Science Can Not Be Defined - 1531 Words

Science Cannot be Defined Calvin Berndt Philosophy of Science Spring 2015 05/03/2015 There are a multitude of conflicting theories that aim to conclude the definition of science, and all of them have underlying issues that make them inadequate in giving a definitive definition. According to Dictionary.com the current prevailing definition of science is â€Å"the systematic study of the structure and behavior of the physical and natural world through observation and experiment (Dictionary, n.d.).† However, the attempted definitions of science are not useless as they are still very useful in understanding what science is and what it means to carry out science, but there is no simple unobjectionable definition. The before-mentioned†¦show more content†¦The example Bauer uses to demonstrate this idea is â€Å"just because the first million swans were white, one could not guarantee that the next one would be white also. (Bauer, 1992)† The very next swan could indeed be black, and because of this possibility the thesis that ‘all swans are white ’ could never be true. David Hume brings up the fact that although we have always perceived the same cause and effect, their connection is not necessarily true. Another example would be that all metals expand when heated. It is true that all of the metals humans have identified to this point in time have been tested to expand when heated, but that doesn’t necessarily mean that we will never discover a metal that doesn’t expand when it is heated in the future. Hume concludes induction cannot be a rational inference, due to the fact that inductions can produce false conclusions from true premises, and ultimately implies the idea that correlation doesn’t necessarily mean causation. These examples demonstrate the idea that no absolute certainty can be derived from the scientific method. Karl Popper expands this idea by saying that what makes something scientific is the concept of it being falsifiable, rather than a method that results in absolute truth (Jogal ekar, 2014). This means that in order for a theory to be scientific it has to have the possibility of being tested to be false or incorrect. According to Popper, this is also

The Importance Of Spaying And Neutering And The Problems...

My topic is on spaying and neutering and the problems those surgeries could make better or even fix completely. Ultimately by spaying or neutering a pet, the owner would be helping lower the overpopulation of animals. A pet owner would also be helping their pet by not allowing them to go through puberty or heat cycles. It is a fact known worldwide that spaying and neutering your pet will help lower the overpopulation of animals. Overpopulation has been a problem for many years now. There are too many stray animals wandering the streets and living in animal shelters awaiting a home that won’t come. The importance of spaying and neutering is to give the pet owner and the pet the best life. Veterinarians perform spaying and neutering†¦show more content†¦Everyone is affected by overpopulation, even those who do not own pets. Human health is at risk from all the stray animals having transmittable diseases such as rabies and the animal waste. There are several different procedures on spaying and neutering pets. Each and every procedure would be performed by a veterinarian who has studied medicine. Discuss all of these procedures with a veterinarian for the best option for each specific pet. Of course, there is the usual spay procedure is a ovariohysterectomy, which is when the ovaries, fallopian tubes, and uterus are removed from a female pet. As a result this makes the female pet unable to reproduce and eliminates the breeding instinct-related behavior and the heat cycle. Also there is the hysterectomy, which is when the uterus and part of the fallopian tubes are removed from a female pet. This makes the female pet unable to reproduce, but the female will still produce hormones because of the ovaries. This may not eliminate the female behaviors related to the breeding instinct. Lastlty, for the female pet there is the ovariectomy, which is when the ovaries are removed from a female pet but the uterus remains. Similarly t o the effects of ovariohysterectomy such as making the female unable to reproduce and eliminates the breeding instinct-related behavior and the heat cycle. The usual neuter procedure is a orchiectomy, which is when the testes are removed from aShow MoreRelatedEssay on Animal Overpopulation 1405 Words   |  6 PagesThe state of companion animals in this country is precarious at best; caught between scientists who subscribe to Descartes’s idea of ‘anima ex machina’ (unfeeling, a living example of biological processes without the status of ‘being’) and the more common phenomenon of people who pamper their pets in ways that most people would envy. For most individuals living in an urban society such as ours, the most common interaction with animals happens within the home – if the animals that we relate to andRead MoreEssay on Prevention of Animal Overpopulation1722 Words   |  7 Pagesnothing to end the horrific crimes of animal deaths in sh elters. It is no secret that this world has become infused with problems that have extended from one side of the globe to the other. Amongst these problems lies a terrible truth: nearly every year, sums of almost eight million cats and dogs have been placed in shelters around the world. Out of these vast numbers, half will be euthanized; that equals to one animal being put down every 8 seconds. Animals that are not adopted are kept in sheltersRead MoreThe Overpopulation Of The Ohio Valley1505 Words   |  7 PagesStates, about 7.6 million companion animals enter animal shelters every year. The overpopulation of these animals and the fact that they have no homes leads to about 40% of them being euthanized (ASPCA). This is a global issue, but the root of it can be found within the local community at The Humane Society of the Ohio Valley. The director of this animal shelter, Kelly Goedel, says that the cause of animal homelessness is primarily the lack of spaying and neutering along with the lack of commitmentRead MoreAdopt and Save a Life Essay1553 Words   |  7 Pagesrealize many animals have a life as this dog has had. Studies show that more animals in the United States are being euthanized every day due to the fact of overpopulation of animals, serious injury or diseases, puppy mills and natural disasters. According to the Humane Society of the United States, they estimated that in 2013, animal shelters care for about six to eight million dogs and cats every year, of this approximately three to four million are euthanized (Pet Overpopulation). This statedRead MoreMission. According To Pets Alive (2016), The Mission Of1019 Words   |  5 Pageshealth and well-being of cats and dogs. To accomplish this, they have partnered with the animal welfare community. This partnership will allow them to provide both life -saving programs and solutions to the problem of overpopulation (Pets Alive, 2016). History According to â€Å"Pets Alive†, in April of 2002, Katrina Sturgeon read a newspaper article that stated that around 70% of the animals brought to the animal shelter in Bloomington were euthanized. Appalled by this statistic, she wrote a letter toRead More Adoption of Animals Essay1808 Words   |  8 PagesAdoption of Animals Anyone, who visits an animal shelter, as I do, sees an extraordinary number of beautiful, affectionate, and desperate dogs and cats. The majority of animals in any particular shelter are dogs, usually adults, for whom there arent enough adoptive homes waiting. A few may have come from responsible breeders, whose owners do not realize that the breeder will take them back,many are those who are lost, and/or from owners who simply got tired of them. Some are pet shop puppiesRead MorePet Overpopulation Epidemic Essay1518 Words   |  7 PagesSpeech Topic: Pet Overpopulation Epidemic General Purpose: To persuade. Specific Purpose: To persuade my audience that the public is to blame for the pet overpopulation epidemic. Central Idea: In order to control the overwhelming population of homeless pets, we need to stop throwing our â€Å"family friends† away. There are 1.5 dogs and cats put to sleep every second 4-6 million dogs and cats are euthanized every year, states the Humane Society. The pet overpopulation epidemic has becomeRead MoreHow to Save an Animal Life Essay1636 Words   |  7 Pageslived a long, healthy life and it was finally his time to go. Many animals today are being taken out of this world in the same manor that Happy did, but the only difference is that it is not their time. This is due to overpopulation in animal shelters. Death in animal shelters due to overpopulation is a problem that I believe could easily be resolved if we were to take more responsibility as pet owners. There are simply too many animals being brought into this world, and not enough space or homes forRead MoreLife Long Friend Make A Good Choice1694 Words   |  7 Pagesbreeders, when in reality our country should be looking at humane societies or a local animal shelter for a new friend. Many of these dogs in the animal shelters are being euth anized each year. Everyone is capable of finding a new dog to take home and love at the shelter. Additionally, the cost to buy a new puppy could be quite expensive especially depending on the type of breed. Even though all these animals in the shelter just want love, affection, along with unconditional attention yet people’sRead MoreChoosing A New Perfect Dog For You And Your Family Sometimes2122 Words   |  9 Pagesor dog breeders when in all reality they should be looking at humane societies or a local animal shelter for a new pet. A big population of the dogs in animal shelters are being euthanized each year. Luckily the percentage is going down. Additionally, the cost to buy a new puppy from a pet store could be quite expensive especially depending on the type of breed the person chooses. Even though all the animals in these different shelters just strive for love, affection, along with unconditional attention

Pros and Cons of Coal Energy free essay sample

The use of coal is much bigger than that. Coal is used to create almost half of all electricity generated in the United States. It is inexpensive compared to other energy sources, which means lower bills for its consumers. Coal is easily stored, transported, and controlled in large volumes A variety of industries use coals heat and by-products. Separated ingredients of coal (such as methanol and ethylene) are used in making plastics, tar, synthetic fibers, fertilizers, and medicines. Coal power produced 49. 7% of the U. S. electricity supply in 2005. According to the Energy Information Administration, the U. S. has demonstrated coal reserves of approximately 496 billion tons of coal (more than any other country in the world), half of which is mine able with existing technologies. Consumption of coal worldwide is expected to increase 71% between 2004 and 2030, driven by developing countries that use coal to generate about 75% of their power. The burning of coal, like that of gas and oil produces carbon dioxide (CO2) the main greenhouse gas, which is linked to global warming. We will write a custom essay sample on Pros and Cons of Coal Energy or any similar topic specifically for you Do Not WasteYour Time HIRE WRITER Only 13.90 / page Coal also contains sulfur dioxide when it is burnt. In the atmosphere, it becomes sulfuric acid by oxidation, an irritant for the lungs and a main component of â€Å"acid rain†. Coal is a fossil fuel that is burnt in order to produce electricity. It produces emissions such as sulfur, nitrogen oxide (NOx), and mercury, which can pollute the air and water. Sulfur mixes with oxygen to form sulfur dioxide (SO2), a chemical that can affect trees and water. Mercury that is released into the air eventually settles in water. This mercury in the water can build up in fish and shellfish, and can be harmful to animals and people who eat them. Now that I’ve told you about the cons of coal as our energy power source, I must tell you that there are changes being made to reduce these risks and ease your minds of ecological concerns. Industry has found several ways to reduce sulfur, nitrogen oxides (NOx), and other impurities from coal. They have found more effective ways of cleaning coal after it is mined, and coal consumers have shifted towards greater use of low sulfur coal. Power plants use flue gas desulfurization equipment to clean sulfur from the smoke before it leaves their smokestacks. There are also technologies that can remove impurities from coal or that make coal more energy-efficient so less needs to be burned. Scientists are also working on new ways to reduce mercury emissions from coal-burning power plants. Reuse and recycling can also diminish coal’s environmental impact. Land that was previously used for coal mining can be reclaimed for uses like airports, landfills, and golf courses.

Mark Twain As Philosopher Essays - Picaresque Novels,

Mark Twain As Philosopher Mark Twain is, according to critics and readers alike, the first great American novelist (Reuben). Throughout his lifetime Twain, born Samuel Longhorn Clemens, held an eclectic mix of jobs, and, wrote a great deal about his experiences and his boyhood. The Adventures of Tom Sawyer (AOTS) and Adventures of Huckleberry Finn (AOHF) are a pair of novels by Twain that: present the new and radical changes in the early 1800s in contrast to the old fashioned ways; mirror Twain's life as a young boy growing up in a one-horse town on the Mississippi River; and, give the reader an idea of his view that the loss of innocence signals the coming of age. Twain was born in 1835 and Tom Sawyer grew up in the 1840s. Around this time, America, especially the North, was undergoing "revolutionary changes in transportation and communication" (Geise 93). The river steam boat was invented in 1807 (Roberts and Kennedy 305) and subsequently took over mass transportation from sailboats using the ocean (Geise). This was a big change from the previous small scale or trans-ocean transport. After the steam boat came the steam train which revolutionised transportation in a similar fashion, and they synergistically opened the West to all people and boosted trade and commerce enormously--not just of the big industrial towns but of the en-route towns and the farms, In 1849, agriculture accounted for over half of the nation's economy, whereas today it is one-fiftieth (Roberts and Kennedy A27). Canals, turnpikes and clipper ships also greatly affected transport and communication between distant places (311). The times were revolutionary in that the old ways of taking dirty, bumpy roads long distances with little profit were over. Another sign of the times was slavery. Racism was widespread during this time period because many large farms and plantations held slaves. Feelings towards slaves in Missouri were not generally sympathetic, and abolitionists were not well accepted because the economy would collapse without the slave based agriculture. Rudyard Kipling wrote at the end of that century "The White Man's Burden," (643) that was taken to mean that blacks must accept their position as underlings. While a false interpretation, it shows that many Confederates and sympathisers held the view that blacks and slaves deserved to be oppressed even after the Civil War (1861-1864). TAOTS accurately reflects the small town economy. The river trade is the centre of all commerce and without it, town life would end. In Chapter Two of TAOTS, Ben Rogers, a local boy, pretends to be a steamboat. This exemplifies how important the boats were to the town. Everything in the town--the mill, the taverns--they all depended on the trade from the river. The town, consisting of a church, a school, a general store, taverns a mill and a docking area for the boats also reflect how important the river really was. The minister's fire and brimstone sermons (35) preach against the evils of drink, gambling and lust, all of which would have been demonstrated by the passing river sailors and conmen. In the AOHF, the town life is not so much the focus of description as river life. But it is the description of the treatment of slaves that truly stands out. Huck was poor, but still he was socially above Jim because he was white and not owned. TAOHF was set a few decades before the civil war so when Huck and Jim escaped down the Mississippi and headed south, they were putting Jim in more peril. When they took on board the King and the Duke these other travelers wanted to turn Jim in. Many non-slave states actually had laws that allowed for the returning of runaway slaves (Geise 109). Both TAOTS and AOHF are accurate in their description of the situation (slave-wise and town-wise) at that time. Mark Twain's views about childhood and the subsequent loss of innocence are a product of childhood experience growing up in Hannibal, Missouri (pop 500), a small town on the Mississippi River. As a young boy, he enjoyed skipping school to go fishing on the nearby island; playing with the off-limits Tom Blackenship (Draper 3713), the son of the town drunk; or spending time with his sweetheart Laura Hawkins (Thayer 5). Twain once had a harrowing experience as a child when he got lost in a local cave with Laura. Living in the small river town, whose only commerce was from the steamboat trade, he witnessed at least four murders (Sanderlin). When he was eleven, his father died (Meltzer 75). He quit school

Fred Page essays

Fred Page essays Fred Page was born on a 3,000-acre farm in Louisiana. He lived there for most of his teenage years. He then moved to Mississippi where he has lived for 52 years. Overall, Mr. Page has had a very long and interesting life. In Louisiana, Freds family was one of the most well off black families of its time. He lived in a great big house and his father worked for many rich people. They raised hogs and cows which was rare for some plantations. When Fred was 12 years old, he was hired to pick cotton. He could pick about a couple hundred pounds of cotton by two oclock. During the picking he would hardly see a white person, unless the overseer was white. When he moved to Mississippi, he began working at the Melrose Plantation at the age of 18. His owner, Mrs. Kelly was very high cultured. She taught him the rich culture ways like they would use in New York, so he would know how the white people would talk to him. Fred was very sophisticated for a young black teenage. When he started working in the State of Mississippi he wasnt quite used to the way a black person was supposed to speak. Everyone you talked to is yas em and no em and yas em giving that high honor, you know. And I started saying yes and no, but I soon got out of that. In a couple of weeks I blended back down to Yas and nosm, that sort of thing. This is very similar to the character Grant in A Lesson Before Dying. Grant was smarter than most of the white people he knew, but sometimes had to act more dumb to give them pleasure or acted smarter to piss them off. He began only giving tours to blacks and everyone gave Freds tours high reviews. When he started giving tours to whites, they asked Mrs. Kelly if he could handle it and they would say, I dont want him talking to my children, while Fre ...

The Longest Words in the German Language

The Longest Words in the German Language The classic longest German word is Donaudampfschiffahrtsgesellschaftskapitn, clocking in with 42 letters. In English, it becomes four words: Danube steamship company captain. However, its not the only super long word in the German language and, technically, its not even the longest. German Spelling Most languages, including English, string smaller words together to form longer ones, but the Germans take this practice to new extremes. As Mark Twain said, Some German words are so long that they have a perspective. But is there really such a thing as the longest German word...  das lngste deutsche Wort? Some of the suggested longest words are artificial creations. They are never used in daily spoken or written German, which is why well look at some words that far surpass our 42-letter title winner mentioned above.   For all practical purposes, this longest-word contest is really just a game. Its more fun than practical and German just happens to offer us some really long words. Even a German or English Scrabble board only has room for 15 letters, so you wont find much use for these. Yet, if youd like to play the longest-word game, here are a few selected items to consider. The 6 Longest German Words (Lange  Deutsche  Wà ¶rter) These words are listed in alphabetical order, with their gender and letter count. Betubungsmittelverschreibungsverordnung(die, 41 letters) Its a mesmerizing word that is rather difficult to read. This lengthy one refers to a regulation requiring a prescription for an anesthetic. Bezirksschornsteinfegermeister(der, 30 letters) This word may be short in comparison to those below, but it is a real word that you might be able to use someday, but even thats not likely. Roughly, it means a head district chimney sweep. Donaudampfschiffahrtselektrizittenhauptbetriebswerkbauunterbeamtengesellschaft(one word, no hyphen)  (die, 79 letters, 80 with the new German spelling that adds one more f in ...dampfschifffahrts...) Even the definition is a mouthful: association of subordinate officials of the head office management of the Danube steamboat electrical services (the name of a pre-war club in Vienna). This word is not really useful; its more of a desperate attempt to lengthen the word below. Donaudampfschifffahrtsgesellschaftskapitn(der, 42 letters) As mentioned, in classic German this is considered the longest word. Its meaning of Danube steamship company captain makes it unusable for the majority of us, though. Rechtsschutzversicherungsgesellschaften(die, plur., 39 letters) This is one you might actually be able to pronounce if you take it one syllable at a time. It means, legal protection insurance companies.  According to Guinness, this was the longest German dictionary word in everyday usage. However, the word below is a longer legitimate and official longest word- in semi-everyday usage, anyway. Rindfleischetikettierungsà ¼berwachungsaufgabenà ¼bertragungsgesetz(das, 63 letters) This hyper word references a beef labeling regulation and delegation of supervision law.  This was a 1999 German Word of the Year, and it also won a special award as the longest German word for that year. It refers to a law for regulating the labeling of beef- all in one word, which is why it is so long. German also likes abbreviations, and this word has one: ReÃÅ"AÃÅ"G. German Numbers (Zahlen) There is another reason why there really isnt a single longest German word. German numbers, long or short, are written as one word. For example, to say or write the number 7,254 (which is not really a very long number), the German is siebentausendzweihundertvierundfà ¼nfzig. That is a single word of 38 letters, so you could imagine what larger and more complex numbers may look like. For this reason, its not difficult at all to make a number-based word that far exceeds any of the other words weve discussed. Longest English Words For comparison sake, what are the longest words in English? Contrary to popular belief, the record holder is not supercalifragilisticexpialidocious (an invented word made famous in the movie Mary Poppins). Just as in German, there is disagreement about which word is actually the longest. Theres little argument, however, that English cant keep pace with German in this department. The English languages two contenders are: Antidisestablishmentarianism  (28 letters):  This is a legitimate dictionary word from the 19th century meaning opposition to the separation of church and state. Pneumonoultramicroscopicsilicovolcanokoniosis  (45 letters):  The literal meaning of this term is lung disease caused by breathing in silica dust. Linguists claim this is an artificial word and that it does not deserve true longest word billing. Likewise, there are many technical and medical terms in English that qualify as long words. However, they are usually excluded from consideration for the longest word game.

Economic issues for a country in Africa Annotated Bibliography

Economic issues for a country in Africa - Annotated Bibliography Example They further stated that the nation’s credit ratings have been decreased by both S&P and Moody’s rating agencies and this has happened for the first time since the apartheid ended. S&P stated that they have decreased their ratings as a feedback to the strikes and the uncertainty of the political conditions of South Africa. The article further asserts that a huge part of the workforce and in accordance to the findings of the Deutsche Bank, 30% of the total mining workers are at strike. The Bank has even made a calculation of decline in the GDP that is targeted to decline by 2.5% if the strikes continue. The mining industry happens to be the backbone of South Africa and it is a huge producer and exporter of minerals such as palladium and platinum. These strikes pose a very significant issue to the economy of South Africa as the 50% of the exports gained by South Africa are backed by mining industry. These strikes are not only impacting the nation in a negative manner, the effect has been experienced by nations that import minerals from South Africa as an example of Anglo American company has been stated that has experienced reduction in production as a result of these strikes. South Africa’s strikes in the mining sector has increased concerns for people all over the world as these strikes have entered different areas of mining such as coal, gold and metal. The most interesting point of this article is that issues that are caused in one nation do not only impact the nation in a negative manner, they even impact other nations around the globe. For example: in this article the strikes conducted by the South African miners have negatively impacted industries that acquire minerals and commodities from South Africa. If the South African strikes continue to exist, they will not only impact the South African economy, they will

Race and Your Community Personal Statement Example | Topics and Well Written Essays - 1250 words

Race and Your Community - Personal Statement Example Admittedly, my community, that is Dallas, is notorious for the existence of racism. It seems that the issue of racism is much higher in this city because of the special nature of the demography here. In this area, all the racial groups are almost equal in size. According to the 2000 statistics, Hispanics are the largest group which constitutes nearly 35.55 % of the population followed by 34.56 percent whites. The presence of blacks is estimated at nearly 25.65%. Asians and other races are the minority, which together contribute less than 5% of the total population. Evidently, this mix of population has a specific role in amplifying racial issues. Due to the almost equal proportion of various groups, both politicians and media are eager to utilize racism as a tool for campaign and publicity. In fact, my neighborhood too is a mix of a variety of races, and the difference is visible in the look of people too. There are people who are black in complexion, some who are rather brown in complexion, and there are whites. The fact is that my society is more mixed than many other cities in the US. The difference is visible in communication too. For example the Hispanics, the Whites, and the Asians are easily distinguished for the way they use language. I have come to know that people like the Asians who first reach the place face serious problems communicating, and this situation leads to poor access to vital social services like medicine and other governmental grants. Admittedly, the leaders within my community are highly aware about the situation, and often, it seems as if they intentionally try to use the matter for easy public attention. Though most people and leaders try to avoid hurting other people by avoiding racial comments, some just make the comments publically. An exa mple is the recent statement made by a Dallas County commissioner to an attorney that â€Å"All of you are white. You can go to hell† (Piggy, 2011). The commissioner claims that he got angry because he found the statement of the attorney racial in nature. Though there are allegations of racism every now and then, from my observation, it seems that the leaders try to avoid any racial discrimination in their dealings. And most of the time, the term ‘racism’ does not arise in public offices and services. However, when there are issues involving people from two different races, the first allegation is racial profiling or racial discrimination. When a black child is punished by a white teacher, the situation is often given the color of racism. Similarly, when a black youth is arrested for drug or looting, the media shows the tendency to present the situation in the light of racism. The frequently made claim by the media is that, or what they try to make people believe is that, blacks are antisocial creatures who live on violence, theft and drugs. Another area where racism is highly visible is peer groups. Admittedly, the youth prefer to remain within groups of their own race. Within such groups, often comments of racial content take birth, and most of the time, it seems that such groups are behind racial attacks and abuses. I feel that the youth, if alone, will not resort to such activities of racism. As White and Perrone (2001) point out, it is the collective identity and the feeling of security within the group that makes them engage in such rare activities of racism. Though it is not always so, there are many whites who prefer to remain within white groups and blacks who prefer to be with blacks, and the main reason behind this situation is not hatred, but fear of getting maltreated by the other that is injected by the society and the media. Thus, though the administration cries ‘no racism’, there is racism in the air, and everyt hing in the society is first analyzed in the light of racism. A catalyst of racism, as already said, is media. It seems to me that media is bent on crating stereotypes and formulas in the social relations. As Rickford (2011) states, while

Van Is a Good Essay Essay Example for Free

Van Is a Good Essay Essay Pie Chart Showing the Occupational Expectation of the Trainees: Government Institutions Pie Chart Showing the Occupational Expectation of the Trainees: Private Training Schools Pie Chart Showing the Total Occupational Expectation of the Trainees: Total 2 2 3 4 5 6 C HAPTER I INTRODUCTION This paper reports the results of a small tracer study and discusses how tracer studies can be used as a planning tool to assist education and manpower planning. To make education institutions more efficient, we need to know what happen to graduates. One way is to conduct tracer studies. Such studies are useful for assessing the labour market performance of graduates and school leavers. Uses of Tracer Studies Tracer studies can be used to achieve the following: †¢ To collect information on how well the graduates are doing in the labour market †¢ To get feedback from the graduates to modify and upgrade the education institutions †¢ To meet the needs of the employers †¢ To make tracer studies as part of labour market information system †¢ To make use of the labour market information to assist policy planners. C HAPTERll METIlODOLOGY AND SURVEY DESIGN. The tracer study questionnaires used include items such as employment status, income, basic demography, waiting time for employment, job changes, attitude to jobs and qualitative feedback on courses and programmes of training institutions. Samples of fresh graduates from Yangon Institute of Technology (YIT), Civil, Government Technical Institute (GTI), Mandalay, Machine Tools, Agriculture High School, Myittha, during the three academic years 1984/85, 1985/86, 1986/87, were first chosen at random and then efforts were made to establish contact with those graduates included in the sample. Next, a few individuals who received training in specific skills like Computer Operations, Tailoring and Auto-repairs run by private establishments were also chosen at random and interviewed. Table 1 Training Institutions Run by the Government No. Training Institutions 1984/85 3 1986/87 Yangon Institute of Technology (Civil Eng. ) Government Technical Institutes (Machine Tools) Agriculture High School (Myittha) 203(10) 244(10) 274(10) 30 17(5) 48(5) 44(5) 15 66(5) 50(5) 24(5) 15 Total 2 Academic Year 1985/86 Sample Size 286(20) 342(2) 315(20) 60 Note: Sample sizes are given within parenthesis. Table 2  Skill Training Course Run By Private Establishments No. Training Institutions 1988 1 2 3 Computing and Typing Tailoring Auto-Workshop Total 5 3 2 Intended Interview 1989 1990 5 3 2 5 3 2 Total 15 9 6 30 Interviewers were sent all over the country to contact the graduates selected for interviewing through local administrative bodies. More than one visit was needed in tracing some 3 respondents for shifts in their places of residents. It is important to note that the sample sizes for private skill-training courses were fixed in advance for the sake of convenience. Table 3 Response Rate by Type of Graduates/Skills Sample Response. Response Rate % B. E (Civil) GTI (Machine Tools) (Mandalay) AHS (Myittha) Computing and Typing Tailoring Auto-workshop 30 15 27 15 90. 0 100. 0 15 15 9 15 15 9 6 6 100. 0 100. 0 100. 0 100. 0 Total 90 87 97. 0 No. Graduates/ Type of Skills 1. 2. 3. 4. 5. 6. C HAPfERill PLACEMENT OF GRADUATES Relationship Between Type of Diplomas/Degrees and Type of Work In our study of graduates from some training institutions, both in the formal and non-formal sectors, we see that some are employed in jobs which are related to their educational training, but a great majority found their jobs do not match with their training or acquired skills. For example, a graduate of Yangon Institute of Technology is found to take up the profession of a primary assistant teacher and another as a self-employed shop-keeper. Also a graduate of an Agricultural High School (AHS), took up the post of a Primary Assistant Teacher (PAT). A tracer study on the graduates of computer centres reveal that most of them take up the course because of the challenge of a new subject and also because the universities are closed for the time being. A currently employed PAT attending the course of computer said that her training and her work does not relate. She has a bachelor degree with physics major and has the feeling that she should be employed in a field in which her training might prove to be useful. Our study reveals that training systems such as computer schools, secretarial, typing schools are currently very popular and heavily attended in the sense that trainees perceived that the new market oriented economic policy would create job opportunities for them. Currently it is seen that such level of employment has not been reached yet. To those who took up computer training a few are fortunate enough to find employment in the same institution. Most of the trainees of typing school usually take it as a pre-course for computer training. Some attended such courses to while away before the university opens. Such reasons clearly bring out the fact that ultimately employers are the ones who decide on employment. Since employers have great influence over labour market the most obvious solution is to place training under the control of a board in which employers receive higher representation. Table 4 Match Between Training and Employment No. Institute (or) Training 1. Institute of Technology Government Technical Institutes Agricultural High School. Computer Training Tailoring Short Hand and Typing Auto-workshop 2. 3. 4. 5. 6. 7. Related 16 Not Related 4 8 11 1 1 2 5 1 1 1 5 Employment and Unemployment Situation of the Graduates The tracer study brought into view that among the 87 respondent, 38 per cent are not employed. Graduates of higher education face unemployment more often than lower education level graduates after completing their studies. A research was conducted in 1977 to find out the prospect that graduates have in securing jobs, the types of occupation where graduates are placed and how graduates have to wait to find jobs. Ten per cent of the graduates from Yangon Arts and Science University, Yangon Institute of Technology, the Institute of Economics, the Institute of Education, the Institute of Animal Husbandry and Veterinary Science. the Institute of Agriculture, under the Department of Higher Education and the Institute of Medicine and the Institute of Dental Science under the Ministry of Health were surveyed. The total response was 41 per cent within two months. Based on a survey regarding the number of applications submitted for jobs, the highest was found among the B. E (Engineering) degree holders and the lowest among the B. Sc degree holders. Six per cent of the graduates who were offered the jobs they had applied for, did not accept the job. The main reasons for this was that either the place of the job was too far out, or there were problems related with the family or the salary was low. Out of the 259 respondents 70 per cent were employed and 30 per cent were not yet employed. B. E. graduates and B. Sc graduates were found to be the largest in number among those not yet employed. In the recent survey in 1990, it was found that more than 59 per cent of the YIT and 53 per cent of the GTI graduates had found employment. For the AHS graduates the extent of employment was about 80 per cent. Among graduates of Computing Typing only 40 per cent and among Tailoring 22 per cent were able to find employment. Auto-workshop trainees are employed cent per cent. This is so because the nature of the job is arduous and young people are not willing take on this type of training highly. Table 5 Type of Training and Extent Employment (Per cent) No. Type of Institution 1. 2. 3. 4. 5. YIT GTI AHS Computing Typing Tailoring Employed Unemployed 59. 0 53. 0 80. 0 40. 0 100. 0 41. 0 47. 0 20. 0 60. 0 0. 0 Occupational Expectation of the GraduatGs The external efficiency of an education system involves relationships between general and vocational education and between schools and work opportunities, what schools and teachers can be expected to do in preparing for future occupations and what may be expected from a combination of learning in and out of school. The relationships are intricate and diverse and can be summarized as follows. 6. First to promote economic growth, it is essential to have a trained labour force equipped to handle technical and managerial problems. This presents a more severe problem for developing countries, where skilled manpower is scarce and enterprises that could provide opportunities both for training and employment is lacking. Secondly the problem of unemployment in the modern sector among graduates and others leaving school. In other words, labour supply versus labour demand. Thirdly policy makers, employers as well as individual tend to consider formal education as passport to jobs in the modern sector. To increase their chances for employment with higher wage, students tend to remain in school as long as possible, sometimes for more years than required by the available jobs. Table 6 Occupational Expectation of the Trainees No. Training Schools 1. 2. AHS No. /1) Industrial Training Centre Government Private JVC 2 3 10 I. /1 +2) 3. 4. 5. Tailoring Computing and Typing Auto-workshop 11. /3+4+5) 9 30% 108 ° 19 69% 22]0 4 7% 25 ° Ill. Total /1 +2+3+4+5) 22 49% 176 ° 19 42% 151  ° 4 9% 33 ° 2 14% 50 ° 13 86% 310 ° 3 6 7 7 2 5 Co-op 7 Figure 1 Pie Chart Showing the Occupational Expectation of the Trainees. Government Institutions J. V. C (15. 7%) Private (0. 0%) Private Training Schools 8 Total Government (49. 0%) Private (42. 0%) In our study on trainees from various institutions, we find that 49 per cent of the trainees wanted to join government departments, 42 per cent the private sector and 9 per cent tlie JVCs. It was found that no one wanted to join the cooperative sector. Among the trainees interviewed, two large groups were identified: those attending government training schools and those attending private and non-formal training schools. It was found that among those attending government training schools 86 per cent wanted to join government departments and 14 per cent preferred the JVCs. Most of the trainees taking private or non-formal training desired to join the private sector and to get government jobs as well. A tracer study of 1976177 graduates also point out that 67 per cent of the ograduates wanted to work with state enterprises or departments; less than 1 per cent the cooperative sector and the private sector and the remaining wanted to take up any type of work or set up their own businesses. This is understandable, for the state had been the major employer prior to the adoption of the market oriented economic policy, but after the adoption of the new economic policy, trainees actually look forward to join the private sector for work. This fact reflected the need for studying the labour market conditions placing more emphasis on employment effectiveness of the private sector. Income is also one of the main factors in the occupational expectation of the graduates. Our study is concerned with the income generation of graduates. Most begin their work with very low pay. For the YIT graduates, the majority had to start work with a monthly income ranging from K 400 K 750. For technical schools it is much lower, between K 200 K 450. Most of them start to work on a flat-payor daily wages basis. Graduates from the non-formal education sector such as auto-workshop workers, typists were found to earn more than university graduates. This is a natural flow of blue collar job workers earning more than white collar job workers in the initial period. The nature of the work is different although there is less wage differential. C HAYfER IV ISSUES AND RECOMMENDATIONS Methodology. Tracer studies are usually ad-hoc exercises covering only a few institutions or types of graduates. To use tracer studies as an aid to education/manpower policy making, graduates from all the institutions or programmes run by the Ministry of Education and other ministries should be analyzed. To encompass all the institutions and programmes, a sample will need to be selected. To be precise, the question should be asked of what size of sample is necessary to reflect the market demand for specific types of educated manpower. Another issue in methodology is formulating questions. At the centre of a tracer study is the questionnaire. In designing questions it should be borne in mind that much of the resulting analysis will be statistical and therefore the answers will need to be in a form where by they can be aggregated. Another important consideration in designing questions is to ensure that their meaning is clear and unambiguous. There are some factors to take into account in the wording of questions. All wording should be simple, direct and familiar. Each question should have a single issue. Operation The result of tracer studies can be useful to and fed into the work of several ministries, particularly Education, Labour and Planning. There should be a permanent tracer study unit in one of the above ministries. The advantage of setting up a permanent tracer study unit is that it can integrate information from tracer studies to national planning purposes. The setting of a permanent tracer study could develop a series of tracer studies right across to education/training system. To improve policy decision on education and manpower, tracer studies need to be organized as part and parcel of the activities of the institution and the information they generate should regularly be fed into the policy-making process. Recommendations. To get up-to date information on the labour market performance of graduates, Tracer Studies should be made regularly. To train more-people who will be in charge of the tracer studies. The person in charge should have, as a minimum, practical experiences of survey work including coding and analyzing cross tabulation. C HAPrER V CONCLUSION As this paper reports the results of a small tracer study and discusses tracer studies as a method of obtaining regular feed-back on the labour market performance of graduates, it can be used as a tool to assist education and manpower planning. Tracer studies provide information on the demand for educated workers. Besides this it has other advantages. First, the surveys are simple to conduct. Secondly, the typical questionnaire used is short and uncomplicated, and can easily be completed by respondents. The third is that almost all of them can be conducted by mail. They are therefore inexpensive to carry out. The survey can be conducted when graduates return for their graduation ceremony. Fourthly the data collected by tracer studies can be easily analyzed and understood by non-economists. Like all techniques to aid policy-making, tracer studies have shortcomings. Some of those shortcomings concern data reliability while others stem from the assumptions that underline tracer studies. I L IST OF WORKING PAPER SERIES 1. Education Data Review and Analysis 1 . 1 1 . 2 Performance Indicators in Higher Education by U Thein Htay (DHE). 1. 3 Performance Indicators in Technical, Agricultural and Vocational Education by U Myat Naing (MERB) and U Nyunt Maung (DTAVE). 1 . 4 Quantitative Review of Education Staff by U Tun Hla and U Myint Thein (DBE). 1. 5 2. Performance Indicators in Basic Education  by U Saw Win (Institute of Economics). Population Projections (1983-2013) by U Nyan Myint (Institute of Economics). The Quality of Education 2. 1 2. 2 Pedagogy by Oaw Nu Nu Win (Institute of Education). 2. 3 Curriculum by U Myint Han (MERB). 2. 4 Student Evaluation by Dr. Khin Saw Naing (Institute of Medicine 2). 2. 5 3. Teacher Quality by Daw 00 Khin Hla (Institute of Education). Student Characteristics by U Maung Maung Myint (DBE). Education and Employment 3. 1 Manpower Demand and Employment Patterns in a Changing Economy by Dr. Thet Lwin (Institute of Economics). 3. 2. A Tracer Study of Recent Graduates: Implications for Education and Manpower Planning by U Kyaw Kyaw (Department of Labour). 3. 3 Linkages between Training Institutions and Employers by U Tun Aye (Ministry of NO. 1 Industry), Daw Myint Myint Yi (Institute of Economics) and U Van Naing (DBE). 3. 4 Education and Work Performance (A Survey of Employers Perceptions) by Daw Hla Myint (Institute of Economics). 3. 5 Non-Formal Education in Myanmar by Daw Win Win Myint (Institute of Economics) and Daw Lai lai Yu (MERB). 3. 6 Labour Market Institutions in Myanmar by U Kyaw Kyaw (Department of Labour). 4. Costs and Financing of Education 4. 1 4. 2 Government Expenditure on Education by Oaw Soe Soe Aung (Institute of Economics) . 4. 3 5. Management and Administration of the Education Budget by Or. Khin Ohn Thant (Ministry of Planning and Finance). Non-Government Expenditure on Education by U Saw Gibson (Yangon University). Education Infrastructure 5. 1 5. 2 A Comprehensive Survey of Education Facilities by U Sein Myint (OBE), U Nyi Hla Nge (Yangon Institute of Technology), Oaw Win Win Myint (Institute of Economics), Oaw Naw Joy Loo (Institute of Medicine 1), U Saw Wynn (OBE) and Oaw Win Win Maw (In.stitute of Medicine 1). 5. 3 The Provision of Infrastructure by U Nyi Hla Nge (Yangon Institute of Technology), U Nyunt Hlaing (Ministry of No. 1 Industry), U Tet Tun (Ministry of Construction) and U Sein Myint (OBE). 5. 4 6. The Construction Sector by U Tet Tun (Ministry of Construction), U Nyi Hla Nge (Yangon Institute of Technology), Oaw Naw Joy Loo (Institute of Medicine 1) and Oaw Win Win Maw (Institute of Medicine 1). Norms and Standards for Education Facilities by U Nyi Hla Nge (Yangon Institute of Technology), Oaw Win Win Maw (Institute of Medicine 1), and U Tet Tun (Ministry of Construction). Organization and Management of the Education System 6. 1 6. 2 The Cluster System for Primary Schools by U Myint Han (MERB). 6. 3 7. Organization and Management of Basic Education by Oaw Hla Kyu (OBE) and U Myint Thein (OBE). Organization and Management of Universities and Colleges by U Saw Gibson (Yangon University) and U Thein Htay (OHE). Special Studies 7. 1 Economics and Business Education by Oaw Cho Cho Thein (OBE). 7. 2 The Teaching of Science and Technology by U Khin Maung Kyi (OBE). 7. 3 The Teaching of English by Or. Myo Myint (Yangon University). 7. 4 Higher Education by U Myo Nyunt (Institute of Education).

the enforcement of EU law :: essays research papers

To answer this question I will firstly explain how EU law became incorporated within the member states I will then explain the various types of EU legislation's in circulation. This is important to define as the various types of methods will involve different enforcement procedures. Finally I will explain how EU law is enforced and the ways EU law will effect the member state and individual businesses. I will summarise my findings at the end of the essay, this will give details of all the key ideas I have ut across. The Schuman proposal began various European countries setting up the European Coal and Steal Community (ECSC), this was signed in 1951 by six countries. The countries that initially set up the ECSC were France, Germany, Italy, Luxembourg, Belgium and the Netherlands. The idea behind the ECSC was the first serious institutional committee in Europe and was the start of integration, the aim was to provide a common market of coal and steel trade. This meant that each of the states would be able to have access to the various qualities of coal and steel that the members had with no tariff of importing and exporting to the various countries. This let the countries concentrate on their more efficient areas leading to specialisation. The ECSC began the beginning of supranational power because the "High Authority could adopt binding decisions" (1) as elements of decision making were carried out by ECSC committees. A supranational power is where the member state has to abide by the de cisions made of the community, as they have power above the national level. When the EC makes a law it is up to the individual state to implement the legislation. An example of EU law which has been passed which has been adopted in the UK is that of the directive 75/117 which states that men and women should receive equal pay. The UK government adopted this directive with the 1975 Sex discrimination Act. There are a number of methods EU legislation is formed for instance regulations, directives and decisions are three different types of EU legislation. I am going to briefly explain these three as the way they will be enforced are different. Regulations have general application that means that all the member states have to adopt the regulation; the member state is expected to adopt the whole regulation. Regulations are directly applicable which means that the "individuals have rights that they can enforce in their own name through national courts"(2).

Americas Cuban Conundrum

MARKETING 505 Assignment #2: Case 5-1 â€Å"America's Cuban Conundrum† ABSTRACT This paper analyzes the key issue that prompted the EU to take the Helms-Burton dispute to the WTO. It will decide who benefits and who suffers from an embargo of this type and it explains the rationale behind the conclusion. It also presents a resolution to the trade situation between the U. S. and Cuba. Finally given the trade relations it determines what type of economic barriers would have to be overcome by a U. S. firm to conduct business successfully in Cuba. Assignment #2: Case 5-1 â€Å"America's Cuban Conundrum† In 1996, President Bill Clinton signed the Cuban Liberty and Democratic Solidarity Act, also known as the Helms-Burton Act. The President’s actions came after Cuban MiGs shot down two U. S. civilian airplanes, killing the four Cuban-American who were on board. † (Green & Keegan, 2011) The Helms-Burton freezes the embargo against trade with and investment with Cu ba. It is a controversial act since it arouses a lot of negative attention on how the U. S. government is manipulating its power to impose its political views on other countries’ economies.The key issue -that prompted the EU to take the Helms-Burton dispute to the WTO -was mainly because they felt that it violated international rules dealing with trade. The EU had a dispute with the U. S. regarding testing agriculture, trade and investments and biotechnology issues which was brought to the attention of the WTO. It felt as if the United States was unlawfully exercising its jurisdiction extraterritorially, in that it was threatening to punish lawful activity such as trade, investment, and tourism carried out by the residents of independent countries. It is no exaggeration to say that U. S. olicy toward Cuba is now rejected and sharply criticized by the rest of the world. Other nations point out, moreover, that while America insists on maintaining its embargo and other pressures against Cuba because of the latter's human rights abuses, the fact is that Washington has close relations with–and extends Most Favored Nation treatment to–the People's Republic of China, Vietnam, Indonesia, and other states that are no more democratic than Cuba and have far worse human rights records, arguing that engagement, trade, and dialogue will do more to bring change in those countries than isolation. (Smith, 1998)It is obvious that the embargo affects a great number of people. To be all fair and neutral it would be necessary to highlight that to a certain extent, both countries are suffering and their benefits are minimal from this embargo. If it was to cease and somehow both countries came to an agreement and decided to put it all in the past and start from fresh, they would both benefit greatly. Production would increase and the producers on both countries would prosper. Cuban resources would be available to the U. S. Market and American producers would imp ort cost effective products that are not available in Cuba.An increase in capital from investors would make the economy run and prosper. A resolution to the trade situation between U. S. and Cuba represent the lifting of abolishment of the embargo. Castro’s regime and that of his successor, his brother Raul, have always talked a good anti-embargo but have not made the least effort to walk the talk, since this embargo may constitute the real reason of their continued hold on power since it gives an â€Å"antagonist† to the Cuban people, to the Cuban progress and it gives them an enemy to blame for the failure of Castro’s socialist revolution.In the U. S. there has been a lot of talk as well. President Obama was in favor of lifting of the embargo in 2004, ever since his views and actions changed, however he continued on implementing new policies that are in favor of establishing some type of relationship with the Cuban community. In 2004 President Obama who was th en a senator said: â€Å"I think it’s time for us to end the embargo in Cuba,† the then-Senate candidate said. And I think that we have to end it because if you think about what’s happening internationally our planet is shrinking, and our biggest foreign policy challenge — and it fits directly into the battle on terrorism and it fits into issues of trade and our economy — is how we make sure that other countries, in developing nations, are providing sustenance for their people, human rights for their people, a basic structure of government for their people that it’s stable and secure so that they can be part in a brighter future for the entire planet. And the Cuban embargo has failed to provide the source of raising standards of living and it has squeezed the innocents in Cuba,† Mr. Obama continued, â€Å"and utterly failed in the effort to overthrow Castro, who’s now have been there since I was born. So, it’s time for u s to acknowledge that that particular policy has failed. † (Speigel, 2009) But in 2008 he declared the opposite: â€Å"I will maintain the embargo,† Mr.Obama said in a speech in Miami on May 23, 2008. â€Å"It provides us with the leverage to present the regime with a clear choice: if you take significant steps toward democracy, beginning with the freeing of all political prisoners, we will take steps to begin normalizing relations. That’s the way to bring about real change in Cuba – through strong, smart and principled diplomacy. † (Speigel, 2009) For change to come about, a major event will have to take place.This situation seems like a non-negotiable on both sides where for the past 50 years they have agreed on disagreeing which isn’t the way to change the lives of all those people suffering and all those people that in one way or another are being affected negatively. In the unexpected event that trade relations resume between the U. S. a nd Cuba, some of economic barriers that would have to be overcome by a U. S. firm to conduct business successfully in Cuba. Firms will always be alert to any risk or uncertainty that can bring about the loss of assets or revenue due to unstable legal processes or corruption.Seizure of Assets for example: â€Å"The ultimate threat a government action to dispossess a foreign company or investor. † (Green & Keegan, 2011. ) Nationalization, expropriation are eminent threats in country with a dictatorship such as Raoul Castro and Fidel Castro. There would have to be some specific amendments to the international law which will pretty much guarantee to the firms and investors, any assets invested in the Cuban Market. Firms and Investors would have to take extra steps in order to protect their selves and clearly recognize the levels of juridical power for Cuba and for the U. S. s well as proactively protect against corruption and piracy which could be a primary issue in Cuba, with a strong intellectual property protection strategy. Any and all can be done once the embargo is lifted, but this would be the first step to greater changes and reforms, without it everything is only assumptions. This paper analyzed the key issue that prompted the EU to take the Helms-Burton dispute to the WTO. It decided who benefits and who suffers from an embargo of this type and it explained the rationale behind the conclusion. It also presents a resolution to the trade situation between the U.S. and Cuba. Finally it has determined what type of economic barriers would have to be overcome by a U. S. firm to conduct business successfully in Cuba. Work Cited Green, M. , & Keegan, W. (2011). Global marketing. (6 ed. ). Upper Saddle River, NY: Prentice Hall. Speigel, L. (2009, April 17). Presiden obama-who once supported, but no opposes, lifting the embargo-seeks â€Å"new beginning† with cuba. Retrieved from http://abcnews. go. com/blogs/politics/2009/04/president-ob-19/ Smith, W. (1998). Our dysfunctional cuban embargo.. (Vol. 42).

A view on Edward Snowden’s case Essay

American security consultant working for NSA on behalf of Booz Hamilton consulting Leaked top secret US and UK govt surveillance programs data to press bcoz of his â€Å"conscience† Charges on him of : espionage, theft of govt property. Leaked it to Guardian. Also, Nuremberg 1945 act says that â€Å"individuals have international duties that transcend national obligations of obedience. Therefore individuals have the duty to violate domestic laws to prevent crimes against humanity† Daniel Ellsberg did similar expose in 1971, releasing US decisions on Vietnam war, and was also awarded in 1978 â€Å"Gandhi Peace Prize†. These â€Å"Pentagon papers† revealed that US knew it was gonna lose the war and that casualties would be higher if it continued. Snowden applied for political asylum in 20 countries but each was indirectly threatened by US. A hero, dissident, traitor, whistle-blower, and a patriot. Snowden made contact with The Guardian in late 2012. Left Hawaii for Hong Kong on May 20, 2013. Iceland was his choice but could not make it coz of fear of being interdicted. A 5min movie Verax was also made showing his successful hiding in Hong Kong 20 applications for asylum. Temporary asylum of 1year in Russia which could be renewed annually. Granted on August 1st Reactions: US National Intelligence chief calls it reckless. Obama dismisses him but still calls him no patriot and Americans were better off without knowing about the surveillance activities. Also, NI chief says it was good coz we needed the debate on secutiry vs privacy since long-time. Bt LAVABIT, an email service provider used by Snowden had to be shutdown after they felt they were going against the grain of being for the people â€Å"What makes us different from other countries is not simply our ability to secure our nation,† Obama said. â€Å"It’s the way we do it, with open debate and democratic process.† If that’s so, then Edward Snowden should be hailed as a hero. There’s simply no doubt that his leaks led to more open debate and more democratic process than would’ve existed otherwise. Obama reluctantly admitted as much. â€Å"There’s no doubt that Mr. Snowden’s leaks triggered a much more rapid and passionate response than would have been the case if I had simply appointed this review board,† he said, though he also argued that absent Snowden, â€Å"we would have gotten to the same place, and we would have done so without putting at risk our national security and some very vital ways that we are able to get intelligence that we need to secure the country.†

New Zealands History, Government, and Geography

New Zealand's History, Government, and Geography New Zealand is an island country located 1,000 miles (1,600 km) southeast of Australia in Oceania. It consists of several islands, the largest of which are the North, the South, Stewart and Chatham Islands. The country has a liberal political history, gained early prominence in womens rights, and has a good record in ethnic relations, especially with its native Maori. In addition, New Zealand is sometimes called the Green Island because its population has high environmental awareness and its low population density gives the country a large amount of pristine wilderness and a high level of biodiversity. History of New Zealand In 1642, Abel Tasman, a Dutch Explorer, was the first European to discover New Zealand. He was also the first person to attempt mapping the islands with his sketches of the North and South islands. In 1769, Captain James Cook reached the islands and became the first European to land on them. He also began a series of three South Pacific voyages where he extensively studied the areas coastline. In the late 18th and early 19th centuries, Europeans began to officially settle on New Zealand. These settlements consisted of several lumbering, seal hunting, and whaling outposts. The first independent European colony was not established until 1840, when the United Kingdom took over the islands. This led to several wars between the British and the native Maori. On February 6, 1840, both parties signed the Treaty of Waitangi, which promised to protect Maori lands if the tribes recognized British control. Shortly after signing this treaty, though, British encroachment on Maori lands continued and wars between the Maori and British grew stronger during the 1860s with the Maori land wars. Prior to these wars, a constitutional government began to develop during the 1850s. In 1867, the Maori were allowed to reserve seats in the developing parliament. During the late 19th century, the parliamentary government became well established and women were given the right to vote in 1893. The Government of New Zealand Today, New Zealand has a parliamentary governmental structure and is considered an independent part of the Commonwealth of Nations. It has no formal written constitution and was formally declared a dominion in 1907. Branches of Government in New Zealand New Zealand has three branches of government, the first of which is the executive. This branch is headed by Queen Elizabeth II who serves as the chief of state but is represented by a governor general. The prime minister, who serves as the head of government, and the cabinet are also a part of the executive branch. The second branch of government is the legislative branch. It is composed of the parliament. The third is the four-level branch comprised of District Courts, High Courts, the Court of Appeal and the Supreme Court. In addition, New Zealand has specialized courts, one of which is the Maori Land Court. New Zealand is divided into 12 regions and 74 districts, both of which have elected councils, as well as several community boards and special-purpose bodies. New Zealands Industry and Land Use One of the largest industries in New Zealand is that of grazing and agriculture. From 1850 to 1950, much of the North Island was cleared for these purposes and since then, the rich pastures present in the area have allowed for successful sheep grazing. Today, New Zealand is one of the worlds main exporters of wool, cheese, butter, and meat. Additionally, New Zealand is a large producer of several types of fruit, including kiwi, apples, and grapes. In addition, the industry has also grown in New Zealand and the top industries are food processing, wood and paper products, textiles, transportation equipment, banking and insurance, mining and tourism. Geography and Climate of New Zealand New Zealand consists of a number of different islands with varying climates. Most of the country has mild temperatures with high rainfall. The mountains, however, can be extremely cold. The main portions of the country are the North and South islands that are separated by the Cook Strait. The North Island is 44,281 square miles (115,777 square kilometers) and consists of low, volcanic mountains. Because of its volcanic past, the North Island features hot springs and geysers. The South Island is 58,093 sq mi (151,215 sq km) and contains the Southern Alps - a northeast-to-southwest oriented mountain range covered in glaciers. Its highest peak is Mount Cook, also known as Aoraki in the Maori language, at 12,349 feet (3,764 meters) above sea level. To the east of these mountains, the island is dry and made up of the treeless Canterbury Plains. On the southwest, the islands coast is heavily forested and jagged with fjords. This area also features New Zealands largest national park, Fiordland. Biodiversity One of the most important features to note about New Zealand is its high level of biodiversity. Because most of its species are endemic (i.e.- native only on the islands) the country is considered a biodiversity hotspot. This has led to the development of environmental consciousness in the country as well as eco-tourism. New Zealand at a Glance Population:  4.9 million (2018 estimate)Capital:  WellingtonArea:  103,737 square miles (268,680 sq km)Coastline:  9,404 miles (15,134 km)Official Languages:  English and MaoriHighest Point:  Mount Cook (Aoraki) at 12,349 ft (3,764 m) Interesting Facts About New Zealand There are no native snakes in New Zealand76% of New Zealanders live on the North Island15% of New Zealands energy comes from renewable sources32% of New Zealands population lives in Auckland References and Further Reading â€Å"The World Factbook: New Zealand.†Ã‚  Central Intelligence Agency, 22 April 2010.â€Å"New Zealand.†Ã‚  Infoplease.â€Å"New Zealand.†Ã‚  U.S. Department of State, Feb. 2010.

How to Make Instant Snow From Boiling Water

How to Make Instant Snow From Boiling Water You probably know that you can make snow using a pressure washer. But did you know that you can also make snow from boiling water?  Snow, after all, is literally precipitation that falls as frozen water, and boiling water is water that is on the verge of becoming water vapor. Its incredibly easy to make instant snow from boiling water. All you have to do is follow the steps below. Materials You only need two things to turn boiling water into snow: Freshly boiled waterReally cold outdoor temperatures, around -30 degrees Fahrenheit The Snow-Making Process Simply boil the water, go outside and  brave the frigid temperatures, and toss a cupful or pot of boiling water into the air. Its important that the water be as close to boiling and the outside air be as cold as possible. The effect is less spectacular or wont work if the water temperature drops below 200 degrees Fahrenheit or if the air temperature climbs above -25 degrees Fahrenheit. Be safe and protect your hands from splashes. Also, dont throw the water at people. If it is sufficiently cold, there shouldnt be a problem, but if your concept of the temperature is mistaken, you could end up causing a dangerous accident. Always be careful when handling boiling water. How It Works Boiling water is water that is at the point of changing from a liquid into water vapor. Boiling water has the same vapor pressure as the air around it, so it has plenty of surface area to  expose to a freezing temperature. The large surface area means its much easier to freeze the water than if it were a liquid ball. This is why it is easier to freeze a thin layer of water than a thick  sheet of water. Its also the reason youd freeze to death more slowly curled up into a ball than if you were to lie spread eagle in the snow. What to Expect If you want to see boiling water turn into snow before you attempt this experiment, you can view a demonstration on the Weather Channel. The video shows a person holding a pot of boiling water and then tossing the scalding liquid into the air. An instant later youll see a cloud of snow crystals falling to the ground. I could watch this all day, the announcer says as she introduces the video, which was shot at  Mount Washington, New Hampshire, the highest mountain in New England. The announcer notes before the video begins that the snow-making folks actually performed the experiment three times- once with a measuring cup, once with a mug, and once with a pot (which is the effort captured in the video). Ideal Conditions In the demonstration video, the temperature of the water was 200 degrees Fahrenheit, and the temperature outside was a frosty -34.8 degrees. The experimenters said that they had diminished success when the water temperature dropped below 200 degrees and when the outside temperature rose above -25 degrees. Of course, if you dont want to go through all of this, and you still want to make snow- or if the temperature outside  is just too warm- you can  make  fake snow  using a common polymer while staying warm and toasty indoors.

Technology Essay Example | Topics and Well Written Essays - 500 words

Technology - Essay Example Therefore, we owe it all to those great scientists and inventors, for making our lives easier and interesting! One of the greatest events that occurred in the history of the human civilisation, that w rought up numerous changes, was the Industrial Revolution. It occured in the late 18th and the early 19th centuries. It started in Britain and gradually, spread across the Western World. The utilisation of iron and coal to generate power paved way for the mechanisation of the textile and transport industry. This helped immensely to increase production, leading to prosperity and growth. The second Industrial Revolution was a consequence of the first one, which is often considered as merged with th second. It took place in the year 1850 and brought some sweeping changes, in terms of electricity generation and utilistaion of the same in the varied industries. The effects of the Industrial Revolution were for the world to see, since life became easier and less stressful! This led to a great spurt in conducting research and experiements in technology, as well. However, the wonder machine, the computer was not in sight, then. Even though Charles Babbage, the Father of Computers had come up with a computer machine in the year 1822, it took more than a century for the computer to gain momentum. This essay reflects and imagines how things would have been, had the computer existed during the Industrial Revolution, with silicon chips and microprocessors making their way into mans life right before the Industrial Revolution! Things would have been turly different! Textile mills and industries would be working on production the way they do today---designing patterns and motifs on the computer and

History Essay Example | Topics and Well Written Essays - 250 words - 86

History - Essay Example Critical thinking is paramount in our day to day lives and provides us with an intellectual approach that facilitates us to choose from the available kind of information what is relevant and applicable in our lives and leave out the rest. The class has enabled me to look at present day as a stepping stone onto the future and that fifteen years from now, the world will have undergone massive transformations ranging from health, infrastructure and the general well being of society (Fea, 2013). Lastly, understanding historical events is essential for effective understanding of the condition of man. This will permit man to build and may also be important in initiating change upon a secured foundation. The most significant determinant of history is time. Time creates history and history exists because of time. And in conclusion, history is situated in time-space, which anchors the cosmos together, framing both the past as well as the present (Fea,

Geopolitics in the Middle East Essay Example | Topics and Well Written Essays - 750 words

Geopolitics in the Middle East - Essay Example Despite the struggles of the advocates of change, with the neo-conservatives envisioning an end for the, sort of chaotic Middle East society, the fight leads to a different scenario than what was expected. Peace is still so elusive, terrorist threats still exist, and instability is still greatly felt. We examined the reasons behind the pitfalls of this political blitzkrieg based on the articles written by Jeffrey Goldberg entitled "After Iraq" and the article "Parties of God: The Bush doctrine and the rise of Islamic Democracy" written by Ken Silverstein. Goldberg generally expressed the fact that the political agenda of the US and its allies are contrary to what Iraq's minority groups and even the rest of the countries in the region are looking for. Silverstein in the other hand expressed the fact that the Bush doctrine and the western democratic system do not fit the Islamic cultures and ways. While the US and the rest of its allies are eyeing to push their own vested interests in the Middle East campaign, the Middle Eastern nations affected by the current transitions in the other hand are looking at it in a different perspective. The war on Iraq was generally aimed at ousting Saddam Hussein, a dictatorial leader, for the major reason of removing a danger to social equality in the region, which Hussein had proven to be a threat, and instate a unified democratic government. . The plans may have been understood plainly by both the allies and Arab compatriots, like the Kurds, Turkey, Egypt, Jordan, and the rest, and laid open for them to work through, but beneath the shadows of their support, the Middle eastern countries, and minority regions within Iraq, are hoping to push through with their own agenda, for their own people, for their leader's interest. Take for example the case of the Kurds. In a personal interview by Jeffrey Goldberg with Abdul Rahman Mustafa, the Kurdish-Iraqi governor of Kirkuk in northern Iraq, considered as domains of the Kurds, it appeared that the Kurds really wanted independence from Iraq (Goldberg 79). The article of Goldberg speak further of other regions in the middle east that wanted to be governed based on their own cultural minority principles such as the "Shiites did not want to be ruled from Baghdad by a Sunni minority"(78), and other minority groups from Egypt, Israel, and other areas want their own sovereignty. In other words, the zeroing in was just superficial. The question is why The answer perhaps lies on the cultural differences between the east and the west, and the way these two civilizations are molded through time. We agree on Goldberg's idea that western democratic strategies don't seem to fit with the Middle Eastern cultures, which are highly diverse. Islam, which is the main religion in the Middle Eastern countries, is a diverse religion with differing principles across different minorities. While the west is generally Christian, although differing in minor teachings, have common core values that has high regard to peace and order. There is no Christian jihad, to cite a particular difference, while the Muslim jihad is primarily exploited by Muslim extremists and used to push their ideals and principles. In the western world, as a common knowledge, governance in civic

Human Carbonic Anhydrase II

Human Carbonic Anhydrase II Human carbonic anhydrase II is one of the fastest studied enzymes known with a variety of roles in reaction catalysis. Its primary function is to catalyze the reversible hydration reaction of carbon dioxide. In addition to carbon dioxide hydration, it is also capable of other latent skills, such as catalyzing esterase activity. The ability of human carbonic anhydrase II to function as a catalyst derives from key residues in and around the active site that play crucial roles in the mechanism. Substitutions to two of those particular key amino acids were performed via Quick-change site directed mutagenesis: H64A and V142D, to investigate the particular role they have in the catalytic active site. Various kinetic experiments and structural analyses were performed on wild-type carbonic anhydrase and the mutants to discern and compare their activity to each other and to literature, including Michaelis-Menten parameters for PNPA hydrolysis, CO2 hydration, and inferring function molecular m odelling. Though the same trends can be seen as the literature, individual values were found to be much lower owing to errors in measurement and equipment. Trends were found to coincide with the mutants known roles in the active site: His64 is the proton shuttle that facilitates proton transfer during the rate limiting step and Val142 participates in the hydrophobic pocket to bind and recruit substrates to interact with the active site. Mutations to both of these sites show that enzyme efficiency and activity strongly decreases. Introduction Human carbonic anhydrase II (hCAII) is a zinc metalloenzyme that catalyzes the following reversible reaction: . The enzyme commonly functions to help shuttle carbon dioxide in red blood cells to rid the body of metabolic waste, and catalyzes the hydrolysis of many aromatic esters [1, 2]. Structurally, a zinc ion is located in the active site, coordinated to 3 histidine residues (H94, H96, H119) and usually a hydroxide ion or water molecule [2]. The mechanism of hCAII proceeds through two major steps: 1) the conversion of carbon dioxide to bicarbonate, and 2) the regeneration of Zn-OH by proton transfer. The active hydroxide that is bound to zinc nucleophilically attacks a nearby carbon dioxide molecule, resulting in a bicarbonate ion binding to zinc [3]. The zinc-oxygen bond breaks to subsequently release a bicarbonate ion, which is replaced with water [3]. The Znà ¢Ã¢â€š ¬Ã¢â‚¬â„¢OH bond is regenerated by a proton transfer to the external buffer, which is facilitated by the His64 residue [3]. The proton transfer step is the rate limiting step of the reaction [3]. The diazole side chain on the histidine residue is what gives it the ability to be a proton acceptor and donor. Mutations in that position (His64) usually result in decreased enzyme activity due to a lack of proton transfer; however the reaction does proceed to a lesser degree without an active His residue, possibly due to its extensive water network in the activ e site forming secondary proton wires [4]. Carbonic anhydrase catalyzes one of the most rapid reactions; it is one of the fastest enzymes studied [1]. Its reaction speed is due, in part, by the amphiphilic nature of the active site [1]. The hydrophobic side is used to bind carbon dioxide, while the hydrophilic patch functions to optimally orient the carbon dioxide molecule for the reaction [1]. The hydrophobic wall forms a well-defined pocket near the zinc-hydroxide and is composed of the following amino acids: Val142, Val121, Leu197 and Trp208. The hydrophilic patch consists of Thr198 and Glu106, which form a hydrogen bond network with the Znà ¢Ã¢â€š ¬Ã¢â‚¬â„¢OH to stabilize and orient it for nucleophilic attack on CO2 [2]. Therefore, any modifications to the hydrophobic pocket would change its structure, and consequently, its catalytic efficiency [1]. In this study, the importance and role of His64 and Val142 to the structure and mechanism of hCAII are determined through site-mutagenesis and subsequent characterization of the new mutants, H64A (His64 Æ’Â   Ala) and V142D (Val142 Æ’Â   Asp) via kinetic and structural analysis. The changes that arise from the substitutions may prove to be applicable to drug synthesis because hCAII is known to be involved in a variety of diseases, for example, Marble brain disease, where mutations in the hCAII gene leads to a deficiency in the enzyme which is an autosomal recessive disease [5]. Studies in hCAII mutations can be used to design folding modulators to suppress misfolding which frequently occurs due to hCAII destabilization [5]. Another major disease involved with hCAII gene is osteopetrosis. The hCAII genes inactivation decreases osteoclast function in bone, and knowledge of hCAII mutations that inactivate the enzyme may lead to better understanding of bone remodelling [6]. Some carbonic anhydrase diseases use inhibitors (CAI) to suppress the hCAII as a therapeutic treatment. Inhibitors prevent hCAII activity by inhibiting either of the reaction steps: the conversion of CO2 which involves V142 in the hydrophobic pocket, or the rate limiting step, proton transfer, in which His64 is crucial. Experimental Procedure Site directed mutagenesis via the PCR-based Quick-change method was performed on hCAII as cited in Woolley (2011) for 10 ng and 20 ng wild-type plasmids (hCA2pET24b from Novagen) [7]. Table shows the sequence of the primers used in the PCR reactions. Products of PCR mutagenesis reactions were run on 0.7% agrose gels to determine size. The gels were run at 150 V in 1X TAE buffer. Red safe dye from Intron Biotechnology was used in the agrose gel instead of ethidium bromide for safety reasons [7]. The standard molecular weight ruler used was a 1 kB DNA ladder from Fermentas. Table : Primer sequences used in mutagenesis of hCAII in the forward and reverse direction for mutants H64A and V142D Mutant Direction Sequence MW (Da) %GC TM ( °C) H64A Forward GGATCCTCAACAATGGTgcTGCTTTCAACGTGGAG 10778 51 67 Reverse CTCCACGTTGAAAGCAgcACCATTGTTGAGGATCC 10709 V142D Forward CTGATGGACTGGCCGaTCTAGGTATTTTTTTG 9868 44 62 Reverse CAAAAAAATACCTAGAtCGGCCAGTCCATCAG 9779 The enzyme, DpnI, was then used to digest methylated DNA (the parent template DNA). The DNA vector that contained the mutation was transformed into supercompetent E.coli turbo cells from New England Biolabs by heat shock [7]. LB-agar plates were prepared to grow the transformed cells containing mutant genes (i.e. H64A and V142D hCAII gene) [7]. Both were injected with Kanamycin to ensure that the culture that grows will have the desired mutation [7]. A miniprep culture was set-up from the LB-agar plate into LB medium to grow one colony for DNA analysis [7]. Restriction enzyme mapping was prepared and XhoI and BglII were chosen, they were used under buffer 3 for optimal efficiency. Plasmid putification was performed using the QIAprep Spin Miniprep Kit, and then the chosen restriction enzymes were carried out and were run on 1% agrose gel [7]. A sample of the purified DNA was sent to an external company (ACGT) for commercial sequencing (Sanger dideoxy type) to verify if the mutagenesis occurred correctly. The sequence was analyzed using the program BioEdit. To determine the level of confidence of the sequencing results, the purified DNA was quantified using UV/Vis absorption via a spectrometer [7]. The concentration was calculated using Ecà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1(260) as 50 ÃŽÂ ¼g/mL. Purified plasmid DNA was transformed into E.coli BL2(DE3) cells to initiate protein expression by heat shock, similar to the transformation into turbo cells [7]. The cells were cultured and a single colony was grown. Once sufficiently grown, ITPG and ZnSO4 were added to induce protein expression [7]. SDS-PAGE was used to confirm protein expression and was analyzed against an unstained protein molecular weight marker by Fermentas. The protein and ladder was stained with coomassie blue [7]. Affinity chromatography was used to purify the mutant hCAII proteins [7]. The matrix used was agrose linked to p-(aminomethyl)benzenesulfonamide, exploiting the tight binding that occurs between hCAII and sulphonamides. Once purified, the protein was dialyzed using a 6000-8000 Da dialysis membrane to replace the elution buffer with protein buffer and removes the matrix from the protein [7]. SDS-PAGE is again used to confirm the protein is still present after purification and to check its approximate molecular weight. It was run for two different amounts of protein, 2 ÃŽÂ ¼g and 10 ÃŽÂ ¼g, and also ran 10ÃŽÂ ¼L of wash fractions from affinity chromatography [7]. Protein concentration was determined by UV absorption at 280 nm in a final concentration of 6M guanidine hydrochloride. From the calculated concentrations, purity of the protein could be assessed via SDS-PAGE. To characterize this purified hCAII protein, a variety of analyses were done. Two types of mass spectrometry (MS) were performed: electrospray ionization (ESI) and matrix-assisted laser desorption ionization (MALDI) [7]. The MS analysis was used to confirm the presence of the mutation in hCAII with intact and digested protein. Protein samples (H64A and V142D hCAII) were not diluted for either of the MS analyses as cited in Woolley (2010). Samples of 10 ÃŽÂ ¼L of stock protein concentrations (37.6 ÃŽÂ ¼M H64A and 3.2 ÃŽÂ ¼M V142D hCAII) were used for analysis of the molecular weight of the intact protein by ESI-MS. Both mutants were then digested by Trypsin Gold (MS grade) from Promega and the resulting fragments were evaluated by ESI-MS as well [7]. A 50 ÃŽÂ ¼L sample was used for each mutant, 40 ÃŽÂ ¼L of the mutant at stock concentration and 10 ÃŽÂ ¼L of the Trypsin Gold. A couple ÃŽÂ ¼L of the digested mutants were saved for MALDI-MS and the rest was used for ESI-MS. Once the molecular weights for each of the digested fragments were determined by ESI-MS, the products were run through a protein database to confirm the identity of the protein and mutations [7]. The 1 ÃŽÂ ¼L of the tryptically digested mutants prepared for ESI-MS, subsequently underwent MALDI-MS. The 1 ÃŽÂ ¼L samples were mixed with a matrix consisting of 1 ÃŽÂ ¼L ÃŽÂ ±-cyano-4-hydroxycinnamic acid (CHCA) and 1 ÃŽÂ ¼L of 0.1% trifluoroacetic acid (TFA) [7]. The entire mixture was pipette onto a MALDI well and was inserted into the mass spectrometer and a MALDI-MS spectra was obtained. Michaelis-Menten kinetics was used to determine the KM and kcat of the p-nitrophenyl (PNPA) hydrolysis reaction [7]. The ionized product from the hydrolysis, p-nitrophenol (PNP) produces a bright yellow colour that was used to follow the rate of the reaction via the Perkin Elmer Lambda UV/Vis spectrophotometer [7]. Various sample concentrations of PNPA were set up to have a final enzyme concentration of 0.2 ÃŽÂ ¼M in protein buffer [7]. The initial rate measurements of each PNPA concentration were taken for wild-type enzyme, H64A mutant, V142D mutant, and a blank with no additional enzyme added (refer to data tables in Enzyme Kinetics I [7]). PNP has a molar absorption coefficient (ÃŽÂ µ) of 1.73ÃÆ'-104 Mà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1cmà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1. This was used to calculate Michaelis-Menten values: Vmax, KM, kcat, and kcat/KM [7]. The ability of hCAII mutants (H64A and V142D) and wild-type hCAII to catalyze the hydration of CO2 was measured. The pH of the solution was measured to track the progress of the reaction because the reaction generates protons. Enzyme solutions were prepared according to table 2 in [7]. The buffer used in the table was 50 mM TRIS buffer (pH 7.8). Additional enzyme samples were prepared for 25 nM of wild-type hCAII and 100 nM of H64A mutant in a final concentration of 22.5 and 29.92 mM imidazole buffer (pH 7.8) respectively to determine chemical rescue of mutant H64A. The pH of the CO2 hydration assay was measured using a pH probe and pH meter at 5 second increments for a total of 90 seconds starting at the beginning of the reaction [7]. The slope of the initial changes in the first 2 points was considered to be the V0 for each enzyme concentration. From the initial velocity, a kcat value can be calculated for each enzyme using the assumption that [S] >> KM, the Michaelis-Menten equati on simplifies to kcat=V0/[E]. The third kinetics experiment used fluorescence to determine the binding constant of dansyl amide (DNSA) and acetazolamide (AZ) (from Sigma-Aldrich) to H64A and wild-type hCAII was performed using the Perkin Elmer Fluorometer [7]. Stocks of 1 mM and 200 ÃŽÂ ¼M of DNSA were prepared from a 21.6 mM DNSA stock by dilution with DMSO. Enzyme stocks were diluted to 0.25 ÃŽÂ ¼M with TRIS buffer to make a 10 mL solution. A 1 mL sample of H64A from stock made was titrated with DNSA in small increments [7]. The fluorometer emissions were taken at 470 nm. AZ titration in competition with DNSA was not able to be completed. The last characterization experiment done was molecularly modelling the hCAII wild-type enzyme, as well as the mutants H64A and V142D. The molecular model of hCAII analyzed was derived by x-ray crystallography and found in the Protein Data Bank (PDB) repository. The wild-type and H64A hCAII structures examined had a PDB code of 1CA2 and 1MOO respectively. At present, no crystal structure has been found for V142D hCAII. The Swiss PDB Viewer program was used to visualize the protein structures. Secondary structures of the proteins were able to be observed. Residues around the metal active site and the Ramachandran plot were explored. Homology between hCAII and other carbonic anhydrase isozymes, hCAIV (PDB code 1ZNC) and hCAI (PBD code H1CB), were also studied by performing an iterative magic fit on the ÃŽÂ ±-carbons and structure alignment for each pair. The root mean square (RMS) between hCAII and the other isozymes were also analyzed to determine conserved and deviated regions in the structures. The binding of cobalt in the hCAII active site was also investigated (PDB code 3KOI). The structural inhibition of hCAII by AZ was also gleaned by structural analysis (PDB code 3HS4). Its mode of inhibition and binding sites were shown through the crystal structure. Lastly, the Swiss PDB Viewer program was used as a tool to theoretically synthesize mutations and compare it to the actual structure as determined by other scientists, for example, by aligning the virtual and crystallized mutations to determine deviations in structure by performing RMS. Results Site-directed mutagenesis PCR. Products from the PCR mutagenesis reactions were examined using 0.7% agrose gel electrophoresis. Two samples of differing amounts of template DNA (10 ng and 20 ng) were used for each mutant (Figure ). Bands were only observed for samples containing 20 ng of the hCA2pET24b DNA template plasmid (Figure ). The size of the bands observed coincides with the size of the plasmid used, 6018 bp. Heat shock transformation and isolation of plasmid. Several colonies were observed after plasmid transformation for both mutants, and 1 colony from each mutant was chosen for restriction enzyme digest with BglII and XhoI. Figure : Electrophoretic run on 0.7% agrose gel of DNA of hCAII mutants from PCR mutagenesis reactions. Lane 1 is the GeneRuler ladder by Fermentas and lanes 10-13 are the following: V142D (10 ng), V142D (20 ng), H64A (10 ng), and H64A (20 ng). As suggested from the gel, the mutants in the 20 ng plasmid was more successful than the 10 ng plasmids in determining relative molecular weights. Both mutants in the 20 ng plasmid show a band at approximately the 6000 base pair mark, which coincides with the number of base pairs in the hCA2pET24b plasmid that was used (6018 base pairs). Quantification of pure plasmid DNA. A 1/20th dilution was carried out on the purified DNA with elution buffer (EB; 0.1 M Tris, 0.4 M KSCN, pH 7). The absorption of the diluted DNA at 260 nm and 280 nm was taken by a UV/Vis spectrophotometer and the relative DNA purity was determined (Table ). The assumption that Ecà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1 260 = 50 ÃŽÂ ¼g/mL for DNA was applied in the calculation of concentrated and diluted concentrations of purified DNA (Table ). Table : Relative DNA purity for mutants V142D and H64A determined by UV/Vis spectrophotometer absorbance at 260 and 280 nm. Calculated concentrations of mutants from absorbance data, where Ecà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1 260 = 50 ÃŽÂ ¼g/mL. Mutant Wavelength, ÃŽÂ » Absorbance Units Relative DNA Purity (A260/A280) Concentrated (ÃŽÂ ¼g/mL) Diluted (ÃŽÂ ¼g/mL) V142D 260 nm 0.3117 1.7852 311.70 15.59 280 nm 0.1746 H64A 260 nm 0.2653 1.7581 265.30 13.27 280 nm 0.1509 Enzyme restriction digest. Purified plasmid DNA of mutants were digested with XhoI and BglII, separately and together in a single and double digest for both mutants. The digested and undigested samples were run on 1% agrose gel, and 2 bands were observed around the 6000 and 7000 bp marker for all 8 samples (Figure , Figure ). The expected length of the bands in the double digest should be 892 bp and 5126 bp (Figure ). Figure : Electrophoresis performed in 1% agrose gel of digested V142D hCAII in lane 1-4. The (1 kB) GeneRuler DNA ladder is shown in lane 5. Lane 1-4 contain the following: V142D plasmid, V142D + XhoI, V142D + BglII + XhoI, and V142D BglII. Double bands are shown at the 6000 and 7000 bp marker for all 4 V142D samples. Figure : Electrophoresis performed in 1% agrose gel of digested H64A hCAII in lane 1-4. The (1 kB) GeneRuler DNA ladder is shown in lane 5. Lane 1-4 contain the following: H64A plasmid, H64A + XhoI, H64A + BglII + XhoI, and H64A BglII. Double bands are shown at the 6000 and75000 bp marker for all 4 H64A samples. Figure : Restriction enzyme cut sites and position of hCAII gene (5072-5854) on the hCAI2pET24b plasmid DNA Sequencing. The mutations for both V142D and H64A in the hCAII gene were successful according to the sequenced DNA result obtained from ACGT. Other mutations in the DNA sequence were observed in both mutants, but since the aligned protein sequence was the same, mutations were likely to be silent mutations due to amino acid redundancies. When sequenced in the forward direction by T7 polymerase, a protein mutation was found (K153N) other than the desired mutation of V142D; however, when sequenced in the reverse direction by T7 polymerase terminator (T7TER), K153N was not observed. Plasmid DNA transformation into E.Coli BL21(DE3) cells. Following transformation into BL21(DE3) cells, colonies were observed for both hCAII mutants (V142D and H64A). A random colony was chosen to be cultured and then was induced to express protein with 270 ÃŽÂ ¼M IPTG and 0.1 mM ZnSO4. SDS-PAGE for protein expression. Protein expression was tested with SDS-PAGE. The expected molecular weight of V142D hCAII is approximately 29.2 kDa and the expected molecular weight of H64A hCAII is approximately 29.1 kDa. SDS-PAGE bands are observed between the ladder markers 25.0 kDa and 35.0 kDa for both mutant proteins (Figure , Figure ). Figure : SDS-PAGE loaded with V142D hCAII proteins to examine protein expression. Samples were loaded in different volumes of protein to ensure gel visualization. Lane 15 contains the Fermentas protein molecular ladder and lane 1-4 contain the following: 1 ÃŽÂ ¼L à ¢Ã¢â€š ¬Ã¢â‚¬â„¢IPTG, 4 ÃŽÂ ¼L à ¢Ã¢â€š ¬Ã¢â‚¬â„¢IPTG, 1 ÃŽÂ ¼L+IPTG, 4 ÃŽÂ ¼L +IPTG. All 4 samples had some form of protein expression between 25.0 to 35.0 kDa. Figure : SDS-PAGE loaded with H64A hCAII protein to examine protein expression. One sample was loaded with 4 ÃŽÂ ¼L of H64A protein and +IPTG in lane 10. Lane 6 contains the Fermentas protein molecular ladder. The one H64A sample loaded showed an expression between 25.0 and 35.0 kDa. Calculation of pure protein concentration and extinction coefficient. Following affinity purification and dialysis, pure protein concentration was calculated from UV absorption measurements at 280 nm and the known extinction coefficient of hCAII as 50070 Mà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1cmà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1 (Table ). The final concentration of the samples of V142D and H64A hCAII were 3.2 ÃŽÂ ¼M and 37.6 ÃŽÂ ¼M respectively. Table : UV absorption measurements at 280 nm of purified protein and the resulting final concentration Mutant Average A280 Protein concentration (ÃŽÂ ¼M) V142D 0.01583 3.2 H64A 0.1884 37.6 SDS-PAGE to assay purity and check approximate molecular weight. Several samples were loaded into the SDS-PAGE for each mutant protein: lysate and wash fractions (collected from affinity chromatography), 2 ÃŽÂ ¼g protein, and 10 ÃŽÂ ¼g protein. For H64A, a visible band was only observed for the 10 ÃŽÂ ¼g sample (Figure ). The band was located between the 35 kDa and 25 kDa markers on the ladder. For V142D, none of the 4 samples resulted in a band on the gel (Figure ). Figure : SDS-PAGE shown for H64A mutant protein. Lane 1 contains the Fermentas protein molecular weight marker. Lane 11-14 contains H64A samples of the following (in order): lysate, wash fraction, 2 ÃŽÂ ¼g protein, and 10 ÃŽÂ ¼g protein. Only the 10 ÃŽÂ ¼g protein had (faint) observable bands located between the 25 and 35 kDa markers. Figure : SDS-PAGE shown for V142D mutant protein. Lane 4 contains the Fermentas protein molecular weight marker. Lane 12-15 contains V142D samples of the following (in order): lysate, wash fraction, 2 ÃŽÂ ¼g protein, and 10 ÃŽÂ ¼g protein. No observable bands are seen for any of the samples. Mass spectrometry. ESI-MS was not successful in analyzing the molecular weight of intact and digested protein of both mutants. A MALDI spectrum was able to be generated for the digested proteins; however, without the digested ESI spectrum to compare to, the peaks from the MALDI spectrum can only be speculatively assigned. Kinetics: Hydrolysis of PNPA. Using the molar absorption coefficient of PNP (1.73ÃÆ'-104 Mà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1cmà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1), the rate of each reaction was determined. The predicted rate was calculated using the Michaelis-Menten kinetics: . The plot of predicted rates and actual initial rates vs. PNPA concentration can be seen in Figure , Figure , Figure for wild-type, H64A, and V142D hCAII respectively. The Vmax and KM values for each enzyme were calculated by minimizing the square difference between the predicted and actual reaction rates, and the kcat was calculated using the equation: (Table ). Table : Calculated Michaelis-Menten parameters for wild-type, H64A, and V142D hCAII catalyzing the hydrolysis of PNPA. Wild-type hCAII H64A hCAII V142D hCAII Vmax (ÃŽÂ ¼M/sec) 1.202 0.812 0.218 KM (mM) 1.280 1.957 8.362 kcat (sà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1) 5.141 ÃÆ'- 10à ¢Ã¢â€š ¬Ã¢â‚¬â„¢3 2.159 ÃÆ'- 10à ¢Ã¢â€š ¬Ã¢â‚¬â„¢2 6.825 ÃÆ'- 10à ¢Ã¢â€š ¬Ã¢â‚¬â„¢2 kcat/KM (Mà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1sà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1) 4.02 11.032 8.162 Figure : Michaelis-Menten plot of initial rate vs. concentration of PNPA added for wild-type hCAII enzyme. Figure : Michaelis-Menten plot of initial rate vs. concentration of PNPA added for H64A hCAII enzyme. Figure : Michaelis-Menten plot of initial rate vs. concentration of PNPA added for V142D hCAII enzyme. Kinetics: CO2 hydration. Initial velocity (V0) values were calculated by measuring the progression of the reaction (via concentration of protons) with time (Table , Table , and Table ). kcat values were then calculated using the same equation as in the hydration of PNPA and averaged for the individual enzymes (wildtype, H64A, and V142D hCAII) in a particular buffer (i.e. TRIS or imidazole). Table : Initial velocity (V0) and kcat values calculated for the hydration of CO2 by wild-type hCAII in TRIS buffer and imidazole buffer. Wild-type concentration (nM) V0 for WT+TRIS (M/s) V0 for WT+Imidazole (M/s) 0 1.3E-08 6.05778E-08 1.5 1.1E-08 N/A 2.5 1.1E-08 5.63E-08 5 2.1E-08 5.16E-08 12.5 5.9E-08 5.63E-08 Average kcat (sà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1) 5.3 ±1.62 12.44 ±9.19 Table : Initial velocity (V0) and kcat values calculated for the hydration of CO2 by H64A hCAII in TRIS buffer and imidazole buffer. H64A concentration (nM) V0 for H64A+TRIS (M/s) V0 for H64A+Imidazole (M/s) 12.5 1.4E-08 6.57E-08 25 1.4E-08 5.8E-08 50 1.7E-08 7.53E-08 Average kcat (sà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1) 0.67 ±0.39 3.03 ±1.97 Table : Initial velocity (V0) and kcat values calculated for the hydration of CO2 by V142D hCAII in TRIS buffer. V142D concentration (nM) V0 for V142D+TRIS (M/s) 12.5 6.2E-09 25 5.4E-09 50 5.5E-09 Average kcat (sà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1) 0.27 ±0.19 Fluorescence detection of ligand binding. DNSA was titrated with H64A hCAII to determine its affinity for the enzyme. The dissociation constant, KD, for DNSA was determined to be 0.086 ÃŽÂ ¼M when protein concentration was 0.25 ÃŽÂ ¼M. Competitive titration of H64A-DNSA hCAII with AZ was attempted, but was not successful as DNSA binding was too tight, making it difficult to be displaced by AZ. Molecular modeling. Literature models of wild-type (PDB code 1CA2) and H64A (PDB code 1MOO) hCAII were analyzed. There is no available structure of V142D hCAII at present. The secondary structure of wild-type is composed of 18 ÃŽÂ ²-sheets (77 residues) and 10 ÃŽÂ ±-helices (42 residues), with the majority of the ÃŽÂ ±-helices falling in the domain of right-handed helices, while very few show left-handed helical properties according to the Ramachadran plot. It also seems that the active site is solely composed of ÃŽÂ ²-sheets, and no ÃŽÂ ±-helices (Figure ). Analyzing PDB structure 3HS4 (AZ bound hCAII), the mechanism as to how AZ inhibits hCAII function can be seen. AZ has 3 binding sites, 2 are novel binding sites and the other provides a mechanism of inhibition. AZ binds the zinc directly at the active site, displacing crucial ligands needed for catalysis. There were some discrepancies found between the crystal structure of H64A [1MOO] as cited on PDB and virtually mutated H64A from wild-type hCAII, resulting in a RMSD (root mean square deviation) of 0.29 Ã… (Figure ). Since no literature structure of V142D is available, no comparison between virtual and crystal structures could be made. Figure : Secondary structure of wild-type hCAII overlain with ribbon to visualize the higher arrangement. Figure : RMSD between H64A hCAII virtually mutated and literature crystal structure. Blues denote the same or similar residues, while reds and oranges indicate completely different amino acids. Discussion Agrose gel results were only visible for samples that contained 20 ng of the plasmid template DNA, rather than the 10 ng plasmid. This may be a result of more amplification during PCR with the 20 ng plasmid, and so would intensely be more visible. Though the 20 ng samples showed bands at the appropriate 6000 bp mark, there was also a faint band that can be seen near the end of the gel. This may be due to non-specific primer annealing. Quantification of DNA purity was done by exploiting the peak absorbances of protein and DNA. DNA maximally absorbs at 260 nm, while protein dominantly absorbs at 280 nm. The purity ratio reports the relative amount of DNA compared to protein present in the sample. The purity of both mutants were approximately 1.8, which is regarded as a relatively pure sample; however, a purity ratio of more than 2.0 would have been ideal. The restriction enzyme digest showed 2 bands (7000, 6000 bp) for all samples, which may have been a sign of poor mixing/ pipetting since the volumes of restriction enzyme were extremely small amounts. If this is the case, only some of the DNA was nicked and some were not, which would result in 2 bands. It was expected that the plasmid sample would have a high band (supercoiled), each of the singly digested samples would have a slightly lower band (nicked), and the doubly digested would show 2 bands that indicated the fragment size of 892 and 5126 bp. Sequencing results showed that a protein mutation occurred when the sample was sequenced in the forward direction by the T7 polymerase. A lysine at position 153 had mutated to glutamine (K153N). However, this mutation was not observed when the T7 polymerase terminator was used to sequence the sample in the reverse direction. A mutation that occurs in one sequencing direction and not the other is usually attributed to sequencing errors, which may be the reason in this case. The SDS-PAGE bands for protein expression coincided with the expected molecular weight for both mutants, which could suggest that the correct proteins were expressed; however, there is a possibility that the proteins expressed could be of similar weight, but completely different. Interestingly, the V142D samples that did not include the protein inducer, IPTG, had a more intense band than the faint ones found for the samples that did include IPTG. This may just be a result of mislabelling. The SDS-PAGE performed to assess purity after the purification process. Mutant V142D had low protein expression as evidenced by its concentration of 3.2 ÃŽÂ ¼M. The V142D mutant should have very low protein expression according to Fierke et al. (1991) because valine at position 142 is uniquely required for maximal expression in E.Coli. It is suggested that by altering position 142, protein stability decreases [2]. Therefore, the protein that was expressed in the previous SDS-PAGE gel may not be V142D hCAII at all. The sample may have been small fragmented contaminant proteins that would have completely run off the gel altogether. However, the low concentration of V142D after purification may also be a major factor in the lack of gel bands observed as well. Unlike V142D, H64A hCAII concentration should not have affected its lack of bands because it was calculated to have had a reasonable concentration of 37.6 ÃŽÂ ¼M. There were some problems loading the samples into the wells; t his could be an explanation as to no observable gel bands. ESI-MS is dependent on concentration because it affects the size of primary droplets [8]. The unsuccessful determination of molecular weight of V142D hCAII may be attributed to its low concentration. The H64 hCAII mutant was also not able to be successfully analyzed with ESI-MS. A possible reason for the failure was that it was not kept on ice while it was not being used. The enzyme may have become inactive and degraded into smaller fragments. This would explain the ESI-MS output obtained for H64A. No definite molecular mass was determined, but the spectrometer did detect a lot of small protein fragments in the sample, all under 1000 amu. The kinetic values obtained from PNPA hydrolysis do not follow similar trends found in literature [2]. The kcat/KM for wild-type hCAII (2500 ±200 Mà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1sà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1) was found to be significantly larger than V142D hCAII (3 ±0.3 Mà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1sà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1) in literature, more than 800ÃÆ'- larger [2]. Experimental calculations yielded kcat/KM for V142D (8.16 Mà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1sà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1) to be about 2ÃÆ'- larger than wild-type (4.02 Mà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1sà ¢Ã¢â€š ¬Ã¢â‚¬â„¢1), which did not follow literature patterns. The literature trends make more biolog