure and it performs as a matrix. The EPS matrix holds the biofilm cells together and protects them from shear forces in fludic environments. The extra cellular matrix for Pseudomonas aeruginosa forms the biofilm development in different stages. The matrix holds the cells together on the surface and forms a critical structural design resists to many environmental changes. Pseudomonas aeruginosa biofilm develops in a five stage multicellular cycles, the process is initiated by the attachment of planktonic (free) cells to a surface. This is followed by the formation of microcolonies. T he microcolonies are seeded and gets dispersed where the cells form the microcolonies comes out to occupy a new surface( Ma. L et al, 2009;Stoodley.P et al,2002). The EPS of Pseudomonas aeruginosa composed of polysaccharides, proteins and nucleic acids. It is the important component of the biofilms. The exo polysaccharide genes lec A , Pil B, Psl A, alginate are responsible for the formation of Pseudomonas biofilms. The locus of the Psl exopolysaccharide encodes bacterial cells to surface and thus maintaining the biofilm structure. The matrix which is anchored by the Psl is not visualised directly during the developmental stages of Pseudomonas aeruginosa biofilm. In addition to the present scenario of exopolysaccharides, extracellular DNA also plays an important part in . P.aeruginosa biofilm matrix (Allensen-Holm M et al, 2006). The extracellular DNA is derived from the chromosomes and it functions for signalling between the cells interconnecting the biofilm matrix. In the biofilm matrix, extracellular cellular DNA promotes cation gradients, release of genomic DNA and induces antibiotic resistance ( Mulcahy H, et al. 2008). Real time PCR Polmerase chain reaction or simply PCR is introduced by Kary mulis in 1980, which really a revolution in the molecular biology. PCR is the standard procedure to amplify small quantities of DNA molecule into large quantities. The method depends on thermal cycling which involves the continuous heating and cooling of reaction for DNA melting and enzymatic replication of DNA. PCR applications employ a general heat stable DNA polymerase such as Taq polymerase enzyme isolated from the bacterium Thermus aquaticus. PCR method is involves three major steps.1) Denaturation 2) Annealing 3) Elongation. In the first step DNA template is denaturated to single strand at high temperature of 940C. In the second step, single stranded DNA template annealed with primers, the temperature is 650C. At 720 C the DNA starts elongation which is third step. DNA polymerase synthesizes new complementary strand to the template DNA strand. Real time polymerase chain reaction or qualitative polymerase chain reaction is a laboratory technique which not only amplifies but also quantifies the target DNA molecule. it helps the user for detection and quantification(absolute and relative) of many specific genes in a present in a given DNA sample. The general principle of polymerase chain reaction (PCR) is also used in the RT PCR. Unlike PCR, RT PCR is a new technology product is detected at the end of the reaction. There are two common methods for the product detection in RT PCR 1) Non -specific fluorescent dyes which interacts with the double stranded DNA molecule 2) Sequence specific DNA probes consists of oligonucleotides that are fluorescently labelled with reporter. The product is detected after the hybridization of the probe and complementary strand. The RT PCR is used to amplify the reversly transcribed cDNA products from the mRNA, which is becoming the effective and powerful tool in the field of molecular biology to quantify and study the gene expression. The RT PCR method is very easy to apply provides the reliable, accuracy and rapid gene expression quantification. The accurate nucleic acid quantification requires mathematical application for data analysis(Michael W.P., 2001). The real time PCR provides the very accurate and reproducible quantification of gene copies (Heid C.A., 1996). Unlike other PCR methods, RT PCR does not require post process sample handling, prevents product contamination, provides faster and accurate assay results. Real time PCR is very accurate and less labour usage than the present gene quantification methods. Nucleic acid sequence quantification has an important role in the biological research. Measuring the gene expression have been extensively used in the biological responses to various external s timuli( Tan et al, 1994; Huang et al,1995). Gene and genome quatification also used for HIV demonstrates the change in the expression level of virus in the disease period( Conner et al. 1993); (piatak et al, 1993). There are two types of quantification methods used in real time PCR: Absolute quantification Relative quantification Absolute quantification determines the input copy number generally by relating the PCR signal to a standard curve. In this method equal input amounts of sample, each sample under analysis amplifies with identical efficiency up to the point of qualitative analysis( Heid et al,1996). We quantitate the unknowns based on a known standard quantity. The unknown samples are compared to the known by creating a standard curve. The input copy number of the PCR signal is related to standard curve in absolute quantification method. Sometimes it is not necessary to know the copy number but it is important to calculate the relative change gene expression (Livak.K.J et al, 2001). The second method is relative quantification, we analyse the changes in gene expression in a given sample relative to another reference sample(untreated control sample). Relative standard again consists of two types of methods they are relative standard curve and comparative Ct method. In the relative quantification the PCR signal is related to the targeted transcript of the samples in the group. The new applications of comparative Ct method is the 2-à ¢Ã‹â€ Ã¢â‚¬  Ãƒ ¢Ã‹â€ Ã¢â‚¬  Ct method which is efficiently useful for the analysis of qPCR data. The 2-à ¢Ã‹â€ Ã¢â‚¬  Ãƒ ¢Ã‹â€ Ã¢â‚¬  Ct method(Livak.K.J et al , 2001): Derivation of 2-à ¢Ã‹â€ Ã¢â‚¬  Ãƒ ¢Ã‹â€ Ã¢â‚¬  Ct method: The equation for the amplification of PCR can be written as: Xn=X0 (1+Ex)n(1) Where Xn is the number of target molecules at cycle n of the reaction. X0 is the initial number opf target molecules. Ex is the efficiency of the target amplicon. n is the number of cycles CT is the threshold cycle, indicates the fractional cycle at which the amount of amplified target reaches a fixed threshold. XT =X0 (1+Ex)CT.X=Kx(2) Where XT is the threshold number of target molecules. CTX is the threshold cycle for target molecule and Kx is constant. A similar reaction for the internal control gene is RT =R0 (1+ER)CTR=KR-(3) Where RT is the threshold number of reference molecules R0 is the initial number of reference molecules ER is the efficiency of the reference molecule. CT.R is the threshold cycle for reference amplicon. KR is a constant. Dividing XT and RT, we get , (XT/ RT )= (X0 (1+Ex)CT.X) / (R0 (1+ER)CT.R )= (Kx / KR ) = K-(4) For real time amplification using Taqman probes the exact values of the XT and RT depend upon the number of factors including the reporter dye and fluorescence properties of the probes, efficiency and purity of the probe, florescence threshold setting. Thus k cannot be equal to one. Assume the efficiencies of target and the reference as equal. ER = Ex=E (X0 / R0) (1+ E) CT.X- CT.R =K(5) XN (1+E)-à ¢Ã‹â€ Ã¢â‚¬  CT =K-(6) Where XN is equal to the normalised amount of target gene (X0 / R0) and à ¢Ã‹â€ Ã¢â‚¬  CT is the difference of the threshold cycles of target and reference (CT.X- CT.R ). The expression can be written as XN=K(1+E) -à ¢Ã‹â€ Ã¢â‚¬  CT -(7) If we divide the whole equation by XN for any sample q by XN as the calibrator(cb) XN.q /XN.cb ={[ K(1+E) -à ¢Ã‹â€ Ã¢â‚¬  CTq ]/[ K(1+E) -à ¢Ã‹â€ Ã¢â‚¬  CTcb ]} =(1+E) -à ¢Ã‹â€ Ã¢â‚¬  Ãƒ ¢Ã‹â€ Ã¢â‚¬  CT Here -à ¢Ã‹â€ Ã¢â‚¬  Ãƒ ¢Ã‹â€ Ã¢â‚¬  CT = -(à ¢Ã‹â€ Ã¢â‚¬  CTq -à ¢Ã‹â€ Ã¢â‚¬  CT.cb) For amplicons which are designed less than 150 bp, primer and magnesium concentration are optimised, the efficiency is nearly equal to 1. Therefore the amount of target gene, normalized to an endogenous reference which is relative to a calibrator, taken as Amount of target =2-à ¢Ã‹â€ Ã¢â‚¬  Ãƒ ¢Ã‹â€ Ã¢â‚¬  CT . When the threshold is set for n standard deviations in the plot , above the mean of the base line emission from cycles 0 to n. The standard deviations is calculated from the data attained from base line of the amplification plot. When the fluorescence exceeds the threshold limit, CT values are determined at that point. The CT values are ten times more than the standard deviation of base line. The point at which amplification plot crosses the threshold can be defined as CT (Heid et al, 1996). Materials and methods Identifying pseudomonas aeruginosa Gram straining: Prepare a small amount of smear on a clear glass slide. Heat fix the smear to slide. Add a drop of crystal violet on to smear allow for 60 seconds. Rinse the slide with water. Add a drop of grams iodine to the smear allow for 60 seconds. Rinse the slide with water. Add 95% absolute ethanol to destain the crystal violet. Rinse again with water. Add a drop of saffronin over the smear, allow for 60 seconds. Rinse with water. Blot dry the slide, do not rub. Observe the smear under microscope. 10. If the colour of the bacteria appears pink it is gram negative, it is gram positive if it appears purple. Rod shaped pink coloured bacteria is clearly observed for gram negative pseudomonas aeruginosa. Shake flask method Prepare media of nutrient broth about 13 grams per litre of distilled water. Mix well and pour 50 ml of media into a 250ml conical flask. Plug the conical flask and wrap the mouth with aluminium foil. The less volumes of media in the flask increases surface area and oxygen transfer rate(OTR). The flasks are autoclaved at 1210 C and 1.5 lbs pressure for 1 hour 15 minutes. Planktonic-contains 50 ml of nutrient broth Biofilm-contains 25ml of PBS with microcarriers plus 25 ml of nutrient broth. Inoculate a loopful of pseudomonas aeruginosa into 50 ml nutrient broth aseptically. Allow the bacteria to grow overnight. Take the optical density at 550nm using spectrophotometer.The amount of the inoculums to be added to the shake flasks for analysis is calculated using the formula below: Amount of sample to add into shake flask = [working concentration X volume(ml)] / stock(OD) obtained After inoculation the flasks are kept in the orbital shaker at 300C. The samples of 1 ml is collected for RNA extraction at particular time intervals at 0,1,2, 4,24 and 48 hours for planktonic. In respect the biofilm samples are collected at time 0,22,24,26,44,46 and 48 hours as their growth is slower than the freely suspended planktonic cells. Then 2 ml of RNA later solution is added to the collected samples for RNA stabilization. The optical density is regularly monitored to check the growth of pseudomonas aeruginosa . Fermentation The fermentation process we perform here is batch fermentation. The bioreactor used for the study is a small scale fermentor from APPLICON BIOTECHNOLOGIES. The process will take place in a 3L fermentor(consists the working volume of 2.5L) for about 24 hours. During the run the criteria of temperature is kept at 300C . The pH is run free, stirrer speed is set at 600rpm for planktonic culture. The fermentor is run free for 24 hours overnight to check the production of foam. The cold finger helps to control the temperature by feeding water from/to the reactor. The o ring is checked for any leakage. The sample port is a tube for sampling using back pressure and forces the liquid into a glass recipient. The inlets for acid, alkali and antifoam are linked to durham bottles, going through the pumps. The durham bottles are blank, because above parameters are not controlled in this process. Air is pumped through a filter. The condenser is connected to water that comes out of the fermentor. Wh en the water is warmer than outside, condensed water reaches the condenser. The gas goes from an outlet connected to the condenser. The dissolved oxygen probe is connected to dO2 cable from the machine. It measures the amount of dO2 in the media. This probe needs to be kept in the buffer, when not in use. The probe should be plugged for 10 to 12 hours before the inoculation inorder to polarize. The stirrer is placed on the top of fermentor. The vessel is heated with the help of heating pad and temperature is detected using temperature probe. The air inlet is set up through the pumps and its volume is controlled with a rotometer in front of the machine. In this case 2.5L of air/min is supplied to the fermentor. For the gas analysis of oxygen and carbon dioxide the gas is passed through a drying column and reaches the monitor which is already set up for calibration. we run two runs of fermentor for both planktonic and biofilm cultures. The parameters such as temperature, aeration for planktonic and biofilm cultures are almost same. The only parameter varies for both is stirring is maintained at 600rpm for planktoni c and 200rpm for biofilm fermentation. Procedure: The fermentor is sterilised and all the valves are sealed with foil before going into the autoclave. The fermentor is run for about 24 hours before adding the inoculum to check any formation of foam. About 20 ml of an overnight culture is added to the fermentor. Sampling :About 7ml of sample is collected for every 20min for planktonic and every 2 hours for biofilm until the exponential phase is reached. The sample collected is preserved in the freezer for different analysis. The sample is assayed for gene expression by adding 2 ml of RNA later solution for RNA stability. The samples are assayed for ammonia, phosphorus, glucose and protease. The fermentation conditions are listed below. RNA EXTRACTION (protocol provided by applied biosystems) Materials required: 2-mercapto ethanol,100% ethanol, 10% SDS(in RNase free water), 0.5  µl/sample. Lysozyme solution: 100  µl/ sample 10mM Tris HCl (PH 8.0)mM EDTA 10mg/ml lysozyme (in RNase free water) Lysis buffer: Before beginning the lysis and homogenization steps, prepare a fresh amount of lysis buffer containing 1% 2-mercapto ethanol for each purification procedure. Add 10 µl 2-mercaptoethanol for each 1ml lysis buffer. Use 350  µl of freshly prepared lysis buffer for 1109 bacterial cells. Lysis and homogenization: Harvest 1109 bacterial cells and transfer them to an appropriately sized microcentrifuge tubes. Centrifuge at 500xg for 5 minutes at 40C to pellet cells. Discard the supernatant. Add 100  µl of prepared lysozyme solution to the cell pellet and resuspend by vortexing. Add 0.5  µl 10% SDS solution vortex to mix well. Incubate the cells in the tube for 5 minutes at room temperature. Add 350  µl lysis buffer prepared with 2-mercaptoethanol. Vortex to mix well. Transfer the lysate to a homogenizer inserted in an RNase free tube and centrifuge at 12,000xg for 2 minutes at room temperature. Remove the homogenizer when done. Purifying the RNA from bacterial cells: These steps are followed to bind wash and elute the RNA from the P.aeruginosa cells sample. Add 250  µl 100% ethanol to each volume of bacterial cell homogenate. Mix thoroughly by vortexing to dispersing any visible precipitate that may form after adding ethanol. Transfer the sample(including any remaining precipitate) to a spin cartridge( with a collection tube. Centrifuge both the spin cartridge and collection tube at 12,000xg for 15 seconds at room temperature. Discard the flow through and re-insert the spin cartridge in the same collection tube. Add 700  µl of wash buffer I to the spin cartridge. Centrifuge at 12,000xg for 15 seconds at room temperature. Discard the flow through and the collection tube. Place the spin cartridge into a new collection tube. Add 500  µl wash buffer II with ethanol to the spin cartridge. Centrifuge at 12,000Xg for 15 seconds at room temperature. Discard the flow through and re insert the spin cartridge into the same collection tube. The steps are repeated once again. Centrifuge the spin cartridge and collection tube at 12,000Xg for 1 minute at room temperature to dry the membrane with attached RNA. Discard the collection tube and insert the spin cartridge into a recovery tube. Add 50  µl of RNase free water to the centre of spin cartridge. Incubate at room temperature for 1 minute. Centrifuge the spin cartridge and recovery tube for 2 minutes at 12,000Xg at room temperature. Store the purified RNA or proceeded for DNase treatment after RNA treatment. (store purified RNA at -800C for long term). Agarose gel electrophoresis: The RNA samples after extraction are checked for the RNA signal. To visualise the RNA 1% agarose gel electrophoresis. The materials required for agarose gel electrophoresis is TAE buffer and agarose. TAE buffer(50x Tris acetate) 242 g of Tris base 57.1 ml glaciel acetic acid 100 ml of EDTA 0.5M PH .8.0 700ml of dH2O . About 20 ml of above solution is mixed with 980ml of distilled water to make it 1X. 1 gram of agarose is added to 100 ml of TAE buffer. The solution is mixed well. The solution is heated until the agarose is completely dissolved and solution becomes colourless. About 2.5  µl of ethidium bromide is added to the solution to visualise the RNA bands. Special care is taken while adding the ethidium bromide because it is bio-hazardous. Then about 5  µl RNA sample is mixed with 1  µl of DNA loading dye. The RNA samples are RUN with the potential difference of 80V with DNA Hyper ladder II as a standard. DNase treatment (protocol and kit provided by applied biosystem) The DNase I treatment is performed to remove any traces of genomic DNA from the eluted RNA. Protocol: Add 10x DNase buffer and DNase I to the RNA sample in the following composition: Amount component 1/9 th volume 10x DNase buffer 4  µl DNase I The components are mixed with gentle pippeting. Then the mixture is incubated at 370C for 90 minutes. DNase inactivation reagent is added to the solution and mixed well, leave at room temperature for 2 minutes. The samples are centrifuged at maximum speed for 1 minute to pellet the DNase inactivation reagent, after that the RNA solution is transferred to a new RNase free tube. The samples are run on 1% agarose gel electrophoresis to check the DNA contamination. We fou