Aural and spatial elements Essay Example for Free

Aural and spatial elements Essay Wertenbaker uses simple and minimalistic production design and uses her actors as her main resource to convey meaning and character especially as they are playing different gender or changing between convict and officer characters throughout. The few props are used to clarify meaning and are important in their own right along with costumes and set. She uses language and music to communicate with the audience and to create a specific atmosphere. Wertenbaker uses the movement or gestures of a character to emphasise a point or show character’s relationships. An example of this is Sideway in the first rehearsal, the stage direction â€Å"walking sideways, arms held high in a grandiose eighteenth century pose† this reinforces the idea of how uneducated the convicts are, as the audience can see Sideway has never seen a theatre production just has an idea of what it would be like. An example where gesture is used to communicate character’s relationships is in Act 1 scene 7 where Harry and Duckling are rowing, the stage directions for Duckling are â€Å"remains morose† and â€Å"doesn’t turn around† these are hostile actions as Harry is desperately trying to get her attention â€Å" Why don’t you look Duckling. † The audience can see that their relationship is unstable. The way Wertenbaker positions her characters on the stage is important in communicating status and character such as in Act 1 scene 11 when Major Ross and Captain Campbell enter, the convicts â€Å"slink away and sink down. † This shows that the convicts are afraid of the officers and they try and get as far away as possible from Robbie and become inconspicuous. The beginning scene has important use of positing, the convict are â€Å"huddled together in semi-darkness. † This gives an idea of how horrible the conditions were for the convicts during transportation. The officers are on deck above the convicts and in better conditions again showing status. This idea is kept throughout the play but gradually changes as the production of the convict’s play takes effect . Costume is used to communicate character, Governor Arthur Phillip’s costume would be grander and more decorative than the other officers and he has a higher status. The convicts would be wearing similar clothes but all slightly different such as Mary’s which would be covering her and suppressing her femininity because she feels guilty about what happened on the ship â€Å"I would have been less of a whore†. Sideway’s clothes would be over the top to show his flamboyant personality . All the costumes also have to be simple to allow quick costume changes as the actors multi-role. Wertenbaker’s set is very simple with only a few props. This is done to represent the bareness of Australia and how they have to create a new colony but have arrived with hardly anything. The set is also kept simple because there are 22 scenes and a complex set would take too long to change each time and therefore slow down the pace of the piece. Props are used sparsely but are very important in showing character’s situations. Example of this are the whipping equipment need in act 1 scene 1, they are used to clearly show the audience how the convicts were treated on the ships and the punishments. The play book is used throughout the play when the convicts are rehearing or talking about the play as this is one of the main themes of Our Country’s Good , however not many other books are used and this emphasises that the convict are uneducated and many can’t read. Ralph’s dairy and the picture of his wife are important props in showing his character. Ralph idolises his wife as something pure â€Å"I might kiss your picture as usual. † He uses his diary to write his thoughts showing he doesn’t feel he can talk to the other officers and is disturbed with the new land and behaviour of the convicts and officers. Food is mention throughout the play â€Å"if I was rich, I’d eat myself sick† but it is never used as a prop this highlights the lack of food in the colony. Wertenbaker uses gun shot sound effects at the beginning of the play in scene 3 Act 1, as the officers are shooting birds this reminds the audience that they have just arrived on a new land and are already destroying its nature and wildlife. The other sound effect Wertenbaker specifies in the play is drumming which occurs at the end In the last scene as the convicts go on stage, this symbolise a parade that a new colony might give as celebration. It also creates excitement for the play. The triumphant music of Beethoven’s fifth symphony is played at the very end as an introduction to The Recruiting Officer, this makes the audience happy and excited and reflects what the convicts would be feeling at that time. Other music Wertenbaker may want to be used during the play, especially the aboriginal scenes is didgeridoo music as this is a native instrument to Australia. The Language Wertenbaker uses shows character, an example of this is Liz’s monologue in Act 2 scene 2 where she use lots of slang â€Å"born under a ha’penny planet† this shows the audience she is uneducated. In contrast to Liz, Arthur is very educated and we can see this due to the literary and historical reverences he mentions when backing up an argument â€Å"In the Meno, one of Plato’s great dialogues. † This is a way that difference in education shows the split between convicts and officers. Language is also used to show change in characters, for example at the beginning Wisehammer is rough and has no hope for the future â€Å"alone, frightened, nameless in this sinking hole of hell†, however as the play progresses we actually see him as being one of the more educated convicts. â€Å"Abject: a man without hope. † In the last scene we see him thinking of the future and with restored hope â€Å"I’ll write a play about justice. † Our Country’s Good would be more suitable for a proscenium arch stage because it communicates the bareness of the new land as no-one is on the other side unlike a transverse stage. Wertenbaker wanted to use a cyclorama to symbolise Australia and this would need to be upstage and therefore would work best in proscenium as it has a back wall. The third reason is the convicts and officers could easily be separated and shown in their groups and more easily commutate status and power. The lighting is very important in staging the play. The lighting would of be harsh blues and yellows to represent the climate of Australia. This lighting is the most effective way of making the audience aware of the different climate and habitat of Australia and how hard it was for the convicts and officers to adapt to the change. The lighting would also be used to show night and day as some of the scenes are during the night, this would create an atmosphere for the piece. Wertenbaker chose to create an obviously theatrical piece of drama instead of a piece of realism because she wanted the audience to see the social message the play contained, how drama can reform and educate people. A theatrical piece is the most effective way to do this as it allows both the actors and audience to see the story from both sides of the colony due to the multi-rolling actors.

Quantitative investigation of immunoglobulins

Quantitative investigation of immunoglobulins Introduction- Quantitative investigation of Immunoglobulins is the basis of the standard laboratory technique within the field of clinical immunology. Immunoglobulins can be measured quantitatively through the use of nephelometry, such measurements are vital in the instance of a suspected immunodeficiency within a patient. For this reason the test is accurate and rapidly measures the amounts of IgM, IgG and IgA proteins within the blood of the patient and from such determine if a number of conditions or disorders are present. The role of such antibodies is within fighting infections and allergies as part of the normal immune response. A disease (or disorder) can be identified through the measurement of such protein levels. IgM for example can appear during an initial infection and then reappear to a lesser extent upon secondary exposure. (Weir, 1978) Nephelometry is usually performed by drawing blood from a vein on the back of the hand or if not possible the inside of an elbow. The needle draws the blood into an airtight vial or tube attached to it. Removal of the needle is followed by sterilisation and covering of the incision site. (Stanley, 2002) Practical Schedule- Nephelometry is an automated system that measures antigen or antibody solution in very limited concentrations by the amount of light scatter. The principle is that when the light comes into contact with the solution it will not be absorbed but scatter away from the main beam and measured at angles between 0-90 from a predefined curve. The subsequent amounts are proportionate to that of the concentration of molecules. As well as dilute solutions there also needs to be a linear correlation between molecules formed and optical density. For this reason several dilutions measurements are recorded and also during the formation of molecules. This process is known as rate nephelometry. When considering this technique it is vital that the relative amount of antigen and antibody must be small enough so that precipitation does not occur but also large enough to allow the formulation of small immune complexes. Immunoprecipitation results are achieved through the use of monoclonal antibodies (MCAs) allowing epitopes to react with the antiserum and MCAs forming immune precipitates with their antigens. Results- Normal results IgG: 560 to 1800 mg/dL IgM: 45 to 250 mg/dL IgA: 100 to 400 mg/dL Evaluation- the automated nature of this technique means that it is both fast accurate with results available within 1-2 hours. Its wide spread use is mainly down to this factor but its simplicity and low sample size and volumes also make it a valued technique with the clinical laboratory setting. (Diamandis et al1996). It is however, as with most techniques not without its draw backs. The usual precautions should be taken as when taking any blood sample. Although rare excessive bleeding, fainting and infection should all be considered as risk factors when taking samples. (Drexel, 14/06/08) The presence of dust particles and other debris can be cause for distorted readings and lead to higher values than expected. This can be addressed through centrifugation of the specimen (Diamandis et al1996). In addition to this air bubbles can cause similar effects on results. To ensure readings are as accurate as possible, the specificity should be at optimum level, as set on the nephelometer and controls should be carried out wherever it is used. (Palmer, 1992) Although this method does determine the amount of each immunoglobulin it does not possess the ability to identify antibodies. Another method that can be used to quantitatively investigate Immunoglobulins in serum, saliva, cerebrospinal fluid (CSF), amniotic fluid, and gastrointestinal juice is Radial Immunodiffusion (Chapel et al1999). This technique allows for the adding of a sample to a well in a gel made up of the antibody specific for the substance being tested for. This then moves through the gel leading to the formation of a visible precipitate around the optimum concentration sample well. The interpretation of such results however is subjective and results are delayed as the process takes several days and as such nephelometry is recommended for greater precision, automation, objectivity and speed and is suitable for large throughput tests (Keogan et al 2006). It is also hard to quantitatively analyse the results using very small samples and a calibration curve. (Chapel et al., 2006). 2. Quantitative Other Serum Proteins-Radial Immunodiffusion Introduction Radial Immunodiffusion acts upon the antigen- antibody complex precipitation reaction. It is used within the field of neurology and oncology. This involves passive diffusion of immunoreactants through an agar matrix. An electrical current is not required for such process to occur as is reliant upon the physiochemical relationships. Practical Schedule Radial Immunodiffusion works via the mixing of antiserum with agar and pouring it on a glass plate to allow it to solidify. The antiserum must be specific for the class of immunoglobulin that is being measured for the technique. The agar mixture is then punctured and the subsequent holes filled using the sera from the test samples. Diffusion of the Immunoglobulins radially causes the formation of precipitate at the point which the number of antibody and antigen are identical. As with previous techniques a calibration curve us used that has been made up from known set of solution to determine the amount of Immunoglobulins present within the sample. Evaluation Accuracy and specificity is the most potent threat to validly of this technique. The fundamental problem is the lack of sensitivity and is not a rapid technique with results taking over 48 hours owing to reaction times. (Chapel, 2002) Whilst it does provide use in the determination of serum proteins quantitatively there are an array of factors that can lead to unreliable results. Temperature of the gel and external environment, molecular size, gel viscosity, reactant concentration and buffer pH highlights a few of the factors that will affect the rate of diffusion but is not exhaustive. (Nakamura et al, 1979) 3. Quantitative other Serum proteins Collection of serum Collect blood in a glass container and allow it to clot at room temperature for an hour. Once the clot has formed loosen the walls of the container to aid retraction. Transfer to 4 degrees and leave overnight if necessary Collect the expressed serum and centrifuge at 150g for 5 minutes to sediment the erythrocytes, and then at 350g for 15 minutes. Transfer the straw coloured serum to suitable containers and heat at 56 degrees for 30 minutes to destroy the heat labile components of complement. (Hay et al., 2002) Qualitative Immunoglobulins Introduction- The stages of diagnosis, determination of immunity and the susceptibility of an individual to many microbial infections, are based upon immunological tests in serum. When blood clots the fluid that remains is known as serum and as such it is rich in Immunoglobulins. Serum however is not easily accessible so other sample sources can be used. The presence of specific Immunoglobulins in urine, saliva and cerebrospinal fluid mean that such bodily fluids, inclusive of others such as semen, can be used instead. Anatomically the most readily available and less intrusive however, as with other bodily fluids, contain low concentrations of IgM and IgG. Semen is abundant with these Immunoglobulins and as such may be perceived as being the most accurate and reliable in any such investigations (PCT, 1987) In Serum- Immunoelectrophresis Introduction- Serum Protein Electrophoresis is a qualitative investigation carried out to test for the presence of monoclonal bands (paraproteins). (Chapel et al., 2002) During electrophoresis, discrete monoclonal bands may appear (M bands). Further investigation is needed in order to determine the immunoglobulin heavy and/ or light chains through immunofixation. This is important when trying to distinguish what sort of Immunoglobulins are present. Determination is achieved through Immunoprecipitation in a gel with anti- sera that is specific for heavy and light chains of the immunoglobulin. Immunoelectrophresis works by separating sera in agarose gel by electrophoresis. Troughs that are parallel to the unfixed electrophoretic strips have specific anti- sera added to them leading to the formulation of precipitin arcs that are clearly visible owing to the process of diffusion. Immunofixation however tends to be more commonly used and as such will be more focused upon within the portfolio. This technique is commonly used in the diagnosis of conditions such as osteoporosis. In the abnormal absence of a heavy chain and an abnormal reaction occurring with the ant- sera that are specific for light chains discrete (M) bands are present. It is also a highlight to the possibility of an IgD or IgE paraproteins although is far less common. If an abnormal reaction occurs with only the heavy chain anti sera it is indicative of a rare heavy chain disorder. It is possibly to quantify individual M bands with the use of a densitometer. This acts by measuring the intensity of the stain taken up by each individual band and as such is the only method at present to be of use in the measurement of paraproteins concentration (Chapel et al1999). Practical Schedule taken from Clinical Immunology. (Chapel et al., 2002) Immunoelectrophoresis- Apply serum samples to an electrophoresis gel at the cathode end alongside a normal serum sample as a control. Apply an electric current for 45 minutes and remove the gel. Use a stain to visualize the bands. Immunofixation-specific antisera to IgG, IgA, IgM and kappa and lambda light chains are then applied to the electrophoresed samples by soaking strips of cellulose acetate in the individual antisera and laying them on the electrophoresis gel. This is then incubated for 2 hours and all the un-fixed proteins are washed off leaving the precipate. Individual monoclonal bands can be quantitatively measured by a densitometer. Results The dark areas indicate monoclonal bands. The picture above shows a positive result for the lambda chain. The presence of monoclonal bands can indicate multiple myelomas or osteoporosis. In this example, the M band is identified as IgG of kappa type. Concentration of the M band is determined using a densitometric trace, as demonstrated in the second image. Evaluation- The presence of air bubbles will distort the formation of protein bands and as such the gel must be degassed. The method detailed above is much quicker and far more sensitive than the singular use of immunoelectrophresis. Its cheapness and low hazard level mean it is a desirable technique in the detection of Immunoglobulins within serum. (Zola et al. 1999) Qualitative Immunoglobulins in Urine- Electrophoresis and Immunofixation Normal physiology of the kidney dictates that protein is usually excreted within the urine in minimal amounts. Higher levels can lead to the suspicion of multiple myeloma that can lead to irrapairable damage to the kidneys as nephritic cells are non replaceable with chronic lymphocytic leukaemia and hypogammaglobulninaemia being suspects. Kidney disorders such as IgA nephropathy may also be a causation of such symptoms. All humans produced excessive amounts of free polyclonal light chains in accompaniment to normal immunoglobulin synthesis with these being secreted into the urine and are detectable in low amounts in all samples (Thompson, 1981). If the normal range of this is exceeded however it is indicative of renal damage. This method is often used in order to detect these small free monoclonal light chains that are also called Bence- Jones Proteins owing to the fact that normal parameters of testing fail to pick them up. (Chapel, 2005). Bence- Jones Proteins are distinguishable by the fact they possess unusual thermal properties, for example they precipitate out of the urine solution at 56 degrees and redissolve upon further heating. (Thompson, 1978) Practical Schedule Determine concentration by ultrafiltration, absorption of water, or by freeze-drying. There are several commercially available kits for determining the concentration of urine. This involves concentrating the urine, then using electrophoresis to determine the presence of monoclonal bands. Then using immunofixation to establish what the monoclonal band is made of. (Chapel et al., 2006) Results Serum protein samples from patients with light chain multiple myeloma and one normal result on the far left.The M protein is seen as a dark dense band localised on the strip, this picture shows the different bands that can be detected. Albumin Decreased with malnutrition and malabsorption, pregnancy, kidney disease (especially nephrotic syndrome), liver disease, inflammatory conditions, and protein-losing syndromes Increased with dehydration Alpha1 globulin Decreased in congenital emphysaema (a1-antitrypsin deficiency, a rare genetic disease) or severe liver disease Increased in acute or chronic inflammatory diseases Alpha2 globulin Decreased with hyperthyroidism or severe liver disease, haemolysis (red blood cell breakage) Increased with kidney disease (nephrotic syndrome), acute or chronic inflammatory disease Beta globulin Decreased with malnutrition, cirrhosis Increased with hypercholesterolaemia, iron deficiency anaemia, some cases of multiple myeloma or MGUS Gamma globulin Decreased variety of genetic immune disorders, and in secondary immune deficiency Increased Polyclonal: chronic inflammatory disease, rheumatoid arthritis, systemic lupus erythematosus, cirrhosis, chronic liver disease, acute and chronic infection, recent immunization. Monoclonal: Waldenstroms macroglobulinaemia, multiple myeloma, monoclonal gammopathies of undetermined significance. (MGUS) Table from lab tests UK online. Evaluation-this method allows the determination of the different proteins in the urine and can be vital in allowing the doctor to work out a diagnosis of the condition. It is relatively simple and reliable however the results can only be read by a skilled worker and owing to its various steps is not as rapid as desired. Results show that different diagnoses are reached depending on which Immunoglobulins are increased in the urine, as indicated in the table above. Qualitative Immunoglobulins in Cerebrospinal Fluid- immunoperoxidase and isoelectric focusing This test allows for the differentiation between IgG and albumin concentrations. This relationship is important to differentiated as IgG is synthesised by lymphocytes within the brain where as albumin is not and is known as the CSF IgG Index that is indicative of this fact as demonstrates how much IgG within the CSF has been synthesised. (Chapel et al2006). Unlike the before mentioned serum where single discrete (M) bands where formed the locally synthesised IgG is often oligoclonal and subsequently cannot be detected by means of electrophoresis of CSF as isnt concentrated. (Roitt et al.. 2002) The only available method for the detection of oligoclonal bands are isoelectric focusing and immunofixation with enzyme labelled antiserum. Investigation and diagnosis of demyelinating disorders such as Multiple Sclerosis is carried out using such tests. (Richard et al 2002) Practical Schedule- Isoelectric focusing and immunofixation with enzyme labelled antiserums. This involves separating the proteins within a pH gradient and transferring them to nitrocellulose membranes that have previously been immunofixed with IgG antiserum to show the specific bands. This can be compared with controls to determine the new bands. (Richard et al., 2002) Results A positive result is where the oligoclonal IgG bands are not found in serums, but, in Cerebrospinal Fluid. These are shown as dense dark bands on the results below. 5-10% of CSF protein tends to be IgG. If a patient has disseminated sclerosis or sub-acute sclerosing panencephalitis then the proportion of IgG in CSF is over 12%. Evaluation This is a relatively modernised method and is approved for use within a clinical setting. The older isoelectric focusing is no longer recommended as it possesses a higher degree of specific (95%) and sensitivity. In addition it is favourable as only requires low concentrations of serum samples and results are available within 2 hours and mostly work on an automated level. (Richard et al.. 2002) Qualitative Immunoglobulins in Saliva- Complement- components Introduction Complement components are large molecular weight proteins. Activation of these usually results in proteolytic cleavage of the molecule into fragments. (Thompson, 1978) Western blotting is used in combination with gel electrophoresis and ELISA and RIAs are used when a whole saliva sample is collected or when there are saliva fractions [Fabian et al., 2007]. Practical Schedule- Gel filtration is carried out on Sephadex G-200. Serum samples of 1.5ml were applied to and 2.5cm diameter, 40 cm length column containing the Sephadex. This is equilibriated with a buffer containing 0.14M NaCl, 0.006M NaH2PO4 and 0.035M Na2HPO at a pH value of 7.3. Fractions of 2.5ml each are collected at a flow rate of 30ml per hour and the protein content of this effluent is measured as UV transmission at 280m µ in an absorbiometer. Results the results are determined by using these filtrated samples and single radial diffusion, a calibration curve is needed to determine amounts. This is created by using standard solutions. (Rose et al., 1997) Evaluation Occur in large amounts in serum can be measured accurately precipitin reaction in gel. Detecting them as antigens however means it cannot be identified as to whether they are active or not. Collecting specimens for complement assays can be difficult as you are to avoid inducing the complement pathway. Care should be taken to avoid false results caused by this when trying to determine the activation that was caused in vivo. Single radial diffusion can be used to determine quantitatively. This test is rapid reliable and easy to carry out and determine results of. (Rose et al., 1997) Complement-breakdown products Complement-C3- Crossed immunoelectrophoresis Introduction The complement system comprises of proteins (which may be membrane bound or present in plasma) that play an important role is host defences [Stanley, 2002]. The system is involved in destroying certain bacteria and viruses, and is also involved in initiating inflammatory response. Complement is also important for opsonisation of foreign materials, facilitation of phagocytosis by leukocytes, and direct cytotoxic reactions [Gaspari Tyring, 2008]. A determinant of the amount of C3 is crossed immunoelectrophresis and has the advantage of differentiating between inactive and active forms of C3. Deficiencies in C3 can lead to systemic infections including sepsis meningitis, pneumococcal and influenza infections. Method First dimension Prepare a 2% agarose solution in the barbitone buffer containing EDTA (ethylene diamine tetra-acetic acid) Pour 3 ml of agarose solution onto the microscope slide and let set. Cut a 1mm well in the slide removing the agarose and filling with the serum sample for the C3 quantification. Apply a potential difference of approx. 150v for 2 hours. Cut a 5 mm wide longitudinal strip containing the sample. Second dimension Prepare 12ml of an anti-C3 solution in 2% agarose solution at 56 degrees. Place the agarose strip at one end of the square glass plate and cover the whole slide with the agarose containing the anti-C3. Place the plate in the electrophoresis tank making sure it is the right way and electrophorese overnight. Wash and stain the precipitin arcs. This method works by using the electric field to separate the complement components. Results Evaluation- as with many of the before mentioned techniques it requires a skilled technician in order to carry out such a test and can edge on the side of time consuming owing to its numerous steps and incubation periods set out in the methodology. (Hay et al.. 2002) Complement- nephritic factor Complement-nephritic factor Introduction nephritic factor is an autoantibody to activated C3, it breaks down C3 in the alternate pathway by cleaving it into two fragments that are inactive forms (C3d and C3c) of the normal version of C3b. It binds and stabilises the alternative pathway C3 convertase (that is present in all sera) in the presence and absence of serum proteins. The alternative pathway C3 convertase blocks inhibitors from acting on and destroying C3 convertase.. The autoantibody (the C3 nephritic factor) reacts in the complement system not by blocking the enzyme active site but instead, block the site where inhibitors limit the action of and destroy the enzyme. Tests to determine the C3 nephritic factor are performed in patients that possess a C3 concentration that is below normal and is unexplained, with normal C4 levels [SAS Centre, 2009]. This is because the presence of the C3 nephritic factor in a patient means that C3 is continuously broken down and depleted. Low levels C3 can be associated wi th kidney disorders or recurrent infections. (Chapel et al., 2006) Practical Schedule-The practical schedule is similar to the before mentioned. It used samples with the suspected nephritic factor and other normal serum samples. They are incubated together and if the nephritic factor is present, it breaks down the C3 in the normal sample. (Chapel et al., 2006) Results-As expected from the similarity in methodology the results are similar to the detection of C3 in the picture above using crossed immunoelectrophoesis. If only inactive forms are present owing to inactivation from nephritic factor than the result is deemed positive. A negative result is when there is no nephritic meaning that none of the C3 has been inactivated. Evaluation-This method is useful in the detection of nephritic factor only and it is not a very direct test as it is carried out to determine the amount of C3. Complement-functional assay CH50 Introduction complement functional assay are the basis for the diagnosis of complement deficiency disorders. They are divided into subcategories dependent upon their relation to another disease. Primary complement deficiencies are genetic based and secondary refer to those that are acquired. Functional assays play a pivotal role within the assessment of the classical, alternative and terminal pathway of complement activation. The most common haemolytic assay used within the laboratory setting is the CH50 assay as it is both the simplest and easiest to carry out. The functional integrity of the classical complement pathway, C1, C2, C3, C4 is measured using CH50 along with total haemolytic complement. This is achieved by measuring the required quantity of serum in order to cause haemolysis of half the quantity that had been stabilised and sensitised red blood cells (Chapel et al..2006). Classical components become activated to lyses sheep erythrocytes that are coated in rabbit anti- sheep E antibodies (Rose.. 1997) Practical Schedule Add to microtiter wells the sera to be tested along with a buffer in different concentrations. Then add the sheep erythrocytes. Cover and incubate at 37dgrees for 1 hour. Then centrifuge and carry out ELISA to detect results. (Rose et al., 2002) Results Evaluation- The method is generally sensitive and reliable providing the specimen is tested quickly and all reagents are kept on ice. (Chapel et al., 2006) The problem arises in availability as they are not widely available and as such functional assays for complement are limited to laboratories that have the equipment (Gaspan and Tyring 2008). As with many immunological techniques the fundamental threat to validity is improper sample collection, this can occur easily in the onsite environment where it can be left to stand for considerable periods of time at room temperature. (Rose, 1997) 5. Microbial Antigens ELISA Introduction By coupling the antigen to an insoluble adsorbent it is possible to detect human antibodies to specific antigens using this technique. Elevated levels of antibody titre remains a reliable indication to the presence and measurement of an active infection within the diagnostic process. ELISAs provide highly sensitive and precise methods for the estimation of biological parameters, with the added advantage that they can handle large numbers of samples that may then be analysed rapidly and are useful in detection of a range of viruses and bacterial infections inclusive of TB and pneumonia and viral antigens. (Chapel et al..2006) Many types of immunoassays can be used to detect and quantitative both antigens and antibodies, but there are differences in the avidity requirements for the antibodies, the signal strengths of the labels, and the amount of background for each of these types of assays. Antibody capture assays are the most appropriate for measuring the titre of the antisera you have generated. ELISAs by definition exploit the use of an enzyme attached to one of the reagent utilized in the test. Subsequent addition of the relevant enzyme substrates/ chromogens cause a colour change: the results can be read both by eye and quantified using specially designed spectrophotometers. The fact that proteins (including antibodies) and carbohydrates can be passively attached to plastics has been exploited in most applications of ELISA. Since one of the components is attached to a solid phase by passive absorption, subsequent reagents can be added, and after a period of incubation, unreacted material can be simply washed away. Such assays are termed Heterogeneous ELISAs. The plastic surface is known as the solid phase and plastic in the form of 96-well microtiter plates has proved highly practical for the following reasons. A large number (96) of sample wells are available in a highly practical from; Multichannel pipets (4,8,12 channels) designed for use with such plates are available, making reagent handling rapid and simple; Test volumes are small (e.g. 50uL, 100uL); Comparative readings of coloured products can be made by eye or by specially designed multichannel spectrophotometers (96 wells are read in 2-5s) The above facts afford the potential to rapidly handle numerous plates, and hence numerous samples may be examined, e.g., 20 plates/person= 1920 sample points/ person. Attachment of reagents also allows great versatility for ELISA since the various components of assays may be used in different combinations and in different phases to investigate their potential. It is difficult to generalise about the potential performance of the various ELISA systems. There is a wide range of configurations available and probably no two scientific groups attempting to perform the same task by ELISA will use identical configurations. Practical Schedule Dissolve antigen in carbonate-bicarbonate buffer. Add 200  µl to each well of a micro-ELISA plate cover and incubate overnight at 4 degrees. Wash so that unbound antigen is removed and fill with casein to block remaining binding sites. Incubate at room temperature for 1 hour. Add 200  µl of test serum and incubate for 2 hours at room temperature in a humid chamber. Wash the plate three times Prepare the peroxidase-antibody conjugate, mix 100microlitres of casein with 1 ml of serum, 100 µl Tween 20 with 50  µl peroxidase-antibody and stir gently. Add 200 µl to each well and incubate at room temperature for an hour. Wash three times. Prepare the substrate solution and add200  µl substrate to each well. Leave in the dark and allow colour to develop. Stop the reaction by adding 50  µl of sodium fluoride to each of the wells. An ELISA reader can then be used to quantify the colour reaction. General Method from Hay et al., 2002. Results A positive result would be characterised by the reaction that causes colour showing the presence of antibodies to the specific type of bacteria highlighted by a dark band. Evaluation On the whole this method remains largely specific and rapid. The major cause of problem is the scientist(s) involved. The main problem is the lack of close- contact training in the fundamentals of ELISA, so that the scientist has the experience to identify and then solve the problems in the use of reagents. The results yielded cannot have the biological implications assessed without general knowledge of several field of science, e.g., epidemiology, immunochemistry, biochemistry and immunology. This however should not be considered too problematic as the ELISA should be a tool for the investigation of specific problems rather than an end in itself. Whilst it in comparison to immunoflourence in the detection of TB and flow cytomentry it may lack sensitivity, it does remain the cheapest and easiest to carry out. (Rose et al..1997) 6. Autoantibodies It is becoming increasingly evident that the presence of tissue auto antibodies is not in itself pathognomic of disease. Improvements in the technique in the last few years have led to increased sensitivity and detection of weak antibodies in sera which would hitherto have been reported negative. As a large series of patients are tested in an increasing number of laboratories previously held views on the specific clinical association of particular antibodies are being revised and reference to early literature may therefore be misleading. Since antigens and antibodies are defined by their mutual interactions, they can be used to quantify each other. At a practical level in a diagnostic laboratory, the functional tests are labour intensive and therefore expensive, and a compromise is usually sought by using immunochemical assays which measure composite of medium to high affinity antibodies and their abundance. The antibody has become the scientists flexible friend! For example, antibodies raised against hormones, serum proteins, cell constituents, cytokines, or even immunoglobulins themselves, allow these parameters to be measured in immunoassays. Immunoassays form the backbone of tests used in the study

COMPARATIVE :: essays papers

COMPARATIVE Homer Winslow and Jules Breton, two men painting the canvas of the nineteenth century. Comparing their art gives birth to numerous differences and unique qualities hidden within their work and lives. Dressing For The Carnival, Homer 1877, and The Weeders, Breton 1868, are fine examples of their careers as artists. "Beyond the aesthetic merits of his work, Breton is significant as the painter whose vision of French rural life best embodies a set of late nineteenth- century ideals: the charm and wholesomeness of rustic ways, the nobility of living close to the soil, the beauty of preindustrial landscape, and the social harmony of the agrarian community." ( Sturges) Breton’s work was unique in content, painting for himself, impressing his personal values to the viewer. Although he did not fit the mold, by producing classical and historical works, there were other artists struggling with expression and values of a newer mind, artists like Winslow Homer. "While he was at work in Petersburg, it became known to a group of fine young fire-eaters that he was consorting w ith the blacks, and they resolved to drive him out of town as a ‘d-d nigger-painter.’ Word had come to him that the place was to be made too hot for him, but he paid no attention to the warning." ( Hendricks) Both Breton and Homer were leaders for impressionism, however, the two works mentioned above vary greatly. Both artists focused on similar subject matter, figures in a scene or landscape. However a closer observation of specific images, narrative, symbols, sources, and process divide the two pieces to separate sides of late eighteen hundreds paintings. Physical elements such as composition, position of figures in space, brush work, color, viewpoint, and surface treatment all contribute to this separation of similar subject matter. The composition of The Weeders is un cropped, fairly balanced and symmetrical. The foreground is bold, the middle ground is expansive and the back ground strong and deep. Our view is that of perhaps a weeder on the field. Homer’s Carnival is cropped and less symmetrical with figures emerging from off the canvas. Less emphasis is placed on use of foreground, in turn creating less depth. Breton’s figures hold much movement and expression, women working the field are crouched close to the viewer. Farther back a woman stands alone, basket full, gaze and body positioned toward the setting sun. The women weeding are bent and tired.

The Tragic Hero of Sophocles Antigone :: Antigone essays

The Tragic Hero of Antigone In Sophocles' Antigone, the question of who the tragic hero actually is has been the subject of a debate for years.   It is unlikely for there to be two tragic characters in a Greek tragedy, and there can be only one in the play Antigone.   The king Creon possesses some of the qualities that constitute a tragic character, but does not have all of the necessary traits. Antigone, however, contains all of the aspects that are required for her to be the   main character.   According to Aristotle's Poetics, there are four major traits, which are required of the tragic character.   The character must be a good and upstanding person.   The character must focus on becoming a better person, must be believable, and must be consistent in his or her behavior. Due to the fact that Antigone represents these four character guidelines, as well as several other protagonist traits, she can definitely be defined as the tragic hero. In order for Antigone to be the tragic character, she first must be a good   and upstanding person.   Antigone is indeed a good-hearted person and has committed no crime up to her decision to give her brother, Polynieces, a   proper burial.   There is no doubt that Antigone is upstanding and a person   of importance in Thebes.   She was scheduled to marry Haemon, the son of   Creon, and was considered a princess.   Aristotle stated that the aspect of a good person was first and most important when creating a tragic character.   The fact that Antigone is a woman makes no difference, because Aristotle   expressly said, "Even a woman may be good.though the woman may be said to be an inferior being."   Aristotle's second rule for determining a tragic character is that the   person must aim at propriety.   The character must work towards becoming a   better person.   Antigone illustrates this second guideline by her effort to   clear her conscious and bring honor to her family by giving Polynieces a   decent burial.   By taking this responsibility, and by denying Ismene's   involvement in her crime, Antigone shows that she has acquired a greater   courage within herself than she had possessed before.   In no way does Creon comply with Aristotle's second guideline.   Throughout the play, he does not   allow himself to see the point of view from other people, such as when   Haemon tries to reason with him, and he neglects the blind prophet,   Tiresias, when he warns Creon of his actions.   The last two expectations of a tragic character are intertwined. According   to Aristotle, the character must be true to life and be consistent in

Monster Essay -- Biography, Kody Scott

Kody Scott, also known as â€Å"Monster† for his viciousness in beating of a man and further crimes, forms a realistic and brutal picture of gang violence in America. Throughout his story, Scott views his gang participation as the only viable means of survival. Killing is done through the necessity to promote oneself in order to become an O.G., or Original Gangster, the pinnacle of gang member status and achievement. The urge to become an O.G. seems to be paramount in Scott's eyes, and he outlines his plan: first he must build his reputation, then his influence as part of his set, and finally as a â€Å"promoter† of the Crips (Shakur, 1993, pp. 14-15). By age eleven Scott's sole desire is to become a gang member of his local set, the Eight Trays. He disregards education, at one point stating how he paid no attention to his middle school teacher, focusing only on the streets and his â€Å"homeboys† as source of lifestyle and adventure (Shakur, 1993, pp. 3-4). He c learly reaches his goal, putting the entirety of his mental and physical being into being a gangster, even though it leads him to a life wreaked by violence and prison sentences. While many factors can be reported to showcase the reasons why an individual would be led to such violence, ethology remains the single most accurate theory in explaining the many facets of gang violence by showing the parallels between animal behavior and the lifestyle of gang members. Ethology also attempts to explain the violence exhibited by humans as corresponding to violence displayed by animals. For predatory animals, violence is necessary for survival and adaptation to the surroundings is vital. Genetically humans are closest to chimpanzees, which â€Å"routinely engage in murder, assassination, rape, raid... ...suit of a promoted street war and personal achievements of status and further violence. Kody Scott's tale of the street war in Los Angeles in which he witnessed and played a dramatic part in is punctuated by the reasons behind his deviant behavior and adherence to a lifestyle focused on violence. By studying the behavioral patterns of animals in ethology, the traits of predatory animals which must fight and guard in order to stay alive is the same intensity of the battle to survive in the case of Monster's reality. Each of Monster's action's proved to be founded in the culture he was born into along with his personal desire to see his acts of violence glorified. Without the drive to commit himself to a rational lifestyle, Kody is swept into the maelstrom of gang-banging, all of its twisted faculties accepted due to an equally twisted and antisocial personality.

The Lost Duke of Wyndham Chapter One

Grace Eversleigh had been the companion to the dowager Duchess of Wyndham for five years, and in that time she had learned several things about her employer, the most pertinent of which was this: Under her grace's stern, exacting, and haughty exterior did not beat a heart of gold. Which was not to say that the offending organ was black. Her grace the dowager Duchess of Wyndham could never be called completely evil. Nor was she cruel, spiteful, or even entirely mean-spirited. But Augusta Elizabeth Candida Debenham Cavendish had been born the daughter of a duke, she had married a duke, and then given birth to another. Her sister was now a member of a minor royal family in some central European country whose name Grace could never quite pronounce, and her brother owned most of East Anglia. As far as the dowager was concerned, the world was a stratified place, with a hierarchy as clear as it was rigid. Wyndhams, and especially Wyndhams who used to be Debenhams, sat firmly at the top. And as such, the dowager expected certain behavior and deference to be paid. She was rarely kind, she did not tolerate stupidity, and her compliments were never falsely given. (Some might say they were never given at all, but Grace had, precisely twice, borne witness to a curt but honest â€Å"well done† – not that anyone believed her when she mentioned it later.) But the dowager had saved Grace from an impossible situation, and for that she would always possess Grace's gratitude, respect, and most of all, her loyalty. Still, there was no getting around the fact that the dowager was something less than cheerful, and so, as they rode home from the Lincolnshire Dance and Assembly, their elegant and well-sprung coach gliding effortlessly across the midnight-dark roads, Grace could not help but be relieved that her employer was fast asleep. It had been a lovely night, truly, and Grace knew she should not be so uncharitable. Upon arrival, the dowager had immediately retired to her seat of honor with her cronies, and Grace had not been required to attend to her. Instead, she had danced and laughed with all of her old friends, she had drunk three glasses of punch, she had poked fun at Thomas – always an entertaining endeavor; he was the current duke and certainly needed a bit less obsequiousness in his life. But most of all she had smiled. She had smiled so well and so often that her cheeks hurt. The pure and unexpected joy of the evening had left her body humming with energy, and she was now perfectly happy to grin into the darkness, listening to the soft snore of the dowager as they made their way home. Grace closed her eyes, even though she did not think herself sleepy. There was something hypnotic about the motion of the carriage. She was riding backwards – she always did – and the rhythmic clip-clop of the horses' hooves was making her drowsy. It was strange. Her eyes were tired, even though the rest of her was not. But perhaps a nap would not be such a misplaced endeavor – as soon as they returned to Belgrave, she would be required to aid the dowager with – Crack! Grace sat up straight, glancing over at her employer, who, miraculously, had not awakened. What was that sound? Had someone – Crack! This time the carriage lurched, coming to a halt so swiftly that the dowager, who was facing front as usual, was jerked off her seat. Grace immediately dropped to her knees next to her employer, her arms instinctively coming around her. â€Å"What the devil?† the dowager snapped, but fell silent when she caught Grace's expression. â€Å"Gunshots,† Grace whispered. The dowager's lips pursed tightly, and then she yanked off her emerald necklace and thrust it at Grace. â€Å"Hide this,† she ordered. â€Å"Me?† Grace practically squeaked, but she shoved the jewels under a cushion all the same. And all she could think was that she would dearly like to smack a little sense into the esteemed Augusta Wyndham, because if she were killed because the dowager was too cheap to hand over her jewels – The door was wrenched open. â€Å"Stand and deliver!† Grace froze, still crouched on the floor next to the dowager. Slowly, she lifted her head to the doorway, but all she could see was the silvery end of a gun, round and menacing, and pointed at her forehead. â€Å"Ladies,† came the voice again, and this time it was a bit different, almost polite. The speaker then stepped forward out of the shadows, and with a graceful motion swept his arm in an arc to usher them out. â€Å"The pleasure of your company, if you will,† he murmured. Grace felt her eyes dart back and forth – an exercise in futility, to be sure, as there was clearly no avenue of escape. She turned to the dowager, expecting to find her spitting with fury, but instead she had gone white. It was then that Grace realized she was shaking. The dowager was shaking. Both of them were. The highwayman leaned in, one shoulder resting against the door frame. He smiled then – slow and lazy, and with the charm of a rogue. How Grace could see all of that when half of his face was covered with his mask, she did not know, but three things about him were abundantly clear: He was young. He was strong. And he was dangerously lethal. â€Å"Ma'am,† Grace said, giving the dowager a nudge. â€Å"I think we should do as he says.† â€Å"I do love a sensible woman,† he said, and smiled again. Just a quirk this time – one devastating little lift at the corner of his mouth. But his gun remained high, and his charm did little to assuage Grace's fear. And then he extended his other arm. He extended his arm. As if they were embarking at a house party. As if he were a country gentleman, about to inquire about the weather. â€Å"May I be of assistance?† he murmured. Grace shook her head frantically. She could not touch him. She did not know why, precisely, but she knew in her bones that it would be utter disaster to put her hand in his. â€Å"Very well,† he said with a small sigh. â€Å"Ladies today are so very capable. It breaks my heart, really.† He leaned in, almost as if sharing a secret. â€Å"No one likes to feel superfluous.† Grace just stared at him. â€Å"Rendered mute by my grace and charm,† he said, stepping back to allow them to exit. â€Å"It happens all the time. Really, I shouldn't be allowed near the ladies. I have such a vexing effect on you.† He was mad. That was the only explanation. Grace didn't care how pretty his manners were, he had to be mad. And he had a gun. â€Å"Although,† he mused, his weapon rock steady even as his words seemed to meander through the air, â€Å"some would surely say that a mute woman is the least vexing of all.† Thomas would, Grace thought. The Duke of Wyndham – who had years ago insisted that she use his given name at Belgrave after a farcical chorus of your grace, Miss Grace, your grace – had no patience for chitchat of any sort. â€Å"Ma'am,† she whispered urgently, tugging on the dowager's arm. The dowager did not say a word, nor did she nod, but she took Grace's hand and allowed herself to be helped down from the carriage. â€Å"Ah, now that is much better,† the highwayman said, grinning widely now. â€Å"What good fortune is mine to have stumbled upon two ladies so divine. Here I thought I'd be greeted by a crusty old gentleman.† Grace stepped to the side, keeping her eyes trained on his face. He did not look like a criminal, or rather, her idea of a criminal. His accent screamed education and breeding, and if he was not recently washed, well, she could not smell it. â€Å"Or perhaps one of those dreadful young toads, stuffed into a waistcoat two sizes too small,† he mused, rubbing his free hand thoughtfully against his chin. â€Å"You know the sort, don't you?† he asked Grace. â€Å"Red face, drinks too much, thinks too little.† And to her great surprise, Grace found herself nodding. â€Å"I thought you would,† he replied. â€Å"They're rather thick on the ground, sadly.† Grace blinked and just stood there, watching his mouth. It was the only bit of him she could watch, with his mask covering the upper portion of his face. But his lips were so full of movement, so perfectly formed and expressive, that she almost felt she could see him. It was odd. And mesmerizing. And more than a little unsettling. â€Å"Ah, well,† he said, with the same deceptive sigh of ennui Grace had seen Thomas utilize when he wished to change the subject. â€Å"I'm sure you ladies realize that this isn't a social call.† His eyes flicked toward Grace, and he let loose a devilish smile. â€Å"Not entirely.† Grace's lips parted. His eyes – what she could see of them through the mask – grew heavy-lidded and seductive. â€Å"I do enjoy mixing business and pleasure,† he murmured. â€Å"It's not often an option, what with all those portly young gentlemen traveling the roads.† She knew she should gasp, or even spit forth a protest, but the highwayman's voice was so smooth, like the fine brandy she was occasionally offered at Belgrave. There was a very slight lilt to it, too, attesting to a childhood spent far from Lincolnshire, and Grace felt herself sway, as if she could fall forward, lightly, softly, and land somewhere else. Far, far from here. Quick as a flash his hand was at her elbow, steadying her. â€Å"You're not going to swoon, are you?† he asked, his fingers offering just the right amount of pressure to keep her on her feet. Without letting her go. Grace shook her head. â€Å"No,† she said softly. â€Å"You have my heartfelt thanks for that,† he replied. â€Å"It would be lovely to catch you, but I'd have to drop my gun, and we couldn't have that, could we?† He turned to the dowager with a chuckle. â€Å"And don't you go thinking about it. I would be more than happy to catch you as well, but I don't believe either of you would wish to leave my associates in charge of the firearms.† It was only then that Grace realized there were three other men. Of course there had to be – he could not have orchestrated this by himself. But the rest of them had been so silent, choosing to remain in the shadows. And she had not been able to take her eyes off their leader. â€Å"Has our driver been harmed?† Grace asked, mortified that she was only now thinking of his welfare. Neither he nor the footman who had served as an outrider were anywhere in sight. â€Å"Nothing that a spot of love and tenderness won't cure,† the highwayman assured her. â€Å"Is he married?† What was he talking about? â€Å"I – I don't think so,† Grace replied. â€Å"Send him to the public house, then. There is a rather buxom maid there who – Ah, but what am I thinking? I am among ladies.† He chuckled. â€Å"Warm broth, then, and perhaps a cold compress. And then after that, a day off to find that spot of love and tenderness. The other fellow, by the way† – he flicked his head toward a nearby cluster of trees – â€Å"is over there. Perfectly unharmed, I assure you, although he might find his bindings tighter than he prefers.† Grace flushed, and she turned to the dowager, amazed that she wasn't giving the highwayman a dressing down for such lewd talk. But the dowager was still as pale as sheets, and she was staring at the thief as if she'd seen a ghost. â€Å"Ma'am?† Grace said, instantly taking her hand. It was cold and clammy. And limp. Utterly limp. â€Å"Ma'am?† â€Å"What is your name?† the dowager whispered. â€Å"My name?† Grace repeated in horror. Had she suffered an apoplexy? Lost her memory? â€Å"Your name,† the dowager said with greater force, and it was clear this time that she was addressing the highwayman. But he only laughed. â€Å"I am delighted by the attentions of so lovely a lady, but surely you do not think I would reveal my name during what is almost certainly a hanging offense.† â€Å"I need your name,† the dowager said. â€Å"And I'm afraid that I need your valuables,† he replied. He motioned to the dowager's hand with a respectful tilt of his head. â€Å"That ring, if you will.† â€Å"Please,† the dowager whispered, and Grace's head snapped around to face her. The dowager rarely said thank you, and she never said please. â€Å"She needs to sit down,† Grace said to the highwayman, because surely the dowager was ill. Her health was excellent, but she was well past seventy and she'd had a shock. â€Å"I don't need to sit down,† the dowager said sharply, shaking Grace off. She turned back to the highwayman, yanked off her ring, and held it out. He plucked it from her hand, rolling it about in his fingers before depositing it in his pocket. Grace held silent, watching the exchange, waiting for him to ask for more. But to her surprise, the dowager spoke first. â€Å"I have another reticule in the carriage,† she said – slowly, and with a strange and wholly uncharacteristic deference. â€Å"Please allow me to retrieve it.† â€Å"As much as I would like to indulge you,† he said smoothly, â€Å"I must decline. For all I know, you've two pistols hidden under the seat.† Grace swallowed, thinking of the jewels. â€Å"And,† he added, his manner growing almost flirtatious, â€Å"I can tell you are that most maddening sort of female.† He sighed with dramatic flair. â€Å"Capable. Oh, admit it.† He gave the dowager a subversive little smile. â€Å"You are an expert rider, a crack shot, and you can recite the complete works of Shakespeare backwards.† If anything, the dowager grew even more pale at his words. â€Å"Ah, to be twenty years older,† he said with a sigh. â€Å"I should not have let you slip away.† â€Å"Please,† the dowager begged. â€Å"There is something I must give to you.† â€Å"Now that's a welcome change of pace,† he remarked. â€Å"People so seldom wish to hand things over. It does make one feel unloved.† Grace reached for the dowager. â€Å"Please let me help you,† she insisted. The dowager was not well. She could not be well. She was never humble, and did not beg, and – â€Å"Take her!† the dowager suddenly cried out, grabbing Grace's arm and thrusting her at the highwayman. â€Å"You may hold her hostage, with a gun to the head if you desire. I promise you, I shall return, and I shall do it unarmed.† Grace swayed and stumbled, the shock of the moment rendering her almost insensible. She fell against the highwayman, and one of his arms came instantly around her. The embrace was strange, almost protective, and she knew that he was as stunned as she. They both watched as the dowager, without waiting for his acquiescence, climbed quickly into the carriage. Grace fought to breathe. Her back was pressed up against him, and his large hand rested against her abdomen, the tips of his fingers curling gently around her right hip. He was warm, and she felt hot, and dear heaven above, she had never – never – stood so close to a man. She could smell him, feel his breath, warm and soft against her neck. And then he did the most amazing thing. His lips came to her ear, and he whispered, â€Å"She should not have done that.† He sounded†¦ gentle. Almost sympathetic. And stern, as if he did not approve of the dowager's treatment of her. â€Å"I am not used to holding a woman such,† he murmured in her ear. â€Å"I generally prefer a different sort of intimacy, don't you?† She said nothing, afraid to speak, afraid that she would try to speak and discover she had no voice. â€Å"I won't harm you,† he murmured, his lips touching her ear. Her eyes fell on his gun, still in his right hand. It looked angry and dangerous, and it was resting against her thigh. â€Å"We all have our armor,† he whispered, and he moved, shifted, really, and suddenly his free hand was at her chin. One finger lightly traced her lips, and then he leaned down and kissed her. Grace stared in shock as he pulled back, smiling gently down at her. â€Å"That was far too short,† he said. â€Å"Pity.† He stepped back, took her hand, and brushed another kiss on her knuckles. â€Å"Another time, perhaps,† he murmured. But he did not let go of her hand. Even as the dowager emerged from the carriage, he kept her fingers in his, his thumb rubbing lightly across her skin. She was being seduced. She could barely think – she could barely breathe – but this, she knew. In a few minutes they would part ways, and he would have done nothing more than kiss her, and she would be forever changed. The dowager stepped in front of them, and if she cared that the highwayman was caressing her companion, she did not speak of it. Instead, she held forth a small object. â€Å"Please,† she implored him. â€Å"Take this.† He released Grace's hand, his fingers trailing reluctantly across her skin. As he reached out, Grace realized that the dowager was holding a miniature painting. It was of her long-dead second son. Grace knew that miniature. The dowager carried it with her everywhere. â€Å"Do you know this man?† the dowager whispered. The highwayman looked at the tiny painting and shook his head. â€Å"Look closer.† But he just shook his head again, trying to return it to the dowager. â€Å"Might be worth something,† one of his companions said. He shook his head and gazed intently at the dowager's face. â€Å"It will never be as valuable to me as it is to you.† â€Å"No!† the dowager cried out, and she shoved the miniature toward him. â€Å"Look! I beg of you, look! His eyes. His chin. His mouth. They are yours.† Grace sucked in her breath. â€Å"I am sorry,† the highwayman said gently. â€Å"You are mistaken.† But she would not be dissuaded. â€Å"His voice is your voice,† she insisted. â€Å"Your tone, your humor. I know it. I know it as I know how to breathe. He was my son. My son.† â€Å"Ma'am,† Grace interceded, placing a motherly arm around her. The dowager would not normally have allowed such an intimacy, but there was nothing normal about the dowager this evening. â€Å"Ma'am, it is dark. He is wearing a mask. It cannot be he.† â€Å"Of course it's not he,† she snapped, pushing Grace violently away. She rushed forward, and Grace nearly fell with terror as every man steadied his weapon. â€Å"Don't hurt her!† she cried out, but her plea was unnecessary. The dowager had already grabbed the highwayman's free hand and was clutching it as if he was her only means of salvation. â€Å"This is my son,† she said, her trembling fingers holding forth the miniature. â€Å"His name was John Cavendish, and he died twenty-nine years ago. He had brown hair, and blue eyes, and a birthmark on his shoulder.† She swallowed convulsively, and her voice fell to a whisper. â€Å"He adored music, and he could not eat strawberries. And he could†¦he could†¦Ã¢â‚¬  The dowager's voice broke, but no one spoke. The air was thick and tense with silence, every eye on the old woman until she finally got out, her voice barely a whisper, â€Å"He could make anyone laugh.† And then, in an acknowledgment Grace could never have imagined, the dowager turned to her and added, â€Å"Even me.† The moment stood suspended in time, pure, silent, and heavy. No one spoke. Grace wasn't even sure if anyone breathed. She looked at the highwayman, at his mouth, at that expressive, devilish mouth, and she knew that something was not right. His lips were parted, and more than that, they were still. For the first time, his mouth was without movement, and even in the silvery light of the moon she could tell that he'd gone white. â€Å"If this means anything to you,† the dowager continued with quiet determination, â€Å"you may find me at Belgrave Castle awaiting your call.† And then, as stooped and shaking as Grace had ever seen her, she turned, still clutching the miniature, and climbed back into the carriage. Grace held still, unsure of what to do. She no longer felt in danger – strange as that seemed, with three guns still trained on her and one – the highwayman's, her highwayman's – resting limply at his side. But they had turned over only one ring – surely not a productive haul for an experienced band of thieves, and she did not feel she could get back into the carriage without permission. She cleared her throat. â€Å"Sir?† she said, unsure of how to address him. â€Å"My name is not Cavendish,† he said softly, his voice reaching her ears alone. â€Å"But it once was.† Grace gasped. And then, with movements sharp and swift, he leaped atop his horse and barked, â€Å"We are done here.† And Grace was left to stare at his back as he rode away.

“if you forget me” pablo neruda Essay

This poem dramatizes the conflict between the poet’s passionate burning desire and unconditional love for a woman, and his stern attitude toward her if she doesn’t feel the same. By demonstrating excellent personification, imagery, and sentimentality, this poet portrays love, compassion, and optimism. Although the poet expresses deep emotion for his love, he also acknowledges that love can be dangerous, and refuses to be damaged. To begin with, while explicating this poem, it was evident to make the assumption that the poet has experienced pain once or twice in his life. The author begins to describe his love for a woman that is so powerful, that it is evident that he would go above and beyond to satisfy her. Arguably, this poem could in fact pertain to the risk, and terror of making oneself available to love and to be loved, which by nature makes oneself vulnerable to incredible hurt and pain. This is the poet’s manifesto in the face of such terror, and he does it with authenticity, vulnerability, and authority. As analyzed by literary analyst Donald S. Walsh, â€Å"This [poem] is vintage Neruda, with all the passion and fickleness of desire. The underlying melancholy is beautifully brought out by the conversational style (a la Mir Taqi Mir) – the conceit would have been metaphysical had it not been for the pain inherent in every verse. † This is love that is hurting, that has been hurt in the past, and yet is open to being hurt again. In the opening stanza of this poem, Neruda underlines his authority with a stern, passive statement: â€Å"I want you to know one thing† (1) this is an obvious attempt to warn the reader that Neruda is in charge of the future conflict. Proceeding to the following stanza, Neruda illustrates the beauty of nature and compares it to the beauty of love by using keen personification and visual imagery. Such as, â€Å"the wrinkled body of the log† (10) and â€Å"as if everything that exists, aromas, lights, metals, were little boats that sail, toward those isles of yours that wait for me. †(12-16) This opening stanza gives the reader the inference that the poet is whole-heartedly committed to this woman. The poet also uses an oxymoron to demonstrate his love in lines (7-9), â€Å"if I touch near the fire the impalpable ash. † Impalpable meaning â€Å"untouchable. † Neruda stresses an excessive amount of sentimentality throughout the beginning of this poem. Proceeding to the third stanza, the poet’s mood completely changes, â€Å"If little by little you stop loving me, I shall stop loving you little by little† this refers back to the first stanza where once again the narrator refuses to be subject to vulnerability. Lines (20-24) stresses the solemnness of the poet by asserting that â€Å"if suddenly you forget [him], do not look for [him] for [he] shall already have forgotten you. † The fifth stanza demonstrates effective use of symbolism by Neruda. â€Å"To leave me at the shore of the heart where I have roots† (29) Neruda is now referring back to the â€Å"isles† of [hers] (16) underlining that the narrator is symbolizing that his body is a tree, and his heart is the roots that have grown deep into the isle (the woman), and have adapted. Neruda then accentuates that if the woman were to leave him at the shore where he has become accustomed to, that â€Å"on that day, at that hour, I shall lift my arms, and my roots will set off to seek another land† (31-35) claiming that the narrator will not wait for her to come back, and he will in fact find another love. Ultimately, Neruda unexpectedly returns to his sentimentality throughout the last stanza, offering optimism. Beginning the stanza with â€Å"but† became very powerful, considering that this was meant to stand-alone; the narrator is offering somewhat of a compromise. The poet then returns to the love entrenched speaker he is sought out to be. â€Å"If each day, each hour, you feel that you are destined for me with implacable sweetness†¦in me all that fire is repeated. In me nothing is extinguished or forgotten. † (36-44) This is where Neruda emphasizes his true heart felt love for this woman, by illustrating that if the feeling is mutual nothing will defer his love. Finally in line (45) Neruda connects this poem with a lasting thought, and possibly a theme. â€Å"My love feeds on your love, beloved† Demonstrating his compassion with a lasting effect. Throughout this poem there are no evident signs of rhythm, or rhyme; although, the reader may view this as very effective. This, in turn, was perhaps intended by the poet because love in most cases, is unpredictable. There was, however, a noticeable pattern when discovering the poet’s mood. The first stanza was a warning, the second, a beautiful picture painted by the poet of love. Neruda continues this pattern with repetitive use of the word â€Å"if† which may be referred to as a presupposition. â€Å"If you suddenly forget me, do not look for me, I shall have already forgotten you†. (20-24) This line can be interpreted as, â€Å"if she forgets him, he will forget her first. † Neruda finally discontinues this pattern by returning to the mood of the second stanza by incorporating beautiful imagery and personification once again. This, in fact insures that the poet is unafraid of the risks of falling in love, but remains completely aware. According to the Poetry Society Of America, â€Å"anyone who knows anything about Pablo Neruda knows that he is considered the ‘poet of love’†. Which is clearly demonstrated throughout this poem. Neruda was effective in creating a love story, one of which can be interpreted in many ways. Neruda painted the reader a vivid picture with the use of personification and the beauty of love. After diagnosing this poem, the tone, form, and consistency of Neruda really fit the overall â€Å"love to be loved† theme.