Pentagon Museum of Art Essay Example | Topics and Well Written Essays - 3250 words

Pentagon Museum of Art - Essay Example Place and time are both texts. So my personal Pentagon Museum of Art is both a place and a long sequence in time. My hiding place is located on the roof of a Midtown Manhattan building. It is a light blue pentagon-shaped construction with a yellow little door as its only entrance. I retreat in that small space of paradise during my off hours. After work in an ad agency as a copywriter, I come over here to relax, to take a nap, to resume writing my first novel, or to take a look at its surrounding white-painted walls each one holding in its surface a masterpiece work of art by Salvador Dal and Pablo Picasso. They are my favorite painters so I placed 5 of their paintings on the five walls of my private Pentagon Museum of Art right on top of a metropolitan apartment building in New York. My Pentagon is an arts Pentagon, not a military one. I find myself at peace with my own self here. It's a metalinguistic feeling that I enjoy to the max. I can lye on the bed. I can write my novel. I can watch closely at the paintings. I can read a book. I can listen to music. My favorite music. My favorite books. Intertextuality. Intratextuality. Extratextuality. All of them merge within my own self. Intertextuality among the paintings. Intratextuality inside my novel. Extratextuality with the surrounding environment that feeds my urgings for creating a different and distinct world of my own. The text becomes a sign, and according to C. S. Peirce the sign can be an icon, a symbol, or an index. Intertextuality can be an icon, intratextuality an index, and extratextuality a symbol. I have decided to make everything in all out of my hiding place. My Pentagon Museum of Art. My little piece of paradise on Earth. Here there is no place for luxury. Everything is common and simple. There is a fridge by the door. Five windows on each side of the pentagon. There is a computer table in the middle of the room with its corresponding PC on top. Five paintings of two master artists of all time: Salvador Dal and Pablo Picasso. There is a small bed by the top vortex, between the two top sides of the pentagon. A nice sound equipment with the Sony trademark right by the side of the computer table. There is an executive rotating chair in front of the computer table. Here everything is interconnected. That's part of the hyperlinking of the objects. One leads to the other. I can go in a matter of seconds from the radio to the computer, from the chair right onto the bed. And I can jump from my bed in order to take a fresh look at one of the Dal's Christs. Texts are interwoven in my mind as well as in this little retreat of comfort and peace. Every text has its rhythm, and I find meaning in the rhythm of this small building with five walls and a quiet sound of the music in my ears. That is Bob Dylan playing with The Traveling Wilburys. At the End of the Line. Yes. At the very end of the line I find a melody filled with harmony in the creativeness of my life. I find meaning in each and every white wall with the paintings hanging like sovereign pieces of wonderful signs. Picasso and Dal share with me their meaningful creatures of their minds. Dylan does his part beautifully singing "Blowing in the Wind". "Like a Rolling Stone" is coming up after that tune. I keep writing on my clone PC. My first novel is taking

Economic Issues Simulation Paper Essay Example for Free

Economic Issues Simulation Paper Essay The Financing of Health Care Economic Issues Simulation Paper Health care system has evolved tremendously in the last few years, with many changes with the health care laws including but not limited to Universal Health Care, many individuals have choices when it comes to their coverage. According to healthcare. gov, in January of 2015, an employer with 50 or more full time employees will have to make an Employer Shared Responsibility Payment if a full time employee gets a lower health coverage premium cost if insurance is purchase in a marketplace. However, employers are not subject to this law if the numbers of employees are lesser than 50 but are still expected to offer coverage for their employees. (healthcare. gov) Employers must make sure that when choosing coverage for their employees, these should be within their needs; within health care requirements as well as inexpensive keeping in mind that lower cost may not necessarily mean better. With many varieties in health care plans such as Preferred Provider (PPO), Point of Service (POS), and Exclusive Provider Organization (EPO); the Health Maintenance Organization (HMO) is the most preferred and utilized group health insurance plan. As a HMO representative of Castor Insurance, health care coverage will be built, including the potential utilization of the services by different enrollees. Castor Collins Health Plan Castro Collins Health Plan is a regional HMO that was founded in 1999. As a HMO, they provide health care services as well as health insurance to individuals in its statewide network of physicians and hospitals utilizing a capitation model to compensate their network of providers. Currently, there are 100,000 enrollees and these numbers are increasing. The responsibility of a Vice President in Strategy and Financial Planning is to interact with new clients and formulating health plans that will suit their needs. With the help of colleagues such as the Chief Financial Officer Helen Fouerman, the Chief Medical Officer Jonathan Wikes, and the Executive Vice President of Planning and Development, Adam Hunter, a plan will be put together that will include pricing and setting insurance premiums. In January of 2006, Castro Collins was approached and met with two groups of people for health insurance coverage. These groups are Constructit and E-editors, neither of them have group employer’s insurance. Constructit have 1000 people and they are willing to pay a maximum of $4000 per person as an annual premium, meanwhile E-editors will pay a maximum annual premium of $4500 per person with 1,600 people. Castor Collins offers three types of health plans: Castor Standard, Castor Enhanced, and the customized plan called Castor Enhanced Minor. The standard plan does not cover pre-existing medical conditions, the enhanced plan, however, cover pre-existing medical conditions and offers more services. Castor Enhanced Minor is a customized plan that is almost equivalent to Castor Enhanced with somewhat lesser services that requires high utilization. Demographics  and Health Care Risk Factors There are 550 men and 450 women employees in Constructit with ages 26 to 45 and 60 percent from this age group ranging from 26 to 42 are married. This means, spouses and children need to be considered in getting health plan. Also, great physical activities are involve within thirty- two percent of the people at Constructit, while 25 percent of the people has moderate physical activity. The remainder which is 43 percent of the people involves activities that are sedentary. There are no major health risks out of the thirty-eight percent an equivalent of 170 men and 210 women in the workforce. Injuries, respiratory system diseases, digestive disorders, migraine and allergic conditions are the major causes of absenteeism in Constructit. Obesity related diseases such as hyperlipidemia, high blood pressure, diabetes, and cardiovascular diseases are moderately high for this workforce that consists of 36 percent of men and 43 percent of women. In E-editors, there are 750 men and 840 women with ages 35 to 54 and most of them are married. For the past ten years, ninety-five percent in the workforce have largely been sedentary; their job involves sitting in front of the computer for long periods of time. Only five percent are required moderate activity. Stress related injury (SRI) and problems with vision were acquired by at least 95 people who had this job for a while. There are no major medical health risks for the 170 men and 182 women (22%) from this group. Respiratory disorder is the 26 percent of the group problem because they are heavy smokers. There are 720 people who are obese in this group because of the heavy sedentary lifestyles, eating habits, and lack of exercise. This also means that there are increased risk of diabetes, hypertension, high cholesterol and cardiovascular diseases. Plan Analysis Based on the plans, I would recommend Castor Standard to Constructit and not to provide insurance to E-editors. Since Castor Standard does not cover pre-existing conditions, the risks of providing this plan are low. The premium that Constructit will be responsible annually is $3,428, with Castor Collins Health Plan earning $3. 43 million. Given the health profile and the expected utilization of services for E-editors, I think that not providing insurance is the right decision. There is a high risk of insuring this group and whatever Castor Collins earns from this group are inadequate to cover those risks. The goal is to maximize earnings. If E-editors are willing to pay a different premium they will be considered by providing an appropriate plan that minimized risks and increased earnings. Risk-averse consumers buy health insurance to avoid losing income or wealth when they are unwell. In other words, consumers pay insurance premium to transfer their risks of medical expenses to the insurance company. The premium that Castor Collins receives is a source of revenue. It is compensation for bearing risk and for bearing expenses such as payment to health care providers. If Castor Collins know that a particular group of enrollees is more susceptible to a particular disorder, its risk for providing coverage for that disorder is higher. However, if, as in the case of Constructit and E-editors, a group of individuals is not willing to pay a higher premium to cover greater risks, Castor Collins may not be able to provide coverage for greater risks. Therefore, while selecting a plan and the services to provide under that plan, Castor Collins has to weigh various considerations – the premiums that enrollees are willing to pay, the risks of providing a particular plan or service, the expected utilization and hence, the costs, of providing various  services in the plan, and the premium Castor Collins needs to charge to maintain profitability.

To Kill a Mockingbird by Harper Lee :: essays research papers

The novel To Kill A Mockingbird by Harper Lee, takes place during the 1930's in Maycomb County, Alabama. Atticus Finch, a white southern lawyer, is appointed to defend Tom Robinson, an innocent black man accused of raping a white woman. Throughout the story Atticus' children learn the meaning of true courage. Although Atticus proves Tom's innocence, the prejudice white jury's verdict is that Tom is guilty. The assumed black characteristics of immorality, dishonesty, shiftlessness and personal squalor are embodied by the white Ewell clan. This is a glaringly obvious fact that the prejudiced Maycomb society refuses to acknowledge.   Ã‚  Ã‚  Ã‚  Ã‚  Three examples of black characters who do not fit his 'stereotype'; are Reverend Sykes, Calpurnia and Tom Robinson. My first example is Reverend Sykes. He is a respected, generous man who runs a clean church and accepts worshipers both white and black. When Calpurnia brings Scout and Jem to the black church, he and the congregation welcome them. This shows that in the eyes of the Reverend, as in the eyes of God their is no prejudice. At the trial, Reverend Sykes makes room for Scout and Jem in the courtroom balcony where the blacks sit. While the trail is going on the blacks show no disrespect for Jem and Scout. After the trial is over, out of respect, the blacks wait until Atticus Finch passes and then they stand.   Ã‚  Ã‚  Ã‚  Ã‚  Secondly is the character of Calpurnia who also does not fit this stereotype. Calpurnia is the housekeeper for the Finchs and also helped raise Scout and Jem. Calpurnia is educated, hard working and well kept. She taught her children to read and Scout and Jem how to write on her own time. All that she had accomplished was done at a time when most Negroes could not read or write.   Ã‚  Ã‚  Ã‚  Ã‚  The last black character who embody the characteristics of immortality, dishonesty, shiftlessness and personal squalor is Tom Robison.. Tom is married with children and works hard to support them with a job. His house and yard are well cared for and he attends church. Not only that, but he stops at the Ewell house to help Mayella knowing that he is putting himself in a compromising position. At the trial, while on the stand, he answers questions in a respectable, dignified manner even though he is being accused of a crime he didn't commit. At one point when he is on the stand, Atticus questions him to tell the jury what Bob Ewell told him and he says, 'Somethin' not fittin' to say - not fittin' for these folk'n chillun to hear-'; (p.

Psychology Project Racism Essay

Racism People see it every day across the world. A group of whites burn down a black , someone gets hurt or murdered from a racial insult, or fights break out at school or in public. These are the extremes of racism. Racism is definitely not a good thing; it’s a power that has taken over through the present daytime. What will the future bring for Racism? What are the main problems of racism? Can we get a hold on racism throughout the world or will we self-destruct in our own hate? Racism is everywhere, as much as today tries to hide it or say that we resolved it years ago. Racism is still all over the place. Definition of Racism: Racism is the belief that a particular race is superior or inferior to another, that a person’s social and moral traits are predetermined by his or her inborn biological characteristics. Racial separatism is the belief, most of the time based on racism, that different races should remain apart from one another. Racism has existed throughout human hist ory: It may be defined as the hatred of one person by another — or the belief that another person is less than human — because of skin color, language, customs, and place of birth or any factor that supposedly reveals the basic nature of that person. Prejudice: If someone is a racist they are of course, prejudiced. Prejudice is when someone doesn’t like someone else for the way they look.. It flies right over their heads – schools, the workplace, our community, our friend’s  house, even our homes. Racist or prejudice jokes are not harmless and it is a reality today. Racist jokes are just the start of it. Many people think the jokes are funny. Maybe they are, but they still hurt the people the jokes are about. Superiority: Superiority is a big part of racism. Every race thinks that they are superior to other ,that they are the best . The power issue may make the difference, that’s one of the main reasons. Each race thinks they are a better race.. Therefore, they fight and create violence to justify who is the greater race. But the reality is, that it doesn’t matter who you are or what color you are. Forms of Racism: †¢Individual Racism takes the form of individual attitudes, beliefs, value s and behaviors. Some examples of racist attitudes are racial prejudice, belittling, jealousy and bigotry. †¢Institutional or Systemic Racism takes the form of the practices, customs, rules and standards of organizations that unnecessarily disadvantage people because of their race, color or ethnicity. †¢Cultural Racism is the cultural values and standards that disadvantage people because of their race, color or ethnicity. . Bio-psycho-social-spiritual phenomenon of racism: Racism is the outward manifestation of an inward system. This system justifies power of position by placing a negative meaning and value on perceived or actual biological/cultural differences. This system gives biological differences, such as skin color, texture of the hair, physical features; or cultural differences, such as language, religion, ethnicity, or accent, a negative value and meaning. In real they are merely biological or cultural necessities. Thus, there is nothing wrong with the color black, brown or yellow. It is not skin color that forms the basis for discrimination, but the negative meaning and value given to the color of skin. â€Å"Color is neutral; it is the mind that gives it meaning.† Neither are women discriminated against because of their gender. If gender were the problem then the solution would also be to have a sex-change operation. But the problem is not gender but value systems.. Women are discriminated against because of the negative meaning and value given to their gender. How one defines a situation depends on how one perceives it. That’s the psycho pattern. The meaning that  people therefore give to their reality, whether or not true, causes people to behave in a manner that makes the original meaning actually come true. The interpretive meaning called â€Å"perception.†. The same goes with the spiritual dimension, the fourth factor. In order to justify such evil use of power, people will appeal to those moral systems which give them a sense of meaning, rightness, and ultimate value in life, their ideological belief systems, which serve as the highest authority in thei r lives, the moral basis of their existence. Like this the bio-psycho-social-spiritual framework works -and how it shapes our world. Common factors of Racism: †¢Color: Prejudice or discrimination in which human beings are treated differently based on the social meanings attached to skin color †¢Religion: Religious discrimination involves treating a person (an applicant or employee) unfavorably because of his or her religious beliefs. Religions, such as Buddhism, Christianity, Hinduism, Islam, and Judaism, but also others who have sincerely held religious, ethical or moral beliefs. †¢Gender: Granting or denying rights or privileges to a person based on their gender. No Boundaries: Victims and perpetrators of racial prejudice are not confined to any period in history, place, skin color, income group or other boundary. No one is immune! But the images we associate with the term racism depend largely on the part of the world in which we live. For instance, in different countries most commonly think of tension between whites and blacks, or increasingly, Hispanics and blacks or whites. Further back, though, other forms existed. Racism –different countries: Racism world-wide is manifested in every continent. In Europe, extreme racist groups have gained more and more acceptability in countries like France, Germany, and Austria. In Australia, the Aborigines have lost much of their land and have been the victim of extreme prejudice. Not only has Apartheid — legalized separation — been practiced in Australia, it has been a well-publicized condition in South Africa. Many African countries have endured long-term civil wars, some beginning shortly after that nation’s freedom from former colonial  countries. In Asia, Cambodians harbor extreme prejudices against the Vietnamese, and Chinese students have been forbidden access to higher education in Malaysia. In the Middle East, antagonism between the Israelis and the Palestinians continues to exist. Racism in Pakistan: In Pakistan people do face problem regarding racism and its done on the following bases. †¢Socio economic status †¢Gender †¢Color †¢Religion †¢Cast †¢Provincial Causes of racism: Major causes of racism common in all societies are: †¢Lack of self love †¢feeling of unworthiness †¢desire to feel superior †¢Fear of loss and displacement †¢lack of understanding †¢false perceptions †¢Ignorance, †¢Xenophobia †¢Brainwashing †¢ upbringing, †¢negative experiences with individuals of a given race, †¢negative images portrayed in media and societal stereotypes, †¢hate †¢ lack of education, †¢distorted thinking †¢ fear †¢Selfishness †¢Environmental Factors Effects of racism: It is said that â€Å" Racism always breaks a country and spoils a society.† The effects that are commonly observed are: †¢Self hate †¢ Suicide †¢Self harm issues †¢Eating disorders †¢Fear of going to school/public places (agoraphobia) †¢Bullying †¢Knife and gun crime †¢Panic attacks †¢Violence †¢hate crimes †¢inequality †¢inhumanity †¢intolerance †¢trade imbalance †¢depression †¢self loathing Racism and future: Today we are living in a load of social issues that we have to deal with and racism is one of them. People blame their faults and disappointments against society and the whole race in which they live in. but anyone can do good or excel if they put their mind to it. The future, when you hear the word – Racism – you start to think about advancing our technology and furthering science. You think of exploring new boundaries and seeing new places. A better world, but a worse world is what we see. The technology may improve as we go deeper into the next century, but what about the children of today that are responsible for the future? Right now, we are only hurting ourselves with racism and hate. We have taken many steps in controlling this disease, but like most diseases, they spread. Does God do favoritism? God is the  Creator of all the races—yellow, black and white (with brown and red). Each has strengths and gifts as well as weaknesses. For our better future it is important to finish this discrimination. If He has created everyone equally then who are we to judge? Who are we to complain? Who are we to discriminate? We being a Muslim, how can we do such a thing? Our religion has a clear concept about equality. It’s clearly written in the Holy book that no one is superior to another. Our God has created everyone equally. Its His choice how and what He do†¦ What Can We Do? â€Å"Racism isn’t born, it’s taugh† Where there is a problem, there is a solution. The only solution that seems to be found is to stop the hate. Dr. Martin Luther King said: â€Å"I have a dream that my four children will one day live in a nation where they will not be judged by the color of their skin but by the content of their character. I have a dream today.† He had a dream.Its up to us that how and what we make a world without racism†¦. Changes: self-awareness†¦Ã¢â‚¬ ¦ Reengineering our attitudes about race means that we must examine ourselves. †¢Closely examine your values and beliefs; why do you think, believe, and behave toward others as you do. †¢Remember, one of the bases of prejudice is ignorance. Try not to ignore but to know other. †¢Measure your words carefully before you speak to others. †¢Try to be a human first . Society is formed by us †¦Ã¢â‚¬ ¦.society changes can be . †¢Do not tolerate prejudiced people, prejudiced talk, or racist action. †¢Give good literature, recommend films, lectures, meetings to your friends and family. †¢Write letters to newspapers, magazines, and TV sponsors giving your opinion on statements and actions that demean people. †¢Organize a cleanup or rebuilding campaign †¢Make an effort to support minority cultural events †¢ Make an effort to teach the value of equality and tolerance For Institutions-Effecting Change: †¢For institutions to change, their culture, vision, values, and ideology must change. All policies must be based around a single ideology. †¢Examine an institution’s deep belief and value systems †¢Hold training seminars in intercultural competence: †¢self-acceptance. †¢Put people to work and interact in interracial learning teams so as to develop positive attitudes and cross-ethnic friendships. †¢Make a commitment and act on it! Things You Can Do in the Schools: This can be done in a school or university setting. †¢sponsor panel discussions †¢awareness activities †¢cultural †¢projects that will promote cultural understanding and inclusiveness. †¢anti-racism education in your school’s curriculum †¢events relating to a variety of cultures.. †¢Ã¢â‚¬ Example: The â€Å"Book of Dreams† To sum it up: Figuring out what to do about the ugly reality of racism in our communities presents a difficult challenge. There is no single or easy set of instructions that will eliminate racism once and for all. Instead, it is something we must continually work against — choosing the strategies and actions that will be most effective in our own individual communities. In this resource, we’ve suggested such strategies and actions, and shown examples of how many of them have been put into practice in real communities. When you start deciding how to reduce racism in your own community, feel free to use any of these ideas, but also feel free to be creative and come up with your own solutions as well. Remember: â€Å"Rainbows aren’t just one color. They’re every single color. Be a rainbow. Enjoy all of the colors in the world.†

The Giant Panda Conservation Project

Although pandas are highly regarded and have a relatively low number of natural predators, they are an endangered species. Humans have severely threatened the panda's existence also so that now there are fewer than 1,600 of them left in the wild. This peaceful creature with a distinctive black and white coat is adored by the world and considered a national treasure in China (WWF, 2013). The bear also is of special importance to the World Wildlife Fund. The panda has been WWF's logo since its founding in 1961. This paper will examine the effectiveness of, and problems related to, the giant panda conservation efforts. The rarest member of the bear family, pandas live mainly in bamboo forests high in the mountains of western China, where they subsist almost entirely on bamboo. They must eat from 26 to 84 pounds of it every day. China’s Yangtze Basin region, which holds the panda’s primary habitat, is the geographic and economic heart of this booming country. An increasing number of roads and railroads are being created throughout the forest, which isolates panda populations and prevents mating. Forest destruction also reduces pandas’ ability to access the bamboo they need to survive. The Chinese government has established more than 50 panda reserves. However, only around 61 per cent of the country’s panda population is protected by these reserves. With the help of conservation organizations and other groups, they have protected more than 45 per cent of the last mountainous regions, equaling over 6,000 square miles of mountain and forest terrain known to be inhabited by wild pandas. They have also set aside corridors of new bamboo for their protection from domesticated animal grazing, and human interference (International, 2013). Chengdu is known for being the hometown of the giant panda. Back in 1987, when it became apparent that pandas were seriously endangered in the wild, the Chinese created the Chengdu Research Base of Giant Panda Breeding. Starting with just six pandas from the wild, they’ve successfully bred more than 100 pandas. The work done at Chengdu and other breeding centers costs millions of dollars a year. Experts believe that of all species in the world, the giant panda is the one species in which the most money is being invested in to save. This has lead some conservationists to argue that too much is being spent to save the giant panda. â€Å"I think we have to make tough choices,† British wildlife expert, Chris Packham, said. â€Å"I think that, ultimately, we have to be pragmatic as well as sentimental. You know, we can't allow our heart to rule our conservation head†¦ and if we channel this much into just one species, then many others which could be far better helped, many others not just species, but communities and ecosystems, could be better protected at the expense of one fluffy, cuddly bear† (Snow, 2013). Packham is in the minority here, but more and more scientists are starting to agree with him. The Chinese government has had much success in educating the public about the future of the giant panda, by setting in place protection against poaching, and even gun control for the reserves. By educating the public, more and more pandas have been brought to the Research Center for recovery and medical care due to injuries or illness. Logging has been banned in the reserves and the bamboo corridors. The roads providing access to the reserves are also being patrolled to help prevent anyone entering without express authority, for the care or study of the environment or the pandas. However, as Packham states, â€Å"Pandas are extraordinarily expensive to keep going. We spend millions and millions of pounds on pretty much this one species, and few others, when we know that the best thing we could do would be to look after the world's biodiversity hotspots with greater care. † His way of thinking is that without a habitat, there is nothing to work with. His suggestion is that we take all the cash we spend on pandas and buy rainforests with it. He realizes that it is the natural response to want to spend money and time on the panda because they are so cute and cuddly. They just pull at peoples' heart strings because of their appearance. The pandas are symbolic of what he refers to as single-species conservation: i. e. , a focus on one animal. This approach began in the 1970s with Save the Tiger, Save the Panda, Save the Whale, and so on, and it is now out of date. His belief is that pandas have had a valuable role in raising the profile of conservation, but perhaps ‘had' is the right word. Panda conservationists say that the panda is a flagship species. But we're also conserving Chinese forests, where there are many other species. As long as this conservation works, Mr. Packham states that he is all for it. But some species are stronger than others. The panda is not a strong species of bear in his opinion. It has gone herbivorous and eats a type of food that isn't ll that nutritious, and so is dying out little by little. It is susceptible to various diseases and, up until recently, has been almost impossible to breed in captivity. This species also has a very limited area of habitation, which is decreasing, since the Chinese population keeps finding ways to use their land. While Mr. Packham's points are valid, it seems he ignores the fact that pandas play a crucial role in the bamboo forests where they roam by spreading seeds and facilit ating growth of vegetation. Also, in the Yangtze Basin where pandas live, the forests are home to wildlife such as dwarf blue sheep, multi-colored pheasants and other endangered species, including the golden monkey, and various birds that are not found anywhere else in the world (WWF, 2013), making the panda a very effective flag ship species, which is something that Packham himself stated was worth preserving. One belief is that large animals can be extremely useful in the cause for conservation. Smaller creatures often don't need a big habitat to live in, so in conservation terms, it's better to strive to save something further up the food chain. By doing this, you are protecting a much larger area, which will also include the smaller animals. Pandas are a good example for the messages you want to put out on habitat conservation. We associate Borneo with the orangutans, the southern oceans with the blue whale, and the North with polar bears. So it is with China and the panda bears (Snow, 2013). Many revere the panda bear and their preservation is important to the Chinese culture. People respond to their situation, which does make a difference. References Benedictus, L. (2009). Should pandas be left to face extinction? | Environment | The Guardian. Latest US news, world news, sport and comment from the Guardian | guardiannews.com | The Guardian. Retrieved April 27, 2013, from http://www.guardian.co.uk/environment/2009/sep/23/panda-extinction-chris-packham International, C. (2013). Threats to Pandas. Conservation International. Retrieved April 28, 2013, from http://www.conservation.org/learn/biodiversity/species/profiles/pandas/pandas/overview/Pages/threats.aspx Snow, K. (2013). Are giant pandas worth saving? – Rock Center with Brian Williams. Rock Center with Brian Williams.

The Secret to Writing a College Admissions Essay That Stands Out from the Crowd

The Secret to Writing a College Admissions Essay That Stands Out from the Crowd If youre in the process of writing a college admission essay, youve already had a taste of the anxiety these types of assignments can cause. While choosing each word carefully, your mind probably spins with a series of questions: Will this essay keep me from getting in? Will they find it boring? Does it stand out from the crowd?Added to that anxiety are the often complex, or just plain strange, questions that many colleges will ask, like Do you believe we are alone? or How do you feel about Mondays? Whether the question is strange or difficult, overly simplistic or not even applicable to your life- here are some quick tips on the secret to writing a college admissions essay that stands out from the crowd of other essays the admissions committee will read along with yours.Less is moreMany colleges will give you a word count maximum but for the ones where a word count minimum is given, dont take that as an invitation to write over 800 words. The admissions committee who will be reading your essay(s) will also be reading a stack of other essays, and will not want to spend more time reading yours than necessary. Keep it concise to maintain their interest without taking up a lot of their time.Uniqueness countsThe key to writing an essay that stands out from the others is to be as unique as possible. While this might be a difficult task for many soon-to-be college freshmen, its worth your time and effort to think of something- anything- that makes you unique compared to the others. Maybe its a travel experience, a family history, a goal, a way that you think or something youve done?Dont cover everythingIf youve led an especially busy life as a high school student, dont try to cover everything youve done. Pick the highlights- the activities that were the most rewarding or the most impressive- and stick to those few things. If you write about 20 different activities within the span of a 500-700 word essay, your writing will inevitably appeared scattered and unorganized . Its simply impossible to write about that many topics with that limited of a word count and keep it organized.Be controversialMany people falsely assume that you should avoid topics such as religion, politics and the like in college entrance essays, but this couldnt be farther from the truth. Although you should avoid soapboxes and topics that might be against school philosophy (if applying to a religious-oriented school), voicing your stance and providing reasonable arguments for it and against it shows that you know how to think logically and coherently about important topics- a trait that you will need to succeed as a college student.Avoid mistakes in grammar, punctuation and spellingWhile this should be obvious, you would be amazed at how many students submit essays with glaring grammar, punctuation and spelling mistakes. The main reason for this is they depend on their word processors spelling/grammar check, which will not catch many of the mistakes that a flesh and blood edi tor would catch. Submitting an essay with these types of mistakes is a guaranteed way to get your application rejection. The admissions committee will believe (and rightly so) that if you have errors on what should be an example of your best work, your daily work in college will have even more errors. When they compare an essay with errors to an essay without them, and have to choose between the two, its obvious which one they will choose. And it wont be yours.Be accurateI am consistently amazed at the number of college admissions essays I receive that refer to particular works or authors, and then get those titles and author names wrong. While a good editor will hopefully catch such errors, its impossible for an editor to know about every topic and every author. For this reason, beyond the mistakes that can be made with spelling, punctuation and grammar, a big mistake that many applicants make is inaccuracy of information. When referring to a particular author who was an influence on your life and choices, be sure to get the name of the author and title of the work right. When discussing theories, research, or any topic for that matter, be sure that you are completely accurate in the context and use of this information. Otherwise, youll seem as if you dont know what youre talking about and are just throwing out information that youve neither studied nor learned.Be descriptiveWhen youre discussing something that youve accomplished or situations in which youve excelled, be descriptive because it lends a sense of credibility and humaneness to what you are saying. This is not to say that you should overload an essay with adjectives and adverbs, but adding details like this will make your writing more exciting and more vivid- two traits that admissions committees love in an essay.Be likeableThis one is perhaps the most important, as long as your grammar, spelling and punctuation are correct. When an admissions committee reads an essay written by a student who has excelled much in their high school years but seems pedantic, stuffy and just plain boring, that student still has a chance of being denied admission. College is as much about social interactions as it is about academics. If you fail to show that you can be likeable and fit in well with the college community, you have missed an opportunity to make your essay stand out from the rest.

Euthyphro †Plato

Euthyphro – Plato The concept that needs to be defined is that of piety/ holiness. Euthyphro is in the verge of prosecuting his father because of the crime he committed. Socrates is surprised by the action of Euthyphro. However, Euthyphro argues that his action is pious.Advertising We will write a custom essay sample on Euthyphro – Plato specifically for you for only $16.05 $11/page Learn More This action makes Socrates to wonder whether impiety and piety knowledge that Euthyphro posses are adequate enough to allow him prosecute his father by not behaving impiously. As a result, they search for piety definition as understood by Euthyphro. The dialogue between the two terminates without getting a clear definition of what piety means (Cohen, N.D). Definition of piety by Euthyphro The definition of piety is offered by Euthyphro in four different perspectives. From the initial dialogue between Socrates and Euthyphro, Euthyphro is in the verge of prosecuting his own father. This is because he murdered a murderer. The underlying philosophy behind the actions of Euthyphro is not understood by Socrates. However, he is keen to learn with an aim of understanding Euthyphro and how he ended holding such a super power in the whole state. Socrates was preparing himself to face trial. As such, he wanted to attain the same respect that had already been achieved by Euthyphro. In their dialogue, Euthyphro understands the various definition of piety. On the contrary, throughout their dialogue, Euthyphro gives out four different piety definitions some of which contradict each other (Brickhouse Smith, 1995). According to Socrates, there are numerous limitations with regards to his definitions and continues to insist for an answer that is satisfactory. His initial definition can be found in the following statement, â€Å"piety means prosecuting the unjust individual who has committed murder or sacrilege, or any other such crime† (Plato, N.D, p.5). This is in r eference to his father’s prosecution for murder charges. Piety is therefore considered as that action which does not maintain the city justice. Therefore, if a single man acts in a disgraceful manner, he needs to face justice by being prosecuted and justice enforced. To emphasize on this point, a reference is made to Kronos the father of Zeus and Zeus himself. Kronos was bound by his son Zeus because he consumed his own children. After being bound, Zeus escaped. Ironically, Kronos managed to castrate his own father because he consumed his own children. However, it is argued by Euthyphro that, in comparison to all gods, Zeus is the most just. Therefore, Euthyphro is trying to appease the gods by being just. Thereby prosecuting the society wrong doers such as in the case of his father, Zeus and his father as presented in the book.Advertising Looking for essay on philosophy? Let's see if we can help you! Get your first paper with 15% OFF Learn More Anothe r definition of piety provided by Euthyphro to Socrates states that, â€Å"the pious is to do what I am doing now to prosecute the wrong doer† (Plato, N.D, p.9). In that case, he prosecutes his own father despite people being against his actions. This definition is not accepted by Socrates because it is only an example given rather than a definition. According to Socrates view, the definition of piety given by Euthyphro is not objective as they are not similar in each and every situation and they are not explanatory. Another definition of piety by Euthyphro is that† The pious is what is dear to gods (6c-8d). This definition is fine to Euthyphro. Nevertheless, Socrates argues that, a lot of things are disagreed upon by gods. For instance, the idea of him prosecuting his father might be okay with Zeus but not appealing to the overthrown Kronos. Socrates main goal From the dialogue between Socrates and Euthyphro, the main goal of Socrates was to have a clear definition of what piety was. However, Euthyphro gave him examples rather than a single definition. In addition to that, Socrates wanted to understand Euthyphro better and how he raised to that powerful position in the state. Socrates was about to face trial and his main objective was to make sure he worn the case by understanding the true meaning of piety. According to my own definition, piety is the reverence and devotion to God and religious practices. For instance, going to church and listening to sermons is a good example of piety. Therefore, Euthyphro was obeying his gods by prosecuting his father. Given the fact that his father had committed a crime, he was supposed to face justice and pay for his sins. Additionally, piety can be perceived as the science of sacrifice and prayer. In essence, piety is acknowledging what needs to be done in sacrifices and prayer in order to please gods. Therefore, individuals should have a clear knowledge of how they can address their gods in times of prayers and sacrifices. Hence, an individual needs to please god. However, according to Socrates arguments, he believes that there is no co-existence between the god-loved class and pious things class. His arguments are based on substituting the two terms† god-loved† and â€Å"pious† in his statements. The substitution result to sentences that are false. References Brickhouse, T. C., Smith, N. D. (1995). Platos Socrates. New York: Oxford University. Press.Advertising We will write a custom essay sample on Euthyphro – Plato specifically for you for only $16.05 $11/page Learn More Cohen, M.C. (n.d). Socrates on the Definition of Piety: Euthyphro 10a- 11b. Retrieved from http://faculty.washington.edu/smcohen/Euthyphro.pdf Plato, M Jowett, B. (n.d.). Euthyphro: Project Gutenberg. Retrieved from gutenberg.org/ebooks/1642

Minmi - Facts and Figures

Minmi - Facts and Figures Name: Minmi (after Minmi Crossing in Australia); pronounced MIN-mee Habitat: Woodlands of Australia Historical Period: Middle Cretaceous (100 million years ago) Size and Weight: About 10 feet long and 500-1,000 pounds Diet: Plants Distinguishing Characteristics: Unusually small brain; primitive armor on back and belly About Minmi Minmi was an unusually small, and unusually primitive, ankylosaur (armored dinosaur) from middle Cretaceous Australia. This plant-eaters armor was rudimentary compared to that of later, more famous genera like Ankylosaurus and Euoplocephalus, consisting of horizontal bony plates running along the sides of its backbone, a noticeable thickening on its belly, and spiky protrusions at the end of its long tail. Minmi also had an unusually small, narrow head, which has led some paleontologists to speculate that its encephalization quotient (the comparative size of its brain to the rest of its body) was lower than that of other dinosaurs of its timeand considering how stupid the average ankylosaur was, thats not much of a compliment. (Needless to say, the dinosaur Minmi shouldnt be confused with the Japanese-born, Caribbean-style singer Minmi, or even Mini-Me from the Austin Powers movies, who are both presumably much more intelligent!) Until recently, Minmi was the only known ankylosaur from Australia. That all changed at the end of 2015, when a team from the University of Queensland re-examined a supposed second Minmi fossil specimen (discovered in 1989) and determined that it actually belonged to an entirely new ankylosaur genus, which they dubbed Kunbarrasaurus, Aboriginal and Greek for shield lizard. Kunbarrasaurus appears to be one of the earliest known ankylosaurs, dating to the same middle Cretaceous time frame as Minmi, and given its relatively light coating of armor, it seems to have only recently evolved from the last common ancestor of both stegosaurs and ankylosaurs. Its closest relative was the western European Scelidosaurus, a clue to the different arrangement of the earths continents during the early Mesozoic Era.

Social impact of the DIAMOND INDUSRTY Essay Example | Topics and Well Written Essays - 250 words

Social impact of the DIAMOND INDUSRTY - Essay Example Diamond stones additionally may be structured under the high weights and temperatures at the site of shooting star sways. The precious stones structured throughout an effect may be generally junior, however a few shooting stars hold star dust, garbage from the demise of a star, which may incorporate jewel gems. One such shooting star is known to hold small precious stones in excess of 5 billion years of age. The bonds are framed when carbon is at high temperatures and under high weights. Characteristic precious stones are made where it counts down in the earth, about 180km underneath the surface, where high temperatures and weights exist commonly. Under the worlds hull is the shelf, which is made up of liquid shake, metals and different materials (Zoellner, 2007). Diamond stones are utilized for cutting glass and off and on again in the coarseness of streamlined sandpaper. Jewels are known for being the hardest characteristic known material on Earth. They must be scratched by an alternate precious stone consequently they are utilized for cleaning devices. Diamond mining or any kind of mining is obviously not reasonable. You burrow an opening; you take stuff out of the gap, and take it someplace else. In the long run, the gap uses up the stuff you were uncovering. That is not economical. In any case as a movement, precious stone mining might be made to help manageability. Firstly, we distinguish that advancement is important to dispense with destitution and to give monetary decisions to northern people groups (Davis, 1999). Incomes from Diamonds stones assume a key part in driving budgetary improvement. They are our single greatest investment affect, and invigorate development and venture. Our associations with governments and groups help us to change characteristic assets into imparted and enduring national fortune. The worldwide Diamonds stone industry utilizes in the ballpark of 10

Laird Plc a Public Owned British Multinational Company Essay

Laird Plc a Public Owned British Multinational Company - Essay Example The solutions to protect the electronic devices of the users from any potential harmful Electromagnetic Interference (EMI) and also safeguard from the heat that is generated during the operation of the device (Laird, 2014a; Laird, 2014d). The company leads in developing, designing and delivering innovative technologies that ensure organizations, applications, and people to stay connected effectively. Laird owns two divisions: Wireless Systems and Performance Materials. The products of Performance Material are distributed to wide markets that include Information Technology and telecom, smartphones, transportation and medical (Laird, 2014c). In this report, the international operation of the company along with competitiveness is examined by performing market and financial analysis. However, the risk associated with the company operations are also studied in order to understand the manner it combats against exchange rate, country, and political risks. The following figure highlights the financial performance of the two main divisions of Laird Plc. The financial performance is evaluated based on the revenue of the divisions over the period 2010- 2013. From the figure, it can be stated that the majority of the revenue during the four years in Laird Plc is generated from Performance Materials. This division accounts for more than half of total revenue that is incurred by the group as a whole. It is observed that the revenue from Performance material division has increased over the years due to the increase in demand for thermal management solutions, shielding materials for electromagnetic interference (EMI) and signal integrity products (SIP) (Laird, 2014c). Despite the growing revenue, the company is encountering challenge because of the highly integrated circuits related to thinner and smaller devices, which are becoming more innovative so as to protect the package devices (Laird, 2014c).

Roe vs Wade Essay Example | Topics and Well Written Essays - 1750 words

Roe vs Wade - Essay Example aised in the two cases were; a) the legality of abortion laws passed by states seeking to criminalize all forms of abortion when interpreted in the letter and spirit of the US Constitution; b) the parameters of the Due Process provision of the 14th Amendment as a safeguard against the right to privacy in abortion; c) whether states had the authority to legislate laws that may prohibit abortion; d) whether the natural termination of Roe’s pregnancy rendered her case, on the same issue, which was pending before the court meaningless; and e) whether the district court was correct in denying injunctive relief. This paper will discuss some of the issues relating to the right of privacy based on the ruling on Roe. In 1880s abortion was legalized in United States specifically to protect the mother’s life from harm. Since the country was facing low birthrate during that particular period, especially among the whites, the government and other human rights agencies were concerned about the negative impacts of the policy. They termed the declining population of whites through abortion as having the same bad effect as suicide since they wanted the women who were born in United States to reproduce. Abortion was had been previously illegal and thus if a woman had to procure an abortion it depended on the race, economic situation, and the location where one lived (Critchlow, 2010). It is notable that poor women who came from other races could not afford it due to the high related cost. In early 1960s, however, the trend somehow changed by virtue of the formation of a group known as the Clergy Consultation Services on Abortion. The group drew its membership from church ministers and rabbis who dealt with illegal abortion since they sympathized with women of color who were disproportionately affected by the problem. The corresponding growth of civil rights and antiwar movements during the period campaigned for and came up with women liberation movement seeking the

Final Essay Example | Topics and Well Written Essays - 1000 words - 2

Final - Essay Example US commission on Civil rights serves as an overall advisor as well as a watchdog and the country’s civil rights division within the department of justice bears the responsibility of enforcing non-discrimination statutes. Nevertheless, the dispersion of responsibilities in agencies with different agendas affected the process of affirmative action implementation. However, all the agencies were concerned with application of non-discriminating policies to the various segments of the US economy despite the rise of various jurisprudence disputes even within their spheres of action; besides administrative actions were related and affected other spheres. By the turn of the twenty-first century, a small number expressed support for racial discrimination after the civil rights movement progressed from obscurity during the civil war to mid-twentieth century activism to the current accepted wisdom. The non-controversial part is the reactive policy that ensures non-discrimination based on race, religion, color, sex and national origin in social, economic and educational affairs. The non-discrimination laws are designed to make sure that individuals are never judged by color of skin, but rather by the content of their character and incase a violation occurs, such individuals are entitled to remedies (Lee, 1999). Positive and Negative Results of Affirmative Action Legislation Affirmative action seems to have a short history though it is a hands-on policy that makes special efforts about employment decisions, entry into college and other public behavior as a means of compensating for past discrimination. Affirmative action bases on the thought that various groups of people even without being discriminated against currently, any individual belonging to such groups are disadvantaged in the workplace and in campuses as a result of the past discrimination aimed at the group. Therefore, affirmative action attempts to level the playing ground for all categories of citizens th ought it emphasizes on disadvantaged groups as opposed to injured individuals. Affirmative action can refer to a court-ordered, remedial programs designed to correct effects of discrimination documented in court. Proponents of affirmative action hold that affirmative action provides long-term cure for discrimination by offering victims chances to show their skills and worth, which eventually changes prejudicial attitudes. Nevertheless, opponents of this controversial program argue that affirmative action never addresses the cause of inequality and the program can create labor market inefficiencies and result in reverse discrimination. Therefore, both sides suggest that effective affirmative action would cause minority employment to increase however; the sides disagree on whether this raise is efficient and on whether it would be sustainable if affirmative action ended (Lee, 1999). Currently there is small opportunity to measure the impact of eradicating affirmative action programs. As Federal, support for enforcement faded and flowed, the Supreme Court ruling in the past decade chipped away at affirmative action making it difficult to confirm whether concurrent changes in minority outcomes are due to affirmative actio

Certification and Standardization of Mediators Essay - 1

Certification and Standardization of Mediators - Essay Example Years later, California added child custody decisions to mediator responsibilities. â€Å"In 1980, California became the first state to mandate all parents with custody or visitation disputes to participate in family mediation prior to a court hearing† (Milner et al., 2004, p.5). By 2004, most states had mandates regarding family divorce disputes, and 38 states had mediation mandates where child custody issues occurred (Tondo, Coronel, & Drucker, 2001, as cited in Milner et al., 2004, p.5). Example One. One of the frontrunners and more controversial figures in divorce mediations was O.J. Coogler, an Atlanta lawyer and author of the book, Structured Mediation in Divorce Settlement (1978). This marriage and family litigator used labor negotiations and social sciences as a guide to build a framework for third-party mediators to use when assisting divorcing couples resolve issues such as property division, alimony, child visitation, and child custody (Milner et al., 2004). Despite establishing the Family Mediation Association (FMA), the bar association harshly rebuked Coogler. â€Å"The bar association declared mediation by non-lawyers to be the unauthorized practice of law and attempted to dissuade lawyers from mediating through the threat of ethical sanctions† (Silberman, 1988, as cited in Milner et al., 2004, p. 5). Example Two. Confidentiality is crucial in court proceedings. In a civil suit involving the still photography and filming of underage girls, the judge ordered mediation in an attempt to circumvent litigation (Zimmerman, 2009). However, the defendant did not take the mediation proceedings seriously. He appeared four hours late, dressed inappropriately, shouted profanities at the court, continually placed his dirty, bare feet on the plaintiff’s table, threatened the plaintiff’s lawyer, and then proceeded to breech confidentiality by telling

Various Assignments Essay Example | Topics and Well Written Essays - 1500 words

Various Assignments - Essay Example In this regard, the procedural fairness provided for in the Bill of Rights is arguably its most important value. Procedural fairness is important because it preserves unto the individual the right to freely participate in political and economic life (Nueborne, p. 83). Moreover, procedural fairness counter-balances the fact that the freedoms and rights contained in the Bill of Rights are not absolute. In other words, there are exceptions to the right to free expression in that the government may curtail freedom of expression in situations where the speaker utters words that can be construed as hate crime or incitement to riot. Thus procedural fairness permits the speaker the right to a fair trial and the right to confront his or her accusers and to defend his or her right to free speech. Procedural fairness recognizes that any government can overstep its power to the detriment of the governed (Nueborne, p. 83). With entitlements to the right to privacy, protection of private property, the right to a fair trial, the right to bail, and so on, the Bill of Rights plays a supervisory role in the constitutional order. ... Institutional enforcement mechanisms are captured by procedural fairness by allowing for judicial review of the exercise of powers by the federal government, state governments and their agents. Even at the state level, state and state agents’ conduct is scrutinized by state judges by reference to the rights and freedoms contained in the Bill of Rights. Thus, as Nueborne puts it, procedural fairness establishes a â€Å"nationwide network† that are â€Å"engaged in constant surveillance of the activities of the governing majority† so that the rights and freedoms articulated by the Bill of Rights â€Å"are respected in everyday life† (Nueborne, p. 84). Procedural fairness is all the more important since the rights and freedoms contained in the Bill of Rights are not expressed in uncertain terms. Many of these rights and freedoms are â€Å"ambiguous† so that they can be open to interpretation (Nueborne, p. 84). Procedural fairness curtails the risk of i nconsistent interpretations of the rule of law in relation to the individual rights and freedoms contained in the Bill of Rights. In promoting procedural fairness judges interpret and apply the Bill of Rights by reference to a system of precedents and thus assures consistency and greater uniformity of rights and freedoms for all (Nueborne, p. 85). In the final analysis procedural fairness ensures that in addition to ensuring that the government treats all individuals equally, procedural fairness also ensures that the government exercise its powers fairly and consistently and that any departure from the fundamental freedoms and rights contained in the Bill of Rights is done so in a way that is only necessary

Certification and Standardization of Mediators Essay - 1

Certification and Standardization of Mediators - Essay Example Years later, California added child custody decisions to mediator responsibilities. â€Å"In 1980, California became the first state to mandate all parents with custody or visitation disputes to participate in family mediation prior to a court hearing† (Milner et al., 2004, p.5). By 2004, most states had mandates regarding family divorce disputes, and 38 states had mediation mandates where child custody issues occurred (Tondo, Coronel, & Drucker, 2001, as cited in Milner et al., 2004, p.5). Example One. One of the frontrunners and more controversial figures in divorce mediations was O.J. Coogler, an Atlanta lawyer and author of the book, Structured Mediation in Divorce Settlement (1978). This marriage and family litigator used labor negotiations and social sciences as a guide to build a framework for third-party mediators to use when assisting divorcing couples resolve issues such as property division, alimony, child visitation, and child custody (Milner et al., 2004). Despite establishing the Family Mediation Association (FMA), the bar association harshly rebuked Coogler. â€Å"The bar association declared mediation by non-lawyers to be the unauthorized practice of law and attempted to dissuade lawyers from mediating through the threat of ethical sanctions† (Silberman, 1988, as cited in Milner et al., 2004, p. 5). Example Two. Confidentiality is crucial in court proceedings. In a civil suit involving the still photography and filming of underage girls, the judge ordered mediation in an attempt to circumvent litigation (Zimmerman, 2009). However, the defendant did not take the mediation proceedings seriously. He appeared four hours late, dressed inappropriately, shouted profanities at the court, continually placed his dirty, bare feet on the plaintiff’s table, threatened the plaintiff’s lawyer, and then proceeded to breech confidentiality by telling

Process vs. Non-Process Research Paper Example | Topics and Well Written Essays - 750 words

Process vs. Non-Process - Research Paper Example Taking a hint from this real life example, the concept of process innovation needs to be analyzed in a broader context. There is no need to say much about the fact that HTML5 is the latest technology that offers much new in the area of extending mobile platform choices to the companies and consumers (Lee, 2012). Yet, the biggest problem in adapting to this new technology is the need to customize and adapt their operations and research initiatives to benefit from the competitive advantage that a shift to HTML5 mobile platforms offers. In that context the HTML5 debate has exposed a hitherto much ignored fact. When it comes to process versus non-process industries, it is a fact that ideally speaking there exists nothing like a non-process industry (Skinner, 1992). The only difference between a process and non-process industry is that in a non-process industry, the operations consist of multiple unwieldy and least synchronized processes, which are least capable of extending a strategic a dvantage to a company against its competitors (Skinner, 1992). However, the other thing that needs to be taken into consideration is that new advances in technology may push a company from being process driven to being a non-process company, in a relative if not an absolute sense. The biggest revolution that the web based products and services have come across is the concept of computing going mobile. This created a dire need for the research in technologies that are compatible with and support mobile computing. In the last 10 years the world of mobile platforms has moved from a domination of the few like Windows Mobile and RIM Blackberry, to an invasion of many new platforms. In that context, there is no doubt that HTML5 stands to be the lowest common denominator, when it comes to developing mobile browsers. Still, many companies are raising a noise about shifting to HTML5 based mobile platforms, because it necessitates the requisite innovations in the processes underlying their pr oducts and services. There are varied reasons why the companies pushed from the status of being process companies to non-process companies owing to a failure to incorporate and adapt to HTML5 in the processes underlying their businesses tend to be hesitant. One important factor is that there exists a schism between the managers and technology personnel governing these companies (Skinner, 1992). While the management driven executives are still sticking to the old paradigms justifying augmentation of sales by resorting to marketing and financial gimmicks, considering the high risk involved in opting for HTML5 oriented process innovations, the technology experts tend to be averse to suggest such changes, fearing possible fallout on their careers. Besides, the requisite process innovations necessitate a long term financial and planning related commitment on the part of the companies, which is difficult to contrive, as evinced by the Facebook experience (Skinner, 1992). Then there are co mpanies which are waiting for their competitors to innovate, while mulling over immense financial savings by adapting to these innovations at a later stage

Isolation of Recombinant Escherichia Essay Example for Free

Isolation of Recombinant Escherichia Essay One technique important in both genetics and biochemistry is the Polymerase Chain Reaction (PCR), first developed in the 1960s, and then automated in 1983. Current PCR technology was not developed until the discovery of thermostable polymerases, specifically Thermus aquaticus (Taq) polymerase (1). The protein Taq polymerase was first isolated from the extreme thermophile T. aqauticus, where extreme thermopiles are bacteria that live in temperatures at or above 45 °C. The Taq enzyme is a member of the DNA polymerase I family (2, 3). The interesting property of Taq polymerase is that it has a temperature optimum at 74-75 °C, allowing it the remain active in temperatures required for PCR double stranded DNA denaturation (3, 1) . The protein has an approximate molecular weight of 6263 kDa when isolated from T. aquaticus, and 94 kDa when isolated from recombinant Escherichia coli, and is still stable at temperatures of 93-95 °C, hence the thermostability of the enzyme ). Taq specifically lacks any proofreading activity in the 3’ to 5’ direction, and therefore has a relatively high error rate of single base mispairings of 1 error per Isolation of Recombinant Taq Polymerase for PCR 9000 nucleotides, as well as a frame shift error rate of 1 per 41,000 basepairs (5, 6). Taq polymerase has an activity that is highly dependent on the environment of which it is in as it is thermostable, and has differing activities at nearly all temperatures up to the point of denaturation. Taq specifically can add up to 1000 base pairs in length on a template in under one minute under typical PCR conditions. The enzyme has a specific activity of 200,000 units mg-1, and can add approximately form 60 nucleotides per second at 70 °C (7). The isolation of Taq is essential for the PCR reaction. The most important reason for Taq being used in PCR is the thermostability at high temperatures (95 °C). This allowed for the process of elongation, annealing, and denaturation to occur without the replacement of new enzyme, and thereby, was more efficient, faster, and cheaper because the reaction could be automated through the use of a machine known as a thermocycler which basically is just a machine able to change temperatures of an isolated environment rapidly (7). Prior to the discovery of Taq, PCR was done using Klenow fragments of E. coli DNA polymerase I at 37 °C. The lack of thermostability required replenishment of enzyme after each PCR cycle (8). One of the initial difficulties of Taq polymerase was the organism in which it was expressed in, T. aquaticus, as it was difficult to culture and produce large quantities of enzyme. E. coli bacteria were engineered to expressed the Taq polymerase gene to allow for retrieval of large quantities of enzyme ). The isolation of the Taq gene involved culturing T. aquaticus and then isolated the DNA of the cells through lysing, proteinase K addition, extracting of aqueous and phenolic phases, dialyzing of extractions, addition of SDS, and then centrifugation of solution to eventually retreieve the DNA of the organism as outlined in Lawyer et al., 1989. With the isolation of the 2401+ BP gene of Taq, the gene was incorporated into a 6.58 kbp plasmid (pLSG1). The gene was inserted 171 bp distal to the lacZÃŽ ± promoter/operator, and 109 bp distal to the BgII site, so the gene expression could be controlled through an inducible promoter. With the pLSG1 plasmid, the vector was introduced to E. coli bacteria to allow for plasmid uptake (4). Other experiments have been conducted towards the purification of Taq from recombinant E. coli. Specifically Engelke et al., 1990 developed a method for purfication of Taq. The E.coli strain 2 DH1 was used for the expression of the recombinant plasmid containing Taq polymerase. The bacteria were grown in 12 Litre batches of Luria Broth; using 1 mL of saturated DH1 culture and 80ÃŽ ¼g/mL of ampicillin. Isopropyl-1-thio-ÃŽ ²-Dgalactopyranoside (IPTG) was added to 0.5mM and the cultures were grown for 16-20 hours. The cells were harvested in 2.4 L of buffer A (50 mM TrisHCL, pH 7.9, 50 mM dextrose, 1mM EDTA) and collected via centrifugation, resuspended in Buffer A with 4mg/mL lysozyme and incubated at room temperature for 15 minutes. Buffer B (10 mM TrisHCl, pH 7.9, 50mM KCl, 1mM EDTA, 1mM phenylmethylsulfonyl fluoride (PMSF), 0.5% Tween 20, 0.5% NP-40) was added and incubated in 180 mL fractions, for 60 minutes at 75 °C in a water bath. The mixtures were centrifuged at 8000 rpm for 15 minutes at 4 °C. Taq then precipitated with polyethyleneimine (PEI) at room temperature, then isolated through centrifugation and suspended in buffer C (20mM HEPES, pH 7.9, 1 mM EDTA, 0.5mM PMSF, 0.5% Tween 20, 0.5% NP-40) containing 0.25 M KCL. PEI eluatents were diluted in 50mM KCL and buffer C and applied to a 150mL BioRex 70 ion exchanger column, and then eluated using 200mM KCL. The protein was dialyzed for 12 hours against two changes of 1 L storage buffer (20mM HEPES, pH 7.9, 100 mM KCL, 0.1 mM EDTA, 0.5 mM PMSF, 1mM dithiothreitol, 50% glycerol. The experiment resulted in 40-50 mg of protein per litre of cell culture (9). The methods used in this experiment differed in certain key aspects. First, Engelke’s experiment made use of a higher concentration of ampicillin. The IPTG was added to the same concentration, but was added after cell growth up to an optical density of 0.700. Instead of a water bath at 75 °C, this experiment made use of an air incubator for the temperature requirements. Engelke’s experiment made use of PEI to precipitate Taq, while this experiment made use of 30g of (NH4)2SO4 per 100mL of supernatant. Buffer C was not used throughout this experiment, and no ion exchange columns were used. The dialysis procedure was done for twice as long with twice as many changes of solution per 6 hours. The changes made from Engelke’s experiment offers a different method for protein precipitation. The method used by Engelke made use of PEI which is an affinity precipitation method versus a salt prec ipitation method. The PEI Isolation of Recombinant Taq Polymerase for PCR method has the major drawback through the lack of selectivity, and can often precipitate nucleic acids as well (10). This is why the BioRex column needed to be used. Ammonium sulfate has the advantage that the precipitation can be controlled based on ionic strength of species involved, as well as has no negative effects on the activity of the target enzyme. Salting out also has the advantage that only native state proteins are precipitated due to the hydrophobicity involved with native state proteins (10). Buffer C was not required for this experiment as no BioRex column was required. This experiment made use of various techniques and methods including: SDS-PAGE, differential centrifugation, Western Blotting, real time-PCR (rtPCR), PCR, agarose gel electrophoresis, and dialysis. Two important techniques were PCR and rt-PCR. PCR does not allow for the quantification of DNA amplicons as it is an end-point PCR, but it does allow for confi rmation of template duplication along with measurement of base pair length. Amplification of primer would confirm the presence of a thermostable DNA polymerase. The following agarose electrophoresis helps to find amplicon size which can tell us the activity of Taq, as well as the specificity, as one template should only return one band in PCR (7). rt-PCR allows for a quantitative assessment of PCR, and therefore the kinetics of the reaction, as it detects the amount of amplicons produced in the reaction. The point at which the standard curve reaches threshold in cycle number gives information on the activity of Taq, as a more active sample of Taq reaches threshold earlier. Melt curve analysis also provides information regards DNA amplicons in solution (11). The purpose of this experiment was the test the methods for the isolation of PCR grade Taq polymerase from recombinant E. coli using differential centrifugation, salting out, and heat denaturation following lysation of cells to potentially improve isolation of Taq from past methods. The presence of Taq will be confirmed through Western blotting, and rt-PCR and PCR reactions along with purity will be assessed through SDS-PAGE. The activity of Taq will be found through rt-PCR and PCR. Finding the most efficient method for the isolation of Taq offers a valuable reagent source for any PCR reactions required. The isolation technique would also be applicable to any thermostable proteins. 3 EXPERIMENTAL PROECDURES Isolation of Taq Polymerase Luria broth (500 mL + 100ÃŽ ¼g/mL ampicillin) was inoculated with 50 ÃŽ ¼L of frozen Taq polymerase expressing E. coli cell stock. Incubation was commenced for 12 hours at 37 °C until the Optical Density had reached 0.700. IPTG (0.5 mM or 0.112g/L culture) was added and the culture was incubated for 12 to 14 hours at 37 °C. The 50mL of cells were then centrifuged (4000 RPM x 15 minutes at room temperature) in an Eppendorf Centrifuge 5810, and 5 mL of buffer A (50 mM Tris-HCl, pH 7.9, 50 mM dextrose, 1mM EDTA) was used to suspend the separated pellet. The solution was then centrifuged again (4000 RPM x 15 minutes at room temperature) in an Eppendorf Centrifuge 5810 and the pellet was once again suspended in Buffer A, with an additional 20 mg of lysozyme added. The reaction was incubated for 15 minutes at room temperature. Following incubation, 5mL of buffer B (10 mM Tris HCl, pH 7.9, 50mM KCl, 0.5% Tween 20, 0.5% NP-40, 1mM PMSF, 1mM EDTA) was added and incubated at 75 °C for 1 hour in a New Brunswick Scientific-Innova 40 incubator shaker series, and shaken by hand approximately every 5 minutes. The solution was then centrifuged (15000 RPM x 10 minutes at 4 °C) in a Thermoscientific Sorvall RC 6+ centrifuge and using a 603s Delta Range 30g of (NH4)2SO4 per 100mL of supernatant (8 mL of supernatant equivalent to 2.4g (NH4)2SO4 ) was added and incubated for 10 minutes at room temperature and shaken on the Innova 40 incubator. The lysate was then centrifuged again (15000 RPM x 10 minutes at 4 °C) in Thermoscientific Sorvall RC 6+ centrifuge and the resultant pellet was suspended in 2mL of buffer A. The solution was then dialyzed in a Spectra/Por membrane tubing set at 6000-8000 Da molecular weight selection in 1 L of storage Buffer (50 mM Tris HCl, pH 7.9, 50mM KCl, 0.1mM EDTA, 1mM DTT, 0.5 mM PMSF, 50% glycerol) for 24 hours at 4 °C changing the buffer every three hours. The dialysis solution was then diluted in a 1:1 ratio of storage buffer and stored at -70 °C until needed. Protein Concentration Determination A Bovine Serum Albumin Bio-Rad assay standard curve was prepared (0 –0.3 mg/mL) using Isolation of Recombinant Taq Polymerase for PCR a 1mg/mL stock solution and an Asys Expert Plus spectrophotometer set at 620 nm. Bio-Rad assay was run in triplicate using 20ÃŽ ¼L of protein dilution and 150 ÃŽ ¼L of diluted Bio-Rad Dye Concentrate. 10x and 100x dilutions of the sample prepared previously were made and 20ÃŽ ¼L were used with 150ÃŽ ¼L of diluted Bio-Rad Dye concentrate. The solutions were incubated for 10 minutes and absorbances were tabulated. sandwich was then assembled with an additional ice block in the transfer apparatus. The apparatus was run at 180mA overnight in a refrigerator and the membrane was then stored in TBST buffer (20 mM Tris-HCl, pH 7.6, 137 mM NaCl, 0.1% Tween 20) and refrigerated. 24 hours prior to the primary antibody (Anti-Taq monoclonal Antibody (8C1)) addition, the membrane was blocked in 1 gram of Carnation nonfat dry milk (5% w/v) and 20 mL of TBST Buffer. The primary antibody in TBST with SDS-PAGE 5% w/v nonfat dry milk at a 1:800 dilution of A discontinuous polyacrylamide gel was antibody was added to the membrane and shaken prepared using a Mini-PROTEAN Tetra Cell for 1 hour at room temperature. The membrane was module. The casting stand was assembled for 1mm then washed three successive times for 15 minutes gel and filled with National Diagnostics 12% with TBST buffer at room temperature. The Resolving Gel (Protogel 2400 ÃŽ ¼L, Resolving Gel secondary antibody (Peroxidase-conjugated Buffer pH 8.8 1560 ÃŽ ¼L, dH2O 1974 ÃŽ ¼L, 30% w/v AffiniPure Goat Anti-Mouse IgG (H+L)) was then APS 21ÃŽ ¼L, TEMED 6ÃŽ ¼L), casted to 1 cm below top applied in TBST with 5% w/v nonfat dry milk at a of glass plate, and then 4% Stacking Gel (Protogel 1:2000 dilution of antibody and shaken for one hour 390 ÃŽ ¼L, Stacking Gel Buffer pH 6.8 720 ÃŽ ¼L, dH2O at room temperature. The membrane was then 1830 ÃŽ ¼L, 30% w/v APS 6ÃŽ ¼L, TEMED 3ÃŽ ¼L) was washed three successive times for 15 minutes with casted on top with a ten well gel comb. The TBST buffer at room temperature. The edges of the electrode set up was then submersed in 1 x Trismembrane were dried with a Kim and next 1mL of Glycine SDS Page Running Buffer. To each 20 ÃŽ ¼L Renaissance Western Blot Kit NEN Life Sciences of sample, 20 ÃŽ ¼L of Laemmli buffer (0.5 M TrisProducts, Cat. No. NEL 101 luminol reagent with HCl, pH 6.8, 4.4% w/v SDS, 20% v/v glycerol, 2% 1mL of oxidizing reagent were mixed together and v/v 2-mercaptoethanol, 10 mg/mL bromophneol then applied to the membrane. The membrane was blue) was added and boiled for 3 minutes and then then imaged with an exposure time of 40 seconds cooled for 5 minutes on ice. To one well 7ÃŽ ¼L of using AlphaEaseFC software. New England BioLabs Inc. Prestained Protein Marker (7-175 kDa) was added. To the following PCR/agarose gel electrophoresis wells 15 ÃŽ ¼L of standard Taq polymerase was added, A master mix for PCR was prepared (1x PCR along with 20ÃŽ ¼L of six different samples, with the buffer minus Mg, 0.2mM dNTP, 1.5 mM MgCl2, fifth being prepared the previous year with the same 0.5ÃŽ ¼M forward primer, 0.5ÃŽ ¼m Reverse Primer, 0.1 method of isolation as outlined previously. The gel ng Template DNA and Nuclease-free PCR water) was run at 200 Volts for 40 minutes, incubated in and 22.5 ÃŽ ¼L of master mix and 2.5ÃŽ ¼L of Taq fixing solution overnight and then stained with Bio- sample, the standard, or the Taq prepared a previous Safe Coomassie Blue for one hour at room year were added to PCR tubes and centrifuged temperature under agitation. The gel was then briefly on a Fisher Scientific Accuspin micro 17 analyzed used AlphaEaseFC software. just briefly using 1.5mL eppendorf tubes with no caps to contain the PCR tube. The PCR tube was Western Blotting then added to T3 Biometra Thermocycler and Using the method described above for SDSdenatured at 94 °C for 3 minutes and then 35 cycles PAGE, a SDS-PAGE gel was taken prior to fixing. of PCR with the denature 94 °C for 45 seconds, The gel was then transferred to transfer buffer anneal 55 °C for 30 seconds, and extension at 72 °C (20mM Tris-HCL, pH 8.0, 150 mM Glycine, 20% for 1.5 minutes. The sample was then incubated at Methanol). Immobilon-P transfer membrane with 72 °C for 10 minutes and then temperature was 0.45 ÃŽ ¼m pore size and Whatman paper were cut to maintained at 4 °C. The samples were then stored at the size of the gel. The membrane was wet with -20 °C until agarose gel preparation. A 1% agarose 100% methanol, then transferred to MilliQwater gel w as prepared through 1.5g of agarose (Sigma and soaked for several minutes. A standard blotting No. A-6877 Type II) to 150mL of Tris-Acetate4 Isolation of Recombinant Taq Polymerase for PCR EDTA (TAE) buffer. The solution was microwave for 1 minute and mixed until in solution. Once cooled to 60 °C, 7.5 ÃŽ ¼L of Biotium Gel Red Nucleic acid stain was added and mixed. The solution was then poured into the electrophoresis tray; a comb was installed, and set at room temperature. One Litre of 1x Tae buffer was prepared through dilution of 50x TAE buffer and then the solution was poured onto the electrophoresis tray to cover the gel in 1mm of buffer. 20 ÃŽ ¼L of PCR product prepared previously and 4ÃŽ ¼L of Gel Red dye were mixed and 20ÃŽ ¼L of each sample, the standard, and Taq prepared the previous year and Invitrogen life Technologies 1 Kb DNA ladder Cat. No. 15615016 was run at 150 Volts, 100 mA for one hour (or until dye reached the bottom of the gel). The bands were then visualized under 300 nm light and fluorescence was measured at 590 nm. The gel was analyzed using AlphaEaseFC software. concentration of the sample Taq was 1.88 + 0.11 mg/mL. The solution of proteins was not pure Taq as confirmed by the SDS-PAGE (Fig. 2) as various proteins created distinct bands (B to K excluding E). The standard Taq revealed only one band (A), indicating band E was most likely belong to Taq, as it was the darkest band in the gel. An analysis of the molecular weights of the bands through electrophoretic mobility (Tab. 3) showed the standard Taq having a molecular weight of 115.2 + 14.6 kDa, and the likely band (E) had a molecular weight of 113.4 + 14.3 kDa. There was a distinct distortion in the bands of the SDS page in all lanes with the exception of the standard Taq and the 2011 Taq (Fig. 3). The distortion is of a smile. The overall gel also has a large distortion, but of a frown. It would appear there was a similar protein to D E and F present in all samples, including the 2011 sample. The standard Taq did not contain the bands. Re al Time PCR The Western Blot (Fig. 4) revealed distinct A master mix for PCR was prepared (1x PCR bands; however, there were more than one band in buffer minus Mg, 0.2mM dNTP, 1.5 mM MgCl2, each lane with the exception of the standard Taq. 0.5ÃŽ ¼M forward primer, 0.5ÃŽ ¼m Reverse Primer, 0.1 Two distinct bands were present in 5, Taq, and 2 (b, ng Template DNA and Nuclease-free PCR water). c). The lanes of * and ? contained several bands To PCR tubes, 22.5 ÃŽ ¼L of Master Mix and 2.5 ÃŽ ¼L also. The overall gel also expressed a slight color of Taq sample or the standard Taq were combined, banding along the solvent front edge which is mixed through vortexing and then centrifuged with shown in both Fig. 3 and 4. The 2011 lane did not a Fisher Scientific Accuspin micro 17 just briefly appear to have any Taq present, as no band was using 1.5mL eppendorf tubes with no caps to distinguished. The entire ladder expressed some contain the PCR tube. The Taq samples were antibody activity. prepared in triplicates. 20ÃŽ ¼L of each sampled were The real time-PCR revealed a threshold reached then transferred to a 96-well PCR plate and then at 20 cycles, with the vast majority occurring at 24 sealed. The well was then placed in a BioRad CFX cycles. The melt curve showed an approximate connect Real Time System using the programing of melting temperature of 81 °C (Fig. 7). enzyme activat ion (95 °C, 30 seconds, 1 cycle), 40 The agarose gel electrophoresis revealed one cycles of Denaturation (95 °C, 1 second) and distinct band at approximately 5883.5 base pairs in annealing/extension (60 °C, 5 seconds), with a melt length. The brightest bands, and therefore the curve of (60-95 °C in 0.5 °C intervals, 3 seconds per highest quantities of Taq enzyme were found in the step, 1 cycle). The samples were then analyzed std., 2 and 4. When the base pairs specific activity using AlphaEase FC software. of the enzyme was calculated it was found to be 834.5 + 63.9 bp/min/ÃŽ ¼g of sample, or 3922.3 + 192.9 bp per minute. RESULTS The results of the Bio-Rad assay on the sample of Taq polymerases diluted to 10x and 100x revealed that the 10x dilution was far to concentrated and fell outside the linear curve of the Bio-Rad assay. The retrieval of protein from the Luria broth was found to be 300.8 + 17.7 mg protein per L of Luria broth. These results (Tab. 1) suggest the protein 5 DISCUSSION Through the analysis made through SDS-PAGE, the MW of the standard Taq was found to be 115.2 + 14.6 kDa and 113.4 + 14.3 kDa. This is different from the accepted literature value of 94 kDa (9). Even with error correction, the prot ein did not fall Isolation of Recombinant Taq Polymerase for PCR within the range of the accepted literature value. In total, the two proteins differ by 23% and 21% without error correction, or 21.2 kDa and 19.4 kDa respectively. In comparison to one another, the two bands have essentially the same molecular weight, indicating whatever error occured in the gel was equivalent on both the standard and the isolated Taq. One explanation for the difference in the molecular weights may be explained through the quantitiy of protein used. The darkest and thickest band ( E, fig. 4) likely belongs to the Taq protein. To get a more defined band, a dilution would be effective in making a higher resolution band (12). The amount of protein isolated per volume of Luria broth was determined to be 300.8 + 17.7 mg per L of Luria Broth. Quite obviosuly, there are issues both with the heating of the gel, and distortion of the bands into â€Å"smiles†. The distoration of the gel likely was caused by unequal heati ng of the gel causing the center of the gel to be hotter than the peripheries, as the walls of the apparatus act as heat sinks (13). The uneven heating can be removed by switching to a lower voltage for a longer period of time (12). The distortion of the protein bands within the individual lanes produced a smile structure. The distortion was likely caused by either an overloading of proteins, which can be solved by dilution of the protein sample, or was due to salt conditions of the loading sample. This step could be fixed through extra steps of dialysis to decrease salt content of the loading sample. (14). One final issue with the SDS-PAGE gel was the distance between bands. The target molecular weight was near 100 kDa, so the concentration of the gel could be decreased to allow for a higher resolution of the higher molecular weight proteins, or allowed to run for a longer period of time (14). A purity assessment of the isolated Taq enzyme can be made through the SDS-PAGE gel (fig. 2). Distinct banding occurs in ten different bands on the Taq lane, with 9 being distinct from Taq protein (E). This highlights that there were infact multiple proteins still present in the Taq solution. This would indicate that the heat shock portion of the methods was insufficient in denaturing all of the proteins in the E. coli, allowing for precipitation upon salting out. This is based on the extra protein banding only occuring for the Taq polymerases prepared for this experiment. A factor that could have also played a role was the incubation at 75 °C was continually 6 interrupted through the need to shake the reaction vessel thereby lowering the temperature of the solution. This was due to mechanical difficulties of the equipment. It would be best to find a working New Brunswick Scientific-Innova 40 incubator shaker series to improve the protein isolation. To decrease the protein impurities, an increased heat cycle could be implemented, as Taq is thermostable at 75 °C, and could sustain structure at that temperature for long durations (7). The ammonium sulfate salting out would be mor e efficent after an increased heat cycle as even fewer native state proteins would remain (10). Another method to decrease impurities would be to add a purification step using another specific property of Taq polymerase. This could be the isoelectric point. This could be done through ion exchange columnsor isoelectric focusing (12). The extra isolation step would significantly decrease the impurities, and increase the specific activity per mg of protein of sample.The impurities were likely a result of other proteins present in E. coli bacteria lysate that were relatively thermostable, as those proteins would be most probable (9). The isolation of Taq can be confirmed through the Western Blotting and PCR reactions (Fig. 4-7), as a distinct band in the Western Blot, and measureable amplicon replication in the PCR and rt-PCR. In the standard of Taq of the Western blot (Fig. 4) there is a distinct band. The same band in the channel containing the isolated Taq can be seen. The band occurs in the same relative vicinity as the Taq molecular weight band in the SDS-PAGE (Fig. 2) so would fit best fit the Taq enzyme. The banding of the blot shows a common band across all lanes that line up with the standard Taq, emphasizing the isolation of Taq. There is a hesitation in confirmation of Taq due to the extra protein banding in the prepared fractions, as these bands were not seen in the standard Taq. The banding would suggest proteins transferred from the gel to the membrane and was still able to bind to the primary antibody or secondary antibody. There are various possible explanations for this. First and foremost, the banding occurred in areas wherever protein was present (ladder and lanes). This would indicate lack of specificity in the primary antibody which is intended to only find full sequence Taq and bind to it (15, 16). Another problem may be due to lack of blocking solution binding to the membrane, or Isolation of Recombinant Taq Polymerase for PCR excessive washing removing blocking solution from the membrane. A final possible explanation may be binding of the secondary antibody to membr ane bound proteins with the exception of casein (the blocking protein used) (15, 17). Antibody specificity can be corrected by finding a new antibody, lack of blocking simply requires longer blocking periods or increased blocking solution concentration, and washing can be minimized to see resultant effect on the membrane. Each of the possible problems with the Western Blot would have to be tested by altering the procedure used above by one method (washing, antibody, blocking solution). The PCR results show template replication through thermocycling, which indicates the presence of thermostable DNA polymerases in the PCR tube. From this, it can be conferred that Taq polymerase was indeed isolated. Further confirmation could be made through further purification of Taq. This could be done through 2-D SDS-PAGE vs Isoelectric point electrophoresis using the isoelectric point of Taq and using the bands emphasized as Taq, and a lower concentration gel (12). Another method would be to analyze the gel bands through other methods such as mass spectropscopy or NMR (18). There wa s distinct differences between three sets of Taq polymerases: the standard, the sample prepared in the previous year, and the sample produced in this experiment. Most distinctly the proteins differ with respect to SDS-PAGE gels. Quite obviously, the purest of the enzymes was the standard Taq, followed by the 2011 sample, and the sample prepared in this experiment. The sample prepared through this experiment had a high amount of a salt concentration and resulted in distorted bands, along with numerous other proteins present in the sample. The enzymes also differed with respect to the Western Blot (Fig. 4). The 2011 sample failed to return 2 ° antibody response, indicating lack of Taq polymerase, or lack of primary antibody binding, while the standard and experimental sample both had representive banding. There may have been excessive blocking or drying of the lane containing the 2011 Taq, as the SDS-PAGE shows a representive band in the region of Taq, that is the darkest band in the lane (15). The protein concentrations as determined through the Bio-Rad assay (Tab. 1, Fig. 1) returned 7 drastically different results. The two protein concentrations differed by 2x concentration. The easiest explanation of thi s result is the 10x dilution was insufficient in reducing the absorbance to within the standard curve. Due to the absorbances being above the standard curve, the results are invalid, as the region in which the curve is linear is up to 0.5mg/mL (19). The 100 x dilution returned a result of 1.88 + 0.11 mg/mL. This coroborates the SDS-PAGE findings as the protein was not excessively overloading the lane. The SDS-PAGE could have been further diluted, but the concentration used was sufficient for the purposes of the experiment. In an analysis of the PCR results (fig. 7), the brightest fluorescence bands occurred in the std., 2 and (4/Taq) lanes. This would indicate the highest activities occuring in these lanes. When compared to the western blot, the darkest banding of regions of Taq (5,?,*) returned the bands with less fluorescence. This result shows that the amount of enzyme may inhibit the PCR reaction as the the bands with the highest recoveries returned the lowest fluorescence. With an assessment of the basepair length, reaction time, and amount of enzyme used, an approximately activi ty of 834.5 + 63.9 bp/min/ÃŽ ¼g of protein, or 3922.3 + 192.9 bp per minute. In comparison to the literature values of the protein, this is slightly above the 60 base pairs per second value, however, that was at 70  °C (7). The rt-PCR returned a consistent melting temperature of 81 °C (Fig. 6)for all amplicon samples indicating the lack of a primer-dimer formation. Threshold was initially reached at 20 cycles (Fig. 5), which an RFU value of approximately 9000. This indicated a high activity of the taq polymerase used, at least above 1.25 Units (20). Both PCR assays agree with one another. There was no primer dimer formation noted on the agarose gel, or the melt curve analysis. There was a high activity of the enzyme sample isolated as found through the bpmin-1 and cycle # of reaching threshold, however, between the two assays, the rt-PCR has the significant advantage of time, and no electrophoresis required. Currently, Taq is widely available and would likely be cheaper to simply purchase commercially. This experiment does however outline a method for thermostable protein isolation which could be used for the more recent and more valuable thermostable enzymes (Pyrococcus furiosus Polymerase) which Is olation of Recombinant Taq Polymerase for PCR are superior to Taq in both thermostability, and error rate due to proofreading ability (21). Overall, the purpose of the experiment was met. Taq was indeed isolated from a culture of recombinant E. coli. This was confirmed through the Western Blotting, and thermostable DNA activity in the PCR and rt-PCR. The purity was assessed and found to be below that of the methods used by Engelke et al., 1990. The purity could be increased through use of a cation exchange column (9). The length of heat denaturation and an automatic heat controlled shaker would help to remove excess proteins and improve purity. The length of dialysis time would need to be increased for less band distortion in SDS-PAGE, and either more selective primary antibody, increased blocking or decreased washing would be required for improved Western Blotting. For further experiments, it is suggested testing the new method modifications, and or implementing recombinant Pyrococcus furiosus Polymerase.

The Work Life Balance In Nestl International Management Essay

The Work Life Balance In Nestl International Management Essay Work/life balance is a very broader term and often referred to finding the right equilibrium between the different aspects and roles in a life of a person. Although there are different views and concepts about work/life balance but it is generally coupled with maintaining an overall balance in different aspects of life. During the last decade technological revolution has stormed all aspects of life and there were different debates about the future of work. Different scenarios predicted a rising trend in un-employment and economic insecurity. Industrial restructuring and emergence of latest technology no only affected the importance of workforce but it was also felt that employees will be treated as superfluous overheads. In this study of work/life balance the main objective will be to examin the ability of employees to handle concurrently the versatile demands of life at Nestle International. Work/life balance demands the right balance and equal amount of time devoted to official as well as personal life. The theory of this concept is not that simple and requires many developments and additional components to be incorporated. There are three different levels of equilibrium that affect the work/life balance. CHAPTER 1 Introduction History of Work/Life Balance: During the 1960s and 1970s, the concept of work/life balance was a new one and was basically used for those working mothers who usually struggle to cop up the requirements of their jobs as well as raising their children. However during the 1980s the importance of work life balance acknowledged all over the world and many multinational companies introduced the policies and measures to secure the rights of the workers and tried to maintain a right balance in their culture. By the end of the decade, work-life balance was seen as more than just a womens issue and male workers also raised their voices. By the mid of the 1990s work-life balance became a fundamental issue for individuals and it became a necessary part of the strategies of all the organizations all over the world. This growing awareness of the central importance of the issue resulted in major growth in attempted work-life solutions during this decade and many new studies and ideas emerged. As for today the situation has changed drastically and many changes have been made in labour law regarding the protection of rights of the workers and these changes in laws also have a significant affect on HRM policies and strategies of the organizations. Why Work/life Balance so Important? Over the past few decades, there has been substantial increase in work which is felt due to a very stiff competition in global markets and ever increasing pressure and demands from the organizations that expects more and more from their employees yet offers a very little security and compensations in return. For many employees today including both male and female their lives are becoming more complex and difficult with an increased burden of family and other personal responsibilities and interests. Therefore, in an effort to retain employees and maximize their performance, it is increasingly important for organizations to recognize this balance. This increased pressure has affected the life of worker at Nestle International in many ways. It is not only harming the social norms but also causing many physical and psychological diseases such as heart problem, weak immune system, stiff muscles, exhaustion and jumpiness etc. Increased working hours means there will be very less time to be spend with family and community. If an employee is not mentally satisfied or balanced, he will not be able to perform his duties with due diligence and care. Now, however, things have taken altogether a different turn. Employees quality of work-life is becoming a burning issue all over the world. For a growing number of employers, human resource challenges are their biggest concern. Leading employers are recognizing that positive work-life outcomes for employees from achieving work-life balance to the satisfactions of challenging work and career development are key ingredients of a successful business strategy. In practice, improving the quality of work life remains a tough challenge. Naturally, many managers see the important links between the quality of work life and organizational performance. Yet they face barriers to progress. So it is necessary to update the case for taking action to improve the quality of jobs and work environments. A happy and well balanced employee will not only perform his daily operation with utmost care but will also be able to avoid different complex social problems. Work/life programs will not only help to boost employees morale, reduce absenteeism, preventing the outflow of valuable information of the organization in difficult economic situations but will also reduce the cost and affect the overall performance of the company. It is very important for the human resource professional to understand the critical issues of work/life balance. Be it employees whose family members and/or friends are called to serve their country, single mothers who are trying to raise their children and make a living. Objectives of the Research To understand the concept of work/life balance. To understand the importance of a good and happy relationship between employer and employee. To evaluate the importance of work/life balance from the perspective of both the employer and employees. To explore the work/life balance at Nestle International. To evaluate the strategies of management at Nestle International and their role in maintaining work/life balance. To find out the solutions of different problems faced by the Nestle management in maintaining work/life balance within the organization. Importance of Work/life Balance from the Perspective of Employer: A balanced culture/environment in any organization will not only be helpful for employees in different ways but it will also provide many tangible benefits to the business. Implementing work/life balance provides an excellent opportunity to consult with staff to review the job design and develop innovative and supportive work practices. This sort of consultation will ensure the needs of individuals are met and a more agile workplace is achieved. It will also lead to a greater level of employee commitment to the success of the business. By minimizing the employees turnover in organization you will not only boost your repute as a good employer but it will also help to achieve the quality standard of products out prevent the outflow of valuable inside information to other organizations because if a skilled worker will leave the organization it will take a hell lot of time any money to replace him with some other suitable person. Importance of Work/life Balance from the Perspective of Employee: The For human being, home is a shadow of heaven on earth where he goes and takes rest but due to imbalance structure between work and life it has become an additional place of work with plenty of office tasks and assignment. Any human being will be able to perform his duties and tasks deliberately only when he/she has a right balance in between his/her official and personal life. A balanced and happy employee is more likely to perform better at work and can provide many tangible benefits for a business. There are new ways to make work work that benefit both employers and employees. In fact, research shows that flexibility is a critical component of workplace effectiveness and that employees who work in flexible and effective work places are more likely to: Be engaged in their jobs and committed to helping the company succeed. Plan on staying with their employers. Be satisfied with their job, which translates to good customer service. Exhibit better mental health and wellbeing. CHAPTER 2 Literature Review Introduction: Nestle is a Switzerland based company, founded by Henri Nestle in 1866. The success story of transforming a single mans idea into a giant corporation started from developing a milk food formula for infants by Mr. Henri in 1866. Soon the Nestle products became a success and more and more businesses wanted to merge or start a partnership with Nestle. In 1905 Henri Nestle joined hands with a Swiss milk company and the companys name became Nestlà © Anglo-Swiss Condensed Milk Co. Then in 1929, Peter-Cailler-Kohler Chocolats Suisses S.A. merged with the company. The name was then changed to Nestlà © Anglo-Swiss Holding Co. Ltd. In December 1947, company acquired all the shares capital of the Alimentana S.A. Company in exchange for fifteen Nestlà © shares and fifteen Unilac shares for each of Alimentana S.A. share, and the name was Nestlà © Alimentana S.A. And then finally, the last name change that the company would endure was in 1977, where it adopted the name Nestlà © SA (www.nest le.com). Today Nestle is one of the biggest giants in the food and beverage market of the world and it is marketing its products in almost 130 countries of the world. The company is manufacturing roundabout 10,000 different products and employing over 250,000 people all over the world. The company is selling almost a billion products everyday. The headquarters of Nestle are situated Vevey, Switzerland, but the operational facilities of the company are spread all over the world. The company not only increasing its size year by year but also increasing its product varieties. Nestlà ©s Greatest Asset are its Workers The slogan of Nestle that its people are their greatest asset is very much true as they care a lot about their people and encourage them to bring out the best in them. At Nestle it is made sure that people benefit maximum from their work at Nestle. What binds together the over 250,000 people across the world is a universal culture and a high degree of loyalty to the Company and to each other. The employees are encouraged to contribute in the company success by introducing new and innovative ideas. Above all, Nestlà © is genuinely international. One simple example is that around 80 different nationalities are represented among the 1,600 people in our Head Office. Each year a thousand people from all over the world come to our Training Centre in Switzerland. The corporate values of Nestle are: Trust Respect Involvement Pride The Concept of Work/life Balance at Nestle International Nestle is following the United Nations Global Compacts (UNGC) guiding principles that insures the human and labour rights and aim to provide an example of good human rights and labour practices throughout our business activities. At Nestlà © it is truly believed that there must be a good and healthy balance between the private and professional life of an employee. This thing not only increases the reputation of Nestle as a good employer but also helps to retain the most valuable people of the organization and reconciles economical essential with welfare. Employees who wish to take part in community welfare programs voluntarily are supported at Nestlà ©. In the same character, Nestlà © encourages flexible and friendly working conditions so that stress factor can be reduced up to the minimum level. Everyone has to find their own balance between work and other activities, but as a company we encourage all our employees to pursue interests outside the workplace. As football is the natural favourite Sports in European region so most Nestlà © units sponsor at least one football team. Nestle supports many cultural activities and home study programs in countries all over the world. One of the biggest evidence of good work/life balance at Nestle is that it enjoys one of the highest percentages of employees retention among the top multinationals of the world. Pre-retirement classes and facilities for retiree activities are available in many countries. Nestle also supports one of the largest retiree club with more than 1,000 members actively participating in events ranging from climbing, skiing and cycling to foreign travel, fitness and computer classes at its headquarters at verve, Switzerland. In addition, several of them take part in local volunteer projects such as delivering meals to other Nestlà © retirees in the region. Role and Strategies of Management in Maintaining Work/life Balance at Nestle One of the greatest debate is who bears the biggest responsibility in maintain equilibrium between official and personal life of a person? But according to the given circumstances the major burden definitely falls on the employer and those who are the part of senior management because these are the people who demand as much as work from their employees and tries to minimize the cost on the verge of a limited work force. Things have changed a lot now because different research works have showed the physical and mental health effects of heavy work load which ultimately affects the productivity and profitability of the company in the long run. At Nestle it is made sure by the management that an appropriate work-life balance exists in organizational culture which means employee will be at less risk for many serious health conditions that result from overwork and stress. Overworked employees not only exposed to many serious physical and mental diseases but also face many problems and issues in their social and family lives. This is a very alarming situation because those employees who spent the most of their time at work are usually error prone. On the other hand those employees who spend most of their time at off-work activities will usually be un-productive and not reliable. Such individuals are found in every office and they are major concern for the management. So it is the responsibility of the Nestle management to address the problems of these individuals. Management should encourage these individual to contribute more towards productivity. If an employee spend four hours working at home and solves more problems by working at home than at office , he should be encouraged by the Nestle management. At Nestle International top management offer different workshops and informative seminars to employees about work-life balance, and should make sure that the policies regarding these issues must be followed strictly. Different technologies have been introduced to employees, which help them in creating the right balance between their lives. Here are some of the strategies used by the Nestle management to enhance work/life balance in organizational culture. Employee Assistance Program (EAP) At Nestle International the introduction of Employee Assistance Programs (EAP) provide easy, free and confidential access to professionally trained counsellors to help the employees to overcome a large variety of professional and personal problems. Conveniences at Work Employees at Nestle International are offered different features in order to make their work convenient and easy such as: Fitness center Mothers lounge On site parking Bonuses at all levels Day care Cafeteria Parental Leave Parental Leaves are available for female and male workers to help them to see off their social responsibilities. Benefits Package Nestlà © offers medical and other visionary benefits packages to meet the health needs of not only their employees but also their families and domestic partners. Benefits of Maintaining Work/life Balance at Nestle International Talented and highly valued employees are the most important corporate resource of any company and it is not very difficult to get such talented employees so in order to retain them a flexible and balanced environment must exist within organization and organizational culture at Nestle fully demonstrate such features. If the employees will be satisfied they will be more focused and confident towards their work and by providing a right balance between the personal and professional lives Nestle ultimately enjoys a constant product quality and customer satisfaction. A balanced employee not only enjoy a happy personal life, avoids different risks relating to physical and mental heath but also add up to the reputation of Nestle as a good employer. By employee retention Nestle not only saves the outflow of very valuable information of the organization but also minimizes the additional cost required to train the new employee to bring him up to the level of expertises of the existing employee. CHAPTER 3 Methodology The main objective of this research is to create an understanding of the importance of work/life balance in the lives of employees as well as in the progress of the organization. General method to collect information for the purpose of conduct a Research Following are some of the method to collect information in order to carry on research. Questionnaire It is a method used to collect the information directly from people. It is consider as a realistic source of information which provide an opportunity for analysis. Moreover it provides more current information as compare to other source. it is considered to be the primary source of information. It also contain different options Interview Interview is another way of collecting data for conduction research. We can conduct different kinds of interview to find out the reasons of any problem. it is a direct face to face communication through which one can receive direct feedback on any issue under consideration. It is also one of the realistic sources of information. Studying the different documents and Survey Reports It is also an effective way to develop an understanding of different topic related documents and surveys which enable us to carry the research in an effective manner. These surveys enable us to develop an understanding on that specific issue in wide scope. Observation Observation is also an effective tool to collect information regarding customer expectations and factors which boost up customer expectation. Observation also helps to collect information regarding customer taste, marketing trends and rival competitive policies Discussion with expertise Carrying on discussions with those who have a vast expertise of understanding customer behaviours and mental situations. Through this we can extract very important information for the real purpose. Case Studies Studying different kind of case study is also enable us to enhance our understanding with the real topic and provide us very valuable information and clears the idea of main concept. The data collected through a questionnaire circulated and interview conducted at Nestle will form the base of this research work. The above mentioned questionnaire will consist of different questions relating to the work/life balance and it will target a group of 20 workers selected through random sampling representing the population as a whole. For the purpose of reference and other historical information different books, journals, articles and websites will be consulted. Time frame for completion of research is as under: No. Task Description Start Date Completion Date 1 Proposal Submission 2 Literature Review 3 Questionnaire Designing 4 Data Collection (Primary Secondary) 5 Data Analysis 6 Drafting Composing 7 Proof Reading 8 Submission