Is Atticus Finch A Good Father Essay - 736 Words

Atticus Finch from the novel To Kill a Mockingbird is a single father to Jem and Scout and a lawyer in Maycomb. He teaches his children about respect and shows it in everything he does. He not only teaches them respect, but also morality and life lessons. He’s upfront with them about how life is and the troubles in it. Because of these three reasons and many more that I haven’t mentioned, Atticus Finch is a good father. Atticus is a good father because he sets an example of respect. He teaches his kids to respect others views and beliefs. Not only does he teach them but he also sets an example of respect even when it is hard to respect a person. â€Å"Mr. Cunningham’s basically a good man†¦ he just had his blind spots along with the rest of us†¦Ã¢â‚¬ ¦show more content†¦Atticus is a good father because he teaches them about morality and life lessons. My last but not least reason why I believe Atticus is a good father is because he is upfront with them and tells them how the world truly is. â€Å"You aren’t really a nigger-lover, then, are you?† â€Å"I certainly am, i do my best to love everybody...it’s never an insult to be called what somebody thinks is a bad name. It just shows you how poor that person is, it doesn’t hurt you.† Atticus is giving Scout a lesson about racism which you usually wouldn’t give a child because they are young. But even though she is young, he is letting her know for the future. Which, gentlemen, we know is in itself a lie as black as Tom Robinsons skin, a lie I do not have to point out to you. You know the truth, and the truth is this: some Negroes lie, some Negroes are immoral, some Negro men are not to be trusted around women—black or white. But this is a truth that applies to the human race and to no particular race of men.† (Lee 232). People t alk so bad about Native Americans just because of the color of their skin but they are no different than any other ethnicity. Jem and Scout know a lot about racism and how the world works than what other children their age knows. I believe that is why Atticus is a good father. Atticus isn’t your typical father but that is what makes him not just an incredible person but an exceptional father to Jem and Scout. He shows the respect that heShow MoreRelatedTo Kill a Mockingbird- Is Atticus Finch a Good Father? Essay examples1803 Words   |  8 Pagesdepicts Atticus Finch as a good father to his children due to his sense of fairness, his teaching, and his honesty. Atticus depicts fairness by treating his children and all others with respect and understanding. He teaches his children important life lessons and prepares them for when they go out into the world, and he sets a good example by always being honest. To be a good father, these are three very important qualities to possess. Atticus Finch’s fairness shows that he is a good father to JemRead MoreThe English Assessment Task For Kill A Mockingbird By Harper Lee1360 Words   |  6 PagesA Mockingbird’ Essay â€Å"Whether Maycomb knows it or not, we’re paying him the highest tribute we can pay a man. We trust him to do right.† To what extent are life values and lessons reflected throughout the novel ‘To Kill A Mockingbird’? The great degree that Harper Lee reflects life values and lessons throughout ’To Kill A Mockingbird’ is elucidated through the teachings of the main adult protagonist, Atticus Finch. Over three summers, Atticus guides his children, Scout and Jem Finch, through theirRead MoreKill A Mockingbird By Harper Lee1304 Words   |  6 Pagessolid ground of good morals and beliefs for the children in the book but also young readers decades later. Children are often influenced by adults in their life whether it has been a positive or negative moral aspect. Children often receive advice and encouragement from their guardians or adult figures. In author; Harper Lee s novel, To Kill A Mockingbird, readers observe three parents who have influenced their children heavily, in this essay readers will look further into Atticus Finch, Mr. RadleyRead MoreWhat lessons does scout learn in to kill a mockingbird1613 Words   |  7 Pagesof this novel is a series of life lessons preparing Scout for the hardships she is going to face in the second part of the novel. Due to the influence of the likes of Atticus, Miss Maudie and Mrs Dubose, Scout goes from a naà ¯ve young girl who thought with her fists rather than her head, into a more mature, empathetic girl. This essay is going to discuss some of the lessons Scout learns and how they impacted the way she became at the end of part one. One of the first lessons Scout learns is to be tolerantRead MoreQuotes To Kill A Mockingbird1096 Words   |  5 Pagesin many different forms. This quote is perhaps one of the biggest themes present in this book. Atticus put this in the mind of Scout after giving her and Jem air rifles. When he instructed Jem to shoot tin cans, Atticus drifted to the possibility of Jem going after birds, saying that it was a sin to kill a mockingbird. When Scout asked Miss Maudie about this incident, she replied with the quote this essay is written upon,  ¨ ‘Your father’s right,’ she said. ‘Mockingbirds don’t do one thing but makeRead MoreAnalysis of the Childhood World of Jem Scout and Dill and Their Relationship with Boo Radley in Part One?917 Words   |  4 Pageselements and some good that together form her idea about the world and the way she thinks reacts to different thing later on in the story and maybe even in the following years after the end of the book . One of the things that older Jean Louise talks about a lot in the book is Arthur ‘Boo’ Radley and his relationship with her, Jem, and Dill. That, basically, is the basis of this essay, together with the analysis of the childhood world of these three kids. Jean Louise Scout Finch is considered smartRead MoreKill A Mockingbird By Harper Lee1686 Words   |  7 Pagesto win† (Lee 101). One of the major lessons of Harper Lee’s novel To Kill a Mockingbird is to always do the right thing. The text is told through the narrator Jean Louise â€Å"Scout† Finch. She lives in the small, old town of Maycomb, Alabama with her brother, Jem Finch and her father, Atticus. Across the street from the Finch lives Arthur â€Å"Boo† Radley who is believed to be a horrible human. Rumor has it he eats squirrels and cats and is locked in the basement of his house for these actions when he wasRead MoreEssay about To Kill a Mockingbird2197 Words   |  9 PagesEnglish essay on To Kill a Mockingbird In To Kill a Mockingbird Atticus finch is presented as a respectable well-known man. Before Atticus Finch there was a customary tradition at the Finchs landing, which has been in place since Simon Finch made it his home and died there. The customary tradition was the men in the family remained on Finchs landing and made their living from cotton. In the twentieth century Atticus Finch went to Montgomery to read law and John Hale Finch, Atticus youngerRead MoreKill A Mockingbird By Harper Lee1751 Words   |  8 Pagesdistinction of good and evil in the morals of human nature. Lee uses the elements of setting, point of view, symbolism, and conflict to help her develop the storyline of the novel. The story is in the point of view of the main character, Scout Finch. The basic summary of the story is that Scout and her brother Jem spend much of their time in their town of Maycomb, Alabama with their friend, Dill. The three children spend their time spying on the town recluse, Boo Radley. Their widowed father, Atticus FinchRead MoreKill A Mockingbird By Harper Lee1270 Words   |  6 Pagescharacter in the book was Atticus Finch, who is a lawyer in Maycomb, Alabama. he has two children, who are Scout and Jem. the symbol of Mockingbird in the book title and the story related to the innocent and oppressed people, which the people judge them without knowing them right, in another world take advantage of someone weaker than you. there are many Mockingbirds in the book, but I want to focus on the most Mockingbirds who are Tom Robinson and Boo Radley in this essay. The book is very interesting

Variations and Characteristics of Mental Retardation Essay...

Mental Retardation is an old fashioned term for people who have problems learning and developing, but is defined as a disability characterized by significant limitations both in intellectual functioning and behaviors in expressing conceptual, social and practical skills in approximately one to three percent of the United States (Sue, Sue, Stanley, 2010). Mental retardation is categorized by the DSM-IV-TR as being a genetic disorder usually first diagnosed in infancy, childhood or adolescence, and is not considered to be a cognitive disorder. The following criteria is required for diagnosis of mental retardation under the DSM-IV-TR, Intellectual functioning under the IQ score of 70, concurrent deficiencies in adaptive behaviors and†¦show more content†¦These features include a long narrow face with a large head, protruding forehead, large ears, flat feet and flexible joints (NIH, 2012). Learning is difficult for people with Fragile X Syndrome because the syndrome affects the ability to think, learn and reason, with problems ranging from mild learning issues to severe developmental disability (NIH, 2012). Other characteristics include behavioral, social and emotional problems including anxiety in new situations, problems making eye contact, attention disorders and aggresive behavior (NIH, 2012). Speech, and language problems also exist with Fragile X Syndrome, including trouble speaking clearly, stuttering, speech may come later than others their age, few may never speak. Lastly, many children with Fragile X Syndrome have sensory issues and can be bothered by bright lights, loud noises, or the way clothing feels on their skin, which can cause the child to act out (Sue, Sue, Stanley, 2010). Down Syndrome is a condition caused by the presecnce of an extra chromosome called trisomy 21 (t21), which is a non sex chromosome and results in mental retardation and distinct physical characteristics (Sue, Sue, Stanley, 2010). Trisomy 21 means that each cell in the body has three copies of the chromosome 21, instead of the normal 2 copies received, one from each parent (NLM, 2012). Most cases of Down Syndrome are not hereditary but occur as random events, affecting the genesShow MoreRelatedFragile X Syndrome : The Most Inherited Form Of Mental Retardation1861 Words   |  8 PagesFragile X Syndrome, commonly known as FXS, is the most inherited form of mental retardation. From a study conducted by Emory University School of Medicine (2015), at least 1 out of every 4,000 males and 1 out of 8,000 females are affected with this abnormality (Emory University School of Medicine, 2015). At present, new information on how to live with FXS are discovered daily. This is helpful to furt her expand the current knowledge and methodologies that are attributed to FXS. There is currentlyRead MoreNutrition For Health And Development1615 Words   |  7 Pagesclinical standards as points of reference. A number of methods and measure may be used. (Raynor and Rudfuf 2000). Growth Retardation: Occurs when a fetus doesn’t develop at a normal rate. It is widely referred to as intrauterine growth restriction (IUGR). Children with IUGR are much smaller than other children of the same gestational age. There are two forms of growth retardation. Symmetrical and asymmetrical. Children of their gestational age. Children with asymmetrical IUGR have a normal sized headRead MoreLearning New Motor Skills For A Chronic Neurological Disorder951 Words   |  4 PagesPhysical characteristics the child may exhibit are delayed movements, difficulty learning new motor skills, problems with balance, and difficulties with skills that involve constant feedback to plan new movements such as printing or handwriting (Missiuna, p. 3). Children with DCD must overcome individual constraints when it comes to any task. They commonly have low muscle mass and low muscle tone, reduced motor control, and lack body awareness (Missiuna, p. 3). As for emotional characteristics, chil drenRead MoreMason was not unlike many children in that according to the most recent Center for Disease Control800 Words   |  4 Pageshave changed the view of autism and people on the autism spectrum (Diagnostic and statistical manual of mental disorders 2013). Although with the DSM-V release in 2013, the terminology associated with these pervasive developmental disorders (PDDs) has changed the symptomatic characteristics of these disorders remain the same. What are the differences in these diagnoses and their characteristics? What makes these children different? How pervasive are these differences? What treatments or cures haveRead MoreHistory of the Patau Syndrome1850 Words   |  7 PagesChildren with the disease are often born with multiple birth defects that are normally associated with T13 but not limited to it, including small eyes, undescended testicles, cleft lip/palate, and they consistently exhibit signs of mental deficiency and/or retardation. Approximately 1 in 10,0 00 newborns are born with Patau Syndrome each year. I choose this rare disease because not only is it unheard about but the present dilemma plaguing the medical field on whether or not a child born with thisRead MoreDisproportionality Researching the Overrepresentation of Minority Students in Special Education3097 Words   |  13 Pagescategories (blind or deaf) and in those cases of mental retardation considered severe and profound do not yield dramatic deviations from proportions that one would expect. Taking this definition into consideration, data has shown that the proportion of minority students in the special education system has varied over the past few decades. African- American children were 2.3 times more likely to be identified by their teachers as having mental retardation than their white counterparts. The overallRead MoreThe Theory Of Culture As A Disability2216 Words   |  9 Pagesmeet those standards, then they are labeled as disabled and different. Throughout the years, African American students have been significantly over-represented in special education, particularly in the categories of emotional disturbance and mental retardation. In Houston, Texas alone, African Americans made up approximately 20% of the city’s total population, and 24% of the student body in Houston Independent School District (HISD). Because African Americans make up a smaller percentage of the populationRead MoreEssay Birth Defects857 Words   |  4 Pagessurface or present in ternally. Birth defects may be life threatening and require surgical correction, or they may interfere with function or appearance. It is estimated that about 3% of all children are born with major defects; minor defects or variations are estimated to occur in 10% to 15% of births. Malformations may be single or multiple. Multiple malformations that occur in a regular recognizable pattern are referred to as syndromes--for example, the FETAL ALCOHOL SYNDROME sometimes observedRead MorePica1443 Words   |  6 Pagesat any point in time. Pica can be associated with mental and stress related disorders. Stress factors such as family issues, pregnancy, poverty, parental neglect, and a disorganized family, are strongly linked to Pica. There are different variations of Pica; it can be genetic, could be desire for the taste, tried once and enjoyed it, or an iron deficiency. It can lead to a â€Å"drunken state† in children, which can result in negative physical and mental development. Pica can lead to nutrition deficienciesRead MoreDevelopmental Disability : A Diverse Group Of Chronic Conditions2023 Words   |  9 PagesDevelopmental disability is a diverse group of chronic conditions that are due to mental or physical impairments. Developmental disabilities cause individuals living with them many difficulties in certain areas of life, especially in language, mobility, learning, self-help, and independent living. Developmental disabili ties are usually identified early in childhood and will probably persist throughout the individual’s lifespan. This paper will identify the various aspects of how developmental

The Mall of America Destroying Life Long Values with Materialism free essay sample

Through his piece â€Å"Enclosed. Encyclopedic. Endured: the Mall of America. † David Guterson shares his experience of the Mall of America as it opened and its effects of the American culture. From sharing statistics about the amount of jobs available, the number of parking spots, or how much cash is dispersed each week from just the ATMs; Guterson allows readers to feel the massive scale of the mall. He shares stories of the people he met and his own views on the mall, and what it says about America and its people’s values. Guterson makes it clear that Americans have become too absorbed with the thoughts of materialistic belongings; and a mall, such as the Mall of America, only makes those thoughts that much worse and destroys the people’s values. Although the points Guterson makes about the crumbling values and presently true and I agree that materialism is a horrible quality for a person to have, the mall in itself is not a bad place. It brings joy to many people and not just through ways of materialism and buying everything your heart desires, but for the sheer pleasure of entertainment. When young children go to the mall with their parents and attend Camp Snoopy or go to LEGOLAND; that is not materialistic in the slightest. All in the entire mall is not the horrible place that Guterson makes it out to be. The mall and its materialistic image have become a major highlight of American society. By allowing mall goers to share their experiences, this is fairly evident. Two women Guterson meets at a restaurant explain how they â€Å"have shopping addictions and live at the mall† (105). These women explain how they feel sorry for other malls because they are so â€Å"small and boring†, as if nothing can compare to the Mall of America. These women are right because the mall is so vast and extravagant nothing else can compare to it; and again they are right because what other malls can say that they have and indoor amusement park?! He later goes to say how the mall is not just a marketplace but rather a tourist attraction, attracting crowds from all over. The mall’s general manager, John Wheeler, is quoted saying that â€Å"I believe we can make The Mall of American stand for all of America. † The mall was said to be greater, and by 1996 was expected to have more visitors than Disneyland. This right here proves that the mall is not a bad thing; have you ever heard a person say that Disneyland corrupted them or destroyed what they believe in? No I don’t think so! So then if the mall is just as great as, or even greater than, Disney it couldn’t possibly be bad. All the more proof that the mall is not a bad place, Guterson talks to the old gentleman, Chuck Brand, who just sits at the mall, outside Camp Snoopy, every day from 10:00 am until his wife picks him up at six. â€Å"I don’t get hassled for hanging out, not shopping. Because the deal is, when you’re seventy-two, man, you’re just about all done shopping† (110). Chuck shows that the mall is so much more than just a place for people to go shopping. That you can just relax, and enjoy the atmosphere. As well, Chuck is right 72 you are pretty much done shopping, which counter acts Guterson’s point of the mall being materialistic. So with all the â€Å"old men†, like Chuck, along with all the little children that absorb themselves in the overwhelming essence of the mall there is no way that it is only a materialistic environment. The Mall of America had become as powerful as a church for some, for example on Valentine’s Day, 1993, â€Å"ninety-two couples were married en masse in a ceremony at the Mall of America† (113). Although church services were being held in the mall some were afraid that it may trivialize religion. According to Reverend Delton Krueger, President of the Mall Area Religious Council, â€Å"A good many of people in churches, feel a lot of the trouble in the world is caused by materialism† (113). I find this hard to believe, some people sure put a lot of stake into their shopping, but it’s hard to find people that would take it to the extent to think that their items and the money they spend would take over their religious views, that just seems a little to unrealistic. Krueger and Guterson are sharing the idea that people are becoming so involved in their shopping and the materialistic things that they are revering these items as form a â€Å"God†. They are forgetting the values that their parents had taught them all their life by making them go to Sunday school, and church every week; to them materialism is â€Å"GOD† they worship who have the best stuff. Still though many thought the trouble lied elsewhere, and with this people and companies around the world planned to build malls exceeding the Mall of America. All it is just a race to see who can have the most immense bizarre to satisfy the most people. As it all becomes more extreme, Americans, and the world in general, are losing all they believe in. It’s all becoming about who has the best stuff, or from the perspective of a teenage girl, who has the cutest clothes. From its vast security systems, to the indoor theme park, and over 520 stores the mall is like a city within itself; closed off from the rest of the world. Guterson writes that the quality and impressiveness of the mall is not comparable to any other. The mall is â€Å"the best of the best† and people visit it because of its modernism. The modernism of the mall is displayed through features like an arcade, the gardens, trees and flowers. The argument of the essay is reasonable in the way people go to spend in the mall. It is true that people are sometimes materialistic. But not all is bad, the mall is fun for children and adults, people of all ages. It doesn’t destroy their life long values but rather allows for sheer enjoyment. The mall is only a past time and nothing more. Yes, it is a representation of American life but not in a demeaning way. Works Cited Guterson, David. â€Å"Enclosed. Encyclopedic. Endured: The Mall of America† The Norton Reader. Eds. Linda Peterson, et al. Shorter 13th ed. New York: Norton, 2011. 102-114. Print.

Global Political of Intellectual Property Rights †MyAssignmenthelp

Question: Discuss about the Global Political of Intellectual Property Rights. Answer: Introduction: The term property is generally used for governing access to and control the land and other material resources as per the philosophical theory of Stanford. According to this theory the philosophers stated that it governs the access to and control of things that are used by individuals. For example use of natural resources, land, manufactured goods and other intellectual products. Everything around is not considered to be property. A property needs to be recognized based on the objects and subjects involved with it. The essential criteria for establishing whether something is property or not, depends on the location of it. A property can be situated on the land as an immovable object[1]. It can also be considered as movable. Therefore, objects that are situated on the land like houses. Immovable objects like cars also fall under the criteria of being property. Property can be classified into public and private. Private refers to that system which allocates particular objects like piece s of land to specific individuals for using and managing[2]. Public property refers to those objects that are used by every individual. As observed from the given case study, the property of a man after he dies will be inherited by his children unless there is a will made by that particular person stating on whom he has given his property to. Xu has entire estate worth $20 million was given to Benji as his wife Kay was deceased. Thereafter, the moment Xu passed away and Moong alleged himself to be the son of Xu. Legally, Benji was suppose to receive the entire estate, which was worth $20 million but now that Moong is claiming to be the son of Xu. This suggests that $20 million dollars will be divided between these two. This can be proved with the help of a DNA test. However, according to the case study, the tissue of the Xus body remained in the local private hospital where his medical treatment failed. As per the rules of the Court, orders are made in respect of a property. The bodily tissue of Xus property belong to the family of Xu. The present family of Xu consisted of his son Benji and his wife Kay. Moong will a lso be entitled to receive the property of Xu once the DNA test is proved with the help of the tissue. Although Xu is dead but even with his remaining tissue, a DNA test can be arranged which will prove if at all Moong is the blood related child of Xu. Therefore, the property of Xu will be handed over at that time when it is proved that apart from Benji if Moong is also the son of Xu. In respect to the property of a person, the court can pass its verdict but the final decision will come down to the time when the DNA reports are out. The bodily tissues of the deceased will not be considered as a property but it can be used for finding out the DNA results. Till the tissue is undergoing the process of the test, it will belong to the natural heir of Xu. This has been observed in the case of Armory v Delamirie(1722) Until the reports are not out, the tissue will belong to Benji who is the legal heir and blood related. If the DNA report states that Moong is also the child of Xu, then both of them will share the entire estate of $20 million. However, the court orders are made based on the nature and situation of the property. A legal proceeding that generally arises among property and boundary disputes are known as an adverse possession. The justification for adverse property defines a situation where a party should show that they have actually used the property. For instance, an adverse possession can claim over a backyard that requires to show the common activities to the type of property which include gardening[3]. When a real property is acquired by adverse possession, justifications should be provided thereafter. Use of this actual property does not imply to situations when the property is used for a full time. Properties can be used during seasonal occasions as well. For a seasonal property, a party needs to prove that it was in use for the relevant season of the year. In case of an adverse possession claim for a breach property, the claimant need not show if it was used in the winter. The concept of adverse possession requires a party to show that the use was open and notorious[4]. This refers to a situation, where it is visible to others especially the record owner of the property. In the provided case study, Andrew James was known as the adverse claimant[5]. In this scenario, a squatter took over a dilapidated terrace in the inner city of Sydney that has abandoned his bid for adverse possession. Andrew James was the person who had moved out of the property worth $1 million that remained the focus of a legal dispute between his neighbor and the local council. Mr. Andrew James was removed by The Supreme Court of NSW as he was the defendant in the proceedings during a directions that was heard at the court. After that particular hearing, the property was vacated. His neighbor Gerard Knapp was bound to take legal action after James had moved out into abandoned house in a bid to take adverse possession. Gerard was faced with a problem where his run down property was claimed to be a structural risk for himself[6]. Such a situation was defined to be a legalized theft along with thousands of properties in Sydney that were bought by the investors of overseas and led into the real lawlessness. Since Mr. Andrews was out of the picture, the court ca nnot allow him for another hearing. Thereafter, Gerard will be summoned in the court and will be claimed for damages from him against Mr. Fuh between the City of Sydney and the lawyer. The merits of this argument states that the claimant Andrew was out of the picture and therefore he did not owe any kind of damages to his neighbor. Andrew got saved from this problem of paying for the damages caused to the neighbor[7]. Andrew James A.K.A Andy Robert had the upper hand in this situation. He had told his neighbors that his wish was to modernize the terrace and rent it out. Andrew was smart enough to think of his profit ratio out of it. The house of Elizabeth Street in Redfern was last sold in September 1991. The Australian and Chinese owner had visited the property only three or four times a year. Sometime in the month of October, Mr. James had lost his bid for adverse possession when the Supreme Court of NSW had removed him as a defendant during the proceedings after the court heard that the property had been vacated. The neighbors on either side of the Elizabeth Street property had opted for the auction of the house. The neighbors did not create much problem when the house was getting auctioned by James and were happy when he was removed as the defendant by the Supreme Court of NSW[8]. James had claimed the rights of the squatters and his intention was to capture or acquire the property and then renovate it. It was an unusual situation at that time. Sometime in June, Mr. Knapp had won a court order for selling off the property sold. The purpose was to improve the cost for extensive damage caused by water through the common wall. This way he got access to the property, he wanted. Through this property, he can manage to get to the common wall. The amount is important, as the squatter had already paid out the unpaid rates. This process-involved risk as the amount was quite high. The neighbors were aware of such condition and therefore they took the decision accordingly[9]. Therefore, the neighbors of the Elizabeth Street property had known the fact that Andrew was a claimant of the adverse possession. The case study stated that the Chinese owner had bought the house but he has not visited or seen in the last nine years after returning to China. Thereafter, Mr. James wanted to take over the property, as no one wanted to. The neighbors opposed in the beginning saying that he was too young but he did try to take the premises. Mr. Andrews was accused of trespassing but he defended himself to the City of Sydney Council stating that only the owner can oppose such a complaint on him. Therefore, he obtained access and started making claim of adverse possession[10]. In Sydney, such an incident was known as legalized theft. In Australia, adverse possession was referred to as the rights of the squatter if a property is occupied by someone for a long time. Due to this, the real owners lose their own rights over it naturally. The City of Sydney Council had summoned the Chinese owner to the court as he had neglected the house[11]. Due to this condition, the neighbors thought that it would damag e their homes as well. The neighbors had requested the Council for putting up that house for public auction[12]. However, the role of City of Sydney Council was to take care of the situation of Mr. James. Mr. Andrew had abandoned his claim for adverse possession since he had acquired that property because the Chinese owner did not visit that property even once in the last 12 years. There were controversies of trespassing the property by the neighbors of Elizabeth Street. He had claimed adverse possession of the property because he had actually used that property. Sometime during 2016, this house was the centre of a bizarre dispute with a squatter who had recognized himself as Andrew James. He had tried taking the ownership by adverse possession. Later he had abandoned his claim for the adverse possession as the Supreme Court of NSW removed him as the defendant[13]. If he had not abandoned the claim then the property acquired by him would have been a disputed one. As an advice for Andrews, it can be said that if he had fought back with the lawyers of the opposite party then he would have won the property. Adverse possession should be claimed only when the owner of the property has granted or passed over the ownership. The purpose of adverse possession is to damage the title of an individual who is not possessed. Therefore, it does not destroy the rights of other persons who have an enforceable interest in the lands. After such a confusion and abandoning the land property, the Council of Sydney had sold the land if the charges imposed on it was not paid for a period of more than five years. Andrew was accused of being a legalized theft in the city[14]. Andrew was bound to abandon the claim also because a claim is usually made against an owner of common law after a period of 12 years. As per the case study, it was observed that Andrews had a strong claim to the property of Elizabeth Street. The City of Sydney Council had defended Andrew and his activities. The City of Sydney Council exclaimed that it has no rights to take the occupation of the property. If the rates of a particular house is not paid then it is the duty of the city to sell the property to recover the expenses that was owed. If not selling completely, the city council might consider the fact of selling it[15]. Since, Andrew had taken care of the property and was residing in it the Council could not impose much allegations on him. The neighbors of the Elizabeth Street opposed and complained about Mr. James for trespassing. According to the Local Government Act, the Council has the right to sell any land on which any charge or rate has remained not paid for more than a period of five years from the date of payable. The City Council of Sydney had made an independent assessment of the structural integri ty of the house that carried out with the access that was provided by Mr. James himself[16]. The issue creating such problems lies between the owner and the person living in the property. The concept gets more complicated than taking adverse possession of the house. The City of Sydney Council could not have defeated Mr. James much. References: Berkowitz, Daniel, Chen Lin, and Yue Ma. "Do property rights matter? Evidence from a property law enactment."Journal of Financial Economics116.3 (2015): 583-593. Bradbrook, Adrian, Susan MacCallum, and Anthony Moore. Australian real property law. Australia, 2002. DeScioli, Peter, and Rachel Karpoff. "Peoples judgments about classic property law cases."Human Nature26.2 (2015): 184-209. Dickens, Bernard M. "Living tissue and organ donors and property law: more on Moore."Organ and Tissue Transplantation. Routledge, 2017. 37-57. Fitzpatrick, Daniel, and Andrew McWilliam.Property and social resilience in times of conflict: land, custom and law in East Timor. Routledge, 2016. Ginzberg, Eli.The institutions of private law and their social functions. Routledge, 2017. Hines, N. William. "Real Property Joint Tenancies: Law, Fact, and Fancy."Iowa L. Rev.51 (1965): 582. May, Christopher.The global political economy of intellectual property rights: The new enclosures. Routledge, 2015. McDonald, Paula, and Paul Thompson. "Social media (tion) and the reshaping of public/private boundaries in employment relations."International Journal of Management Reviews18.1 (2016): 69-84. Mossman, Mary Jane, and Philip Girard. Property law: cases and commentary. Emond Montgomery Publications, 2014. Murphy, Richard S. "Property rights in personal information: An economic defense of privacy."Privacy. Routledge, 2017. 43-79. Paul, Ellen Frankel.Property rights and eminent domain. Routledge, 2017. Rabin, Edward, et al. "Fundamentals of Modern Property Law." (2017). Roness, Paul G. "Types of state organizations: Arguments, doctrines and changes beyond new public management."Transcending new public management. Routledge, 2017. 77-100. Williamson, Claudia R. "Praise for Property."Journal of Private Enterprise32.4 (2017): 83-94.

Macbeth Vs. Lord Of The Flies Essays - Characters In Macbeth

Macbeth Vs. Lord Of The Flies Macbeth vs. Lord of the Flies The human being, according to evolution, has evolved from animals to become a superior animal form that has the ability to decipher right and wrong, have recognizable feelings, and at the same time have that animalistic characteristics wild beasts have. The authors of Macbeth and Lord of the Flies attempt to exhibit the idea that human beings are totally capable of acting like wild beast. The authors explain to us that different factors bring out our animal, or beast, instincts. Envy, fear, and jealousy are factors, which contribute to our regression as savages. Human beings thrive on competition, so, such factors can change the way we think. These authors use characters like Jack and Macbeth as examples of human being self corruption through letting our natural call of the wild come through. Leadership can never come in second place and is only held by the first. These characters, after realizing the idea of kill or be killed, will do anything, even acts of inhumanity, to advance and come out on top. This with the combination of other factors simply blind folds our minds into thinking that we must always come out victorious. All the values and mores that society has taught us have been wiped away simply with the illusion, internal animal characteristics, that we must strive, in any way we can, to be in the position of leadership. Physical examples like Jack killing Piggy and the slaying of Banquo by Macbeth are models of how people can be temporarily blinded by jealousy, envy, and fear. Authors, trying to prove that human beings see their own eventual downfall by using such devices as apparitions and ghost to remind us we are human. Apparitions haunted Macbeth, in the same way our conscious come into play when we know we have violated the values set up by society. This realization of doing wrong only makes some humans to more unexpected things. In a way, they become frightened of returning to human values. This is actually the beast in us that does not want to be repressed. In lord of the flies we see that this animal or if you want to call it savagery, in human beings, can be visible in even innocent children. Fear drives many of these children to act very different sometimes even aware that one would never do such things. This fear was brought on by the illusion that a stronger more dominant beast was on the island as well. Jacks animal instinct is brought out with the feeling of being second to Jack and most of the supposed beast. This is why he insists on hunting rather then working on a plan to be rescued. Jack will not tolerate being in second. Sometimes these feeling can not be pushed away and only seem to worsen. This is seen by the fact that Jack is no longer hunting to survive but for the thrill of knowing he has the power to kill that which is less then him. His animalistic characteristics only seem to get worse. Going from killing to survive, killing to know you have the power, and finally the murder of human beings. We see that Jack is now making excuses for the quick high of know he is number one and in total control. He kills Simon claiming he thought it was the beastie. Truly the fact that they dance around like savages and never stop think is prove that the beast in Jack has finally taken over his mind. The authors of Lord of the Flies and Macbeth attempt to show us that hope is not lost. Obvious devices as apparitions doubling as our conscience mind display that human beings can over come the regression to savagery and to face the fact that we all have a simple yet powerful dark side Shakespeare

Tumor-Stroma Interaction †Biology Research Paper

Tumor-Stroma Interaction – Biology Research Paper Free Online Research Papers Tumor-Stroma Interaction Biology Research Paper Abstract Matrix metalloproteinases (MMPs) are metal-dependent endopeptidases that play pivotal roles in tumor disease progression. In many solid tumors, MMPs are indeed produced by tumor stromal cells, rather than by tumor cells. This expression pattern is, at least in part, regulated by tumor-stroma interaction via tumor cell-associated extracellular matrix metalloproteinase inducer (EMMPRIN). In vitro, recombinant EMMPRIN dose-dependently stimulated MMP-1 production by primary human fibroblast cells. Interestingly, in addition to stimulating MMP expression, EMMPRIN also induced its own gene expression. To further explore this potential positive feedback regulatory mechanism, we generated human breast cancer cells expressing different levels of EMMPRIN. Coculture of EMMPRIN-positive tumor cells with fibroblast cells resulted in a concomitant stimulation of MMP-2, MMP-9, and EMMPRIN production. This induction was EMMPRIN dependent, was further enhanced by overexpression, and was reduced by antisense suppression of EMMPRIN expression in tumor cells. Increased expression of membrane-associated EMMPRIN was accompanied by an MMP-dependent generation of a soluble form of EMMPRIN representing a proteolytic cleavage product lacking the carboxyl terminus. On the basis of these findings, we propose a model in which tumor cell-associated EMMPRIN stimulates MMPs, as well as EMMPRIN expression in tumor stroma. Increased MMP activity in tumor local environment results in proteolytic cleavage of membrane-associated EMMPRIN, releasing soluble EMMPRIN. Soluble EMMPRIN in turn acts in a paracrine fashion on stroma cells that are both adjacent and distant to tumor sites to further stimulate the production of MMPs and additional EMMPRIN, which consequently contributes to tumor angiogenesis, tumor growth, and metastasis. Introduction Degradation of basement membrane by matrix metalloproteinases (MMPs) is one of the most critical steps in various stages of tumor disease progression, including tumor angiogenesis, tumor growth, as well as local invasion and subsequent distant metastasis (1–3). MMPs are a family of more than 25 endopeptidases that are metal dependent and share a common modular domain structure. MMPs are overproduced in tumor local environment. Collectively, these enzymes are capable of cleaving all of the extracellular matrix components of the parenchymal and vascular basement membranes, normally mechanical barriers to tumor cell migration and invasion. High levels of MMPs have been correlated with increased tumor invasion capacity both in vitro and in vivo (4, 5). Clinically, elevated levels of MMPs detected in the body fluids of cancer patients correlate with adverse prognosis in various cancers (6, 7). Imbalances in the production of MMPs and their endogenous inhibitors, tissue inhibitors o f metalloproteinases (TIMPs), result in tumor angiogenesis and metastasis (2, 3, 8–12). Interestingly, recent studies of clinical tumor specimens provided evidence that most MMPs are generated by peritumoral fibroblasts in the stromal compartment, instead of by tumor cells themselves. Peritumor stromal cells are indeed responsible for the production of tumor-associated interstitial collagenase (MMP-1), stromelysin-1 (MMP-3), stromelysin-3 (MMP-11), and gelatinase A (MMP-2), in breast, colon, lung, skin, and head and neck cancers (13–16). The induction of MMP production is, at least in part, mediated by tumor-stromal cell interaction via a tumor cell surface glycoprotein, extracellular matrix metalloproteinase inducer (EMMPRIN; 17, 18). EMMPRIN was originally purified from the plasma membrane of cancer cells as a glycoprotein of Mr 58,000, and was designated tumor collagenase stimulating factor (TCSF) because of its ability to stimulate fibroblast synthesis of collagenase-1 (MMP-1; 19, 20). Subsequent studies further demonstrated that EMMPRIN also induced fibroblast synthesis of MMP-2, MMP-3, as well as the membrane-type 1 MMP (MT1-MMP) and MT2-MMP that function as endogenous activators of MMP-2 (17, 21, 22). Therefore, EMMPRIN functions as an upstream modulator of MMP production in tumor local environment. EMMPRIN-positive tumor cells stimulate neighboring fibroblast cells to express MMPs and therefore facilitate tumor invasion and metastasis. This rationale for the function of EMMPRIN has been supported by results from several clinical studies that demonstrated high levels of EMMPRIN expression in tumor compartments as compared to peritumoral stromal tissues. These tumors include the lung (16), breast (16), bladder (23, 24), and glioma (25). Examination of EMMPRIN expression in these clinical samples by a variety of means, including Northern blot, in situ hybridization, and immunostaining, revealed that in most cases, EMMPRIN is expressed by tumor cells, but not by the neighboring stromal cells. In contrast, MMPs are expressed by peritumoral stromal cells. The biological significance of increased expression of EMMPRIN in tumor cells was investigated by in vitro studies using recombinant EMMPRIN or native EMMPRIN purified from tumor cells. EMMPRIN has been shown to stimulate the expression of various MMPs produced by fibroblasts (17, 18). The induction occurred at the transcription level and is at least in part mediated by a mitogen-activated protein kinase (MAPK) p38 kinase signaling pathway (18). The role of EMMPRIN in tumor growth and metastasis was directly illustrated using EMMPRIN-overexpressing human breast cancer cells. MDA MB 463 cells are normally slow-growing cells when they are implanted into nude mice. However, when these cells were transfected with EMMPRIN, they adopted a more aggressive growth pattern, with both accelerated growth rate and metastatic phenotypes (26). To further study the interplay between tumor cell-associated EMMPRIN and fibroblast cells, we used recombinant engineering to generate MDA MB 231 human breast cancer cells expressing different levels of EMMPRIN. We discovered a novel positive feedback regulatory mechanism controlling the interaction between tumor and fibroblast cells. Our results demonstrate that EMMPRIN positively regulates its own gene expression, in addition to stimulating MMP expression. Furthermore, we also detected generation of soluble EMMPRIN in the coculture of tumor and fibroblast cells that occurs via an MMP-dependent cleavage. On the basis of these new findings, we propose a new model in which the interplay between tumor and stromal compartments is regulated by both tumor cell surface EMMPRIN, as well as by soluble EMMPRIN, generated from MMP-dependent cleavage. Results Induction of MMP-1 Expression by Soluble Recombinant Human EMMPRIN in Primary Fibroblast Cells The stimulatory effects of EMMPRIN on cultured fibroblast cells were studied using a soluble recombinant EMMPRIN protein (srEMMPRIN) corresponding to the extracellular portion of the molecule. srEMMPRIN was added at different concentrations to primary normal human fibroblast cells in culture. Serum-free media conditioned by these cells were collected at 1 or 3 days after EMMPRIN stimulation. MMP-1 production in response to EMMPRIN stimulation was determined by assaying MMP-1 activity secreted into the conditioned medium. EMMPRIN dose-dependently stimulated MMP-1 production by normal human lung fibroblast (NHLF) cells (Fig. 1 ). The stimulation of MMP-1 production by srEMMPRIN also occurred on the transcription level as determined by TaqMan analysis (Fig. 2A ). FIGURE 1. Induction of MMP-1 expression by soluble recombinant human EMMPRIN in human primary lung fibroblast cells. srEMMPRIN dose-dependently stimulated MMP-1 production by NHLF cells. Ten thousand NHLF cells (passage 1) were plated in each well of a 96-well cell culture plate. Cells were challenged with different concentrations of EMMPRIN. After 1 day and 3 days, conditioned medium was collected from each well and MMP-1 activity was determined. Points, mean values of duplicates from three independent experiments; error bars, SDs. FIGURE 2. Induction of EMMPRIN expression by soluble recombinant human EMMPRIN in primary fibroblast cells. Primary normal human fibroblast cells of the lung were challenged with 5 Â µg/ml of srEMMPRIN. A. Total RNA was collected from untreated cells or at 4, 24, and 48 h after EMMPRIN treatment. EMMPRIN and MMP-1 gene expression level was determined by quantitative TaqMan analysis and was expressed as percentage to that in untreated cells, which was designated as 100%. B. Western blot analysis of EMMPRIN expression in fibroblast cells in response to stimulation with recombinant EMMPRIN. Cell lysate containing 50 Â µg of total protein was loaded into each lane, G361 high EMMPRIN-expressing melanoma cells (lane 1), fibroblast cells treated with EMMPRIN (lane 2), and unstimulated fibroblast cells (lane 3). Positive Feedback Regulation of EMMPRIN Gene Expression in Primary Fibroblast Cells Interestingly, when we examined the expression level of EMMPRIN in fibroblast cells after stimulation with 5 Â µg/ml EMMPRIN, we observed a substantial increase in EMMPRIN gene expression in these cells. As determined by quantitative TaqMan analysis, a 2.2-fold increase in EMMPRIN gene expression was induced at 4 h after cells were stimulated with EMMPRIN (Fig. 2A). EMMPRIN gene expression continuously increased to 4.6-fold at 24 h and 3.7-fold at 48 h after treatment. On EMMPRIN stimulation, MMP-1 gene expression level also increased (Fig. 2A). This induction occurred more rapidly and transiently, with a peak expression of a 3.3-fold increase at 4 h, and dropped to near pretreatment level at 48 h. When EMMPRIN protein expression in fibroblast cells treated with EMMPRIN was examined with Western blot analysis, we observed a strong increase in EMMPRIN protein expression (Fig. 2). In contrast, only minimal levels of EMMPRIN protein were detectable in unstimulated fibroblast cells. Tak en together, these results suggest that in addition to increased expression of MMP-1, fibroblast cells also respond to EMMPRIN stimulation by up-regulating EMMPRIN gene expression. Generation of MDA MB 231 Tumor Cells Expressing Different Levels of EMMPRIN To further explore the potential regulatory mechanism of EMMPRIN gene expression in fibroblast cells, especially in the context of tumor-fibroblast cell-cell interaction, we used recombinant DNA engineering technology to generate cancer cells that express different levels of EMMPRIN. As described in Materials and Methods, MDA MB 231 human breast cancer cells were transfected with expression vectors harboring the full-length coding region of human EMMPRIN in either sense or antisense orientations. Stably transfected cells were cloned and EMMPRIN expression level in these cells was determined by Western blot analysis. As shown in Fig. 3 , EMMPRIN protein expression in one of sense transfected cell clones S1-3 was approximately 2-fold higher as compared to that of wild-type control cells. In contrast, EMMPRIN protein expression was substantially inhibited in two antisense transfected cell clones, AS1-5 and AS2-5, to only 47% and 62% of that in wild-type cells. Changes in EMMPRIN protein expression in these engineered cells were also independently confirmed by ELISA analysis (Fig. 4 ). In addition, changes in EMMPRIN protein expression also occurred on cell surface in sense and antisense transfected cells, respectively, as determined by fluorescence-activated cell (FAC) analysis (data not shown). FIGURE 3. Generation of MDA MB 231 tumor cells expressing different levels of EMMPRIN. A. Western blot analysis of EMMPRIN protein in cell lysates. Samples containing 20 Â µg of total protein from each cell type were loaded in duplicates, MDA MB 231 wild-type cells (lanes 1 and 2), S1-3 cells (lanes 3 and 4), AS1-5 cells (lanes 5 and 6), and AS2-5 cells (lanes 7 and 8). A single EMMPRIN protein band of approximately Mr 52,000 was detected in all of the samples. B. Quantitative determination of EMMPRIN expression by scanning densitometry. EMMPRIN Western blot was scanned and the pixel density of each band was determined by scanning densitometry. EMMPRIN expression level in each cell type was expressed as the percentage of density of that in wild-type cells (lane 1), which was designated as 100% error bars, SDs. FIGURE 4. Stimulation of both cell-associated and soluble EMMPRIN expression in coculture of tumor cells and fibroblast cells. EMMPRIN protein concentration was determined by ELISA analysis in total cell lysate (A) or in conditioned medium (B) of breast cancer cells expressing different levels of EMMPRIN either in monoculture or in coculture with NHDF cells. All concentrations were normalized to total protein amounts. WT, wild-type MDA MB 231 human breast cancer cells; S1-3, sense transfected EMMPRIN-overexpressing cells; AS1-5 and AS2-5, antisense transfected cells. Columns, representative of two independent experiments; error bars, SDs. We next used these cells with different levels of EMMPRIN protein expression to address the role of tumor cell-derived EMMPRIN in regulating its own expression during tumor-fibroblast cell interaction. Stimulation of Both Cell-Associated and Soluble EMMPRIN Expression in Coculture of Tumor Cells and Fibroblast Cells In monoculture of tumor cells or fibroblast cells, only low levels of EMMPRIN protein were detected in cell pellets or conditioned medium by ELISA analysis. Transfection with sense or antisense expression constructs resulted in substantial changes in cell-associated EMMPRIN expression, with an increase from 3.32 ng/ml in wild-type to 7.2 ng/ml in sense transfected S1-3 cells, and a decrease to 0.85 and 1.43 ng/ml in antisense transfected AS1-5 and AS2-5 cells, respectively (Fig. 4A). Only low levels of soluble EMMPRIN were detected in conditioned medium of these cells in monoculture. Transfection of tumor cells with sense and antisense expression constructs resulted in only minor changes in soluble EMMPRIN protein levels (Fig. 4B). The expression of cell-associated EMMPRIN was readily induced when tumor cells were cultured together with fibroblast cells. In coculture, EMMPRIN was detected at 10 ng/ml as compared to 0.8 ng/ml in fibroblast cells and 3.32 ng/ml in wild-type tumor cells in monoculture (Fig. 4A). This increase in EMMPRIN expression in coculture was mediated by tumor cell-associated EMMPRIN. When EMMPRIN expression in tumor cells was inhibited by antisense transfection in AS1-5 and AS2-5 cells, the increase in EMMPRIN expression stimulated by coculture was concomitantly reduced to minimal levels (Fig. 4A). It is known that coculture of EMMPRIN-positive tumor cells with fibroblast cells stimulates MMP expression (22, 27). These MMPs can in turn cleave cell surface proteins to generate soluble proteins. We therefore postulated that there may be an increase in soluble EMMPRIN protein level in conditioned medium of these cells in coculture. In monoculture, the levels of soluble EMMPRIN were extremely low in conditioned medium of fibroblast cells (0.31 ng/ml) and wild-type tumor cells (0.53 ng/ml; Fig. 4B). However, coculture of tumor cells with fibroblast cells induced the release of substantial amounts of soluble EMMPRIN, 7.48 ng/ml by wild-type and fibroblast coculture and 10.39 ng/ml by S1-3 and fibroblast coculture (Fig. 4B). Similar to cell-associated EMMPRIN, the increase in soluble EMMPRIN expression was also correlated with the degree of EMMPRIN expression in tumor cells. Inhibition of EMMPRIN expression by antisense transfection virtually abolished the generation of soluble EMMPRI N in coculture (Fig. 4B). Taken together, these results demonstrate a positive feedback regulatory mechanism of EMMPRIN gene expression during tumor-fibroblast cell-cell interaction. In addition to stimulating EMMPRIN expression and MMP expression, tumor cell-associated EMMPRIN also mediates the release of soluble EMMPRIN during tumor-fibroblast cell interactions. MMP-Dependent Generation of Soluble EMMPRIN in Coculture of EMMPRIN-Positive Tumor Cells and Fibroblast Cells Western blot analyses using antibodies against different regions of EMMPRIN were performed to further characterize soluble EMMPRIN generated in the coculture of tumor cells and fibroblast cells. Antibodies against the extracellular portion of EMMPRIN molecule detected a single band of approximately Mr 52,000, consistent with the size of intact protein, in total cell lysates. Meanwhile, this same antibody detected a slightly smaller molecule in the conditioned medium from coculture, indicating a loss of a portion of EMMPRIN protein (Fig. 5A ). Interestingly, antibodies specific to the carboxyl terminus of EMMPRIN only detected EMMPRIN in total cell lysates, but failed to detect any EMMPRIN protein in conditioned medium (Fig. 5A). Taken together, these data suggest that soluble EMMPRIN secreted into coculture medium of tumor and fibroblast cells is likely a cleaved product of the full-length transpanning form of membrane EMMPRIN. FIGURE 5. Soluble EMMPRIN in coculture of EMMPRIN-positive tumor cells and fibroblast cells- proteolytic cleavage products generated in an MMP-dependent fashion. A. Western blot analyses of cell lysates or conditioned medium of coculture of wild-type MDA MB 231 human breast cancer cells and fibroblast cells. Membranes with 50 ng recombinant EMMPRIN extracellular domain (lane A), cell lysates of 50 Â µg total protein (lanes B and D), and conditioned medium of 50 Â µg total protein (lanes C and E) were blotted with antibodies that recognize extracellular domain (lanes A–C) or intracellular carboxyl terminus (lanes D and E). B. Soluble EMMPRIN concentration in conditioned medium from coculture of tumor cells and fibroblast cells was determined using ELISA analysis. 1,10 phenanthroline was included in the study from 0.1, 0.5 to 1.0 Â µM to inhibit MMP enzymatic activity. WT, wild-type MDA MB 231 human breast cancer cells. Columns, representative of two independent experiments; error bars, SDs. , NHDF; , WT; , WT + NHDF; , WT + NHDF + 0.1 Â µM 1,10 PA; , WT + NHDF + 0.5 Â µM 1,10 PA; , WT + NHDF + 1.0 Â µM 1,10 PA; *, P < .05 compared to WT + NHDF. Because the generation of soluble EMMPRIN occurred concomitantly with increased production of MMP in coculture of tumor and fibroblast cells, we speculated that the release of soluble EMMPRIN might be mediated by MMP-dependent proteolytic cleavage. To further explore the mechanism of soluble EMMPRIN generation, 1,10 phenanthroline (1,10 PA) a general metalloproteinase inhibitor, was used at noncytotoxic concentrations to inhibit the enzymatic activity of MMPs. As shown in Fig. 5B, inhibition of MMP activity by 1,10 PA resulted in significant decreases in the level of soluble EMMPRIN in a dose-dependent fashion. These results demonstrate that the generation of soluble EMMPRIN stimulated by tumor fibroblast cell-cell interactions is the result of proteolytic cleavage of membrane-associated EMMPRIN by increased MMP activities in coculture. Discussion It is now known that the microenvironment of the tumor-host interface plays a proactive role during malignant disease progression, including the transition from carcinoma in situ to invasive cancer, tumor cell proliferation, and tumor cell dissemination and metastasis (28). On the host side, the stromal compartment is comprised of a variety of cell types, including immune cells, inflammatory cells, muscle and myofibroblast cells, and vascular cells. Tumor cells express molecules, either secreted or presented on cell surface, to interact with surrounding stromal cells. In the current study, we have demonstrated that EMMPRIN exists in both soluble and membrane-bound forms. Generation of soluble EMMPRIN resulted from tumor-fibroblast cell interaction in an EMMPRIN-dependent fashion, and was mediated by MMP proteolytic activities. These novel findings suggest a positive feedback regulatory mechanism of EMMPRIN expression that occurs at tumor-host interface. EMMPRIN is normally expressed as a glycoprotein on tumor cell surface that migrates at an apparent molecular size of Mr 58,000 (19). The domain structure of EMMPRIN consists of an extracellular portion which contains two immunoglobulin-like domains, a transmembrane domain, and a short cytoplasmic domain (19). It has been suggested that tumor cell-associated EMMPRIN exerts its MMP stimulatory effects on fibroblast cells via close cell-to-cell contact. However, the production of soluble EMMPRIN by tumor cells has recently been indicated in several studies (29, 30). In these studies, tumor cells affect fibroblast cells in the absence of physical contact in an EMMPRIN-dependent fashion, resulting in increased MMP-2 expression and activation (29), or enhanced cell migration or invasion (30). In addition, soluble full-length EMMPRIN has been purified from concentrated culturing medium of breast cancer cells, presumably secreted in shed vesicles (31). However, secreted soluble EMMPRIN in ca ncer cell monoculture is estimated to account for only 2–3% of EMMPRIN produced by these cells (31). In the present study, we have demonstrated the release of a substantial amount of soluble EMMPRIN into cell culture medium of cocultured tumor and fibroblast cells. Because fibroblast cells respond to EMMPRIN stimulation in producing increased levels of EMMPRIN, it is likely that the soluble EMMPRIN detected in coculture is derived from both tumor and fibroblast cells. In addition, the soluble EMMPRIN generated in coculture lacks the intracellular carboxyl terminus as determined using an antibody against the intracellular region of the molecule. In contrast, this carboxyl terminal region was preserved in cell-associated EMMPRIN. More interestingly, generation of soluble EMMPRIN in coculture could be efficiently blocked via inhibiting MMP activity. Therefore, this soluble EMMPRIN likely represents the proteolytic product of membrane-associated EMMPRIN. The generation of soluble EMMPRIN at the interface of tumor-host interaction may have profound biological significance. The soluble EMMPRIN could diffuse away from local tumor invasion site and stimulate stromal cells at distant sites. We and others have shown that soluble EMMPRIN consisting of only the extracellular portion of the molecular is functional in stimulating MMP expression in fibroblast and endothelial cells (17, 32). Increased MMP activity in the stromal compartment in turn promotes tumor invasion and angiogenesis. The receptor on fibroblast cells that is responsible for EMMPRIN-mediated stimulation of MMP production has remained elusive (17, 21). It has been suggested that EMMPRIN may serve as its own counter-receptor in cancer cells, thus stimulating MMP via a homophilic interaction (33). One could speculate that EMMPRIN on fibroblast cells could also serve as counter-receptors for tumor cell-associated EMMPRIN in stimulating MMP expression. However, this hypothesis is not supported by the fact that only extremely low levels of EMMPRIN expression could be detected in normal fibroblast cells. Our finding of a novel positive feedback regulatory mechanism of EMMPRIN expression provides an alternative explanation for the potential role of EMMPRIN as its own counter-receptor in fibroblast cells. When fibroblast cells were exposed to EMMPRIN stimulus, EMMPRIN expression is up-regulated in these cells. Newly synthesized EMMPRIN could then be presented on the cell surface and serves as the counter-re ceptor for EMMPRIN-dependent signaling between tumor and fibroblast cells. This novel regulatory mechanism is supported by recent findings of EMMPRIN mRNA expression in peritumoral fibroblasts in ovarian carcinoma (34). However, this homophilic interaction likely only accounts for EMMPRIN-mediated tumor-host interaction in some tumors because EMMPRIN expression has not been detected in the stroma of many other tumors including melanoma (35), or lung and breast cancer (16). In these samples, despite of a lack of EMMPRIN expression in stromal tissue, MMP expression is stimulated in stromal tissues adjacent to EMMPRIN-positive tumor cells, suggesting that there might be an unknown receptor(s) on stromal cell surface mediating these interactions. In conclusion, we have identified a novel positive feedback regulatory mechanism of EMMPRIN expression in fibroblast cells and discovered a soluble form of EMMPRIN. These findings suggest an amplification of EMMPRIN-mediated, MMP-dependent signaling events at the tumor and host interface, as well as at distant sites (Fig. 6 ). FIGURE 6. Proposed model for signal loops between EMMPRIN and MMP and tumor cells and host cells. Tumor cells use cell surface EMMPRIN to initiate contact with surrounding fibroblast cells (1), signaling fibroblast cells to synthesize MMPs (2). MMPs secreted by fibroblast cells subsequently cleave cell surface EMMPRIN and generate soluble EMMPRIN (sEMMPRIN; 3). These sEMMPRIN molecules, in turn, act on cells either in local tumor environment or diffuse away to act on distant cells (4) to further stimulate MMP and EMMPRIN expression (5) and augment the migration and invasion potential of tumor cells (6). Materials and Methods Cell Culture MDA MB 231 human breast carcinoma cells were purchased from American Type Culture Collection (Manassas, VA) and were cultured under recommended conditions. Briefly, cells were cultured in DMEM containing 10% fetal bovine serum (Life Technologies, Inc., Grand Island, NY). NHLF or normal human dermal fibroblast (NHDF) cells were obtained from Clonetics (Walkersville, MD) and were cultured in fibroblast growth medium containing 1 Â µg/ml human recombinant fibroblast growth factor (hFGF), 5 mg/ml insulin, 50 mg/ml gentamicin, and 50 mg/ml amphotericin B, at 37Â °C, 5% CO2. For coculture experiments, 100,000 cancer cells (MDA MB 231, MDA MB 231 S1-3, MDA MB 231 AS1-5, or AS2-5) were cultured together with 200,000 NHDF cells in a six-well cell culture plate in complete DMEM culturing medium. After 24 h, culturing medium was replaced with fresh serum-free DMEM and cells were cultured for 2 days. Culture medium was then replaced with 1.0 ml of fresh serum-free DMEM. Conditioned medium was collected 3 days later and secreted EMMPRIN protein concentration was determined using ELISA. Cells were lysed with 200 Â µl lysis buffer [50 mM Tris (pH 7.8), 150 mM NaCl, and 1% NP40] to determine cell-associated EMMPRIN concentrations. Cell Treatment and MMP-1 Activity Assay Recombinant EMMPRIN corresponding to the extracellular domain of human EMMPRIN protein was produced in NSO cells (RD Systems, Minneapolis, MN). MMP-1 activity in serum-free medium conditioned by fibroblast cells treated with different amounts of recombinant EMMPRIN protein was quantitatively determined using an MMP-1 Activity Assay Kit according to product manual (RD Systems). Briefly, MMP-1 contained in 150 Â µl of standards or samples was captured by anti-MMP-1 antibodies immobilized on the bottom of assay wells. Captured MMP-1 was subsequently activated by 4-aminophenylmercuric acetate (APMA). MMP substrate added into each well was cleaved by activated MMP-1 and the resulting fluorescence was determined using SpectraFluor Plus Plate Reader (TECAN, Research Triangle Park, NC) with the following parameters: excitation wavelength at 320 nm and emission wavelength at 405 nm. Generation of Sense and Antisense Expression Constructs and Establishment of Stable Transfectants To study the function of EMMPRIN in mediating tumor and stromal fibroblast cell interaction, the cDNA sequence corresponding to human EMMPRIN open reading frame (ORF) was PCR amplified and subcloned into pcDNA3.1TOPO vectors (Invitrogen, Carlsbad, CA). The EMMPRIN open reading frame was cloned in both sense and antisense orientations for cytomegalovirus promoter-driven expression in mammalian cells. Transfection of MDA MB 231 human breast carcinoma cells was performed using Effectene transfection reagents according to manufacturer's recommendations (Qiagen, Valencia, CA). Stably transfected cells were selected using Genicitin at 1000 Â µg/ml (Life Technologies). Individual transfectant colonies were established by cloning antibiotic-resistant cell colonies. Effects of sense and antisense transfection on EMMPRIN protein expression were determined using Western blot analysis and ELISA. RNA Sample Preparation and TaqMan Quantitative Reverse Transcription-PCR Total RNA was extracted from cells using RNeasy Kit (Qiagen) following manufacturer's instructions. Five micrograms of total RNA were used in the reverse transcription (RT) reactions which were carried out in a 50-Â µl total volume using Superscript First-Strand Synthesis System for RT-PCR kit following manufacturer's instructions (Invitrogen). Briefly, RNA samples were mixed with 0.5 Â µg/Â µl Oligo(dT), 0.4 mM dNTP mix in a total volume of 25 Â µl and were incubated at 65Â °C for 5 min, followed by quick cooling on ice for 2 min. The tubes were spun briefly and added with 5 Â µl 10x RT buffer, 10 Â µl 25 mM MgCl2, 2 Â µl 0.1 M DTT, 1 Â µl RNaseOUT, 1 Â µl (50 units) of Superscript II RT, and 6 Â µl nuclease-free water. The tubes were gently mixed and incubated at 42Â °C for 50 min. The reaction was terminated at 70Â °C for 15 min followed by quick cooling on ice for 2 min. The tubes were spun briefly and 1 Â µl of RNase H was added and incubated for 20 min at 37Â °C before proceeding to amplification of the target gene. Real-time PCR analyses were performed in 50 Â µl volume containing 2x TaqMan Universal PCR Master mix (Perkin-Elmer, Norwalk, CT), appropriate amounts of cDNA or genomic DNA, and 200 nM of TaqMan primers and 400 nM of TaqMan probes. Primer pairs and probes used in the study were designed using Primer Express software (Perkin-Elmer), with sequences listed in Table 1. Probes were labeled at the 5'-end with fluorescent reporter dye Fam and at the 3'-end with fluorescent quencher dye Tamra by Synthegen (Houston, TX) to allow direct detection of PCR products. Real-time PCR amplification and detection were performed in 96-well optical plates using ABI 7900HT sequence detector (PE Biosystems, Norwalk, CT). PCR conditions included thermal cycles of 30 s of 95Â °C for denaturing, 30 s of 60Â °C for annealing, and 60 s of 68Â °C for elongation. Relative gene copy numbers were calculated using a standard curve generated using PCR standards, serially diluted human genomic DNA (Clontech, Palo Alto, CA). Protein Electrophoresis and Western Blot Analysis Stably transfected cells were lysed in cell lysis buffer containing 50 mM Tris (pH 7.8), 150 mM NaCl, and 1% NP40. Protein concentration of cell lysate or conditioned medium was determined using the MicroBCA method (Pierce, Rockford, IL). Equal amount of proteins was loaded onto 4–15% gradient gels and was separated by SDS-PAGE under reducing conditions. Resolved proteins were electrophoretically transferred to nitrocellulose membranes (Invitrogen). The membranes were blocked with 5% low fat dry milk in TBS-T [10 mM Tris (pH 7.2), 50 mM NaCl, 0.5% Tween 20] for 1 h at room temperature, followed by incubating with primary antibody at 4Â °C overnight. Anti-EMMPRIN monoclonal antibody purchased from Research Diagnostics Inc. (Flanders, NJ) recognizes the extracellular portion of the protein. C-19 polyclonal antibody from Santa Cruz Biotechnology (Santa Cruz, CA) was raised against peptides of the carboxyl terminus of EMMPRIN protein. Blots were extensively washed with TBS-T and incubated with 1:5000 dilution of horseradish peroxidase (HRP) conjugated secondary antibody (Vector Laboratories, Burlingame, CA) diluted in TBS-T for 1 h at room temperature. Labeled proteins were visualized with Western Blotting Luminol Reagent (Santa Cruz Biotechnology). Pixel density of protein bands on Western blot was determined using Un-Scan-It software (Silk Scientific Corporation, Orem, UT). EMMPRIN ELISA EMMPRIN protein concentration in conditioned medium was determined using EMMPRIN ELISA. Briefly, 96-well Nunc Immuno Plates (Nunc, Denmark) were coated overnight at 4Â °C with 50 Â µl of anti-EMMPRIN antibody (RD Systems) at 5 Â µg/ml in buffer A (pH 7.4, 20 mM Tris-HCl, 150 mM NaCl, 1 mM CaCl2, 1 mM MgCl2, and 10 Â µM MnCl2). After one wash in buffer A, nonspecific binding sites were blocked with 150 Â µl of blocking buffer [3% bovine serum albumin (BSA) in buffer A] for 1 h at 37Â °C. Wells were then rinsed two times with binding buffer (buffer A with 0.3% BSA and 0.2% Tween 20). Either recombinant human EMMPRIN or conditioned medium was added into each well and was incubated at 37Â °C for 2 h. After three washes with binding buffer at RT, plates were blocked with 150 Â µl of blocking buffer (3% BSA in buffer A) at 37Â °C for 1 h. Fifty microliters of 1:5000 diluted biotin-conjugated anti-EMMPRIN antibodies (RDI-147, Research Diagnostics) were added into each well and incu bated at 37Â °C for 2 h. After three washes with binding buffer, 50 Â µl of horseradish peroxidase-conjugated streptavidin 1:10,000 diluted in binding buffer were added into each well, and were incubated at 37Â °C for 1 h. After three washes with binding buffer at RT, 50 Â µl of developing buffer (1 mg/ml O-phenylenediamine, 0.1 M citric acid, 0.2 M sodium phosphate, 0.01% H2O2) were added to each well and incubated at RT until color develops. Colorimetric reactions were stopped by adding 25 Â µl of 4 N H2SO4. ELISA data acquisition was performed using VersaMax Tunable MicroPlate Reader (Molecular Devices, Sunnyvale, CA) at 490 nm. Data were analyzed using Softmax Pro 3.1 software (Molecular Devices). 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Strategic Deterrence Essay Example | Topics and Well Written Essays - 1750 words

Strategic Deterrence - Essay Example Clearly analyzing the effect of the multipoint contact on firms strategic moves change according to the level of competition of the rival firm. Strategic deterrence may sound somewhat negotiating between the companies or the firms but ideally speaking it really reduces the risk of incurring losses. Not only that it also helps reduce excessive manpower used if deterrence is not abided. The concept of Strategic deterrence all begins with the primary intention of saving the market from multi market competition battle, which can ultimately bring stability among multi markets. Strategic deterrence is not accepted the moment the multi market contact start to encroach the others market but it is just an understanding that too mutually agreed up activity, which after incurring substantial amount of losses from the companies involved. This situation comes when firms start loosing customer base, profit on sales and the business tending to break-even point, which is generally referred as no profit and no loss point. This position comes when the situation is from a very long time. Though Strategic deterrence is mutually agreed concept is in deed an unwilling act on behalf of the companies involved. It ensures no loss concept but never promises profits out of the deterrence. Strategic deterrence may be a positive attitude but is accepted only by small or medium companies but not by large multi national companies. Its implications on the multi markets are not only largely understood but are but are being analyzed to overcome the disadvantages due to the market encroachment. The markets un touched by this phenomenon is the monopoly market which stands high on sales and profit yet not letting any one step into its market. This is the only market, which does not... The researcher states that today, many companies may compete in some other field, but try to avoid competing in the respective fields. This deterrence is only to avoid incurring losses, unnecessary from his competitor. The competitor may be a weak party, but still would be unwilling to compete. This strategy is primarily to avoid losses during industry shake out for any particular product. It is also stated that some firms compete while others forbear from competing with their ‘multipoint’ rivals — those faced in more than one market or market segment. According to the study it is clear that multi point contact may effect only within the same products or services but not between the strategic groups. There are many speculations that whether multipoint contact deters competition depends on strategic objectives. These predictions are often supported by estimates made by market exit rates. There are many indications and proofs that weak competitors can out survive st rong competitors due to forbearance. This essay aims to presents such proofs. The researcher then concludes that he had a concern that is deeply rooted towards competitive dynamics perspective support the mutual forbearance and resource dissimilarity theories of competitive behavior. Hence, from the essay detailed discussion the concept of mutual forbearance and strategic deterrence is clearly analyzed, theorized and to the possible extent gave enough validations which can bring these concepts and implications to a better understanding.