Determination of Glucose Concentration Using Trinder Method

Determination of Glucose Concentration Using Trinder Method The Trinder method is used to determine glucose concentration only, (Lott et al, 1975). This method was first described by Trinder in 1969 thus named after him, (Lott et al, 1975). It uses an enzyme glucose oxidase for the first reaction and peroxidase for the second reaction thus the name of the enzyme Glucose oxidase/peroxidase (GODPOD), (Meiattin, 1973). Enzymes are biological or any chemical catalysts that speed up a reaction without it being used up, (Jan, 2010). It functions to catalyse a reaction by lowering the activation energy of the reaction. Activation energy is the energy needed to initiate the reaction. It is a point of high energy and requires more energy than the substrates. An enzyme also contains an active site for the substrate to catalyse the reaction. Its efficiency depends on the concentration of the substrate and conditions like temperature or pH, (Hames et al, 2005). The Trinder method, is based on two sequential enzymatic reactions, the first one involves the oxidation of glucose to gluconic acid and H2O2, (Casabnon et al, 2005). This reaction is catalysed by the enzyme Glucose oxidase. Then, the H2O2produced is quantified by a chromogenic reaction with peroxidase (POD), as the enzyme that catalyses the reaction with the reduced dye, (Casabnon et al, 2005). The oxidised dye changes colour to pink or red according to the glucose concentration. The colour formed is stable at room temperature for at least two hours after development, (Anon, 2010). The main advantage of this method is that it is very specific. It doesnt target other sugars except glucose. It is also simple straight forward and easy to manipulate. Its results are very reliable and specific, (Bauninger, 1974). Its final products are stable as they are not reactive at room temperature. Other colorimetric methods to identify glucose include, oxidation of glucose in the presence of Cu+2 to give Cu2O, and different types of Chromatography, (Casabnon et al, 2005). Glucose can be detected with o-toledine or other amines, (Casabnon et al, 2005). The experiment had to check for the specificity of the assay thus other carbohydrates were assayed. These were galactose, fructose, maltose and ribose. Maltose is a disaccharide which is made up of two glucose molecules joined together by a glycosidic bond. Galactose is a major sugar found in milk, (Hames et al, 2005). Its structure consists of six carbons with a glycosidic bond to join the next glucose molecule, (Berg et al, 2007). Fructose is abundant in fruits. It is a monosaccharide with six carbons as its structure. Ribose is a pentose sugar molecule with 5 carbons. It is mostly abundant in the nucleotides. The aim of the experiment was to determine glucose concentration in different concentrated solutions and unknown solutions. The specificity of the assay was to be determined by application of the assay on different sugar molecules. A standard curve was to be drawn from the absorbencies acquired from the spectrophotometer at 515nm Materials 12 Test tubes 2 Long pipette tips 1ml of 0.5mM Fructose 1ml of 0.5mM Maltose 1ml of 0.5mM Galactose 1ml of Unknown Glucose 1ml of 0.5mM Ribose 5ml of 0, 5mM Glucose 5ml of Distilled water 7ml of 0.1% Phenol 20ml of GODPOD Reagent 9 pipette tips 8 Disposable Cuvettes Black Marker Stop watch P100 Pipette Automatic pipette Recoding paper and pen Spectrophotometer at 515nm Water bath at 37oC Test tubes rack Blotting paper Method The test tubes were marked T1 to T6, for those that had to be inoculated with glucose and S1 to S6, for those that had to be inoculated with different types of sugars. They were arranged in order of concentration on the rack. One row was left out for agitating the test tubes. Inoculation commenced by transferring glucose into different test tubes T1being the least concentrated. 0.5mM of Glucose was transferred using a P100 pipette and not changing the tip. 0.2cm3 was inoculated into T2, 0.4 cm3 into T3, 0.6 cm3 into T4, 0.8cm3 into T5 and 1.0 cm3into T6. Distilled water was then inoculated into the test tubes using a different tip to avoid cross contamination. One centimetre cubed was inoculated into T1, 0.8 cm3 into T2, 0.6 cm3 into T3, 0.4 cm3 into T4 and 0.2 cm3 into T5. There was no water inoculated into the last tube T6. Phenol was then inoculated into all the twelve test tubes. It was transferred using a different tip to avoid cross contamination. One centimetre cubed of different sugars were inoculated in specific S tubes. 0.5mM of Galactose was inoculated into S1, 0.5mM of Glucose was inoculated into S2, Glucose unknown was inoculated into S3, 0.5mM of Fructose was inoculated into S4, 0.5mM of Maltose was inoculated into S5 and then 0.5mM of Ribose was inoculated into S6. These transfers were done with different tips for different sugars. One and half millilitres of GODPOD reagent was then inoculated into all the test tubes using an automatic pipette and a long pipette tip. The test tubes were then agitated on the rack and incubated in the water bath for forty minutes. The temperature was constantly checked during incubation. After forty minutes, the solutions changed colour from colourless to light pink according to the concentration. These different solutions were then read on a spectrophotometer in a cuvette. The spectrophotometer was zeroed at first then absorbencies of Glucose and other sugars were read and recorded. A cuvette was wiped on the soft side to minimize absorbencies caused by contamination. These different absorbencies were recorded on a table. Discussion The reactions of glucose with the GODPOD were slow due to the fact that the enzyme was stored in ice thus it took long for the reaction to take place. The enzymes structure was disrupted because it was kept in cold thus it took time to equilibrate with the conditions. The reason why GODPOD was slow to react was because it is sensitive to its environment, (Teal et al, 1985). Enzymes are sensitive to pressure, temperature and pH. This added to the fact that the colour produced was not very dark because the enzyme was adjusting to the conditions. The enzyme also didnt denature because it was kept in ice at 4oC not in the heat above 40oC. On the graph, the points that are not on the line of best fit might have appeared because there might have been a competitive inhibitor thus the reaction didnt go on well owing to reduced absorbance. The inhibitor might have been so because of cross contamination. The same pipette might have been used to transfer the solutions thus cross contamination. The specificity of the enzymes might have caused the other solutions not to produce reliable results. Enzymes are sensitive to pH, (Jan, 2010). The pH of the test tube might have been so low or higher than the required thus some of the points are not in the line of best fit. This might have been avoided by carrying out the experiment repeatedly and then getting average values. When the enzyme was applied to the other sugars, there was no absorbance at all because the enzyme is specific to one substrate thus it didnt catalyse the reaction of other sugars and GODPOD. There might have no absorbance because these sugars might have their own wavelengths they absorb the light. This might have been avoided by scanning the various wavelengths and determine the exact wavelength. The unknown glucoses were determined by the use of the graph. The line of best fit was used to determine the glucose concentration. An equation was used also to determine the concentrations. The other points not on the line of the best fit might be there because the transitional state might have been great, thus when thirty minutes had passed, the enzyme had not gone past the transitional stage. This might have been characteristic of the colours produced according to the concentration of the different solutions. The unknown glucose solutions showed to have the same concentration as the stock solution. It might have the same compounds and properties as the stock solution. The results of the experiment were according to the literature values. This was highlighted by the absorbencies of stock solutions and different sugars. The specificity of the reaction was achieved. In conclusion, the aims of the experiment were achieved by obtain reliable data and results. The standard curve showed the absorbencies of the different solutions and unknowns.

Social and Political Discrimination Against Women

Social and Political Discrimination Against Women The world is facing many problems that are demolishing its unity, forcing the process of making a better world harder to achieve. One of the most devastating problems yet to be solved is the womens rights against discrimination. Discrimination, according to Cambridge Universitys dictionary is the act of treating a person or particular group of people differently, especially in a worse way from the way in which you treat other people, because of their skin colour, religion, sex. Discrimination against women is a type of gender discrimination. According to the Australian Office of Anti-Discrimination Commissioner (OADC) gender discrimination occurs when someone is treated less favourably than another person because of his or her gender (Justice, 2009). Womens discrimination is a series problem, it is just not a discrimination against a minority (with all do respect to all minorities). It is impossible to realize our goals while discriminating against half the human race Kofi Annan[1]. Annan described discriminating against women discriminating against half of the human race which rely on how important the role of women in the progressing of the human race. The problem of discrimination against women was officially addressed to the world through the first couple of years of the establishing of the United Nations (UN). Women inscribed their identity as holders of rights in the founding documents of the UN-the UN Charter (1945) and the Universal Declaration of Human Rights (1948) the Convention of Civil Rights to Women (1948) and the Convention on the Political Rights of Women (1952) (Jain Sen, 2005, pp. 12-13). Many countries and nations have issued legislations against discrimination, and specifically against gender or sex discrimination. One of the most notified acts against discrimination is the Sex Discrimination Act 1975 by the British Parliament. Many countries and nations have acts against discriminating women; however, women are still being discriminated b efore the law (Franciscans). Discriminating against women is not just discriminating against a person it is discriminating against families as well; who would want the mother, the sister or the wife to be discriminated at work, at school or at club under no reason but solely because of the gender. The purpose of this essay is to examine the act of discrimination against women and demonstrate on how women suffer in the social life, the political arena and the workplace, and the education. Moreover this essay will adjudge the previous solutions to overcome the problem of discriminating women due to their gender held previously decades ago by countries or nations, and why they did not reap what they have sown. Ultimately, it will propose a solution on how to surmount the problem of discriminating women, because women should not be discriminated due to the prejudice against their gender. Women play the part of half the human race if they were less competent or reliable than men and do n ot deserve equality God would not let them share every role with men. Womens discrimination is much far from being a local issue in a certain part of the world. Women suffer from discrimination, violence and sexual harassment because women most of the times are thought to be less competent than men because of their physical structure. Women are not only discriminated in the developing countries because of the lack of sophistication, women are also being discriminated in the developed countries. Discrimination against women in the UK is deeply ingrained, a government report concludes as cited in (Barriers still in womens way, 2005). The UK, one of the most important and developed countries that plays a major role in the world issues, has a discrimination problem against women which is described by a governmental report to be deeply ingrained or firmly held that it is not likely to be changed. Thus it is a problem that needs more that attention to be solved. According to the United Nations Development Fund for Women (UNIFEM), Data shows that discriminato ry practices against women exists and dominates in almost all parts of the world (2008). Therefore, we do not need each developed country telling a one that is not that the developed country is free from discrimination or it is in very small ratio that discrimination hardly can be found. Statistically according to figure 1[2], the ratio between the Middle East and North Africa (MENA) and the developing countries does not gap critically; life expectancy is more in MENA than in the developing countries while it decreases in the other areas. The world needs all countries and nations help because it is not a one nation problem it is a problem that we all command. Women social life is covered by many kinds of discrimination. Women in social life suffer from many daily-life appellations, especially in the Middle-East other than the West. In the Middle-East women are classified by their relationship status other than by her contribution to the community. While searching for life partner men get repelled from divorced women. In the process of searching, most of the men search for virgin girls whether those men have previous relation or not. Divorced women are treated different than married women. Whereas single-mothers most of the time do not marry after their first marriage because men do not want extra burden or responsibility. On the other hand single-fathers easily can get married most of the time as there is no social norm against them to re-marry. Single mothers do not refuse to marry, however, they are refuted by the men how are seeking marriage. Divorce is more costly for women than men. The most common impact of divorce on women is the f inancial insecurity it creates, increasing the possibility of poverty for them and their children. Data show that after divorce, women experience a 73 percent loss in their former standard of living and men experience a 42 percent rise (Headlee Elfin, 1996, p. p.52). Not only divorce leave women socially downgraded it also leaves her in economical insecurity, and if a women is supporting a child the disaster will be doubled. According to Clarke-Stewart Brentano, divorced women make only five new friends in the first year of divorce due to the emotional damage of divorce (2006, p. 70). Women are more likely to have blighted social life than men after divorce. Nevertheless, society does not welcome divorcees leaving women in a dark corner of the society. Politically, women suffer from a great impact from discrimination. Women have been discriminated in the political arena, as the society gives the women politicians less creditability than men due to some ideas stuck in the minds that women are more likely to be secretaries other than being the boss. The media has a huge impact on such negative ideas, giving the women always as the soft secretary that has a model body shape and a voice that rhymes with music. According to Abdel-Wahabs film, he sketched a wife being a CEO in a governmental institution, while the employees of that institution do not believe that a woman can hold such a sophisticated position (1966). In that film the female CEO proves to her husband and to her employees at the end of the film that she is as competent as any male CEO that has ever held this position. This is the kind of media that needed to be seen today, not the ones that weaken the image of women and strengthens the stereotyping of them in politics and in workplace. According to Constance B. Morella a US Republican Congresswoman, who represents the Seventh District in Maryland, in politics (once elected) there is equity in terms of salary, but not in terms of leadership. Women are excluded from many issue areas and commissions where they might serve as cited in (Headlee Elfin, 1996). In the US congress the government cannot give smaller salary to women, of course or it would be contradicting its own policies out in the public, nonetheless, they do not give women the right of leadership as they might provide help to their country, the US- her country- deny the womens help in leadership. Unlike men, women pursue politics for the sake of issues and morals not for career advancement. The rejection of women being in the political arena gave them the opportunity to be more active volunteers than men, which gave them more than enough experience to successfully enter the political arena (Headlee Elfin, 1996, p. 26). Men do not have an e xtra brain that makes them excel in politics and likely women do not, hence, they are equal and should have equal political opportunities. Women went to work thus, affecting the mens jobs and the economy mainly because of money. Women worked because they did not have husbands as unmarried or divorced, or they had husbands who were in low-pay jobs. In 1994, 59 percent of married women were working for pay which increased by 19 percent from 1970 (Headlee Elfin, 1996, p. 3). As of discrimination in the work place, statistically, pregnant women suffer widespread discrimination at work, figures show, with almost one in 14 mums-to-be denied opportunities for promotion and one in 50 demoted (Pregnant Women; Discrimination at work, 2006). Pregnant women takes the largest piece in the pie chart according to discrimination, some of the pregnant women do not even get paid for maternity leave, while, some of them do not get promoted and some get demoted. Bibliography Abdel-Wahab, F. (Director). (1966). My Wife is CEO [Motion Picture]. Barriers still in womens way. (2005). Evening Post (South Wales) , p. 5. Clarke-Stewart, A., Brentano, C. (2006). Divorce : Causes and Consequences. New Haven: Yale University Press . Franciscans, I. (n.d.). Discrimination Against Women. Retrieved December 21, 2009, from Franciscans International: http://www.franciscansinternational.org/issues/women/discrimination Headlee, S., Elfin, M. (1996). Cost of Being Female. Westport: Greenwood Publishing Group, Incorporated. Jain, D., Sen, A. K. (2005). Women, Development, and the UN : A Sixty-Year Quest for Equality and Justice. Bloomington: Indiana University Press. Justice, T. D. (2009, September 4). Anti-Discrimination Commissioner: Gender Discrimination. Retrieved December 21, 2009, from Tasmania Online Your gateway to Tasmania: http://www.antidiscrimination.tas.gov.au/publications/gender_discrimination Pregnant Women; Discrimination at work. (2006). Advertiser, The (Adelaide) , 15. UNIFEM. (2008). Justice / Field Stories / Discrimination Against Women : UNIFEM : Progress Of The Worlds Women 2008/2009. Retrieved December 21, 2009, from United Nations Development Fund for Women: http://www.unifem.org/progress/2008/media/POWW08_Report_Full_Text.pdf

Morning Sickness :: essays research papers

Why do they call it morning sickness if I feel nauseated all day long? About three-quarters of all pregnant women have nausea (and sometimes vomiting) during their first trimester. For some, it's worse in the morning and gets better over the course of the day, but "morning sickness" can strike at any time. The nausea usually starts around 6 weeks but can come on as early as 4 weeks for some women, and it usually tapers off around 13 weeks as you start your second trimester, although queasiness can come and go throughout your pregnancy. What causes nausea and vomiting during pregnancy? No one knows what causes nausea during pregnancy, but it's probably due to some combination of the many physical changes taking place in your body. Some possible causes include: †¢ Rapidly increasing levels of hCG, estrogen, and other hormones during early pregnancy No one knows why they may contribute to your nausea, but the timing is right: Nausea tends to peak around the same time as your levels of these hormones do. †¢ An enhanced sense of smell and sensitivity to odors. It's not uncommon for a newly pregnant woman to find that she's now overwhelmed by the smell of a bologna sandwich from four blocks away, for example, and that certain aromas instantly trigger her gag reflex. This, too, may be a side effect of rapidly increasing estrogen in your system. †¢ A tricky stomach. Some women have a gastrointestinal tract that's more sensitive to the changes they're undergoing in early pregnancy. One study even found an association between h. pylori (a gastrointestinal bacteria that can cause ulcers) and severe nausea during pregnancy, although no one knows why the two may be related. You may have heard that morning sickness can be caused by a B vitamin deficiency. While taking a vitamin B6 supplement does seem to help ease nausea in many pregnant women, that doesn't mean they have a vitamin deficiency. In fact, at least one study has shown no significant differences in the levels of B6 in women with morning sickness and those without it. No one knows why B6 is helpful. Some researchers also believe that stress and emotions may also play a role in morning sickness, but it would be hard to say whether it's stress causing the nausea or the other way around. Are some pregnant women more likely than others to have nausea?

A Typical American Wedding Essay -- Personal Narrative Writing

A Typical American Wedding Marriage. Are there any specifications to when someone gets married? When is it the appropriate time to get married? All these questions swarm my mind day after day as I envision my wedding. I don't believe those questions have a correct answer, but can only be answered by that person who is considering marriage. I on the other hand seem to answer those questions every day and develop a new answer everyday. As of right now I have been with my boyfriend, Forrest, for 3 years and two months. According to my stipulations of when to become engaged, I believe it should have already happened. We have each discussed it numerous times. My mom has subscriptions of wedding magazines that seem to arrive just as I walk into the door to visit, and my niece already refers to him as her uncle, yet I do not have a ring! I have thrown out every hint to him, from brochures of ring settings, to bringing up subjects of other friends who are engaged, but still no proposal. As the holidays and my graduati on approach, I anticipate the idea of an engagement. Although it may seem that I am eager to expedite this memorable event, it is also obvious to point out that there are many arrangements and a lot of stress that is tacked on to planning a wedding. First, I will have to acknowledge that I will have to deal with my family who has been sitting on pins and needles waiting for this day, possibly more so than myself, and of course there is the strategic plans. Since I have been waiting for this day, I have talked with my cousin's fiancà ©e, Lyssa, who has gone through this very similar delay of a proposal. I had to listen to her gripes and now she gets to listen to mine. But fortunately her time has come and she has gone from ... ... Two words of advice for brides-to-be. First, Be shacked up for at least a year because living with Kevin makes it so easy to marry him. I know what I am in for and I am marrying him with no doubts! Second, Have a lengthy engagement to make sure your wedding includes every detail you want. You'll have time to save needed money for all the extras and you'll have time to make everything happen. Last, I am adding a word for the grooms. If you are shacked up don't even think about surprising your girl with a ring, just do it within a year or your engagement will be an ultimatum." Personally I believe weddings are beautiful, but after witnessing many conflictions that Lyssa has gone through, I have learned that there are other aspects to planning a wedding. But in the end it is all worth it, and I anticipate watching the actual wedding of Kevin and Lisa on February 2nd.

An Ethical Theory Applied to Business Organizations Essay -- Business

The object of this essay is to establish whether there is an ethical theory that can be successfully applied to business organizations. In order to answer this question, it is necessary first to define the major ethical theories, which are utilitarianism, deontology and virtue ethics, before determining whether there are any other options. After that, the ethical needs, problems and limitations of work organizations will have to be examined so that the different theories can be evaluated in this context. It will also be important to draw a distinction between the terms â€Å"accurate† and â€Å"useful† as these actually result in two different questions the answer to which need not necessarily be the same. Another essential part of this discussion is the more general question why there should be ethics in work organizations in the first place, and to what extent ethical behavior is feasible in the business world. Hopefully this will provide a framework within which the answer to the essay question can be included. To begin with, the â€Å"traditional† approaches to solving moral problems will be defined, namely those of act-utilitarianism and deontology. These are traditional not because they have existed longer than virtue ethics, but because for a long time they have been the approaches most commonly used as a basis for trying to solve ethical problems. After looking at these two moral theories, the approach of virtue ethics regarding moral dilemmas will be investigated in order to compare differences and advantages as well as problems. Utilitarianism is a form of consequentialism developed by Jeremy Bentham and John Stuart Mill. It adopts the consequentialist view that it must be right to make the world a better place and therefore the right action is the one that produces the best consequences. Utilitarianism takes this one step further and defines the action that will produce the best consequences as the one that produces the most happiness. Act-utilitarianism holds that each case should be judged separately in terms of which possible action would maximise happiness; this means that there are no rules to follow, but the theory seems to provide a decision-procedure to act upon in that one acts upon a principle of justice. Intuitively this seems to be an appealing solution to ethical dilemmas. To maximise happiness makes utilitarianism a noble ideal. Furthermore... ...usiness Ethics, Vol.19, No.3, April II 1999 Cicero: Selected Works. Penguin Classics, Harmondsworth - England 1971 Donaldson, D. and Davis, E.: ‘Business Ethics? Yes, But What Can it Do for the Bottom Line?’ Management Decision, Vol 28, 6, 1990 Legge, K.: ‘Is HRM ethical? Can HRM be ethical?’ in Parker, M. (ed.): Ethics and Organisations. Sage Publications - London 1998 Madsen and Shafritz Essentials of Business Ethics. Penguin Books 1990 McDonald, G.: ‘Business Ethics: Practical Proposals for Organisations.’ in Journal of Business Ethics, Vol.19, No.2, April I 1999 McNamara, C.: ‘Complete Guide to Ethics Management: An Ethics Toolkit for Managers.’ The Management Assistance Program for Nonprofits 1999 (http://www.mapnp.org/library/ethics/ethxgde.htm) McNaughton, D.: Moral Vision. Blackwell - Oxford 1988 Norman, R.: The Moral Philosophers. Clarendon Press - Oxford 1983 Pearson, G.: Integrity in Organisations. McGraw-Hill Book Company - London 1995 Richards, R.: ‘Cicero and the ethics of honest business dealings.’ The Online Journal of Ethics 1997 Townley, P.: ‘Business Ethics: Commitment to Tough Decisions.’ Vital Speeches, pp. 208-211, 1992 (Jan)

Branding Yoga Essay

I think that’s why there are more and more people are encouraged to practice yoga and yoga has become a power brand which is able to compete with other industry. The yoga brand is managed systematically with branded styles of teaching, branded yoga clothing, branded training programs, branded instructors and etc. There are two branded styles of yoga which are Bikram style and Tara stiles. They are two different styles which appeal to two different sorts of people. Firstly, Bikram Choudhury who is the founder of Bikram yoga in America patent his approach to traditional yoga style. In the process of branding yoga, Bikram obtained the patent for his book, trademarked his company’s name and actively fighting for the copyright of his style. When branding a style of yoga, the legal rules which involves patent, trademark and patent intend to protect the founder’s right and disseminate the gains of the innovation so that it can be recognized by the people with the brand identity. The brand is also supported by Indian government which avoid others following Bikram’s branded poses even though the government only considered the yoga was part of the India’s traditional knowledge ( Deshpande, Herman and Lobb 2011). Additionally, the branding strategy in terms of the Bikram ‘s classical style of yoga is based on the inherent value and adds the more substantive value but still consistent and adherence to the addition (Cohen 2013). Secondly, Tara stiles is distinctive from Bikram’s style which develop the brand in a very fresh and personal way like the founder Stiles said â€Å"making yoga cool†. Stiles broke the rules and added many creative values to the brand such as music in order to maintain the relevance in the society especially the younger generation (Girard 2013). A more modern yoga style get the people more interested in and got popular soon by the promotional videos via different channels of communication in modern society. Stiles’s yoga is considered as authentic yoga since it is created as distinctive, exciting, attractive with personal values. By thinking the reason of success of Stiles’s success, the key is how Stile manage the brand by fitting it in American society. According to the feature of the liberal society in America, Tara promote a natural style not rigid with a certain style which perfectly stay relevance to the social values. Furthermore, the brand communication through the YouTube, Facebook, Ipad app strength brand identity and brand influence. In conclusion, by analyzing the two styles of yoga, I learned that the values adding to the brand is very important for branding experience. The brand needs to be relevant to the features of the society and the brand needs to be protected through the legal regulation. Managing brand is a long process which requires both adherence to the tradition and innovative modification. Desphande R. , Herman, K. and Lobb, A. 2011, Branding Yoga, Harvard Business School, Harvard. Cohen, M. 2013, Branding a style of Yoga, yoga journal, viewed 17 May 2013, <http://www. yogajournal. com/for_teachers/1683#comment> Girard, K. 2013, Branding Yoga: Good Business or Blasphemy? , Forbes, Harvard Business School

kingdom of matthias Essay

Starting in the early 1600’s English settlers began migrating to the Americas in search for liberty and the right to govern themselves in what they deemed a Christian manner. Thomas Morton, was one of the first to write about the Indians of New England. He condemned their religious beliefs claiming it was similar to devil like worship. (VOF,4) The Native Americans were seen to be living a much happier life even without religion, law and a king. This daunted Christian and they did not understand how this could be. It’s written that they claim their prosperity was full of evil and they would lead a happier life once bought to Christianity.(VOF,6) I feel this is the last America sees of true religious freedom. In reality, religious liberty existed in very few parts of the Atlantic world in during this time period. Most nations outlawed religious groups that rulers deems dangerous or disruptive. Among all the colonies, one that demonstrated religious freedom in a higher sen se would be Maryland. Cecilius Calvert was a Catholic who wished to demonstrate that Protestants and Catholics could live in peace, something that was not seen in Europe. In 1649 Maryland established an Act Concerning Religion, which introduced religious tolerance, saying that anyone who troubled a Christian due to their religion would be punished.(VOF,27) I feel this allowed others to safely demonstrate â€Å"religious freedom†, but this is just one state among the colonies. During this time it was rare to have this type of true freedom. Like many settlers, the puritans came to American in search of liberty, and to govern themselves in what they deemed a Christian manner.(VOF,29) This is where Religious freedom loses the sense of freedom, because the puritans created their own definition of the word freedom. John Winthrop explains that freedom to them meant obedience to God’s laws and the law of rulers such as himself.(VOF,29-30) Basically you can see that they are imposing their moral stan dards on society as a whole, denying anyone other than themselves of true religious freedom. Unlike what Maryland was trying to implement, this colony believed in follow their religion, or be punished, much like a wide array of colonies. A prime example of this would be the trial of Anne Hutchison in 1637. Hutchison was accused of expressing opinions that did not match up to those in authority.(VOF,33) The trial gives examples of how if you stray against what the government deems Gods word, then you would be punished. Hutchison argues that she is reciting what she believes God told  her is his word, and Governor John Winthrop, one of her accusers, argues that this is another religion and its punishable. (VOF,35-39) Because Hutchison followed her own beliefs she is banished from their society, clearly being stripped of her natural right of religious freedom. Based on examples like this, what religious freedom really meant was, â€Å"do as I say or be punished†. You can see this start to shift as time goes on, you can see people start to realize the damage this is putting on society and settlers, such as Roger Williams, begin to branch out and start new movement. Williams founded Providence, Rhode Island, and then came to publish a letter claiming that no one should be forced to follow any particular religious belief, but they should still obey the government in charge of civil matters. (VOF,40) This is what religious freedom should have been from the beginning.