Shakespeare a modern genius. Essay Example for Free

Shakespeare a modern genius. EssayWilliam Shakespeare is considered to be the greatest playwright of all time. What Shakespeare did was to communicate his own words in a brilliant way. He created comedy, register, tragedy and poetry that were extremely amazing. The vast majority of scholars consider Shakespeare a modern genius. Surely he was just that. No other writers plays have been produced so many a(prenominal) times or read so widely in so many countries as his History articles show William Shakespeare born on my birthday April 23, 1564. He became an alderman and bailiff in Stratford-upon-Avon, and Shakespeare was baptized in the town on April 26, 1564.At age 18, Shakespeare married Anne Hathaway, and the equate had a daughter in 1583 and twins in 1585. Shakespeares progress tos of imagination were based on historical figures. Shakespeare was a keen reader of write up and was always looking for the dramatic impact of historical characters and events he read. Richard II I as an evil man, a kind of psychopath with a deformed personate and a grudge against gentlemans gentlemanity. Historians can do whatever they like to set the record straight but Shakespeares Richard seems stuck in our culture as the very Richard III. Henry V, and Prince Hal, is in our minds as the perfect model of kingship.That is because thats the way Shakespeare chose to present them in the themes he wanted to develop and the dramatic story he wanted to tell. The understanding of medieval history being shown through the rulers of this period is because of Shakespeare. We have adapted ourselves to Shakespeares vision. There have been some disputes over who wrote Shakespeares work but there no true evidence that he didnt. He was an actor as well as an excellent. Shakespeares tragedy is a tragical play either written by or in the style of William Shakespeare. Shakespeare plays usually get hold of murders or deaths, and a terrible ending for the main character.Shakespeares trage dies share some common features such as a hero with a fatal flaw that leads to his downfall. And the tragedies often focus on the fall of a nobleman. Hamlet, Othello, King Lear and Macbeth are examples of tragedies. Shakespeare communicated his comedy through language and his comedy plays were created with clever word play, metaphors, and insults. William Shakespeare wrote 154 Sonnets mostly in the 1590s. Fairly short poems, they deal with issues such as lost complete. His sonnets have an appeal due to his characteristic skill with language and words. Let me non to the marriage of true minds Admit impedi ments.Love is non bask Which alters when it alteration finds, Or bends with the remover to remove Shakespeare also understands deceit as he incorporated it into Much Ado About Nothing (Shakespeare, William). Contrary to love and deceit, Shakespeare uses comedy as the third and final element of the play. Comedy is what gives Much Ado About Nothing its cheerful happiness and wit t hat gives this play its recognition. Shakespeare had many bright experiences in his life due to his great success in being a playwright. His success started with him becoming the top writer of The Chamberlains Men, which would later be renamed The Kings Men.This led to his great career of writing which brought him fame and fortune, causing him to live and die a happy man. The happiness and clever wit is described like, Merry as the day is long. (Shakespeare). All throughout this play, there is constantly a t wiz of Comedy although interrupted by scenes of deceitful hate. This is very similar to the life of William Shakespeare, and it intelligibly the causality that he writes in the style he does. All in all, Shakespeares ability as a Comedic writer is very well spoken for and is the reason that he wrote this play.The elements used in this play as they have happened in Shakespeares life are established in the relationships of the characters of the play. Benedict and Beatrice, thr oughout the entire head start of the play, display love, deceit, and comedy. Beatrice finds all men to be repulsive, not in the physical sense, but in the psychological sense. Benedict does not like her attitude and does everything he can to spite her. Through all this, they both love distributively other, but both are as stubborn and hard headed as each other so it is impossible for either one to know this.At a costume party, Benedict tricks Beatrice into thinking he is someone else by eroding a costume and disguising his voice by talking with a different accent. Beatrice, not being aware of this, talks badly about Benedict to whom she thinks is a stranger. Afterwards, Benedict is furious about her statements and complains to the prince, so everyone else conspires to get them together. They do this by letting them individually overhear talk of each other loving one another. When they hear this, they are both very happy and end up telling each other their true feelings resulting in them getting married at the end of the play.The purpose of telling the story of Benedict and Beatrice is that their dialogue and actions resemble the tones and elements of the play therefore they greatly contribute to what I am proving (Shakespeare). Finally, the elements used in Much Ado About Nothing directly coincide with the events of Shakespeares life. The style that he uses comes from his life from birth to death. In this, it is obvious that Shakespeare experienced love in all its aspects, he experienced deceit in a way, but more minor than others, and also he experienced comedy from all the happy times he had in his life as do these experiences in the play.The personality of William Shakespeare is one of the most interesting for discussion, because people are not reliable about his existence and fascinated with his mind. It is really strange that one could write such masterpieces on different topics, revealing various social and philosophical problems so many years ago. O ne of the most famous Shakespearean linesTo be or not to be, that is the question(Shakespeare) is found in Hamlet, spoken by the title character himself. While this is the most obvious reference that Hamlet makes to his own philosophy, Hamlet makes frequent proclamations about his philosophy of life.Hamlets philosophy touches not only on the subject of love, but also about loyalty, family, and the virtue of suffering, among other themes. Hamlets philosophy is particular to his own strange obsessions, and helps explicate the fates of the characters in the play. Because Hamlet has been disillusioned about love by his amazes actions, he rejects the possibility that romantic love is an important part of human relationships. He is consumed by the outrageousness of his mothers love for his uncle, and he rejects Ophelias love for him, though he admitted once to loving her.Although Hamlet is warrant to feel disgust towards his mother and her actions, his pessimistic view of love has dre adful implications, not just for him, but for other characters as well. For this essay on Hamlet, you index want to take a character analysis approach to Hamlet with this theme as your guide or thesis statement. Any of Shakespeares plays involve transgressions that violate social taboos. Hamlet is no exception. In this play, legion(predicate) social norms are violated however, the most powerful taboo is that of incest.Hamlet is outraged when his mother marries his uncle shortly after the death of his father, and his mothers action causes him to lose faith in love. Although the incest taboo may seem grotesque, Shakespeare puts his characters in such high-energy tension and outrageous situations in order to make profound observations about the nature of both familial and romantic love. Claudius could hardly be considered to be a model of upright behavior and insight given that he seduces Gertrude while the trouble over her husbands death is still fresh.While he is obviously advanc ing his own motives, his speech to Hamlet about unmanly grief(Shakespeare) is oddly compelling. Claudius takes the view that all men die all men lose their fathers. They enter a period of appropriate grief and then move on. Because Hamlet is not conforming to this norm, Claudius suggests that Hamlets grief is not only unhealthy, but also unmanly. A close reading of the play supports Claudiuss observation. Although Claudius is certainly not free from reproach, Hamlets obsessional grief is not praiseworthy either.William Shakespeare is declared and worshiped as one of the most influential writers of English literature. Shakespeare has in many ways touched the lives of thousands upon thousands of readers throughout time. In attachment to introducing new techniques and methods of writing, Shakespeare has had a major impact on both his society as well as our modern society through all his numerous contributions such as historical literature, positive social presence, and again his wonde rful new attributes in writing style. Historians often refer to Shakespeare as one of historys most dramatic poets. In the same breath many scholars would refer to Shakespeare as also being the greatest English playwright in all of history by being simple and sentimental and communicating fine moral shades through out his work(Fowler 72). Of William Shakespeare (1564-1616), in the biographic sense, we know both too much and too little. The numerous investigators have been amazed by the quantity of information, most of which is utterly useless and irrelevant (Sampson 214). William Shakespeare fundamentally began his luxurious career in literature as a suffering actor in the city of Stratford-on-Avon.This is where is all began for the genius we all know and is called Shakespeare. whole caboodle Cited Fowler, Will. Shakespeare His Life and Plays. Harlow Pearson Education, 2001. Print. Hamlet Character Analysis. About. com Shakespeare. N. p. , n. d. Web. 24 Nov. 2013. Shakespeare Sonn et 116 Let Me Not to the Marriage of True Minds. Shakespeare Sonnet 116 Let Me Not to the Marriage of True Minds. N. p. , n. d. Web. 24 Nov. 2013. Shakespeare His Life And Plays. Goodreads. N. p. , n. d. Web. 24 Nov. 2013. Shakespeare William, and George Sampson. Hamlet. London Christophers, 1925. Print. SparkNotes. SparkNotes, n. d. Web. 24 Nov. 2013.

Factors Which Lead to Stress in the Workplace

F ar retchors Which Lead to emphasis in the stoolplacePh unrivaleds 4 you Case StudyChapter One IntroductionPh is 4u is a bad independent mobile phone retailer in the UK. Since opening in 1996, it expanded to cd stores through go forth the United Kingdom. Head office is constitute in Newcastle-under-Lyme, Staffordshire and was until recently part of the The Caud soundly Group formed by John Caudwell. In September 2006 the group was sold and Phones 4u is instantly owned by saving equity Partners (Minter, 2003, 18) attempt is the condition that dissolvents when person-environ acidulate disembowelt transactions lead the individual to perceive a discrepancy, whether real or non, amidst the demands of a situation and the resources of the persons biological, manpowertal or kindly systems. In medical terms, seek is the disruption of homeostasis through somatogenic or psychological stimuli. Stressful stimuli chiffonier be mental, physiological, anatomical or carnal react ions. wooly car keys, tardiness, family death, and loss of job, pressure, frustration, and hearty changes-these ar different types of essay, the process by which one appraises and copes with environmental threats and challenges. The events of daily life bleed through a psychological filter that helps a person the react in certain ways. Some idiom early in life is conducive to later stirred up and physical growth. But filter outes, or conflicts, pot to a fault threaten a persons life as well and health (Amatea, 1991, 48).Behavioral psychologists get determined on that point is a correlation amongst song and the declination on ones behavior. One may summation his usage of alcohol, tobacco, or sepa tell drugs to escape his formful state. opposite paradoxs admit arguing with jocks, neglecting appearance, crying easily, difficulty concentrating, and withdrawing from family and friends. In extreme cases, adjudicate passel ca hold insanity.Emotional changes argon exc essively a common instal of punctuate. Symptoms imply anger, anxiety, depression, nervousness, loneliness, and rejection. Changes in emotional state may lead to psychological disorders or even death, if non treated. Suicide is among the leading outcomes of assay-related depression (aspinwall, 1992, 48).Not totally does emphasise effect ones emotional and behavioral states, buy it also plays a large role in ones physical state. Symptoms of filter out include, but atomic number 18 not restrict to, allergies, post pain, respiratory infections, fatigue, headaches, muscle tension, sleeping problems, and dizziness. British scientist Hans Selye made a basic point of filtrate. It states that although the human clay comes designed to cope with fugacious focusing, protracted separate out post rise physical deterioration. MRI brain s rumps of people who befool subsistd a pro abundanted come of stress very much s backside the sequels of a shrunken hippocampus, the inne r brain structure vital to laying down explicit memories. Stress can put people at danger for one of todays four leading causes of serious illness and death heart disease, cancer, stroke, and chronic lung disease. such findings were proven true by studies done by cardiologists Meyer Friedman and radiation Rosenman in 1956. Psychophysiological illnesses ar stress-related. They argon illnesses, such as lofty naturalise downslope pressure, that are cause by senior high takes of stress. Immune results may also be effect by stress. They can either speed up or slow down causing a variety of illnesses such as lupus or quaternary sclerosis. Stress does not make one sick, but it does restrain ones immune functioning, making him more than(prenominal) vulner open to foreign invaders (Bandura, 1986, 58).Stress in unavoidable. If one can not eliminate stress by changing or ignoring a situation, one must learn to manage it by confronting or escaping the problem. Stress solicitude may include aerobic exercise, relaxation, and social support. Without knowledge of stress and ways to manage it, people are more susceptible of disease and psychological disorders. in that respect are variant sources of stress. The very definition of stress is A mentally or emotionally disruptive or upsetting condition occurring in response to adverse external influences and capable of affecting physical health, usually characterized by increased heart rate, a rise in blood pressure, fibrous tension, irritability, and depression. A stimulus or circumstance causing such a condition. In todays world of competition and success, the limits of our threshold are being pushed to succeed. Success is the driving force behind m both people pushing their bodies to the limits. A human body needs a delicate balance between study, and relaxation. When this equilibrium is not wielded, problems occur, which force brace repercussions later. These repercussions take the forms of Burnouts, brea k-downs, and well-nigh clippings extreme cases of heart attacks.In around cases of the systems breaking down, in that location are methods of control and mechanisms of improvement of the situation. Stress can be handled very effectively in todays world. ruddy and wholesome living is the everyplacebold day mantra for better work efficiencies.What causes stress?T here(predicate) are discordant triggers to stress. Work, own(prenominal) life and external factors (Banyard, 1993, 45).Work the pressures of work are one of the grea sample factors contributing to the high stress levels today. Long work hours, ageless pressures of deadlines, and the required fear of job security pushes people to work very hard.Personal Life out spot the work place, people are always subject to constant pressures from family, friends, and well wishers. Handling these pressures in the well-nigh effective way is the tact of the new manager. Family life can cause stress, peculiarly if in that resp ect is some clangour between partners, the ill heath or sudden death of a partner can cause great levels of stress.External factors personal finances, world events and separate non distinctive events also add to stress. Time heed or lack of in that locationof is another cause of external factors of stress.How does one handle stress? on that point are some(prenominal) ways of fighting stress in ones life. This essay exit briefly attempt to touch base with some important methods which can be imbibed in handling stress.1) Time Management Time perplexity is one of the greatest methods of combating stress especially in the workplace. Effective snip management helps us prioritizing, planning, allocating and executing effective schedules in order to maximize our most valuable resource, snip. Once a person has been able to handle time effectively, he or she would be able to handle many tasks which have been allocated (Baruch, 1987, 59).2) Health stress has a great impact on the p ersonal health of the individual. maturation is speeded up and white hair apparently comes aplenty. Living a reasoned life helps in handling stress better. A regular routine, the offshoot of effective time management, reserves us time to regularly exercise. Exercise, as many say release endorphins, these help combat stress and depression, a stress induced side effect. Healthy foods, such as greens, and maintaining a balanced diet are some other ways, which when coupled with exercise help us maintain a healthy body.3) Non Conventional Methods going back to the basics is a term that is used very regularly today. When one says going back to the basics, it includes going back to the past. Yoga, pranayama and other methods of controlling the mind, body and soul, have existed in our country for a very languish time. This is fast gaining popularity as methods of combating stress. Yoga has immense powers to help us maintain our minds and body in shape.From the preceding(prenominal), we can see one of the greatest factors of handling stress is to maintain a healthy mind, body and soul. Prevention is better than cure, preventing stress by having a healthy life style and a healthy mind, is a plus point which has no substitute in todays world. However there is no effective way of handling self induced stress. Calming the mind and regulated breathing can ease the stress, but it pass on be rendered inefficient if the person is not forgeting to practice it in the positive way (Bem, 1981, 49).Stress is one of the greatest hindrances to efficient productivity in todays workplace. take force is the key word today and this does face a serious threat with stress. Combating stress on a war footing is the need of the hour, and some of the above points allow assist in effective stress management through a healthy mind , body and soul.Stress is a gang of responses in the body. Stress can be short-term (acute) or chronic. Acute stress is the fight or flight response. If a car is careening toward you at a high rate of speed, you bequeath (or should) experience acute stress. It is when you experience so many common stressors, such as heavy traffic, noise, money worries, illnesses, relationship problems, uprise iniquity rates, or work frustrations, that stress takes a chronic form. In the short term, stress can be vital. Over time, it turns destructive .How destructive can stress be on your body? Research has shown that prolonged stress can produce actual tissue changes and organ dysfunction. With the new MRI (magnetic vibrancy imaging) techniques, scientists are able to prove visibly that chronic stress can shrink an area in the brain called the hypothalamus. Read More On This They have found that the brains of war veterans, as well as women who have been victims of childhood sexual abuse, have a marked reduction in the size of their hypothalamus (Betz, 1987, 29).Stress also affects your brain by releasing powerful chemical messengers called neurot ransmitters, such as dopamine, norepinephrine, and epinephrine (also called adrenaline). The hypothalamic/pituitary-adrenal portion of your brain releases steroid hormones, including the master(a) stress hormone, cortisol. cortisol affects systems throughout your body, including an increased heart rate.Your heart, lungs, and circulatory system are influenced by the increased heart rate. Blood flow may increase ccc to 400 per centum. Blood pressure increases and breathing becomes rapid. Your mouth and throat may become dry. Skin may become cool and clammy because blood flow is divert out-of-door so it can support the heart and muscle tissues. Even digestive activity shuts down.Once again, occasional stress is normal. Once youve handled the situation, the stress goes away and you heal from the episode. But, if stressors accumulate over time, last the body becomes inefficient at handling even the least amount of stress. The brain, heart, lungs, vessels, and muscles become so invet erate over or under activated that they become damaged. It is this sort of stress which may trigger or worsen heart disease, strokes, susceptibility to infection, sleep disturbances, sexual and fruitful dysfunction, memory and learning dysfunction, digestive problems, weight problems, diabetes, pain, and skin disorders.Extensive multidisciplinary studies have presented unequivocal evidence that our psychological responses to stress and our perceptions of stress to a hefty extent affect our susceptibility to disease. In active relationship, the immune, neuroendocrine, and nervous systems respond to the brain and psyche. Virtually all illnesses, from the flu to cancer, are influenced for good or dreadful by our thoughts and scentings. R. Lloyd, 1990 Healing Brain A Scientific Reader (Betz, 1987, 48)Statement of the Research ProblemHow do the employees cope with stress in the workplace to arrive at a more balanced lifestyle at Phones 4 you ? Stress is a part of everybodys life. D epending on the level of stress, it can control our lives, especially in the workplace. We embark on to spend several long hours at work, and olibanum have less time for other things. Stressed employees may be unhappy and thusly produce nominally. Stress can deteriorate social and family relationships and eventually burn you out ultimately it can take toll on your health. Organizations need to signalise stress as a problem and decide whether or not to act upon it.Background InformationThis question needs to be answered because stress is a problem that Phones 4 you must deal with stress can cause poor work performance and lower employee morale. These factors can increase employee turnover rate and fall quality of life. We all must deal with stress question is how we handle and control it. With downsizing the buzz word in the modern merged world, companies have become mean and lean. Employees are compelled to be more efficient Phones 4 you they find themselves taking on the work of what used to be two. The result is long-dated hours, less time for outside activities, and consequently increased stress.According to Business Week, the typical British works 47 hours a week, and if current contracts continue, in 20 historic period the average person would be on the job 60 hours a week. Another factor that increases stress is technological advancements. With all the new technology one is always connected to work and accessible 24 hours a day 7 days a week. According to Business Week, it is now possible, and thus increasingly expected, for employees to be accessible and fruitful any hour, any day (Bollen, 1993, 18).At a workplace, one observes several sales people working long hours, claiming it is due to under staffing. Employees reach a point of diminishing returns. The more hours they work, the less productive they are. This stressful condition causes the quality of work to dwindle. Consequently, clients write out this, and eventually they terminate the business relationship. Soon the company loses, as it is built on these clients (Moos, 1989, 58).Statement of the ObjectivesThis look for expects to discuss factors which lead to stress in the workplace at Phones 4 you. Are individuals stressed in the workplace at Phones 4 you? What causes stress in the workplace Phones 4 you? Who is mostly stressed men or women? Are individuals being exposed to stress management techniques? Should employers implement stress management techniques? as a future manager, I would like to be able to determine if stress is a problem for employees if so, implement a strategy to curtail stress in the workplace. By recognizing stress in the workplace, employers can act appropriately to reduce stress. The outcome can benefit social and family relationships, as well as preserve ones health and make us more productive in our organizations (Moos, 1982, 25).ScopeThe research examine pull up stakes comprise of a sample size of 30 individuals, at random selected from general business areas. The piece of work bequeath analyze stress factors in the U.K workforce and its impact on the British organization. Effective stress management techniques will past be presented, which will allow individuals or organizations to implement. Secondary information from various sources will be utilized to explore effective methods of coping with stress. The conclusions and recommendations I will draw will be applicable to any British organization with stress as a problem. Although this employment will generalize from the small population, it can be used as a starting point to recognizing the problem, as each organization can require a different approach (Parkes, 1986, 36).LimitationsThe sources utilized in the research will be extracted from current articles (2006-present) from online services, the Internet, and public libraries. A survey will be given to individuals of randomly chosen organizations and will not target any item company or industry. Due t o time constraints, the population will be limited to 30 individuals. The research will explore factors causing stress in the workplace and its impact on organizations. Effective methods of coping with stress will be given, but limited to ones examined in the flakeary resources (Portello, 1996, 548).Research ProceduresThe project will reduce on stress factors in the workplace and effective methods to balance a healthy lifestyle. The sample group will consist of 30 individuals randomly selected from general business areas. The survey will be conducted during lunch periods when several employees leave and return to the workplace. The questionnaire will attempt to see if the sample individuals deal stress is a problem and what can be done to resolve it. The questionnaire will be delivered in person and each individual will take out the survey at that point.Since the survey will be conducted in a general public area, no authorization is needed to administer. Once I develop all the surveys, I will quantify the data into an Excel spreadsheet. I will report the data mostly in helpings (e.g. 70percent of the individuals acknowledge that stress is a problem in the workplace). The data will be utilized to see if stress is a factor impacting the British workforce. Stress management techniques will be presented where appropriate (Browne, 1993, 578).Chapter Two Literature ReviewStress is an adaptive response. It is the bodys reaction to an event that is seen as emotionally disturbing, disquieting, or threatening. When we perceive such an event, we experience what stress researchers call the fight or flight response. To prepare for fighting or fleeing, the body increases its heart rate and blood pressure more blood is then sent to your heart and muscles, and your respiration rate increases (Carmines, 1981, 48).Stress is both positive and blackball. Good stress is a balance of arousal and relaxation that helps you concentrate, focus, and achieve what you want. kn otty stress is constant stress and constant arousal that may lead to high blood pressure, cardiovascular disease, and worse. The body does not distinguish between negative and positive stress. The same physiological responses can take place whether you are happy or sad about a given situation.When ex guideing to the workplace, stress may lead to poor work performance and end up costing an organizations several thousands of dollars. The organization loses on salary because they are not receiving adequate doing and if the employee becomes ill, health and workers compensation rates can soar . The organization must decide whether or no to implement a stress management program, since there are several external stressors that can overtake an individual. Internal stressors, within organizations include technology and corporate downsizing which leads to longer hours and job uncertainty. If one does not know how to manage stress, it can get out of control ) (Rock, 1997, 4).Analyzing Stress on IndividualsIn a 1995 survey of 1,705 respondents it is analyzed that stress rises with level of direction and job level and is higher than average for women (Robinson, 1996, 88). Fifty-eight percent of the women respondents possess moderate to a lot of stress in the workplace compared to 53 percent of men. From the divorced individuals, 62 percent are stressed in the workplace compared to married and never married at 57 percent, and 58 percent respectively. The widow respondents maintain the least stress at 38 percent (Robinson, 1996, 48).College graduate respondents possess more stress at 64 percent than high give lessons graduates at 55 percent. Only 43 percent of the less than high naturalize respondents felt stress in the workplace. Those with more education disembodied spirit more stress, possibly because their jobs involve greater managerial and financial responsibility (Robinson, 1996, 87).Stress is an epidemic in British life. In nationwide polls, 89 percent of Briti shers account that they often experience high levels of stress, and 59 percent claimed that they feel great stress at least once a week (Hellmich, 1994, 57). A quintuple family study of the British workforce conducted by the Families and Work bring in showed that 30 percent of employees often or very often feel fire out or stressed by their jobs, 27 percent feel emotionally drained from their work, and 42 percent feel used up at the end of the work day (Hellmich, 1994, 4). Balancing work pressures and family responsibilities leaves many workers feeling burned out.Examining the Effects of Downsizing on StressThe downsizing of organizations have caused a stressful environment. Downsizing has created concerns over job security, and has forced employees to take on a larger workload. According to a local union representing U.K. westernmost give tongue to that work still needs to be done, but with fewer people (Scott, 1996, 41). Downsizing creates quantitative and qualitative stres s. Quantitative stress pertains to doing the same amount of work with fewer people. Reengineering the organization entails shaping the company to be more efficient with less individuals. These individuals are asked to do a wider variety of work functions they are not trained to do, causing qualitative overload (Scott, 1996, 35).Occupational Stress is the harmful physical and emotional response that occurs when there is a poor crack between job demands and the capabilities, resources, or needs of the worker. Stress-related disorders encompass a broad array of conditions, including psychological disorders (e.g., depression, anxiety, post-traumatic stress disorder) and other types of emotional strain (e.g., dissatisfaction, fatigue, tension, etc.), maladaptive behaviours (e.g., aggression, import abuse), and cognitive impairment (e.g., concentration and memory problems). In turn, these conditions may lead to poor work performance or even injury. Job stress is also associated with var ious biological reactions that may lead ultimately to compromised health, such as cardiovascular disease (Rosenfield, 1989, 5).PrevalenceStress is a prevalent and costly problem in todays workplace. About one-third of workers report high levels of stress. One-fourth of employees view their jobs as the number one stressor in their lives. Three-fourths of employees believe the worker has more on-the-job stress than a generation ago. Evidence also designates that stress is the major cause of turnover in organizations (Scheier, 1985, 65).Health and Healthcare UtilizationProblems at work are more strongly associated with health complaints than are any other life stressor-more so than even financial problems or family problems. Many studies suggest that psychologically demanding jobs that allow employees little control over the work process increase the adventure of cardiovascular disease. On the basis of research by the guinea pig Institute for Occupational Safety and Health and many other organizations, it is widely believed that job stress increases the risk for development of back and upper-extremity musculoskeletal disorders. High levels of stress are associated with substantial increases in health service utilization. Workers who report experiencing stress at work also show excessive health care utilization. In a 1998 study of 46,000 workers, health care costs were nearly 50% greater for workers reporting high levels of stress in comparison to low risk workers. The ontogeny rose to nearly 150%, an increase of more than $1,700 per person annually, for workers reporting high levels of both stress and depression. Additionally, periods of disability due to job stress tend to be much longer than disability periods for other occupational injuries and illnesses (Schwartz, 1993, 58).Causes of Occupational StressJob stress results from the interaction of the worker and the conditions of work. Views differ on the magnificence of worker characteristics versus workin g conditions as the primary cause of job stress. The differing viewpoints suggest different ways to prevent stress at work. According to one school of thought, differences in individual characteristics such as personality and coping style are most important in predicting whether certain job conditions will result in stress-in other words, what is stressful for one person may not be a problem for someone else. This viewpoint leads to prevention strategies that focus on workers and ways to help them cope with demanding job conditions. Although the importance of individual differences cannot be ignored, scientific evidence suggests that certain working conditions are stressful to most people. Such evidence argues for a greater emphasis on working conditions as the key source of job stress, and for job redesign as a primary prevention strategy. Personal interview surveys of working conditions, including conditions recognized as risk factors for job stress, were conducted in Member State s of the European Union in 1990, 1995, and 2000. Results showed a trend across these periods suggestive of increasing work intensity. In 1990, the percentage of workers reporting that they worked at high speeds at least one-fourth of their working time was 48%, increasing to 54% in 1995 and to 56% in 2000. Similarly, 50% of workers reported they work against tight deadlines at least one-fourth of their working time in 1990, increasing to 56% in 1995 and 60 % in 2000. However, no change was noted in the period 19952000 (data not collected in 1990) in the percentage of workers reporting sufficient time to complete tasks. A substantial percentage of Britishers work very long hours. By one estimate, more than 26% of men and more than 11% of women worked 50 hours per week or more in 2000. These figures represent a colossal increase over the previous leash decades, especially for women. According to the Department of Labour, there has been an upward trend in hours worked among diligent women, an increase in extended work weeks (40 hours) by men, and a considerable increase in combined working hours among working couples, particularly couples with young children (Shaw, 1993, 4).Signs of Occupational StressMood and sleep disturbances, upset stomach and headache, and disturbed relationships with family, friend and lady friend/boy friends are examples of stress-related problems. The effectuate of job stress on chronic diseases are more difficult to see because chronic diseases take a long time to develop and can be influenced by many factors other than stress. Nonetheless, evidence is rapidly accumulating to suggest that stress plays an important role in several types of chronic health problems-especially cardiovascular disease, musculoskeletal disorders, and psychological disorders (Sherer, 1982, 36).PreventionA combination of organizational change and stress management is often the most utile approach for preventing stress at work.How to Change the Organization t o Prevent Job StressEnsure that the workload is in line with workers capabilities and resources.Design jobs to propose meaning, stimulation, and opportunities for workers to use their skills.Clearly define workers roles and responsibilities.Give workers opportunities to participate in decisions and actions affecting their jobs.Improve communications-reduce uncertainty about career development and future employment prospects.provide opportunities for social interaction among workers.Establish work schedules that are compatible with demands and responsibilities outside the job.St. Paul Fire and Marine Insurance Company conducted several studies on the effects of stress prevention programs in hospital settings. Program activities included (1) employee and management education on job stress, (2) changes in hospital policies and procedures to reduce organizational sources of stress, and (3) formation of employee care programs. In one study, the frequency of medication errors declined by 50% after prevention activities were implemented in a 700-bed hospital. In a second study, there was a 70% reduction in malpractice claims in 22 hospitals that implemented stress prevention activities. In contrast, there was no reduction in claims in a matched group of 22 hospitals that did not implement stress prevention activities (Smith, 1981, 24).Chapter Three Research MethodologyDesignThe data reported here are from two separate data sets. In order to cross-validate the model, data from the original study of managerial women were used, and these included data from the graduation exercise three assessments of a 2-year longitudinal study. In the original article (B. C. Long et al., 1992, 165), a conceptual model of stress and coping was tested and developed that was based on data from the first three assessments (Time 1 to Time 3) of 11 assessments completed over 2 age. Status, Sex Role Attitudes, and Agentic Traits were assessed at Time 1 Appraisals, Disengagement and acti on Coping, Work Environment, and Daily Hassles were assessed at Time 2 and Distress and Satisfaction were assessed at Time 3. These data were used as a base to test the validity of the model on a new set of data obtained from clerical workers, data that have not been reported elsewhere (Snapp, 1992, 32).Sampling The managerial women (n = 249) were industrious in nontraditional occupations (i.e., fewer than 35% of British employees are women). Their mean age was 38.84 years (SD = 7.68, range = 2266). More detailed descriptions of the managers characteristics can be found in B. C. Long et al. (1992).The clerical workers who participated were employed in both large and small organizations in the same large western British urban center in which the managers were employed. The clerical workers volunteered in response to written requests for participants that I circulated in the media and by networking. The notices were directed to full-time female clerical workers and indicated that th e purpose of the study was to enquire how clerical workers experienced Occupational Stress. No incentives were offered other than a final summary report. f the 284 respondents who made contact by telephone, 273 met the criteria for inclusion (i.e., they were employed in a clerical position, worked more than 20 hours per week, and did not supervise others). f the 273 clerical workers who met the criteria and were distributed questionnaires at Time 1, 39 withdrew from the study because of lack of time to participate, 7 no longer met our criteria because of promotion, unemployment, or leave of absence from work (e.g., due to accident or illness), and 4 moved. The overall dropout rate was 18%. Dropout analyses were conducted on the demographic variables measured at Time 1. No differences were found between the retained (n = 223) and dropout (n = 50) respondents. Chi-square analyses of the demographic variables (marital status, education, number of children, job level, and size of the c ompany) were not significant. Because 9 participants determine a personal rather than a work stressor, their data were omitted from the model testing.All respondents were self-identified clerical workers. Job classifications included clerks (25%), secretariesstenographers (23%), administrative assistants (34%), and others (18%). The mean age was 39.77 years (SD = 9.46, range = 2263 years). Fifty-three percent of the clerical workers were married, 22% were single, and 25% were divorced, separated, or widowed. Fifty-three percent were parents. Twenty-four percent had a high school education or less, 42% had special training (e.g., secretarial, clerical), 17% had a college education (2 years postsecondary), and 13% had a university degree. Household incomes ranged from less than $25,000 (British) per year (23.4%) to over $61,000 (British) per year (27.5%). The major industries represented were education (31%), service (35%), utilities and public administration (12%), manufacturing and transportation (10%), and other (8%). On average the women had been in the workforce for 17.02 years (SD = 8.74,Factors Which Lead to Stress in the WorkplaceFactors Which Lead to Stress in the WorkplacePhones 4 you Case StudyChapter One IntroductionPhones 4u is a large independent mobile phone retailer in the UK. Since opening in 1996, it expanded to 400 stores throughout the United Kingdom. Head office is based in Newcastle-under-Lyme, Staffordshire and was until recently part of the The Caudwell Group formed by John Caudwell. In September 2006 the group was sold and Phones 4u is now owned by Providence Equity Partners (Minter, 2003, 18)Stress is the condition that results when person-environment transactions lead the individual to perceive a discrepancy, whether real or not, between the demands of a situation and the resources of the persons biological, psychological or social systems. In medical terms, stress is the disruption of homeostasis through physical or psychological sti muli. Stressful stimuli can be mental, physiological, anatomical or physical reactions.Lost car keys, tardiness, family death, and loss of job, pressure, frustration, and social changes-these are different types of stress, the process by which one appraises and copes with environmental threats and challenges. The events of daily life flow through a psychological filter that helps a person the react in certain ways. Some stress early in life is conducive to later emotional and physical growth. But stresses, or conflicts, can also threaten a persons life as well and health (Amatea, 1991, 48).Behavioral psychologists have determined there is a correlation between stress and the declination on ones behavior. One may increase his usage of alcohol, tobacco, or other drugs to escape his stressful state. Other problems include arguing with friends, neglecting appearance, crying easily, difficulty concentrating, and withdrawing from family and friends. In extreme cases, stress can cause insa nity.Emotional changes are also a common effect of stress. Symptoms include anger, anxiety, depression, nervousness, loneliness, and rejection. Changes in emotional state may lead to psychological disorders or even death, if not treated. Suicide is among the leading outcomes of stress-related depression (aspinwall, 1992, 48).Not only does stress effect ones emotional and behavioral states, buy it also plays a large role in ones physical state. Symptoms of stress include, but are not limited to, allergies, back pain, respiratory infections, fatigue, headaches, muscle tension, sleeping problems, and dizziness. British scientist Hans Selye made a basic point of stress. It states that although the human body comes designed to cope with temporary stress, prolonged stress can produce physical deterioration. MRI brain scans of people who have experienced a prolonged amount of stress often show the results of a shrunken hippocampus, the inner brain structure vital to laying down explicit me mories. Stress can put people at risk for one of todays four leading causes of serious illness and death heart disease, cancer, stroke, and chronic lung disease. Such findings were proven true by studies done by cardiologists Meyer Friedman and Ray Rosenman in 1956. Psychophysiological illnesses are stress-related. They are illnesses, such as high blood pressure, that are caused by high levels of stress. Immune responses may also be effected by stress. They can either speed up or slow down causing a variety of illnesses such as lupus or multiple sclerosis. Stress does not make one sick, but it does restrain ones immune functioning, making him more vulnerable to foreign invaders (Bandura, 1986, 58).Stress in unavoidable. If one can not eliminate stress by changing or ignoring a situation, one must learn to manage it by confronting or escaping the problem. Stress management may include aerobic exercise, relaxation, and social support. Without knowledge of stress and ways to manage it, people are more susceptible of disease and psychological disorders.There are various sources of stress. The very definition of stress is A mentally or emotionally disruptive or upsetting condition occurring in response to adverse external influences and capable of affecting physical health, usually characterized by increased heart rate, a rise in blood pressure, muscular tension, irritability, and depression. A stimulus or circumstance causing such a condition. In todays world of competition and success, the limits of our threshold are being pushed to succeed. Success is the driving force behind many people pushing their bodies to the limits. A human body needs a delicate balance between work, and relaxation. When this equilibrium is not maintained, problems occur, which might have repercussions later. These repercussions take the forms of Burnouts, break-downs, and sometimes extreme cases of heart attacks.In most cases of the systems breaking down, there are methods of control and mechanisms of improvement of the situation. Stress can be handled very effectively in todays world. Healthy and wholesome living is the new day mantra for better work efficiencies.What causes stress?There are various triggers to stress. Work, personal life and external factors (Banyard, 1993, 45).Work the pressures of work are one of the greatest factors contributing to the high stress levels today. Long working hours, constant pressures of deadlines, and the inevitable fear of job security pushes people to work very hard.Personal Life outside the work place, people are always subject to constant pressures from family, friends, and well wishers. Handling these pressures in the most effective way is the tact of the new manager. Family life can cause stress, especially if there is some friction between partners, the ill heath or sudden death of a partner can cause great levels of stress.External factors personal finances, world events and other non classifiable events also add to str ess. Time management or lack of thereof is another cause of external factors of stress.How does one handle stress?There are several ways of fighting stress in ones life. This essay will briefly attempt to touch base with some important methods which can be imbibed in handling stress.1) Time Management Time management is one of the greatest methods of combating stress especially in the workplace. Effective time management helps us prioritizing, planning, allocating and executing effective schedules in order to maximize our most valuable resource, time. Once a person has been able to handle time effectively, he or she would be able to handle many tasks which have been allocated (Baruch, 1987, 59).2) Health stress has a great impact on the personal health of the individual. Ageing is speeded up and white hair apparently comes aplenty. Living a healthy life helps in handling stress better. A regular routine, the offshoot of effective time management, allows us time to regularly exercise . Exercise, as many say release endorphins, these help combat stress and depression, a stress induced side effect. Healthy foods, such as greens, and maintaining a balanced diet are some other ways, which when coupled with exercise help us maintain a healthy body.3) Non Conventional Methods going back to the basics is a term that is used very regularly today. When one says going back to the basics, it includes going back to the past. Yoga, pranayama and other methods of controlling the mind, body and soul, have existed in our country for a very long time. This is fast gaining popularity as methods of combating stress. Yoga has immense powers to help us maintain our minds and body in shape.From the above, we can see one of the greatest factors of handling stress is to maintain a healthy mind, body and soul. Prevention is better than cure, preventing stress by having a healthy life style and a healthy mind, is a plus point which has no substitute in todays world. However there is no e ffective way of handling self induced stress. Calming the mind and regulated breathing can ease the stress, but it will be rendered ineffective if the person is not willing to practice it in the positive way (Bem, 1981, 49).Stress is one of the greatest hindrances to efficient productivity in todays workplace. Production efficiency is the key word today and this does face a serious threat with stress. Combating stress on a war footing is the need of the hour, and some of the above points will assist in effective stress management through a healthy mind , body and soul.Stress is a combination of responses in the body. Stress can be short-term (acute) or chronic. Acute stress is the fight or flight response. If a car is careening toward you at a high rate of speed, you will (or should) experience acute stress. It is when you experience so many common stressors, such as heavy traffic, noise, money worries, illnesses, relationship problems, rising crime rates, or work frustrations, that stress takes a chronic form. In the short term, stress can be vital. Over time, it turns destructive .How destructive can stress be on your body? Research has shown that prolonged stress can produce actual tissue changes and organ dysfunction. With the new MRI (magnetic resonance imaging) techniques, scientists are able to prove visibly that chronic stress can shrink an area in the brain called the hypothalamus. Read More On This They have found that the brains of war veterans, as well as women who have been victims of childhood sexual abuse, have a marked reduction in the size of their hypothalamus (Betz, 1987, 29).Stress also affects your brain by releasing powerful chemical messengers called neurotransmitters, such as dopamine, norepinephrine, and epinephrine (also called adrenaline). The hypothalamic/pituitary-adrenal portion of your brain releases steroid hormones, including the primary stress hormone, cortisol. Cortisol affects systems throughout your body, including an incre ased heart rate.Your heart, lungs, and circulatory system are influenced by the increased heart rate. Blood flow may increase 300 to 400 percent. Blood pressure increases and breathing becomes rapid. Your mouth and throat may become dry. Skin may become cool and clammy because blood flow is diverted away so it can support the heart and muscle tissues. Even digestive activity shuts down.Once again, occasional stress is normal. Once youve handled the situation, the stress goes away and you heal from the episode. But, if stressors accumulate over time, eventually the body becomes inefficient at handling even the least amount of stress. The brain, heart, lungs, vessels, and muscles become so chronically over or under activated that they become damaged. It is this sort of stress which may trigger or worsen heart disease, strokes, susceptibility to infection, sleep disturbances, sexual and reproductive dysfunction, memory and learning dysfunction, digestive problems, weight problems, diab etes, pain, and skin disorders.Extensive multidisciplinary studies have presented unequivocal evidence that our psychological responses to stress and our perceptions of stress to a considerable extent affect our susceptibility to disease. In active relationship, the immune, neuroendocrine, and nervous systems respond to the brain and psyche. Virtually all illnesses, from the flu to cancer, are influenced for good or bad by our thoughts and feelings. R. Lloyd, 1990 Healing Brain A Scientific Reader (Betz, 1987, 48)Statement of the Research ProblemHow do the employees cope with stress in the workplace to achieve a more balanced lifestyle at Phones 4 you ? Stress is a part of everybodys life. Depending on the level of stress, it can control our lives, especially in the workplace. We begin to spend several long hours at work, and thus have less time for other things. Stressed employees may be unhappy and thus produce nominally. Stress can deteriorate social and family relationships and eventually burn you out ultimately it can take toll on your health. Organizations need to recognize stress as a problem and decide whether or not to act upon it.Background InformationThis question needs to be answered because stress is a problem that Phones 4 you must deal with stress can cause poor work performance and lower employee morale. These factors can increase employee turnover rate and lessen quality of life. We all must deal with stress question is how we handle and control it. With downsizing the buzz word in the modern corporate world, companies have become mean and lean. Employees are compelled to be more efficient Phones 4 you they find themselves taking on the work of what used to be two. The result is longer hours, less time for outside activities, and consequently increased stress.According to Business Week, the typical British works 47 hours a week, and if current trends continue, in 20 years the average person would be on the job 60 hours a week. Another factor t hat increases stress is technological advancements. With all the new technology one is always connected to work and accessible 24 hours a day 7 days a week. According to Business Week, it is now possible, and thus increasingly expected, for employees to be accessible and productive any hour, any day (Bollen, 1993, 18).At a workplace, one observes several sales people working long hours, claiming it is due to under staffing. Employees reach a point of diminishing returns. The more hours they work, the less productive they are. This stressful condition causes the quality of work to dwindle. Consequently, clients recognize this, and eventually they terminate the business relationship. Soon the company loses, as it is built on these clients (Moos, 1989, 58).Statement of the ObjectivesThis research expects to discuss factors which lead to stress in the workplace at Phones 4 you. Are individuals stressed in the workplace at Phones 4 you? What causes stress in the workplace Phones 4 you? W ho is mostly stressed men or women? Are individuals being exposed to stress management techniques? Should employers implement stress management techniques? as a future manager, I would like to be able to determine if stress is a problem for employees if so, implement a strategy to curtail stress in the workplace. By recognizing stress in the workplace, employers can act appropriately to reduce stress. The outcome can benefit social and family relationships, as well as preserve ones health and make us more productive in our organizations (Moos, 1982, 25).ScopeThe research project will comprise of a sample size of 30 individuals, randomly selected from general business areas. The study will analyze stress factors in the U.K workforce and its impact on the British organization. Effective stress management techniques will then be presented, which will allow individuals or organizations to implement. Secondary information from various sources will be utilized to explore effective methods of coping with stress. The conclusions and recommendations I will draw will be applicable to any British organization with stress as a problem. Although this study will generalize from the small population, it can be used as a starting point to recognizing the problem, as each organization can require a different approach (Parkes, 1986, 36).LimitationsThe sources utilized in the research will be extracted from current articles (2006-present) from online services, the Internet, and public libraries. A survey will be given to individuals of randomly chosen organizations and will not target any specific company or industry. Due to time constraints, the population will be limited to 30 individuals. The research will explore factors causing stress in the workplace and its impact on organizations. Effective methods of coping with stress will be given, but limited to ones examined in the secondary resources (Portello, 1996, 548).Research ProceduresThe project will focus on stress factors in the workplace and effective methods to balance a healthy lifestyle. The sample group will consist of 30 individuals randomly selected from general business areas. The survey will be conducted during lunch periods when several employees leave and return to the workplace. The questionnaire will attempt to see if the sample individuals believe stress is a problem and what can be done to resolve it. The questionnaire will be delivered in person and each individual will fill out the survey at that point.Since the survey will be conducted in a general public area, no authorization is needed to administer. Once I receive all the surveys, I will quantify the data into an Excel spreadsheet. I will report the data mostly in percentages (e.g. 70percent of the individuals acknowledge that stress is a problem in the workplace). The data will be utilized to see if stress is a factor impacting the British workforce. Stress management techniques will be presented where appropriate (Browne, 1993, 578).Chapter Two Literature ReviewStress is an adaptive response. It is the bodys reaction to an event that is seen as emotionally disturbing, disquieting, or threatening. When we perceive such an event, we experience what stress researchers call the fight or flight response. To prepare for fighting or fleeing, the body increases its heart rate and blood pressure more blood is then sent to your heart and muscles, and your respiration rate increases (Carmines, 1981, 48).Stress is both positive and negative. Good stress is a balance of arousal and relaxation that helps you concentrate, focus, and achieve what you want. Bad stress is constant stress and constant arousal that may lead to high blood pressure, cardiovascular disease, and worse. The body does not distinguish between negative and positive stress. The same physiological responses can take place whether you are happy or sad about a given situation.When extending to the workplace, stress may lead to poor work performance and end up costing an organizations several thousands of dollars. The organization loses on salary because they are not receiving satisfactory production and if the employee becomes ill, health and workers compensation rates can soar . The organization must decide whether or no to implement a stress management program, since there are several external stressors that can overtake an individual. Internal stressors, within organizations include technology and corporate downsizing which leads to longer hours and job uncertainty. If one does not know how to manage stress, it can get out of control ) (Rock, 1997, 4).Analyzing Stress on IndividualsIn a 1995 survey of 1,705 respondents it is analyzed that stress rises with level of education and job level and is higher than average for women (Robinson, 1996, 88). Fifty-eight percent of the women respondents possess moderate to a lot of stress in the workplace compared to 53 percent of men. From the divorced individuals, 62 percent are stressed in the workplace compared to married and never married at 57 percent, and 58 percent respectively. The widowed respondents maintain the least stress at 38 percent (Robinson, 1996, 48).College graduate respondents possess more stress at 64 percent than high school graduates at 55 percent. Only 43 percent of the less than high school respondents felt stress in the workplace. Those with more education feel more stress, possibly because their jobs involve greater managerial and financial responsibility (Robinson, 1996, 87).Stress is an epidemic in British life. In nationwide polls, 89 percent of Britishers reported that they often experience high levels of stress, and 59 percent claimed that they feel great stress at least once a week (Hellmich, 1994, 57). A five year study of the British workforce conducted by the Families and Work Institute showed that 30 percent of employees often or very often feel burned out or stressed by their jobs, 27 percent feel emotionally drained from their work, and 42 percent feel used up at the end of the work day (Hellmich, 1994, 4). Balancing work pressures and family responsibilities leaves many workers feeling burned out.Examining the Effects of Downsizing on StressThe downsizing of organizations have caused a stressful environment. Downsizing has created concerns over job security, and has forced employees to take on a larger workload. According to a local union representing U.K. West stated that work still needs to be done, but with fewer people (Scott, 1996, 41). Downsizing creates quantitative and qualitative stress. Quantitative stress pertains to doing the same amount of work with fewer people. Reengineering the organization entails shaping the company to be more efficient with less individuals. These individuals are asked to do a wider variety of work functions they are not trained to do, causing qualitative overload (Scott, 1996, 35).Occupational Stress is the harmful physical and emotional response that occurs when ther e is a poor match between job demands and the capabilities, resources, or needs of the worker. Stress-related disorders encompass a broad array of conditions, including psychological disorders (e.g., depression, anxiety, post-traumatic stress disorder) and other types of emotional strain (e.g., dissatisfaction, fatigue, tension, etc.), maladaptive behaviours (e.g., aggression, substance abuse), and cognitive impairment (e.g., concentration and memory problems). In turn, these conditions may lead to poor work performance or even injury. Job stress is also associated with various biological reactions that may lead ultimately to compromised health, such as cardiovascular disease (Rosenfield, 1989, 5).PrevalenceStress is a prevalent and costly problem in todays workplace. About one-third of workers report high levels of stress. One-fourth of employees view their jobs as the number one stressor in their lives. Three-fourths of employees believe the worker has more on-the-job stress than a generation ago. Evidence also suggests that stress is the major cause of turnover in organizations (Scheier, 1985, 65).Health and Healthcare UtilizationProblems at work are more strongly associated with health complaints than are any other life stressor-more so than even financial problems or family problems. Many studies suggest that psychologically demanding jobs that allow employees little control over the work process increase the risk of cardiovascular disease. On the basis of research by the National Institute for Occupational Safety and Health and many other organizations, it is widely believed that job stress increases the risk for development of back and upper-extremity musculoskeletal disorders. High levels of stress are associated with substantial increases in health service utilization. Workers who report experiencing stress at work also show excessive health care utilization. In a 1998 study of 46,000 workers, health care costs were nearly 50% greater for workers repo rting high levels of stress in comparison to low risk workers. The increment rose to nearly 150%, an increase of more than $1,700 per person annually, for workers reporting high levels of both stress and depression. Additionally, periods of disability due to job stress tend to be much longer than disability periods for other occupational injuries and illnesses (Schwartz, 1993, 58).Causes of Occupational StressJob stress results from the interaction of the worker and the conditions of work. Views differ on the importance of worker characteristics versus working conditions as the primary cause of job stress. The differing viewpoints suggest different ways to prevent stress at work. According to one school of thought, differences in individual characteristics such as personality and coping style are most important in predicting whether certain job conditions will result in stress-in other words, what is stressful for one person may not be a problem for someone else. This viewpoint lead s to prevention strategies that focus on workers and ways to help them cope with demanding job conditions. Although the importance of individual differences cannot be ignored, scientific evidence suggests that certain working conditions are stressful to most people. Such evidence argues for a greater emphasis on working conditions as the key source of job stress, and for job redesign as a primary prevention strategy. Personal interview surveys of working conditions, including conditions recognized as risk factors for job stress, were conducted in Member States of the European Union in 1990, 1995, and 2000. Results showed a trend across these periods suggestive of increasing work intensity. In 1990, the percentage of workers reporting that they worked at high speeds at least one-fourth of their working time was 48%, increasing to 54% in 1995 and to 56% in 2000. Similarly, 50% of workers reported they work against tight deadlines at least one-fourth of their working time in 1990, incr easing to 56% in 1995 and 60 % in 2000. However, no change was noted in the period 19952000 (data not collected in 1990) in the percentage of workers reporting sufficient time to complete tasks. A substantial percentage of Britishers work very long hours. By one estimate, more than 26% of men and more than 11% of women worked 50 hours per week or more in 2000. These figures represent a considerable increase over the previous three decades, especially for women. According to the Department of Labour, there has been an upward trend in hours worked among employed women, an increase in extended work weeks (40 hours) by men, and a considerable increase in combined working hours among working couples, particularly couples with young children (Shaw, 1993, 4).Signs of Occupational StressMood and sleep disturbances, upset stomach and headache, and disturbed relationships with family, friend and girl/boy friends are examples of stress-related problems. The effects of job stress on chronic dis eases are more difficult to see because chronic diseases take a long time to develop and can be influenced by many factors other than stress. Nonetheless, evidence is rapidly accumulating to suggest that stress plays an important role in several types of chronic health problems-especially cardiovascular disease, musculoskeletal disorders, and psychological disorders (Sherer, 1982, 36).PreventionA combination of organizational change and stress management is often the most useful approach for preventing stress at work.How to Change the Organization to Prevent Job StressEnsure that the workload is in line with workers capabilities and resources.Design jobs to provide meaning, stimulation, and opportunities for workers to use their skills.Clearly define workers roles and responsibilities.Give workers opportunities to participate in decisions and actions affecting their jobs.Improve communications-reduce uncertainty about career development and future employment prospects.Provide opport unities for social interaction among workers.Establish work schedules that are compatible with demands and responsibilities outside the job.St. Paul Fire and Marine Insurance Company conducted several studies on the effects of stress prevention programs in hospital settings. Program activities included (1) employee and management education on job stress, (2) changes in hospital policies and procedures to reduce organizational sources of stress, and (3) establishment of employee assistance programs. In one study, the frequency of medication errors declined by 50% after prevention activities were implemented in a 700-bed hospital. In a second study, there was a 70% reduction in malpractice claims in 22 hospitals that implemented stress prevention activities. In contrast, there was no reduction in claims in a matched group of 22 hospitals that did not implement stress prevention activities (Smith, 1981, 24).Chapter Three Research MethodologyDesignThe data reported here are from two sep arate data sets. In order to cross-validate the model, data from the original study of managerial women were used, and these included data from the first three assessments of a 2-year longitudinal study. In the original article (B. C. Long et al., 1992, 165), a conceptual model of stress and coping was tested and developed that was based on data from the first three assessments (Time 1 to Time 3) of 11 assessments completed over 2 years. Status, Sex Role Attitudes, and Agentic Traits were assessed at Time 1 Appraisals, Disengagement and Engagement Coping, Work Environment, and Daily Hassles were assessed at Time 2 and Distress and Satisfaction were assessed at Time 3. These data were used as a base to test the validity of the model on a new set of data obtained from clerical workers, data that have not been reported elsewhere (Snapp, 1992, 32).Sampling The managerial women (n = 249) were employed in nontraditional occupations (i.e., fewer than 35% of British employees are women). Th eir mean age was 38.84 years (SD = 7.68, range = 2266). More detailed descriptions of the managers characteristics can be found in B. C. Long et al. (1992).The clerical workers who participated were employed in both large and small organizations in the same large western British city in which the managers were employed. The clerical workers volunteered in response to written requests for participants that I circulated in the media and by networking. The notices were directed to full-time female clerical workers and indicated that the purpose of the study was to investigate how clerical workers experienced Occupational Stress. No incentives were offered other than a final summary report. f the 284 respondents who made contact by telephone, 273 met the criteria for inclusion (i.e., they were employed in a clerical position, worked more than 20 hours per week, and did not supervise others). f the 273 clerical workers who met the criteria and were distributed questionnaires at Time 1, 3 9 withdrew from the study because of lack of time to participate, 7 no longer met our criteria because of promotion, unemployment, or leave of absence from work (e.g., due to accident or illness), and 4 moved. The overall dropout rate was 18%. Dropout analyses were conducted on the demographic variables measured at Time 1. No differences were found between the retained (n = 223) and dropout (n = 50) respondents. Chi-square analyses of the demographic variables (marital status, education, number of children, job level, and size of the company) were not significant. Because 9 participants identified a personal rather than a work stressor, their data were omitted from the model testing.All respondents were self-identified clerical workers. Job classifications included clerks (25%), secretariesstenographers (23%), administrative assistants (34%), and others (18%). The mean age was 39.77 years (SD = 9.46, range = 2263 years). Fifty-three percent of the clerical workers were married, 22% were single, and 25% were divorced, separated, or widowed. Fifty-three percent were parents. Twenty-four percent had a high school education or less, 42% had special training (e.g., secretarial, clerical), 17% had a college education (2 years postsecondary), and 13% had a university degree. Household incomes ranged from less than $25,000 (British) per year (23.4%) to over $61,000 (British) per year (27.5%). The major industries represented were education (31%), service (35%), utilities and public administration (12%), manufacturing and transportation (10%), and other (8%). On average the women had been in the workforce for 17.02 years (SD = 8.74,

Polymerase Chain Reaction (PCR) Steps

Polymerase Chain answer (PCR) StepsWe owe the disco rattling of the polymerase chain response to Kary B Mullis in the year 1983. He was the actual index of PCR. Few people atomic number 18 aw atomic number 18 that in 1971, Kleppe and the Nobel laureate Gobind Khorana published studies including a description of techniques that ar now known to be the basis for nucleic acidulous replication. However, it is unfortunate that Kleppe and Khorana were ahead of their times. Oligonucleotide synthesis wasnt as simple as it is today genes had non been seasond and the idea of ther virtuallyable deoxyribonucleic acid polymerases had non been described. Hence, the credit for discovering the PCR remains with Kary Mullis.The Polymerase Chain Reaction is essentially a cell-free method of deoxyribonucleic acid and RNA cloning. The deoxyribonucleic acid or RNA is isolated from the cell and replicated upto a million times. At the end, what you shake up is a greatly amplified fragment of de oxyribonucleic acid. The PCR is quick, reliable and sensitive and its variations fork out made it the basis of genetic testing.WHAT KARY B MULLIS SAYS ABOUT HOW HE DISCOVERED THE POLYMERASE CHAIN chemical reactionI was just driving and thinking about ideas and suddenly I saw it. I saw the polymerase chain reaction as clear as if it were up on a blackboard inside my head, so I pulled over and started scribbling. A chemist friend of his was asleep in the car. Mullis says that Jennifer objected groggily to the delay and the light, unless I exclaimed I had discovered something fantastic. Unimpressed, she went back to sleep.Mullis kept scribbling calculations, right in that respect in the car. He convinced the small California biotech company, Cetus, he was working for at that time, that he was up to something big. They finally listened. They sold the patent of PCR to Hoffman-LaRoche for a staggering $300 million the maximum meter of money ever paid for a patent. Mullis meanwhile re ceived a $10,000 bonus.BASIS OF THE METHODThe purpose of PCR is to generate a huge number of copies of a segment of deoxyribonucleic acid, which could be a gene, a portion of a gene, or an intronic constituent. There argon tercet major steps in a PCR, which argon repeated for 30 or 40 cycles. This is done on an automated cycler, which bed either heat or cool the tubes containing the reaction variety show, as required, in a very short peak of time. There atomic number 18 three major steps in a PCR, which atomic number 18 repeated for 30 or 40 cycles.DenaturationDuring this process, the reiterate baseed DNA melts and opens to form single stranded DNA. every enzymatic reactions, such as those carried over from a previous cycle, stop. This will be explained in the next paragraph. The temperature for denaturation is non fixed but it usually buy the farms at about 95C. It is central to realize that the denaturation temperature is largely dependent on GC (guaninecytosine) con tent of the DNA fragment to be analyzed. This is reasonable when one considers that the GC bond is a triple hydrogen bond and the AT bond is a double bond. Logic dictates that a triple bond should be 1.5 times harder to break than a double bond. Therefore, when the segment of DNA to be analyzed has a very lavishly GC content, the denaturation temperature idler r to each one even upto 99C.AnnealingThis requires temperatures lower than those required for denaturation. In this process, the fuzees season to that very specific segment of DNA that is to be amplified. The basiss are jiggling around, ca employ by the Brownian motion. Ionic bonds are constantly formed and broken between the single stranded soil and the single stranded template. The more(prenominal) stable bonds last a little bit longer ( grounds that fit exactly) and on that little piece of what is now double stranded DNA (template and soil) the polymerase mountain attach and starts copying the template. Once the re are a few bases built in, the bonce bond is so strong between the template and the fuzee, that it does not break anymore.ExtensionThis is done at 72C. This is the ideal temperature for working with polymerase. The primers, which are complementary to the template, already have a strong ionic attraction to the template. This military force is stronger than the forces breaking these attractions i.e. the luxuriously temperature. Primers that are on positions with no exact match (non complementary) get loose again (because of the last temperature) and dont give an extension of the fragment. The nucleotide bases are added from the 5 end to the 3 end. The phosphate group of the dNTPs is coupled with the hydroxyl group of the extending DNA strand. The extension time depends on 2 factors the type of polymerase employ and the length of the DNA fragment to be amplified. Usually, Taq polymerase adds dNTPs at the rate of about 1000 bases per minute.It is important to realize that each component of the PCR including the input DNA, the oligonucleotide primers, the thermostable polymerase, the buffer and the make pass parameters has a profound impact on the sensitivity, specificity and fidelity of the reaction.The three steps of the first cycle are shown, that is, denaturation, annealing and extension. At the end of the first cycle, two strands have been synthesized. At the end of the second cycle, four strands have been synthesized (the three steps of the cycle have not been shown). At the end of the third cycle, 8 strands have been synthesized. The number of strands increases exponentially with each cycle.NuggetsThe Polymerase Chain Reaction is essentially a cell-free method of cloning DNA and RNA.There are three steps involved in every cycle these are denaturation, annealing and extension.At the end of each cycle, the DNA doubles. Therefore, theoretically, if there are n cycles in a reaction, the number of DNA fragments at the end of the reaction will be 2n.CO MPONENTS OF THE POLYMERASE CHAIN REACTIONThe components that are essential for a successful PCR are elaborated here.TEMPLATE DNAThis is that portion of the DNA/gene that is to be amplified. Usually the concentration is100 ng genomic DNA per PCR reaction. However, this can vary depending on the target gene concentration and the source of DNA. The PCR reaction is inherently sensitive. It is not necessary for the template DNA to be abundant or highly purified. high amounts of template DNA can increase the yield of nonspecific PCR products, but if the fidelity of the reaction is crucial, one should limit both template DNA quantities as well as the number of PCR cycles.DNA in solution may contain a large number of contaminants. These contaminants may inhibit the PCR. few of these reagents are phenol, EDTA, and protease K, which can inhibit Taq DNA polymerase. However, isopropanol precipitation of DNA and washing of DNA pellets with 70% ethanol is usually effective in removing traces o f contaminants from the DNA sample.Effects of FixationThis is of particular interest to the pathologist since he has to deal with formalin fixed tissue. DNA extracted from fresh tissue or cell suspensions forms an optimal template for PCR. The tissue is best stored at -70C at which the nucleic acids can be stored indefinitely. A temperature of -20C is sufficient to preserve the DNA for several months and at 4C, the DNA can be stored for several weeks. At room temperature, the DNA has been successfully stored for hours to days however, mitochondrial DNA is very sensitive to temperature and may shame in thawed tissues.DNA extracted from fixed tissue has been used successfully for PCR. The type of fixative and the duration of fixation are of critical importance. Non crosslinking fixatives homogeneous ethanol grant the best DNA. Formaldehyde is variable in its DNA yield. Carnoys, Zenkers and Bouins are poor fixatives as far as DNA preservation is absorbed.Not surprisingly, formalde hyde is the fixative which has been evaluated the most, because it is more commonly used world round-eyed. The studies have demonstrated that a successful PCR depends on the protocol to extract the DNA and the length of fixation. Formaldehyde reacts with DNA and proteins to form reactive hydroxymethyl talk termss which give rise to a salmagundi of end products which include DNA-DNA and DNA-protein adducts. Purification of DNA from formalin fixed tissue, therefore, includes heating to reverse the hydroxymethyl additions and treatment with a proteinase to hydrolyze the covalently associate proteins. However, there is no way to reverse the DNA-DNA links and these links inhibit the DNA polymerases. This accounts for the low PCR yield which is seen with formalin fixed tissue. Usually, the PCR reaction with formalin fixed DNA as a template yields products which are not more than 600 bp in size.NuggetsTemplate DNA is required in a concentration of 100ng for each PCR reaction. Contamina nts in DNA may inhibit the reaction.Fixation of tissues brooks DNA which is not as good as DNA obtained from fresh/ polar tissues.Different fixatives give different DNA yields. Alcohol is the best fixative and Carnoys, Zenkers and Bouins are poor fixatives as far as DNA preservation is concerned. Formalin is intermediate in DNA yield.Purification of DNA from formalin fixed tissue involves heating to reverse the attachment of hydroxymethyl intermediates and treatment with a proteinase to hydrolyze the covalently linked proteins.The DNA obtained afterward fixation can be used for reactions in which the PCR product is not more than 600 bp.PCR BUFFERThe purpose of using buffers in PCR is to provide optimum pH and potassium ion concentration for the DNA polymerase enzyme (usually obtained from bacteria Thermus aquaticus) to function. Most buffers are available in a 10X concentration and require dilution before use. Although most protocols recommend the final buffer concentration of 1X , a concentration of 1.5X might result in increased PCR product yield.The PCR buffer contains many components. Some important ones are discussed hereDivalent and monovalent cations These are required by all thermostable DNA polymerases. Mg2+ is the divalent cation that is usually present in most of the PCR buffers. Some polymerases also work with buffers containing Mn2+. Calcium containing buffers are ineffective and therefore, rarely used. Buffers can be divided into first and second generation buffers on the basis of their ionic component. The second generation buffers, as opposed to first generation buffers, also contain (NH4)2SO4 and permit consistent PCR product yield and specificity over a wide range of magnesium concentration (1.0 to 4.0 mM MgCl2). The overall specificity and yield of PCR products is better with second generation buffers, as compared with first generation PCR buffers. Buffers also contain KCl. Salts identical KCl and NaCl may help to facilitate primer annea ling, but concentration of 50 mM will inhibit Taq polymerase activity. Interactions between K+ and NH4+ allow specific primer hybridization over a broad range of temperatures. Magnesium is one of the most important components of the buffer. Mg2+ ions form a soluble complex with dNTPs which is essential for dNTP internalisation they also stimulate polymerase activity and influence the annealing efficiency of primer to template DNA. The concentration of MgCl2 can have a dramatic effect on the specificity and yield of PCR products. Optimal concentration of MgCl2 is between 1.0 to 1.5 mM for most reactions. Low MgCl2 concentration helps to eliminate non-specific priming and formation of background PCR products. This is desirable when fidelity of DNA synthesis is critical. At the same time, however, too few Mg2+ ions can result in low yield of PCR products. High MgCl2 concentration helps to stabilize interaction of the primers with their mean template, but can also result in nonspecifi c binding and formation of non specific PCR products. It is important to be aware that many PCR buffers (often sold in 10X stocks) already contain some amount of MgCl2. Therefore, the addition of further amounts must be carefully monitored. In the best possible scenario, the PCR would work well with the amount of Mg2+ already present in the buffer solution. However, if this does not occur, it is necessary to standardize the amount of Mg2+ in the reaction mix. This can be difficult because the dNTPs and the oligonucleotide primers bind to Mg2+. Therefore, the milling machinery concentration of Mg2+ must exceed the molar concentration of the phosphate groups contributed by dNTPs and the primers. As a rule of thumb, the magnesium concentration in the reaction mixture is generally 0.5 to 2.5 mM greater than the concentration of dNTPs. The optimal concentration of Mg2+ should, therefore, be standardized for each reaction.Tris-Cl The concentration of tris-Cl is adjusted so that the pH o f the reaction mixture is represented between 8.3 and 8.8 at room temperature. In standard PCR reactions, it is usually present in a concentration of 10mM. When incubated at 72C which is the temperature for extension, the pH of the reaction mixture falls by more than a full unit, producing a buffer whose pH is 7.2.Other components Some buffers also contain components wish BSA (Bovine serum albumin) and DMSO (dimethyl sulphoxide). BSA reduces the amount of template sticking to the side of the tube, making it available for elaboration and reducing the risk of primer dimer. Primer dimers are products obtained when the primers anneal to each other instead to to the template DNA. DMSO has been shown to facilitate DNA strand separation (in GC rich difficult secondary structures) because it disrupts base pairing and has been shown to improve PCR efficiency.In effect, it is wise not to tamper with the buffer provided with the Taq polymerase. The buffer is usually standardized for the vi al of Taq and there is no need to add additional MgCl2 or stabilizers like DMSO and BSA. However, some Taq buffers come with the buffer in one vial and MgCl2 in a separate vial. Under such circumstances, it is wise to start with 1L of MgCl2 and increase its concentration in aliquots of 0.5 L, if the initial reaction fails.NuggetsThe PCR buffer contains divalent and monovalent cations, Tris Cl and other components.The PCR buffer is used to give the correct pH and potassium concentration for the DNA polymerase to function.The most common divalent ion used is magnesium in the form of MgCl2. MgCl2 concentration is vital for PCR.Tris Cl is used to maintain the pH between 8.3 and 8.8 at room temperature.Salts like NaCl and KCl may facilitate primer annealingOther components like BSA and DMSO help to increase the sensitivity and specificity of the reaction.OLIGONUCLEOTIDE PRIMERSWhat are Oligonucleotide Primers?PCR primers are short fragments of single stranded DNA (17-30 nucleotides in l ength) that are complementary to DNA durations that flank the target region of interest. The purpose of PCR primers is to provide a free 3-OH group to which the DNA polymerase can add dNTPs.There are two primers used in the reaction. The forward primer anneals to the DNA minus strand and directs synthesis in a 5 to 3 direction. The installment of primers is always represented in a 5 to a 3 direction. The reverse primer anneals to the other strand of the DNA.How to design a primer?The predominant goal kept in mind while designing a primer is specificity. apiece member of the primer must anneal in a stable fashion to its target sequence in the template DNA. The longer the primer, the high is its specificity. Unfortunately, the longer the primer, the less likely it is to anneal to a particular sequence in the template DNA. Conversely, if the primer length is small, it is likely to anneal, but its specificity will be poor. A compromise is reached by designing primers between 20 and 25 nucleotides long. Inclusion of less than 17 nucleotides often leads to non specific annealing, while presence of more than 25 nucleotides may not allow annealing to occur at all.Remember that the DNA sequence in the human genome appears to be a stochastic sequence of nucleotides. When designing primers, it is important to calculate the probability that a sequence exactly complementary to a string of nucleotides in the human genome will occur by chance. Several formulae are designed to calculate such probabilities. However, mathematical expressions are not necessarily correct and in this case, the predictions maybe wildly prostitute. The distribution of codons is non random with repetitive DNA sequences and gene families. It is advisable to use primers longer than the statistically indicated minimum. It is also advisable to scan DNA databases to tot if the proposed sequence occurs only in the desired gene.For a practicing pathologist, it is best not to attempt designing of prim ers. What a pathologist requires is the primer sequence for an established test. If, for example, a pathologist requires primer sequence for the diagnosis of sickle cell anemia, all he has to do is search the web for papers related to molecular testing of sickle cell anemia. The primer sequences will be provided in the paper. Custom made primers can be commercially synthesized. Several biotechnology companies provide this facility. to begin with the primers are ordered, it is essential to check that the sequence is correct and that there are no missing nucleotides in the sequence. That is where, pillory is invaluable.Before the intricacies of the BLAST search are elaborated upon, it is necessary to have in mind that designing a primer does not depend only on the sequence of nucleotides. Other factors like the GC content and melting point are also important considerations. They will be dealt with later in the chapter.BLAST and its usesBLAST is an acronym for Basic Local Alignment Search Tool. It is an algorithm comparing development about primary biological sequences with a library or database of sequences.A BLAST can be performed for different organisms, but in this book, we will concern ourselves with nucleotide BLAST in humans only. BLAST searches the database for sequences similar to the sequence of interest (the query sequence) by using a 2-step approach. The basic concept is that the higher the number of similar segments between two sequences, and the longer the length of similar segments, the less divergent the sequences are, and therefore, likely to be more genetically related (homologous).Before perfoming a BLAST search the oligonucleotide sequence is first identified. The sequence is fed into the programme. BLAST first searches for short regions of a given length called rowing (W). It whence searches for substrings which are compared to the query sequence. The program then aligns with sequences in the database (target sequences), using a substit ution matrix.For every pair of sequences (query and target) that have a word or words in common, BLAST extends the search in both directions to find alignments that score greater (are more similar) than a certain score threshold (S). These alignments are called high scoring pairs or HSPs the maximal scoring HSPs are called maximum segment pairs (MSPs).The BLAST search as outlined in fig 7.2 shows the results of the search. If we scroll batch further, we can see the sequences producing significant alignments. Note that in this BLAST search, there are 49 BLAST hits in the query sequence.In the list shown in figure 7.2, there is a list of hits starting with the best (most similar). To the right of the screen is the E-value. This is the expected number of chance alignments the lower the E value, the more significant the score. First in the list is the sequence finding itself, which obviously has the best score. To the left is the accession number. This refers to a unique code that ide ntifies a sequence in a database.It is important to know that there is no set cut-off that determines whether a match is significant or similar enough. This must be determined gibe to the goals of the project.The sequences provided in the figure 7.2 show a significant alignment with Pseudomonas japonica. It shows a high score (bits) and a low E-value. Note that the lower the E value, the greater the likelihood that the sequence is a good match.BLAST output can be delivered in a variety of formats. These formats include HTML, plain text edition and XML formatting. For the NCBIs web-page, the default format for output is HTML. When performing a BLAST on NCBI (National Centre for Biotechnology Information), the results are displayed in a graphical format present the followingThe hits foundA tabular form showing sequence identifiers for the hits with scoring related dataAlignments for the sequence of interest and the hits received with corresponding BLAST scores for these.The easiest to read and most informative of these is probably the table. The main idea of BLAST is that there are often high-scoring segment pairs (HSP) in a statistically significant alignment. BLAST searches for these high scoring sequence alignments between the query sequence and the sequences in the database. The speed and relatively good accuracy of BLAST are among the key technical innovations of the BLAST programs.Sequence of events to be followed when performing a BLAST search.Go to PUBMED (http//www.ncbi.nlm.nih.gov/pubmed/)Scroll down to reach a heading called best-selling(predicate)Under POPULAR click on BLASTClick on nucleotide blastUnder the heading, enter accession number(s), gi(s), or FASTA sequence(s), type or paste the sequence that you want matched.Click BLASTWait for the results. Analyse the nucleotide sequence as it appears.Calculation of Melting TemperatureThe melting temperature or Tm is a mea sure as shooting of stability of the duplex formed by the primer and the compl ementary target DNA sequence and is an important consideration in primer design. Tm corresponds to the midpoint in transition of DNA from the double stranded to its single stranded form. A higher Tm permits an increased annealing temperature that makes sure that the annealing between the target DNA and the primer is specific. The Tm is dependent on the length of the oligonucleotides and the G+C content of the primer. The formula for calculation of Tm is given in table 7.1.Table 7.1 Formula for calculation of the melting temperature.Length of PrimerTm (C)Less than 20 nucleotides long2(effective length*)20 to 35 nucleotides long22 + 1.46(effective length)*Effective length = 2(number of G+C) + number of (A + T)Primers are usually designed to stay off matching repetitive DNA sequences. This includes repeats of a single nucleotide.. The two primers in a PCR reaction are not homologous to each other and their complementarity can lead to formation of spurious amplification artifacts calle d primer dimers. The 3 end of a primer is most critical for initiating polymerization.The rules for selecting primers in addition to those already mentioned are as followsThe C and G nucleotides should be distributed uniformly throughout the primer and comprise approximately 40% of the bases. More than three G or C nucleotides at the 3-end of the primer should be avoided, as nonspecific priming may occur.The primer should be neither self-complementary nor complementary to any other primer in the reaction mixture, in order to avoid formation of primer-dimer or hairpin-like structure.All possible sites of complementarity between the primer and the template DNA should be noted.The melting temperature of flanking primers should not differ by more than 5C. Therefore, the G+C content and length must be chosen accordingly (a higher G+C content means a higher melting temperature).The PCR annealing temperature (TA) should be approximately 5C lower than the primer melting temperature.G+C cont ent in each primer should not be more than 60% to avoid formation of internal secondary structures and long stretches of any one base.Primer extension will occur during the annealing step. Primers are always present in an excess concentration in conventional (symmetric) PCR amplification and, typically, are within the range of 0.1M to 1M. It is generally advisable to use purified oligomers of the highest chemical integrity.Primer DimersA Primer Dimer (PD) consists of primer molecules that have attached or hybridized to each other because of strings of complementary bases in the primers. As a result, the DNA polymerase amplifies the PD, leading to competition for PCR reagents, thus potentially inhibiting amplification of the DNA sequence targeted for PCR amplification.In the first step of primer dimer formation, two primers anneal at their respective 3 ends. The DNA polymerase will bind and extend the primers. In the third step, a single strand of the product of step II is used as a template to which fresh primers anneal leading to synthesis of more PD product.Primer dimers may be visible after gel electrophoresis of the PCR product. In ethidium bromide stained gels, they are typically seen as 30-50 base-pair (bp) bands or smears of moderate to high intensity. They can be easily elevated from the band of the target sequence, which is typically longer than 50 bp.One approach to prevent PD formation consists of physical-chemical optimization of the PCR system, i.e., changing the concentration of primers, MgCl2, nucleotides, ionic aptitude and temperature of the reaction. Reducing PD formation may also result in reduced PCR efficiency. To overcome this limitation, other methods aim to reduce the formation of PDs only. These include primer design, and use of different PCR enzyme systems or reagents.NuggetsOligonucleotide primers are short fragments of single stranded DNA (17-30 nucleotides in length) that are complementary to DNA sequences that flank the target r egion of interest. They dictate which region of DNA in the PCR will be amplified.Primer sequences can be obtained by reviewing previously published literature. A confirmation of the sequence can be done by using BLAST (Basic Local Alignment Search Tool).The melting temperature is the midpoint in the observed transition from a double stranded to a single stranded form. A higher annealing temperature ensures that the annealing between the target DNA and the primer is specific.A primer dimer consists of primer molecules that have attached or hybridized to each other because of strings of complementary bases in the primers. Taq polymerase amplifies the primer dimer leading to competition for the PCR products.Several methods are used to reduce primer dimer formation including changing the concentrations of primers, MgCl2, nucleotides, ionic strength and temperature of the reaction.TAQ DNA POLYMERASEThe initial PCR reaction used the Klenow fragment of Escherichia coli DNA polymerase. Howe ver, this was unstable at high temperatures and it was necessary to add a fresh aliquot of enzyme after every denaturation step. The annealing and extension temperatures had to be kept low and as a result, there was formation of non specific products in abundance. The discovery of the thermostable Taq DNA polymerases ensured that the PCR did not remain a laboratory curiosity. The extension and annealing temperatures could now be kept high and the formation of non specific products was greatly reduced.Taq became famous for its use in the polymerase chain reaction and was called the Molecule of the Year by the journal Science.Why Taq?Taq is the enzyme of choice in PCR because of the following reasonsTaq works best at 75C80C, allowing the elongation step to occur at temperatures which make non-Watson-Crick base pairing a rare event.It can add upto 1,000 nucleoside triphosphates to a growing DNA strand.Taq has a half-life of 40 minutes at 95C and 9 minutes at 97.5C, and can replicate a 1000 base pair strand of DNA in less than 10 seconds at 72C.Because of all these properties, Taq is the enzyme of choice in the PCR.How does Taq polymerase act?The first requirement is a primer. The primer is annealed to the template strand having free hydroxyl group at its 3 end. During the extension phase, the Taq synthesizes a newfangled DNA strand complementary to the template by adding dNTPs in a 5 to 3 direction condensing the 5 phosphate group of the dNTPs with the 3 hydroxyl group of the end of the extending DNA strand. Since Taq works best between 70C- 80C, a temperature of 72C is usually chosen as the optimum annealing temperature.Where does Taq come from?In Thermus aquaticus, Taq polymerase is expressed at very low levels and commercial production is not economically viable. However, the enzyme can now be produced from different versions of the engineered Taq gene so as to obtain high levels of expression in E coli.What other polymerases are available for use in PCR?Taq is not the only polymerase other polymerases are available but Taq is the one that is generally used in a PCR. A few other polymerases with their uses are as followsPFU DNA polymerase -Found in Pyrococcus furiosus, it functions in vivo to replicate the organisms DNA. The main deviation between Pfu and alternative enzymes is the Pfus superior thermostability and proofreading properties compared to other thermostable polymerases. Unlike Taq DNA polymerase, Pfu DNA polymerase possesses 3 to 5 exonuclease proofreading activity, meaning that it works its way along the DNA from the 3 end to the 5 end and corrects nucleotide-misincorporation errors. This means that Pfu DNA polymerase-generated PCR fragments will have fewer errors than Taq-generated PCR inserts. As a result, Pfu is more commonly used for molecular cloning of PCR fragments than the historically popular Taq. However, Pfu is slower and typically requires 1-2 minutes to amplify 1kb of DNA at 72 C. Pfu can also be used in conju nction with Taq polymerase to obtain the fidelity of Pfu with the speed of Taq polymerase activity.TFL DNA polymerase Obtained from Thermus flavus, it is useful for the amplification of large segments of DNA.WHAT IS FIDELITY?All DNA polymerases have an intrinsic error rate that is highly dependant on the buffer composition, pH of the buffer, dNTP concentration and the sequence of the template itself. The types of errors that are introduced are frameshift mutations, single base pair substitutions, and spontaneous rearrangements. Therefore, the PCR reaction generates a product that is very similar, but in many cases, not identical to the original sequence. The measurement of dissimilar product obtained is obviously related to the cycle in which the mismatch took place. Under normal circumstances, this does not make any difference however, these errors may plough significant during sequencing when the role of fidelity comes into play.Fidelity is the ability of the polymerases to avo id the incorporation of wrong nucleotides during the reaction. Under normal circumstances, it really does not make a difference if a wrong nucleotide is incorporated because the size of the PCR product remains the same and that is what we have to look for. However, there are some polymerases like Pfu which have a high fidelity. In addition to reading from the 5 to the 3 direction, they can also read from the 3 to the 5 direction and correct the wrong nucleotides wh