English Literature Lesson 2 Essay Example | Topics and Well Written Essays - 1000 words

English Literature Lesson 2 - Essay Example responses to the plays 1992 New York production, adding that "one can expect few other reactions when Carol is such a viper."1 However that is an individual view. Mamet employs language that offers different points of view that we shall analyze. Mamets presentation of the conflict between a professor and his female student is marked by ambiguous discourse, troubling physical contact, and subsequent charges of sexual harassment. Oleanna, which unfolds in three acts, takes place entirely in the office of a male college professor in his forties who receives three visits from a twenty-year old undergraduate named Carol. In the first of these visits, Carol seeks help from John because of her difficulties in his class on higher education. Carols questions, which are often cut off in midstream by his responses, are also interrupted by periodic phone calls, all of which involve Johns impending purchase of a house for himself and his family. Having just received word of his being considered for tenure, John feels secure enough to begin negotiating the purchase of a new home. Towards the end of the first act, as each character becomes more frustrated with the other, an exasperated John makes the mistake of offering Carol an "A" in the class, if she will just meet him in his office a few times during the semester to discuss her problems. He does this because he "likes" Carol and thinks they might be "similar." She pr otests, only to have John tell her to "forget about the paper," for "what is The Class but you and me?" (21, 25, 26). John then punctuates his lesson on theories of higher education with an indecorous analogy about how "the rich copulate less often than the poor. But when they do, they take more of their clothes off" (32). Not surprisingly, Carol continues to profess a lack of understanding for the subject (35-37), even though she has started to ask some rather penetrating questions and is on the brink of telling her professor a secret that she has "never told

Auteurism Essay Example | Topics and Well Written Essays - 4000 words

Auteurism - Essay Example However, nobody else believed the rain would fall as Noah had prophesied and hence all the people drowned except Noah’s family. In the Aronofsky’s film ‘Noah’, action starts by showing Noah being haunted by frightening visions that indicate that all humanity was faced with divine destruction as a punishment for their sin (Stasukevich, 2014). Another character was ‘Methuselah’ who was more of an advisor to Noah and was acted by Antony Hopkins. Accompanying Noah and Methuselah in the movie was his wife ‘Naameh’ acted by the Jennifer Connelly, and Noah’s children. Noah is portrayed building the Ark with only his family while the rest of the people watch in delusion. Methuselah is depicted to be living in a cave in a mountainous area. The people were led by Tubal Cain in rebellion and since they finally ignored God’s warning, they faced judgment. Tubal Cain was acted by Ray Winstone (Stasukevich, 2014). The film does not clearly display who the hero is between Tubal Cain and Noah. It is because Tubal Cain emerges sharply in action with strong influence over the people, only to perish with them in water. At some point, the rebelling people referred to as watchers, capture Noah and his family and put them in a pit. Therefore, Tubal Cain’s role in the film appears to be an anti-climax while the less action-oriented actor Noah emerges as the hero. Unlike in the real bible story, Noah is seen in a battle with Tubal Cain. The animals enter the Ark in a systematic manner, almost showing divine direction and Noah’s ability to control their behavior. According to the cinematographer Mathew Libatique, Aronofsky prefers image’s motion that is controlled, moves naturally, and almost flows uniformly with the film background (Stasukevich, 2014). Libatique explains this emphasizing the reason why he had to use handheld camera to shoot Aronofsky’s film.

Analysis of VP28 Gene in White Spot Syndrome Virus Infection

Analysis of VP28 Gene in White Spot Syndrome Virus Infection Temporal analysis of VP28 gene in White spot syndrome virus infected fresh water crabs Chiin Nei Chinga, Mansi Parihara, R. Sudhakaranb* aSchool of Bioscience and Technology, VIT University, Vellore – 632 014, Tamil Nadu, India. *bAssistant professor, SBST, VIT University, Vellore – 632 014, Tamil Nadu, India. ABSTRACT Aquaculture is the farming of aquatic organisms such as fish, crustaceans, molluscs and aquatic plants. Disease emergence is a concern in wild fisheries due to environmental pressures, the direct impact of human activities and the risk of pathogens spread from aquaculture. Common viruses are Taura syndrome virus (TSV), white spot syndrome virus (WSSV), and the necrotizing hepato pancreatitis bacterium (NHP-B). The White Spot Syndrome Virus is the most economically devastating viral pathogen to global shrimp aquaculture production and has been proposed to be capable of infecting all decapod crustaceans. WSSV is an enveloped ellipsoid virus, which belongs to the genus Whispovirus of the family Nimaviridae. VP28 is one of its major envelope proteins, and plays a crucial role in viral infection. In this study, the proteins of the infected crab were purified using SDS page and then Western Blotting was performed to extract that particular protein. The VP28 protein will appear as specific bands in the blot. Keywords:Â  Aquaculture, White spot syndrome virus, VP28, SDS-Page, Western blotting. 1. INTRODUCTION White spot syndrome virus (WSSV) is one of the major shrimp pathogen that causes a high mortality rate of 90-100% within 3-10 days of infection (Lightner, 1996). Natural WSSV infections have been found in captured and cultured specimens of the mud crab. WSSV is an enveloped ellipsoid virus, which belongs to the genus Whispovirus of the family Nimaviridae. In addition, WSSV can also infect a wide range of hosts including both decapod and non-decapod animals with more than hundred species described to date. So far, the genome from three different WSSV isolates has been sequenced. Sequence analysis showed that WSSV contains approximately 500 putative open reading frames (ORFs), most of which have no homology with any known genes or proteins in public databases. Till now, more than 50 structural and non-structural proteins were identified. Tools such as polymerase chain reaction (PCR) and nested PCR, originally developed by Lo etal., have been widely used and recommended by the Office of International Epizootics (OIE) to be used as standard diagnostic methods for the detection of WSSV. Despite their excellence in specificity and sensitivity, these methods were not suited in some circumstances due to their complications, the requirement of thermal cycler, time-consuming, and labor-intensive. Moreover, the classical agarose gel electrophoresis with ethidium bromide staining, following the visualization under the ultraviolet (UV) transilluminator required to analyze the result of PCR products. Western blotting identifies with specific antibodies proteins that have been separated from one another according to their size by gel electrophoresis. The blot is a membrane of nitrocellulose or PVDF (polyvinylidene fluoride). The gel is placed next to the membrane and application of an electrical current induces the proteins in the gel to move to the membrane where they adhere. The membrane is then a replica of the gel’s protein pattern, and is subsequently stained with an antibody. Therefore, these features could be limited their applications, particularly in the resource-limited areas and non-laboratory environments such as at the pond or station sites. 2. MATERIALS AND METHODS 2.1. Tissue homogenate preparation: Gills, muscle, hepato pancreas and Head soft tissue from the crab infected with WSSV were homogenized in 1:10 suspension with NTE Buffer. It was then freeze and thaw for three times then centrifuged at 5000 rpm for 5min. The supernatant were collected separately in a tube and stored at -20Â °C. This supernatant was used for protein analysis. 2.2. Protein estimation: Lowry’s method was performed for the estimation of protein in order to know its concentrations (Lowry et al., 1951). 2% sodium carbonate in 0.1N sodium hydroxide (Reagent A) 0.5% copper sulphate in 1% potassium sodium tartrate (Reagent B) Alkaline copper solution: Mixed 50 ml of solution A and 1 ml of B prior to use (reagent C) Folin Ciocalteau reagent (Reagent D) Protein standard stock: 50 mg of bovine serum albumin (fraction V) was weighed and dissolved in distilled water and made up to 50 ml (1 mg/ml) Working standard- diluted 1ml of stock solution to 5 ml with distilled water. (200 Â µg of protein/ml). 2.3. SDS-PAGE: SDS-PAGE is currently used to determine the molecular masses, and also to investigate the subunit composition and the domain structure of proteins. TAE Buffer (1X) was poured in tank which maintain the temperature and pH of the gel. Gel was prepared and protein sample is mixed with bromophenol blue which act as a tracking dye. Then it was loaded into the well and electric current of 50V is applied into the tank which is later on increase to 100V when the dye contacts the resolving gel. When the dye reach the end of the resolving gel it was taken out and stain with the help of Coomassie Brilliant Blue R-250 (methanol-50 ml, distilled water 40 ml, glacial acetic acid 10 ml) for 1 hour and de-stain the gel by immersing in the de-staining solution (65 ml of distilled water, methanol 10 ml and 25 ml of glacial acetic acid). 2.4. Western blotting: Transferring of the protein into a membrane The membrane can be either nitrocellulose or PVDF. PVDF requires activation with methanol for one minute and is then rinsed with transfer buffer before preparing the stack. After stacking, the electro transfer is carried out for 90 minutes at a constant current of 150 mA set by maintaining the voltage at maximum. Transfer to the membrane and checked using Ponceau Red staining before the blocking step. (PVDF cannot be stained). After ponceau staining, wash the nitrocellulose membrane with TBST for 1 min at room temperature. Blocking and Antibody incubation Block the membrane for 1 hour at room temperature in blocking buffer (10 % milk in TBST). After blocking, incubate the membrane in primary antibody (1:1000 dilutions) in 2.5% milk in TBST overnight at 4Â °C. Wash the membrane for three washes using TBST for 5 minutes each. Incubate the membrane with secondary antibody (1:2000 dilutions) in 2.5% milk in TBST at room temperature for 1 hour. Wash the membrane in three washes of TBST, 5 minutes each. Add ECL reagent (Solution A and B in 1:1 ratio) on to the membrane and spread it thoroughly on to it using a pipette. Remove the excess reagent and place the membrane on to the x-ray cassette and expose to x ray film in the dark room. An initial 10 sec exposure should indicate the proper exposure time because due to the kinetics of the detection reaction, signal is most intense immediately following incubation and declines over the following 2 hrs. 3. RESULTS AND DISCUSSION 3.1. SDS-PAGE: Fig.1 SDS-PAGE which shows different bands under white light. Lane 1- Gill; Lane 2- HST; Lane 3- Muscle; Lane 4-Hepatopancreas. The polypeptides of purified virus were resolved by SDS-PAGE using discontinuous buffer system. In 12.5% acrylamide gel, the purified virus yielded different polypeptides base on their molecular mass in which lower molecular weight will elute faster as compared to those having higher molecular weight, which were Visualized on Coomassie brilliant blue R 250 staining. 3.2. Western blotting: Fig 2. Western blotting Viral polypeptides separated by SDS-PAGE were electrophoretically transferred to nitrocellulose membrane (0.22 ÃŽ ¼m) in transfer buffer (192 mM glycine, 25 mM Tris and 20% methanol, pH 8.3), at 0.8 mA/cm2for 1 h. it was later on view by using film to visualize the relative abundance of proteins Discussion By performing SDS-PAGE, (fig.1) we can determined the different molecular weight of polypeptide. The difference in the number of the polypeptide may be attributing due to the difference in concentration of the resolving and other physical conditions. The western blot analysis of proteins of semi-purified white spot syndrome virus separated on 12.5% gel (fig.2) showed different number of immunogenic proteins based on its molecular weight. Out of these, those proteins, which stained more intensely, were considered as major immunogenic proteins. However, further studies are needed to establish its immunogenic nature and feasibility for its use as vaccine. Because electrophoretic separation of proteins is always carried out under denaturing conditions, the problem of solublization, aggregation and co-precipitation of target proteins with adventitious proteins are eliminated. REFERENCES [1] Lowry, O. H., N. J. Rosebrough, A. L. Farr and R. J. Randall (1951): Protein measurement with the Folin phenol reagent. J. Biol. Chem., 193(1), 265-275. [2] van Hulten, M. C., J. Witteveldt, S. Peters, N. Kloosterboer, R. Tarchini, M. Fiers, H. Sandbrink, R. K. Lankhorst and J. M. Vlak (2001): The white spot syndrome virus DNA genome sequence. Virology, 286(1), 7-22. [3] Seetang-Nun Y, Jaroenram W, Sriurairatana S, Suebsing R, Kiatpathomchai W (2013)Â ­: Visual detection of white spot syndrome virus using DNA-functionalized gold nanoparticles as probes combined with loop-mediated isothermal amplification. Mol Cell Probes, 27(2):71-9. [4] Maurice Pagano (1999): Application of electrophoresis and related methods, such as western blotting and zymography to the study of some proteins and enzymes. Analytica Chimica Acta, 83(1–2): 119–125. [4] Lightner, D. V. (1996): A Handbook of Shrimp Pathology and Diagnostic Procedures for Diseases of Cultured Penaeid Shrimp. World Aquaculture Society.

Americas Flawed Constitution :: American History

America's Flawed Constitution Right from the beginning of it’s creation the constitution of the United States has been a shaky document. The very basis for it being there was in fact illegal. The story of American politics starts with the Declaration of Independence. This document was brilliantly written by Thomas Jefferson and compacted all of the great ideas of enlightenment into one short easy to read paper. The declaration stated all of the ideals the new American nation would strive for. A constitution was needed as a way in which to fulfill those goals. The articles of confederacy were created as that constitution. However, they were weak, because no state wanted to give away any of their powers, and so the articles eventually failed. That is when the modern day constitution was starting to form. The Articles of Confederacy stated that in order to change any part of the document all thirteen states must agree to the change. Therefor a meeting was called so that they could amend the failing articles. However, representatives from two of the states did not show up. Even though not all states were represented the meeting started and the first vote was to totally throw away the Articles of Confederacy. The constitution wasn’t formed yet and it was already a flawed document. Because not all states were represented when the articles required it, the constitution was an illegal document. The delegates working on the constitution new that they needed a stronger document, because the articles proved too weak, but it still needed to please all of the states. This was impossible. So what ended up happening was the new ducocument became more and more vague. The only way to create a document that would pass was to make a document which didn’t really solve any problems but make each state believe that there problems would be fixed. This was accomplished by making it so that it was too vague to offend anybody but you could read into it. This made for a document that would be seriously flawed because people would be able to read into it too much. It could not work. The Constitution of the United States of America was too vague to work. The way the constitution was written it gave power to four parts: the congress, the executive branch, the judicial branch, and the states. Because it was so vague it did not really define which powers went where (with a few exceptions).

Living Together Without Married

What are the advantages and disadvantages of living together without marriage There are many advantages and disadvantages in living together before marriage. Today there are many couples living together before marriage. Sometimes these kinds of relationships ‘living together before marriage' end up with success and sometimes they are unsuccessful. Some of the advantages of living together before marriage are such as getting to know your partner, learning about one's abilities if he/she can satisfy your expectations and more.Also, there are some disadvantages in living together before marriage and they are such as religious and family values, parenting problems and more. I think there are more advantages then disadvantages in living together before marriage, because sometimes disadvantages in this kind of relationship are avoidable. One of the advantages of living together before marriage is getting to know a person that you might marry with. It is important for a person to know almost everything about the other person that he/she is going to get marry with.However, it can’t be accomplish without living together for a while before getting married. People need to know how a person is handling his/her life from all aspects such as behavior, mental, financial and others before a person decides to get married with. This cannot be completed in a few days, therefore, it is important to live together for a while before deciding whether to get married or now. First of all,cohabitation will be a trial period before marriage.We encounter with divorces in our daily lives because of couples who do not know enough about each other. They do not know other's home life or disturbing behaviours during the relationship. These are realized when they get married and start to live together. In addition some marriage decisions are given in 2 or 3 months after they met first. At this period,having an assumption about each other is very difficult. Cauples can be aware of o ther's special life and behaviours in cohabitation.They can make a step to a strong marriage with living together before marriage. Secondly,in cohabitation there will be financial independence and no legal obligations for spouse. When the couples start to live together,they will just share rent,money of pills and basic needs of home such as shopping,cooking and cleaning. Except for these,they will continue to their daily lives like living alone. When woman want to buy new shoes,she does not need to think what her partner will say. She buys them and does not have obligation for her spouse.Couples will be independence their personal needs. At this point,cohabitation is more alluring than marriage. Finally,it is argued that living together without marriage is against moral standarts at the society. Although there are many people who think so,I believe that moral values can not affect how people make a decision. They do not live for maral values. They live for their happiness. If we get married immediately with our partner who we fall in love without information about her/him , a divorce is inevitable.At this point,moral values do not help to save our marriage. To sum up,cohabitation is a experiment before marriage for couples who want to get married at the end or a life style for couples who love each other without legal obligations and regulations. For most people cohabitation might seem against to moral values or lack of legal regulations. However,I firmly believe that happiness is more important than these values and legal regulations. Therefore,we should give up to integrate and affect people's decisions. They have to live how they want. Living Together Without Married What are the advantages and disadvantages of living together without marriage There are many advantages and disadvantages in living together before marriage. Today there are many couples living together before marriage. Sometimes these kinds of relationships ‘living together before marriage' end up with success and sometimes they are unsuccessful. Some of the advantages of living together before marriage are such as getting to know your partner, learning about one's abilities if he/she can satisfy your expectations and more.Also, there are some disadvantages in living together before marriage and they are such as religious and family values, parenting problems and more. I think there are more advantages then disadvantages in living together before marriage, because sometimes disadvantages in this kind of relationship are avoidable. One of the advantages of living together before marriage is getting to know a person that you might marry with. It is important for a person to know almost everything about the other person that he/she is going to get marry with.However, it can’t be accomplish without living together for a while before getting married. People need to know how a person is handling his/her life from all aspects such as behavior, mental, financial and others before a person decides to get married with. This cannot be completed in a few days, therefore, it is important to live together for a while before deciding whether to get married or now. First of all,cohabitation will be a trial period before marriage.We encounter with divorces in our daily lives because of couples who do not know enough about each other. They do not know other's home life or disturbing behaviours during the relationship. These are realized when they get married and start to live together. In addition some marriage decisions are given in 2 or 3 months after they met first. At this period,having an assumption about each other is very difficult. Cauples can be aware of o ther's special life and behaviours in cohabitation.They can make a step to a strong marriage with living together before marriage. Secondly,in cohabitation there will be financial independence and no legal obligations for spouse. When the couples start to live together,they will just share rent,money of pills and basic needs of home such as shopping,cooking and cleaning. Except for these,they will continue to their daily lives like living alone. When woman want to buy new shoes,she does not need to think what her partner will say. She buys them and does not have obligation for her spouse.Couples will be independence their personal needs. At this point,cohabitation is more alluring than marriage. Finally,it is argued that living together without marriage is against moral standarts at the society. Although there are many people who think so,I believe that moral values can not affect how people make a decision. They do not live for maral values. They live for their happiness. If we get married immediately with our partner who we fall in love without information about her/him , a divorce is inevitable.At this point,moral values do not help to save our marriage. To sum up,cohabitation is a experiment before marriage for couples who want to get married at the end or a life style for couples who love each other without legal obligations and regulations. For most people cohabitation might seem against to moral values or lack of legal regulations. However,I firmly believe that happiness is more important than these values and legal regulations. Therefore,we should give up to integrate and affect people's decisions. They have to live how they want.

Character Education and Me Essay

After reviewing all of the materials for the course, I am convinced that character education is something that must have a presence in schools. Students of all grade levels can benefit from learning how to become civilized members of society. I agree with Lickona (1992) as far as the family being the first and foremost teacher of morals. As adults, we continue to reflect on our morals and virtues with hope of continually improving ourselves, especially when we become parents. I as well as many other new parents ran out to get a parenting manual to assist me in raising my children the best I could. Character education should be incorporated into the lessons daily in context. As the School Family video showed us, it should not be done through bribing, rewarding and punishing students without a lesson involved. After the school adopted the Conscious Discipline program, it appears that they have students use their words to talk out their issues with one another. I feel this has a better impact on the students in the long run. They will be better able to handle conflict in the future because of the lessons their teachers have helped them to understand. Kohn (1997) explained the side of character education with which most teachers are familiar. He explains, much like the Fern Creek video states, that students are most frequently drilled and rewarded through the character education programs. I do not necessarily agree that the values should be taught out of context. As a language teacher, I can tell you first hand that rote memorization does not show understanding of a concept. In addition, any person who has raised a preschooler can tell you that one of the first questions a child asks is â€Å"Why? † Children do not learn from simply being corrected or told what to do. They learn when they can explain why something is the way it is. Good character is completely developed by knowing ones morals and wanting to abide by the virtues by which we all live. If more parents were sensitive to the information that Lickona discussed in the Raising Good Children video, schools would have an easier time with character education. I agree that parents and schools need to form a partnership. Parents typically do not line their children up and drill right or wrong. They teach their children virtues in context. If my son steals his brother’s toys and a fight breaks out, I am not likely to say â€Å"that’s not right! † and end the conversation there. Young children are more likely to stop the bad behavior if they know why it was wrong or hurtful. I would explain to my son, â€Å"It hurts your brother’s feelings when you take his toys. Do you like your feelings hurt? Please ask him next time. † Incorporating the â€Å"do unto others† ideal does not make an impact unless the child can relate directly to the situation. I have noticed the times I had forgotten the explanation of why to stop; my son would repeat the misbehavior a short time later. Many of the virtues I have learned have come from my mother and my grandmother, since they were around me the most as a child. Through gentle reminders from these women I learned a set of standards to live by, my virtues. By knowing this information and using the good moral judgment that my family taught me, I have developed my character. Some children come to our classrooms without the opportunity to learn the way I did. Therefore, I need to be sensitive to their needs and help them to focus on how to change their behaviors. By treating students fairly and with compassion on a consistent basis, hopefully I can encourage students to intrinsically want to have good character.