Complement Serum Activity by Lysing Sheep Erythrocytes

Complement Serum Activity by Lysing Sheep Erythrocytes Introduction The Immune system is a series of complex processes which has evolved to protect the body from attack by foreign pathogens. These pathogens are able to enter our body through the skin or lining of the internal organs. The immune system is able to protect us from intracellular and extracellular organisms as well as from ourselves, stopping malignancies and autoimmune diseases from spreading in our bodies (Bastian, 1993). There are two lines of defence, the adaptive (specific) and innate (non-specific immunity), though both are united in their goal to destroy pathogens they have different ways to tackle this. Innate immunity is the 1st line of defence while adaptive immunity is the 2nds line and thus takes longer to act (Clancy, 1998). The complement system is part of the immune system and can be bought into action by the adaptive system if required. Complement is a group of proteins working together within the immune system; once stimulated by one of many triggers, proteases begin to c leave protein in the system, bringing a cascade of enzyme reactions in order to fight off foreign pathogens and activate the inflammatory response. Within the complement cascade there are many proteins that play a role but C3 is a protein critical to the effector functions of the system (Abbas, 1994). There are many paths for immune mediated lysis and the one we will be looking at is intravascular haemolyse and occurs when the complement has been triggered through the classical pathway. When the antibody binds to the antigen on the surface of the erythrocyte, a complement component triggers the membrane attack complex to form pores in the cell membrane resulting in cell lysis (Chapel, 1990). The intensity and speed at which cells lyse is dependent upon the rate at which the complement cascades to enable complete cell lysis. Experiments like these are able to provide us with an understanding of how the complement immune system functions. It can also increase our understanding of autoimmunity and perhaps lead to ways in which the effects of immunity can be prolonged or inhibited according to the disease. Systemic Lupus Erythematosus (SLE) is an autoimmune disease, in which complement is analysed, as getting SLE is dependent upon the gene which is responsible for producing MHC, a component used in haemolysis (American, 1993), patients with other immunological disorders can require their complement activity to be monitored and thus this assay would be able to show how efficiently the complement component of the immune system is working to defend their bodies. Aims To determine complement serum activity by lysing sheep erythrocytes To determine the volume of complement required for 50% lysis. Materials 20 Cuvettes 1.0ml 20 test tube plastic disposable Automatic pipette 200-1000 ÂÂ µl 6 tips Automatic pipette 0-200 ÂÂ µl 6 tips Water bath at 37Â °C Spectrophotometer Test tube rack Centrifuge Ice bucket Ice Method Wash 4ml of erythrocyte suspension three times with barbitone saline solution. Prepare a 6% stock solution of erythrocytes In one test tube mix 3.0ml of sheep anti-erythrocyte antiserum, diluted 1/50 3.0ml of the 6% SRBC Mix and gently by capping and inverting several times Incubate at 37Â °C for 15min in the water bath, mix every 5min. Set up the test tubes on ice in duplicates and label Add the reagents in order as shown in table 1 below Incubate the tubes for 60 minutes at 37Â °C mixing gently every 15minutes Place the tubes on ice and then centrifuge at 200g for 10 minutes at 4Â °C Remove the samples and put into cuvettes and read the absorbance at 541nm, with ammonia solution as blank record the results in a table. Results Discussion When carrying out the experiment raw data was recorded, and presented in table 1. However the results obtained during the practical were not used as the erythrocytes lysed before complement was added and therefore complement activity could not be observed as adding complement to lysed cells is not able to produce results, therefore the ideal data provided was used and analysed. From table 1 it is clear that absorbance levels increased as serum volume increased, this is due to the fact that as volumes of complement increase more red blood cells are lysed which in turn allows haemoglobin to be let out, this is of a dark colour and as more cells are lyses the darker the resulting sample will be, and so the absorbance as read the spectrophotometer will increase. After the guinea pig serum has been mixed with the sensitised erythrocytes, it produces anti-body coated cells with complement attaching to the antibody, and activating this attracts the MAC molecules to take action and lyse the cell (Kuby, 1994). Following the pattern seen in table one table 3 shows a progressive % lysis of cells as the volume of serum is increased, however for the 100% lysis an ammonia buffer was used to ensure that all cells are lysed during the experiment. Further to this graph 1 produced a sigmoid curve, from which it was possible to estimate CH50. However calculating the 50% lysis from this graph is not very accurate. Thus a log graph 2 was constructed, with the use of van Krogh equation to determine the actual value of 50% lysis. The equation was provided by the lecturer. Van Krogh equation: x= k[ y ]1/n 100-y Where: x= amount of complement (ml of undiluted serum) y= proportion of cells lysed k=50% unit of compliment n=inclination of graph (ideally 0.2) This resulted in table 4 giving a volume of 133.5 CH50/ml. However when calculating CH50 the x values were all in the negative. Moreover, it was not possible to compare data sets obtained against ideal data as the experiment did not yield results due to lysis of erythrocytes before complement was added. This could have occurred due to improper pipetting, handling or transporting of the cells as shaking them too much could have lysed them due to shock, as the cells were sensitized and thus prone to quick lysis. Further to this it was reported by Inglis, et al, 2007, that the use of erythrocytes from different sheep can yield inaccurate results and thus produce different CH50. Although there are many inaccuracies present within the experiment, it also gives scope to further improve the method as well as explore other area of the subject at hand such as factors which affect the performance of complement like temperature or PH. This assay is a good way to measure the activity of the immu ne system within patients, such as patients with LSE as mentioned earlier, other patients with low immunity can also be tested to see how the complement system is or isnt aiding their recovery, thus steps can be taken by medical professionals to either boost or monitor the progress of the patients immunity as fundamentally the immune system is required to work at its optimum to keep humans and animals from dying of disease(Inglis,et al, 2007). Conclusion Overall this experiment has shown how complement is important in aiding white blood cells to lyse foreign bodies. Though in the experiment carried out the blood cells lysed before complement was added the method was presented and the ideal set of data, showed what results should have been obtained. Also the hypothesis that as the complement concentration increases so will the absorbance proved positive.

Philoaophy Of Law :: essays research papers

  Ã‚  Ã‚  Ã‚  Ã‚  In comparing the issues surrounding the distribution and depiction of pornographic and racist materials, very few differences, if any, can be derived from the two. Besides the obvious differences in which one form appeals itself to the adult community and the other to the racist community, the two extremes, nonetheless, fall under a much broader category. They are both recognized and valid forms of speech, and as such are equally entitled to the same constitutional protection provided by the freedom of speech clause of the First Amendment as are various other legitimate forms of speech.   Ã‚  Ã‚  Ã‚  Ã‚  In the situation provided before us, we are asked to determine whether an individual should possess the right to distribute racist films graphically depicting whites verbally abusing, beating, and urinating upon blacks. My immediate response to the question would undoubtedly argue that such morally offensive material should not be allowed constitutional protection. The mere mentioning of such a proposition strikes anger at the heart of moral conscience. But, my moral convictions are not, nor are anyone else’s for that matter, sufficient grounds to deny anyone their First Amendment right to freely engage in the distribution of such material if they so desire to do so. Moreover, the First Amendment clearly dictates that Congress shall make no law... abridging the freedom of speech. Thus, as a long established and highly empowered legal doctrine, it must ultimately be respected by the government to the fullest extent.   Ã‚  Ã‚  Ã‚  Ã‚  The First Amendment do es not state, nor does it imply, that only specific forms of speech which are morally just shall be free of governmental interference, while other forms believed to be offensive to social morality, such as pornography or racist films, shall not enjoy such a privilege. If that were to be the actual case, â€Å"freedom of speech,† which has long been revered by our nation as one of the fundamental liberties of American history, would further cease to exist. All that would have to be proven to restrict speech would be that the message being expressed contains the slightest mention of morally offensive content. Fortunately, however, the freedom of speech clause grants people the power to convey their opinions in the manner which they deem fit. Thus, if the owner of a video store chooses to sell videos in which African-Americans are repeatedly verbally and physically abused, then ultimately his right to do so must be respected.

Ethics/Legal/IEP Essay -- essays papers

Ethics/Legal/IEP Essay In the treatment of those individuals labeled as disabled, be it physically, mentally, or a combination thereof, we are faced with many challenging ethical questions. The first and foremost of these is, what is ultimately, â€Å"the right thing to do.† While we may or may not agree on the answer to this question, as well as those that follow, many advances have been made in our society to deal with disabled individuals in the most humane and ethical way possible. What is the right thing to do? Is there an ultimate resolution? In 1975, Public Law (PL) 94-142 was passed. This law has several parts, each pertaining to a different aspect of discrimination, or nondiscrimination, of disabled persons. This law was passed in hopes of avoiding not only the horrific scenarios found in Nazi propaganda films such as Selling Murder, but further to guarantee all disabled individuals a fair chance at life: to live to the fullest extent of their capabilities and fulfill their potentials. In the film, Selling Murder, Nazi officials convey to the public that disabled people are a threat to the world community, and that they are living a â€Å"life without existence.† It goes further and says that not only should those who are disabled be quarantined and banished from society, but for their own sake they should be sterilized, and ultimately killed. Labeling the mass deaths of disabled persons â€Å"mercy killings,† the film explains how they, the disabled, are unable to function as â€Å"real† human be ings, and that if they had the mental facilities to make the choice, they would choose death over such a menial existence. Showing gruesome photos of the severely physically disfigured, the propaganda seeks not only to convince, but to scare the masses into agreement. On the other hand, films such as Regular Lives demonstrate the ability of disabled individuals not only to survive, but to flourish and better their communities through their own efforts. It demonstrates the necessity to include disabled students in any and all ways possible in order to help them adapt to a social environment as normal as is possible. PL 94-142 states that all disabled students are guaranteed a FAPE (free and public education). With this comes the formulation of an IEP (individualized education plan). This grants students a goal guided objective list for their education. The ... ...ther teacher or administrator to explain evaluation results to those involved. Why does it matter? What is the relevance/necessity of making moral judgments when formulating a course of action? Disabled people have been discriminated through time. They have been played as Jesters for entertainment, banished as havens for evil spirits, and massacred simply for being different. They have been subject even to sterilization, seen as people unfit to breed. We are forced to ask ourselves- why then, does it matter? Hardman states on page 52, â€Å"Social justice and equality for all people should be of paramount importance in the design and delivery of all curricula.† Fair and equal treatment of disabled persons matters because they, like all other citizens of this country, are people. They are born in this country with the guarantee that they will be granted â€Å"life, liberty, and the pursuit of happiness.† With this in mind, it is our moral and ethical obligation to â€Å"hold these truths to be self evident, that all men are [indeed] created equal.† This country was founded on these beliefs, and as we continue our progress forward with laws such as PL 94-142, we support this creed.

Aubrey Beardsley and Oscar Wilde :: Salome Plays Essays

Aubrey Beardsley and Oscar Wilde Oscar Wilde claimed to have discovered Aubrey Beardsley, when he asked him to illustrate his Salome. However, many people have claimed the same thing. Author Robert Ross on the other hand, thinks that Beardsley really started with the men with whom his work will always be associated. The men he worked with on the Yellow Book. (Aubrey Beardsley, p.14). Aubrey was born on the twenty-first of August 1872, in Brighton England. He was a quiet reserved child of an upper middle class family. He showed as a child very little caring for his lessons. However, he always showed an aptitude for drawing. Beardsley’s father through very unfortunate circumstances lost his inherited fortune. Beardsley at this time suffered from Tuberculosis; this was what eventually caused his death. His mother also became ill and was unable to take care of both him and his sister. Therefore, they were sent off to live with an old aunt. Their lives there was lonely and Aubrey developed a taste for reading as well as drawing. His aunt placed him in a boarding school where he indulged in his talent by drawing caricatures of his teachers. In July 1888 he left the school and started working in an architect’s office. Beardsley wanted to enter the art world. He accomplished this in an incident, which became famous. It occurred when he was invited to see th e studio of painter Sir Edward Burne-Jones. The artist was impressed by the drawings in Beardsley’s portfolio, and recommended that he attend night classes at the Westminster School Of Art. This was the only formal training Beardsley had ever had. Ian Fletcher author of Aubrey Beardsley by Ian Fletcher claims that Beardsley is not an impressionist, nor an expressionist, but essentially eclectic. " He had no facility, no admiration for nature-pantheism, the superstition of the cultivated classes. (Aubrey Beardsley by Ian Fletcher, p.23). Much of Beardsley’s work does connect directly with literary texts. "Beardsley is indeed much concerned with the reader or viewer, but hardly in the humble facilitating mode of the average illustrator and reader is the precise word. Yet, he does mediate between author and reader, not conducting word into image, but bringing to light rather what implicit, forbidden, or subversive elements of a text so disconcerting the author and forcing the reader to become a voyeur by recognizing in himself what he condemns in others".

Gatsby, Corruption of Wealth

Cold Cash Turns to Cold Hearts The 1920s’ was a hectic time period for many young and wealthy people living in New York City. Many entrepreneurs took advantage of these economic opportunities and profited greatly. Although, some took their wealth in strife, to benefit others, some people misused their new power. In the novel The Great Gatsby, F. Scott Fitzgerald shows how money and power are often abused. Gatsby never understood the genuine value of hard work. While Tom and Gatsby are talking, Gatsby exclaimed, â€Å"After that, I lived like a rajah in all the capitols of Europe† (Fitzgerald 65). Gatsby was constantly living the life of extreme leisure and luxury, having no responsibilities, and no problems. It is almost as if Gatsby is the â€Å"king† of West Egg, with his huge palace and nice cars. As an article states, Gatsby had â€Å"his white suit, silver shirt, and gold tie-his palatial house, his grand parties, his fancy automobile, his hydroplane, and his library of real books. †(Pauly) With endless amounts of materialistic items and fake friendships what is there to live for? Although Gatsby tells people that he had â€Å"inherited† his wealth, Nick believes he made the money a different and immoral way. When Nick and Gatsby travel to the city one afternoon, they meet a shady man who claims to be good friends with Gatsby. At first, Nick mistakes him for a businessman but, Gatsby explains â€Å"Meyer Wolfsheim? No, he’s a gambler†¦ He’s the man who fixed the World Series back in 1919† (Fitzgerald 78). Gatsby appears to be an honest man, but there is a shady and crafty to side to him as well. A quote from an article proclaimed, â€Å"Jay Gatz, whose ill-gotten wealth is acquired solely to gain acceptance into the sophisticated, moneyed world of the woman he loves, Daisy Fay Buchanan† (Telgen). When people picture a sleazy gangster they see someone who is dirty, probably poorly dressed. However, Thomas H. Pauly’s article demonstrates, â€Å"Jay Gatsby effectively overturned the dated assumption that gangsters were lowlifes from the Bowery and replaced it with an upscale figure who was enviably wealthy and fashionably stylish. † Judging someone based on their appearance can have its repercussions. The squalid acts Gatsby would do just to get rich is mind-boggling, the money can bring out the most deceptive people. Some people may speculate that money can buy happiness, friends, and security. However, in the end Gatsby ended up with nothing, when he sadly discovered that money cannot buy true love. When people obtain a large amount of money, they either spend it wisely and profit or â€Å"blow† it, spending on superficial possessions they do not need. Unfortunately, with obvious disregard for morals, ethics and even the law, there is a corruption of wealth and power in the novel The Great Gatsby and in modern society as we know.

Children and Happiness Essay

What makes children happy? ‘That’s easy,’ you might say, ‘a new toy, an ice cream or a trip to the amusement park.’ While these things bring an immediate smile to many children’s faces, they are probably not the things that will keep them happy day by day, or turn them into happy adults. The basic ingredients that make children happy during childhood seem to be the same ones that help them to become happy adults: a secure relationship with parents gives the base to confidently explore the world and develop a sense of mastery and recognition, all important components in the recipe for happiness. However, in the short term, the new toy might provide a smile too! Man has to pass through many stages of life, since he is born until he dies. He passes through infancy, childhood, boyhood, youth, middle age and old age. Childhood continues from the 6th to the 12th year of age. Boyhood continues from the 12th to the 19th year of age. Youth lasts from the 20th to 35th year of age. Middle age lasts from the 36th to the 55th year of age. Old age continues from 56th year until death. Of all the stages of life childhood is the most important. Because it is the formative period of life. It is also the first conscious stage of life. Important of childhood: Childhood is the formative period of life. Human character takes its shape in childhood. Man gets deep and lasting impressions in his childhood. Because at this stage men’s mind is soft, receptive and plastic. It is just like the potter’s clay. So human character should be properly shaped in childhood. What should be done in childhood: Childhood is one of the growing stage in man’s life. The human beings grow up to 24th year of age. So, the health of the child should be properly looked after. He should be given balance diet. He should be encouraged to from healthy habits like early rising. He should be taught to be neat and clean, to do exercise regularly, to eat, sleep, read and play at habits. He should be kept happy and cheerful. All these will help him to grow healthy in body and mind. Proper and appropriate food should be provided to the child’s mind. He should be given the most carefully chosen books to read. he should be given children’s magazines. He should be given interesting and instructive pictures. At this stage, records should be kept of the child’s inclination, aptitude, knack, mood, temperament, interest, abilities and his latent qualities. On the basis of these records, the future of the child should be planned. The child should be trained on the basis of this plan. Care of parents and teachers: In the modern system of society and education in India, nobody takes the responsibility of the child’s welfare. The parents think that it is a responsibility of the teachers, because the child stays in the school for six hours a day. The teachers think that it is the responsibility of the parents, because the children belong to their parents, and in future the children will come to any use only to their parents and families. But in the interest of society, nation, country and above all mankind all have responsibility for the welfare of the children. Conclusion: It is the foremost duty of the Government to see to the welfare of the children. But in India we find that a lot of children in schools are dying by food-poisoning in the Government Mid-day meal system. The school buildings are defectively constructed. So the buildings breakdown and many children die underneath. the milk-powder that comes to Panchayats and Block offices is said to be put to black-marketing. Government should look to all these affairs. Childhood is the most important period in a men’s life. So, the Government should take utmost care of it.

My relationship with God

As a start for developing a closer relationship with God, I decided that I wanted to try daily devotionals and Bible reading. I considered finding a devotional guide, but instead decided to begin by simply reading the Bible for myself. I thought about finding a daily devotional guide, but chose instead to start my devotional readings by simply reading the New Testament and letting God speak to me as He determined not as some other person might have been inspired.My decision was partially based on a need for this to be a personal journey and a knowledge that many people who attempt to read the Bible begin at the beginning and get discouraged when they reach the histories of the Books of Law. And, as a Christian, I believe the importance of the New Testament speaks for itself. This was a particularly difficult task for me. Choosing a time to read regularly and coupling it with prayer for understanding and meditation to determine the meaning of the scriptures was very difficult.I discov ered that the first thing in the morning was not a good time for me to try to read my Bible. Too often, I was overly tempted to hit the snooze button and often did not find my concentration level where it should be to really understand the scriptures, even though most of it was familiar territory. I then tried to do my devotional reading at the end of the day and found that too was a bad idea. Some days exhaustion seemed to bull me away from my studies in an untimely manner and I decided that too was a bad time. Instead I chose to set aside an hour every morning just before lunch.This seemed to be the best time for me to be able to devote my attention to the Bible and not be distracted. I set an alarm to let me know when I could be done and then tuned out all distractions. In the three weeks, I managed to read through all of the gospel of Matthew and get started in Mark without feeling like it was a chore. As the time wore on, I found myself looking forward to my daily devotional mo re than I had in the beginning and found it was easier to maintain once I found the appropriate time. Intercessory prayer as a devotional tool also appealed to me.Like many churches, the church I grew up in had a prayer chain devoted to praying for those in need. Remembering the peace and joy that came from praying for another person without any goal or personal motivation lead me to choose intercessory prayer Practicing this devotional method reminded me of the need to be more worried about others than I am about myself. I thought this was going to be my favorite devotional form, but I found that it wasn’t. Too often it felt like I was pushing my will onto God instead of allowing His will to work through me.The other two devotional forms I chose were praise singing and random good deeds. These are the two forms of devotion which I believe had the biggest impacts on my life. I chose to do my devotional singing while in the car. I turned off the radio and began singing hymns i n the car and immediately noticed a change in my attitude. I was no longer as willing to spout off angrily at other drivers, no longer feeling the need to drive unreasonably fast and no longer impatient in traffic. Changing my heart through song made my daily commutes easier and relieved tension in my life.I found myself with an sincerely improved attitude and a greater desire to praise God for the miraculous things he was working in my life and int eh world around me. The author says that the practice of doing good deeds simply for the sake of doing them is a good thing because it reinforces the image that we want non-believers to have of Christians. I found that the biggest impact it had on me was that this is a manner I believe Christians should act in. By committing daily random acts of kindness I felt better about myself.I felt that I was behaving more the way that Christ would want me to behave. My random acts of kindness included letting others go first in the grocery line an d putting coins in parking meters. Finally, last week, I also purchased a small quantity of stuffed animals and placed them randomly on the windshields of cars at the doctor’s office. I believe that the act may have helped just one other person to get through a rough day and that is what God has called each of us to do. These actions are things that I will continue.I believe that the daily Bible readings will be the most difficult to consistently maintain, but I also believe it may be the most important for my continued spiritual growth. The praise singing and random good deeds are easy to maintain because they have an immediate uplifting effect. The Bible reading requires the biggest commitment, but a better understanding of God’s purpose for me should be found in His word. I believe that the intercessory prayer is also going to continue to be a part of my life, though I am interested in determining how I can best pray without feeling like I am placing demand on my Lo rd.