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.

Effect Size and Classroom Management Essay

Teachers play various roles in a typical classroom, but surely one of the most important is that of classroom manager. Effective teaching and learning cannot take place in a poorly managed classroom. If students are disorderly and disrespectful, and no apparent rules and procedures guide behavior, chaos becomes the norm. In these situations, both teachers and students suffer. Teachers struggle to teach, and students most likely learn much less than they should. In contrast, well-managed classrooms provide an environment in which teaching and learning can flourish. But a well-managed classroom doesn’t just appear out of nowhere. It takes a good deal of effort to create—and the person who is most responsible for creating it is the teacher. We live in an era when research tells us that the teacher is probably the single most important factor affecting student achievement—at least the single most important factor that we can do much about. To illustrate, as a result of their study involving some 60,000 students, S. Paul Wright, Sandra Horn, and William Sanders (1997) note the following: The results of this study will document that the most important factor affecting student learning is the teacher. In addition, the results show wide variation in effectiveness among teachers. The immediate and clear implication of this finding is that seemingly more can be done to improve education by improving the effectiveness of teachers than by any other single factor. Effective teachers appear to be effective with students of all achievement levels regardless of the levels of heterogeneity in their classes. If the teacher is ineffective, students under that teacher’s tutelage will achieve inadequate progress academically, regardless of how similar or different they are regarding their academic achievement. p. 63) [emphasis in original] Researcher Kati Haycock (1998) uses the findings of this study and others conducted by William Sanders and his colleagues (e. g. , Sanders & Horn, 1994) to paint a dramatic picture of the profound impact an individual teacher can have on student achievement. The point is illustrated in Figure 1. 1, which depicts the differences in achievement between students who spend a year in class with a highly effective teacher as opposed to a highly ineffective teacher. According to Figure 1. 1, tudents in the classes of teachers classified as the most effective can be expected to gain about 52 percentile points in their achievement over a year’s time. Students in the classes of teachers classified as least effective can be expected to gain only about 14 percentile points over a year’s time. This comparison is even more dramatic when one realizes that some researchers have estimated that students will exhibit a gain in learning of about 6 percentile points simply from maturation—from growing one year older and gleaning new knowledge and information through everyday life (see Hattie, 1992; Cahen & Davis, 1987). The least effective teachers, then, add little to the development of students’ knowledge and skill beyond what would be expected from simply growing one year older in our complex, information-rich society. Figure 1. 1. Impact of Teacher Effectiveness on Student Achievement Sanders and his colleagues, who gathered their data from elementary school students in Tennessee, are not the only ones to document dramatic differences in achievement between students in classes taught by highly ineffective versus highly effective teachers. Haycock (1998) reports similar findings from studies conducted in Dallas and Boston. I have come to similar conclusions in my work, although I have taken a very different approach from that used in the studies that form the basis for Haycock’s conclusions. Whereas the studies conducted in Tennessee, Dallas, and Boston were based on data acquired from students over time, I used a research process called meta-analysis to synthesize the research on effective schools over the last 35 years (see Marzano, 2000a, 2003b). That approach enabled me to separate the effect on student achievement of a school (in general) from the effect of an individual teacher. Figure 1. 2 illustrates my findings. Figure 1. 2. Effects of a School vs. a Teacher on Student Entering at 50th Percentile To understand the impact that a teacher can make, let’s consider each of the five scenarios in Figure 1. 2. (For a detailed discussion of how the computations in Figure 1. 2 were derived, see Marzano, 2000a. ) As depicted in Figure 1. 2, if a student begins at the 50th percentile in mathematics, let’s say, and attends an average school and has an average teacher, her achievement will still be at the 50th percentile at the end of about two years. The student has learned enough to keep pace with her peers. But what happens to that student if she attends a school that is considered one of the least effective and is unfortunate enough to have a teacher who is classified as one of the least effective? After two years she has dropped from the 50th percentile to the 3rd percentile. She may have learned something about mathematics, but that learning is so sporadic and unorganized that she has lost considerable ground in a short time. In the third scenario, the same student is in a school classified as most effective, but she has a teacher classified as least effective. Although the student entered the class at the 50th percentile, two years later she leaves the class at the 37th percentile. In contrast to the two previous scenarios, the fourth presents a very optimistic picture. The student is not only in a school classified as most effective, but also is in the class of a teacher classified as most effective. She enters the class at the 50th percentile, but she leaves at the 96th percentile. The fifth scenario most dramatically depicts the impact of an individual teacher. Again, the student is in a school that is considered least effective, but she is with a teacher classified as most effective. The student now leaves the class at the 63rd percentile—13 percentile points higher than the point at which she entered. It is this last scenario that truly depicts the importance of individual teachers. Even if the school they work in is highly ineffective, individual teachers can produce powerful gains in student learning. Although the effect the classroom teacher can have on student achievement is clear, the dynamics of how a teacher produces such an effect are not simple. Rather, the effective teacher performs many functions. These functions can be organized into three major roles: (1) making wise choices about the most effective instructional strategies to employ, (2) designing classroom curriculum to facilitate student learning, and (3) making effective use of classroom management techniques. The first role deals with instructional strategies and their use. Effective teachers have a wide array of instructional strategies at their disposal. They are skilled in the use of cooperative learning and graphic organizers; they know how best to use homework and how to use questions and advance organizers, and so on. Additionally, they know when these strategies should be used with specific students and specific content. Although cooperative learning might be highly effective in one lesson, a different approach might be better in another lesson. Some general strategies that have a good research â€Å"track record† in terms of enhancing student achievement have been detailed in Classroom Instruction That Works: Research-Based Strategies for Increasing Student Achievement (Marzano, Pickering, & Pollock, 2001). The second role associated with effective teaching is classroom curriculum design. This means that effective teachers are skilled at identifying and articulating the proper sequence and pacing of their content. Rather than relying totally on the scope and sequence provided by the district or the textbook, they consider the needs of their students collectively and individually and then determine the content that requires emphasis and the most appropriate sequencing and presentation of that content. They are also highly skilled at constructing and arranging learning activities that present new knowledge in different formats (e. g. stories, explanations, demonstrations) and different media (e. g. , oral presentations, written presentations, video presentations, Web-based presentations, simulations, hands-on activities). The third role involved in effective teaching is classroom management. This, of course, is the subject of this book. The following chapters detail and exemplify the various components of effective classroom management. Before delving into classroom man agement, however, it is important to note that each of these three roles is a necessary but not sufficient component of effective teaching. That is, no single role by itself is sufficient to guarantee student learning, but take one out of the mix and you probably guarantee that students will have difficulty learning. Nevertheless, a strong case can be made that effective instructional strategies and good classroom curriculum design are built on the foundation of effective classroom management. As Long and Frye (1985) note in their book, Making It Till Friday: A Guide to Successful Classroom Management, it is a myth to believe that . . . ffective teachers can prevent all discipline problems by keeping students interested in learning through the use of exciting classroom materials and activities. The potential for problems exists beyond academics. Students experience difficulties at home which spill over into the classroom; students experience problems with peers during class breaks and in the classroom which often involve the teacher; and students experience mood changes which can generate problems, to name just a few. (pp. 3–4) Similarly, in their ynthesis of the research, Edmund Emmer, Julie Sanford, Barbara Clements, and Jeanne Martin (1982) note that At all public school grade levels, effective classroom management has been recognized as a crucial element in effective teaching. If a teacher cannot obtain students’ cooperation and involve them in instructional activities, it is unlikely that effective teaching will take place †¦ In addition, poor management wastes class time, reduces students’ time on task and detracts from the quality of the learning environment.

Rumored Buzz on Topics for Essay Writing in English Exposed

Rumored Buzz on Topics for Essay Writing in English Exposed When prior brainstorming is finished, you can begin drafting your essay. Hence, if you'd like to balance your academic and family pressure side-by-side, you can just become completely free essay samples so you may have a simple idea of writing proper essays. Hunting for the argumentative essay tips for college do not forget that the chosen theme should attract you in the very first spot. You don't need to acquire super technical with legal argumentative essays, but don't forget to do your homework on what the recent laws about your favorite topic actually say. Even a quick plan is much better than no plan whatsoever, and will begin to provide the writer a feeling that completing an essay on such topic is well in their grasp. Taking into account that lots of persuasive essays concern controversial topics, before writing, you might want to sit down and think of what your opinion on the topic actually is. Topics for Essay Writing in English Explained Realising an essay isn't supposed to be perfect the very first time you write that, really helps some people. Questions people live longer and longer, the notion of cloning human beings essay order to offer spare parts is getting to be a reality. Some questions feel that it is going to improve people's lives in lots of ways. If you're writing about social problems, you will usually be writing about the way the topic a ffects larger groups of people, like how poverty affects a whole generation. When you choose a specific topic, step one is to develop into an authority in the area. Then list all the questions which you'd like answered yourself. There are many questions written each year you may discover your practice answering many questions on various topics. If you have difficulty developing a topic, then you ought to take some opportunity to visit with your professor. If you're interested in your topic, learning about it'll be more pleasurable and you'll write with increased passion, so select your topic thoughtfully. If you're applying at more than 1 college, it is reasonable use a topic that will get the job done for all your choices so you just have to write one great essay. Many students realize that they just should talk through a number of topic ideas until they make their final selections. After the work on your paper is already in process, bear in mind you should be prepared for the upcoming discussion. Some inertia with writing can be because of deficiency of knowledge. If you don't have a time for that, contact us and we'll write If you're hating argumentative essay writing, you have plenty of company. Unfortunately, in regards to writing papers on various assignments, there isn't any perfect collection of all of the s trategies and options. The Foolproof Topics for Essay Writing in English Strategy In case the latter is true, the topic should be chosen dependent on its depth and controversial nature. Because your term paper is all about what you've learned and what more you wish to learn, you need to clearly show your knowledge together with evidence of one-of-a-kind thought. Discussion essays consider each side of an argument. The Little-Known Secrets to Topics for Essay Writing in English Dissertation writing is not uncommon to all degree courses in most universities all around the world. Writing a superb persuasive research paper is a difficult undertaking. Dissertation writing tends to be quite challenging and time consuming to the majority of the students. When research papers are assigned, it's vital to pick a topic which fits with the requirements from the training course. If you're assigned to compose a research paper your main task is choosing international small business research paper topics. Otherwise, a friend who's knowledgeable about science topics are sometimes a great resource, too. Next, select two topics which you would remove. Our physics topic suggestions can be utilised to create unique tips for your own research papers. If You Read Nothing Else Today, Read This Report on Topics for Essay Writing in English As a writer you can't ever underestimate the demand for research work. You won't even notice the way the hours of research pass, as you'll be too absorbed with the new intriguing facts that you discover in the procedure. For instance, researching a wide topic like business management'' is difficult since there can be hundreds of sources on all facets of business administration. Lots of the medical issues that people are resume template objective lines in the present world are the result of the simple fact that we've got an extre mely sedentary way of life.

Drug Abuse On The United States Essay - 1344 Words

Drug Abuse on the Rise in the Community â€Å"In 2014, nearly two million Americans either abused or were dependent on prescription opioid pain relievers (â€Å"Injury Prevention and Control: Opioid Overdose,† 2016.)† The CDC is now saying that the United States is in an epidemic. The purpose of this paper is to explore the increasing drug use in the United States over the last decade. The primary focus is on the administration of opioid analgesics and the addicting mechanisms of these medications, and what they are used for. It will talk about the deaths related to these medications and the health effects, and the other types of drugs these medications can lead to, such as heroin. This paper will also talk about the costs related to opioid analgesics in the hospitals and in the community, and what we as health care providers can be doing to help stop this epidemic. Drug Abuse on the Risk in the Community Opioid analgesics, also known as narcotic analgesics, and are used as pain relievers that act on the central nervous system; these drugs can have very addicting mechanisms involved. â€Å"According to the centers for disease control and prevention, prescription drug abuse has reached â€Å"epidemic† proportions, deaths from prescription drugs, particularly opioid analgesics, now vie with automobile accidents as the most common cause for accidental death (Herzberg, Guarino, Mateu-Gelabert, Bennett, 2016).† Throughout the last decade there have been more deaths and addictionShow MoreRelatedDrug Abuse And The United States1645 Words   |  7 PagesNixon launched the war on drugs stating, â€Å"America’s public enemy number one in the United States is drug abuse. In order to fight and defeat this enemy, it is necessary to wage a new, all out offensive† (Sharp, 1994). 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