Wednesday, May 6, 2020
Coke Behind the Image Essay - 2786 Words
Coke: Behind the Image Almost everywhere I go, I see advertisements for ââ¬Å"The Real Thing.â⬠Even during my trip to Mozambique, a country stricken with poverty where the people rarely speak English, native children knew how to ask for their favorite drink, available just down the road at the street vendorââ¬Ës hut. Created in 1885 by pharmacist John Pemberton, Coca-Cola has evolved from a tonic peddled by traveling salesmen to cure headaches and hangovers, to one of the most recognized and most widely enjoyed beverages of all time (Oliver 14). On average, North Americans drink at least one serving of Coke per day (CSPI sc1). With over two million dollars spent every year on advertising and with veryâ⬠¦show more contentâ⬠¦Coca-Cola argues many points in support of its marketing and image. Especially in the years following WWII, Coke has worked hard to present an image associated with having a good time. Since the late 1800s, Cokeââ¬â¢s advertising has shown people in beautiful places and in beautiful clothes, having a beautiful time and enjoying their beautiful lives--even in times of war and economic recession. While wars, riots and scandals rage in this country, Coke has been ââ¬Å"holding forth to the world a picture of itself as pleasant and rewarding--the kind of place most people would like for it to beâ⬠(Watters 216). In Cokeââ¬â¢s more recent advertising, Coke continues its image of fun and beauty, showing teens enjoying a sunrise and at a rave dance party in the woods. However , these more current commercials were in response to two other adds that were considered ââ¬Å"not funnyâ⬠and ââ¬Å"mean spiritedâ⬠by public opinion polls (Sampey par 1-4). Those ads showed family members and friends physically fighting with each other because someone had forgotten to bring Coke. Along with the claim that it facilitates a good time, Coke also claims that it wants to give everyone a voice, minorities included. With the Vietnam war raging in the 1970s, Coke featured a commercial that showed different ethnicities from around the world holding hands and singing a song, including the words, ââ¬Å"Iââ¬â¢d like to buy the world a Cokeâ⬠and ââ¬Å"Iââ¬â¢d like toShow MoreRelatedAdvertising And Its Effect On Advertising888 Words à |à 4 Pagesto a more sexual image, or something more creative that requires intelligence to create and understand. First, by taking a sip of the Pepsi advertisement, I noticed the horizontal advertisement has a gentle light blue background. The Pepsi logo sits in the corner, not drawing much attention to itself. In the center of this ad is a Pepsi can. Next to the Pepsi can is a red can with a silver curved thick line going up the side from the bottom of the can to the top, resembling a Coke can. The differenceRead MoreCoca Cola Advertising Essay1448 Words à |à 6 Pagesthem effective. The specific advertisement that is being discussed is one in which a man on a beach is enjoying a cold Coca- Cola. He reaches down to grab his drink to discover that it was no longer where he left it. He did not see the penguin hiding behind him that took the cold Coca-Cola for itself. This advertisement uses logos, pathos, and ethos to persuade its audience to buy Coca-Cola and is an effective advertisement. Coca Cola draws its viewers in by displaying colorful, unique adsRead MoreMarketing Strategy Of Gatorade And Coke977 Words à |à 4 Pagesthem in their everyday lives. Gatorade has decided to revamp their internet marketing strategy to gain an online presence and get the upper hand on their competitors. The competitors we are going to analyze and critique are PowerAde, Vitaminwater and Coke. I identified the competitors by looking at the sports drink and soda industries to see what companies are Gatoradeââ¬â¢s main competition and what strategies they can use to get ahead of these 3 prominent competitors. The first competitor Iââ¬â¢m going toRead MoreCoca-Cola Company901 Words à |à 4 Pageswent hiding and sixty five received death threats . Union alleged Coca Cola to be behind all these activities Breach of Confidence in Employees In the year 2006 , three Coca Cola employees were charged for trying to sell trade secrets and confidential information to the competitor PepsiCo . PepsiCo had taken an ethical decision by informing the Coca Cola about the offer . This has increased the brand image of PepsiCo but questioned Coca Cola s ability to maintain confidence among its employees Read MoreCoca Cola Vs. Pepsi Cola Essay1174 Words à |à 5 PagesAmerica. While having emphasis on its refreshing cool taste and convenience, these ads created a certain attitude with its audience causing the rise in popularity and sales. During this same time Pepsi was also on the rise competing with Coca- Cola. Both Coke and Pepsi used similar strategies but Coca- Cola gave itself a slight edge through innovative technique and the use of ââ¬Å"lifestyleâ⬠advertising. Coca- Cola really did create a refreshing beverage but created an ad campaign using refreshing, excitingRead MoreCoca Col A Symbol Of American Culture1599 Wor ds à |à 7 PagesCoca-cola is truly a symbol of American culture, however everyoneââ¬â¢s favorite soft drink may not be so sweet once you know the full history. Coca-Cola was founded in 1886. Itââ¬â¢s inventor was a pharmacist named John Pemberton. Pemberton originally sold Coke as a pharmaceutical drug in the form of syrup containing cocaine and wine. It was intended to cure morphine addiction, however soon after Pemberton finalized his formula the campaign for prohibition began and he was forced to replace the wine withRead MorePepsi Of Coca Cola And Pepsi1259 Words à |à 6 Pagesrival those of Coca-Cola and Pepsi. The longevity and prosperity of these two competitors is enough to inspire countless businessââ¬â¢ to model their own companies after these icons. This paper will detail some of the business aspects that have helped Coke and Pepsi to thrive, as well as considering what mistakes have been made in their pursuit of success. Learning from the successes and failures of other companies can help a new business to know which roads of marketing, expansion, etc. to exploreRead MoreMarketing Strategy Of Coca Cola1342 Words à |à 6 PagesSimilar to everything else in our world, advertisements continue to change year after year and era after era. When thinking of common brands with impressive advertisements and campaigns, Coca-Cola is one that we are introduced to at a young age. Coke has been advertising their brand for over one hundred years. This brand continues to relate to many consumers by modifying and creating advertisement campaigns, which relate to current trends. Two advertisements in particular test the theory of howRead MoreCoca Cola Vs. Pepsi1312 Words à |à 6 Pagescompany controls 42% of the soft drink market in comparison with Pepsi who owns 30% (ââ¬Å"Coke Vs. Pepsi: By The Numbers.â⬠). With market power, Coke and Pepsi have accomplished manipulating prices and controlling profits for ages leading towards their success. Coke and Pepsi are two economic giants that sell similar products in the cola industry and market power keeps them both competitive. A brief history about Coke starts with when it was founded at a drug store by a pharmacist named John Stith PembertonRead MoreThe Case Of Food And Beverage Companies947 Words à |à 4 Pageshuge number of people donââ¬â¢t examine the products they buy. Companies use millions of dollars to advertise, using celebrities to present their products with images that are not close to the reality. This is the case of some food and beverage companies such as McDonaldââ¬â¢s, Carlââ¬â¢s Jr. and Coca Cola. These three companies promote and advertised images of people enjoying food, clowns encouraging children to eat junk food, and polar bears making believe that families have a perfect life drinking coca cola
Cosmetology School Free Essays
Every year thousands of people who have a passion for fashion, style and beauty and who enjoy making other people and themselves look amazing and feel appealing enroll in cosmetology schools. Cosmetology school is a place for creative, sociable, ambitious and constantly developing individuals. There are a lot of various directions in the program of a cosmetology school, so every person is free either to choose the subject he or she likes, or to take up several of them: nail technician, esthetician, hair styling, permanent make up specialist, massage therapist, etc. We will write a custom essay sample on Cosmetology School or any similar topic only for you Order Now The sphere of cosmetology is multisided and very perspective and rewarding nowadays and there are a lot of reasons why attending a cosmetology school may be beneficial. The most important advantage of having a cosmetology school education is that you will hardly ever be unemployed (unless you will wish to). People would always like to look and feel beautiful, so you will always have enough opportunities to make them be and feel such. The only task you will always have to fulfill is to be always aware of the latest style and fashion trends and the newest techniques, and take any chance to cultivate your creativity and collect precious experience in your job. Another advantage of this career is that you are free to choose time and even place where you are eager to perform your skills. You may make weekends whatever days of week you want and you will never have to get up early in the morning(unless you have a yoga-class), because most of stylists prefer starting their working day in the afternoon, during the lunch break of their clients, or even in the evening, after normal business hours. And even if your working place is in the salon, it is likely that you can set the schedule convenient for you. People say: ââ¬Å"If you want to succeed in business, the best way for you is to make your hobby your own businessâ⬠ââ¬â a cosmetologist career is the best example to reflect this view. Working with clientsââ¬â¢ hair, nails or makeup may be extremely exciting. You meet many different people with different views as for style and fashion and all of them want you to bring their dreams to reality. During this process you practice not only as a cosmetologist, but also as psychologist. And the best reward is a grateful smile on the face of your client. It is also necessary to point out that if you are creative and skillful enough and enjoy trying new provocative styles, it is possible that you will be asked to style the hair and makeup of models and celebrities, or even well-known politicians. You will even have a possibility to work for big companiesââ¬â¢ hair shows, such as Joico or Paul Mitchell, or for some prominent modeling agencies, theatres or magazines. You can make various astonishing acquaintances and may be you will be well-known among very important people. In a field of cosmetology your incomes depend on nothing else but type, amount and quality of your own work. The more clients you are eager to have a day the more money you may earn. By the way, if to work hard and make a number of personal clients, there is a possibility of expanding your career by becoming a salon manager or even by opening your own salon. I am hardworking and willful, confident of my abilities and skills, so I hope to open my own spa salon some day. I know that my skills and natural charm and sociability will help me to build a base of clients. Moreover I am planning to open my own blog or even a site, where I will give necessary advice on fashion, style, hair design and nail art and tell people about the newest style tendencies. Probably I will face a lot of competitors on my way, but I am not afraid of that. People like to have their own specialist whether in hair style, makeup or manicure, thatââ¬â¢s why, they are searching for the ââ¬Å"rightâ⬠person, who would meet their needs. I am sociable and flexible enough to meet the needs of different kinds of people and I am conscious that people will notice that. Attending a cosmetology school can let one build a fun, flexible and very rewarding career. This job allows flexible working hours, high incomes, guarantee of being employed, plenty of room for professional and personal growth, and acquaintance and experience of interaction with various people. It is necessary to keep up with changing style and fashion tendencies to succeed in this field. In order to keep the clientsââ¬â¢ interest a stylist should always be in a process of self-improvement and self-education, as well as he should always find common language with clients. A cosmetology education provides a person with an opportunity of expanding his or her career by becoming a salon or spa manager or even an owner of such. Although it will take a lot of efforts to do this and there are a lot of competitors on the way, I am sure that some day my goal of owning a salon will be achieved. I believe that cosmetology school education will help me to take the first step to my dream and once I will be proud to call my job ââ¬Å"my hobbyâ⬠. How to cite Cosmetology School, Papers
Coca Cola Demographics and Psychographics in Market Segmentation
Question: Discuss about theDemographics and Psychographics In Market Segmentation. Answer: Introduction Research problem-market segmentation strategies need to be evaluated for effectiveness. This research will ascertain whether demographic variables are effective for market segmentation. Limitations of the report- the report is dependent on secondary sources of information which may have subjective opinions. Therefore care should be given in when implementing the recommendations. The sources and methods used-This report utilized secondary sources including academic underpinnings journals and internet sources. Report Organisation-this report will first evaluate the advantages and disadvantages of using demographics as a base for market segmentation. Discussions Advantages of using demographics as a basis for market segmentation The first advantage of using demographics in market segmentation is that the demographics data is easily available for use and it is cost effective. Most secondary data including the census data are expressed in demographics. This makes it easy to access this form of data and apply it in identifying the target market. The census data is readily available and the costs involved in accessing this kind of data are low (Piercy, et al. 2011). Secondly, demographics are easily measurable when compared to other segmentation factors. Considering when other variables like lifestyle and personalities are used in market segmentation their demographic characteristics have to be ascertained in order for the true characteristics of a market segment to be known (Bruwer, et al. 2007). Therefore the use of demographical data in deciding which market segment should be targeted is vital even when other factors have been considered. This set of data offers an opportunity to examine the population of different locations and develop products that suit their needs (Tigert, et al. 1971). For example, it is not enough to give the percentage of people who use Moleskines product range but it will be important to know their age, their gender and income ranges. Third, the application of demographics in segmentation will lead to customer loyalty and retention. When the customer needs and wants are carefully evaluated and their trends studied, an organisation will channel its marketing campaigns towards those needs and wants (Sessler 2011). If the customers are satisfied, they will become loyal and they will tend to use the products more often. An example is the Moleskines product range. The company has engaged its customers in buying its non-digital products through incorporation of the products in the digital systems. For instance the MSK 2 is one of the products that customers can use to submit their art, hacks and videos on the company website. Besides, these clients can access various templates for the creation of their journals (Koenke 2006.). Fourth, demographic data reveals the ongoing trends that give a clear signal indication of business for marketers. For instance, demographics can reveal the income levels of a population and that can show the purchasing power of that population and products can be developed to suit the income levels of such a population (Barry, et al. 2009). Finally, the use of demographics in market segmentation can save the business the costs that may be incurred in expansion. Since the population characteristics can be defined and the future predicted, the organisation can save on costs that can be incurred on further research and the losses that can be incurred through unsatisfied customers. Disadvantages of Using Demographics as a Basis for Market Segmentation First, one major limitation with the use of demographics is that it is one dimensional and does not facture in product differentiation. For instance, demographics can give details on a certain product for example the aftershave but it cannot give information about a particular aftershave brand. And who exactly uses the brand (Shiffman 2013). Secondly, demographic data is only descriptive and his limits the understanding of the consumers themselves since the data available is general and generalization cannot be used in the development of products that can be used by such a population (Sessler 2011). Thirdly, demographic variable require that other variables are used in the development of a market segment. This is due to the weakness that is posed by only being descriptive of the data and not being detailed on the brand types that are preferred by different demographic segments (Shiffman 2013). Lastly, demographic data is readily available and competitors can easily utilize it to copy the products that an organization may be selling in a particular market. This is a great disadvantage because the competitors may produce better products which can lead to loss of customers. Can demographics and psychographics be used together to segment markets? In the analysis of whether demographics and psychographics can be used together in a segment market, it is important to describe what is contained in psychographics. Psychographic segmentation is related to research on the psychological needs and wants of consumers (Narang 2010). This form of research is also referred to as lifestyle analysis. This has been an important tool that has been used in determining the most promising market segments. The promotion of such products as Colgate, KFC fast foods, Lux soap and many others relies on psychographic research which is used in capturing the potential customers profiles. Consumer interests, activities and opinions are some of the psychographic measures that can be used in the development of a market segment. Besides, the profiles which can be used when se factors are the ones that can be used in coming up with consumer-based products (Kumar,et al. 2008). Psychographic and demographic can be used complimentarily. That is, when the demographic and psychographic variables are applied together, there is sufficient information that can be used in the development of products for a particular market segment. In marketing, information is important and the more information is available the more the marketing activities will be effective since the customer needs will be met. For a brand to be successful in the market, it needs to satisfy the customer needs fully. Organizations should invest more in market research in order to come up with the products that are needed by the market. It is not possible for the organization to produce products that will satisfy the customer needs if they do not conduct sufficient research on what those customers needs are (Simons, et al. 2016). Using both demographic and psychographic data is also important because this technique develops a deep understanding of what needs to be put into the market. Demographics help in developing the quantities needed while psychographics will develop the needed qualities. For a market to be fully satisfied both quantity and quality have to be adhered to. A product may be of the required quality and the market demand is so high and if the quantities cannot satisfy the demand then there may be a crisis that may lead customers away from the companys products. Also, a product may be abundant in the market yet it is not of the required standard and quality according to psychographic profiles. This also may tarnish the companys reputation and customers will avoid such products ((Shiffman 2013).). Moleskine is an example of a brand that fully utilizes the concept of market segmentation. This organisation used to produce notebooks and it is a popular brand name in this industry. The digital era has really affected this company but still it has managed to survive the tides that come with digitalization. Since the digital era came with a wave that could reduce the need to write in notebooks, the company did an analysis of their customer desires and lifestyles. Since these customers have a heavy presence in the online platforms and social media, the company developed products that can be used in the digital era. By doing this the company has survived through a challenge that many other companies could not survive through (Spence,et al. 2007). Recommendations When applying market segmentation as a strategy in marketing, demographic data is important and it should be used together with psychographic data. The combination of these sets of data makes it easy to identify and present products to the market will fulfil the customer needs and wants.A critical analysis of lifestyles and the [population is important in this strategy and therefore I recommend that the two sets of data should be used complimentarily for effectiveness. Conclusion Market segmentation is a marketing strategy that is effective in penetrating and managing a market. This is done through profiling the market needs and developing products that suit such a market. The use of demographics in market segmentation is important in determining the different customer characteristics. However, there are some limitations which need that a combination with other variables should be done in order to make sure that market segmentation is done in a way that will lead to customer and market satisfaction. When the market is satisfied, the brand easily becomes successful in such a market because it will be trusted and customer loyalty will be developed. Reference List Barry, James, and Weinstein Art. 2009. "Business psychographics revisited: from segmentation theory to successful marketing practice." Journal of Marketing Management 25, no. 3-4: 315-340. Business Source Premier, EBSCOhost (accessed September 15, 2016). Bruwer, Johan, and Li, Elton. 2007. "Wine-Related Lifestyle (WRL) Market Segmentation: Demographic and Behavioural Factors." Journal of Wine Research 18, no. 1: 19-34. Business Source Premier, EBSCOhost (accessed September 15, 2016). Koenke. Teri. 2006. "Destroying Demographics: The New Art of Strategic Customer Communications." U.S. Banker 116, 22-23. Business Source Premier, EBSCOhost (accessed September 15, 2016). Kumar, Vishal, and Sarkar, Amitava. 2008. "Psychographic segmentation of Indian urban consumers." Journal of the Asia Pacific Economy 13, no. 2: 204-226. Business Source Premier, EBSCOhost (accessed September 15, 2016). Mostafa Mohamed M. 2009. "Shades of green: A psychographic segmentation of the green consumer in Kuwait using self-organizing maps." Expert Systems with Applications 36, no. 8: 11030-11038. Academic Search Premier, EBSCOhost (accessed September 15, 2016). Narang, Ritu.2010. "Psychographic segmentation of youth in the evolving Indian retail market." International Review of Retail, Distribution Consumer Research 20, no. 5: 535-557. Business Source Premier, EBSCOhost (accessed September 15, 2016). Piercy, Niall, Campbell, Collin, and Heinrich, Daniel. 2011. "Suboptimal segmentation: Assessing the use of demographics in financial services advertising." Journal of Financial Services Marketing 16, no. 3/4: 173-182. Business Source Premier, EBSCOhost (accessed September 15, 2016). Sessler Jerrod. 2011. "Demographic Segmentation: Social Media In-A-Box." Franchising World 43, no. 9: 50. MasterFILE Premier, EBSCOhost (accessed September 15, 2016). Shiffman Leon. 2013. Consumer Behaviour, 6th Edition. Pearson/Australia, 2013-09-01. VitalBook file. Simons, Dirk, and Zein Nicole. 2016. "Audit Market Segmentation The Impact of Mid-tier Firms on Competition." European Accounting Review 25, no. 1: 131-154. Business Source Premier, EBSCOhost (accessed September 15, 2016). Spence, Jacqui, and Abratt Russel. 1997. "Use of psychographics in consumer market segmentation: The South African experience." South African Journal of Business Management 28, no. 2: 33. Business Source Premier, EBSCOhost (accessed September 14, 2016). Tigert, Douglas, Lathrope Richard Michael, and Bleeg. 1971. "The Fast Food Franchise: Psychographic and Demographic Segmentation Analysis." Journal of Retailing 47, no. 1: 81. Business Source Premier, EBSCOhost (accessed September 15, 2016). Wilson-Jeanselme, Muriel, and Reynolds Jonathan. 2006. "The advantages of preference-based segmentation: An investigation of online grocery retailing." Journal Of Targeting, Measurement Analysis For Marketing 14, no. 4: 297-308. Business Source Premier, EBSCOhost (accessed September 15, 2016).
Friday, April 24, 2020
Pernicious Anaemia Causes and Curing Procedures
Introduction Pernicious anaemia refers to a form of anaemia, which results from failure of the body to absorb adequately absorb vitamin B-12. Vitamin B-12, being an important nutrient in the body is absorbed in the small intestine from foods rich in vitamin B-12. Lahner and Annibale (2009) explain that vitamin B-12 binds itself to intrinsic factor, a parietal-cells-secreted protein, forming a complex, which is readily absorbed by the small intestine.Advertising We will write a custom report sample on Pernicious Anaemia: Causes and Curing Procedures specifically for you for only $16.05 $11/page Learn More Irvine (1965) submits that when the aforementioned intrinsic factor is absent from the body, perhaps because of autoimmunity of genetic issues, vitamin B-12 is reduced in the victimââ¬â¢s body, leading to pernicious anaemia. Further, victims of autoimmune disorders are more exposed to pernicious anaemia because their antibodies attack parietal cells, leading to abnormal secretion of the intrinsic factor, which in turn leads to poor vitamin B-12 absorption and consequently, pernicious anaemia (Toh, Van Driel Gleeson, 1997). Therefore, the pathophysiology and pathogenesis of pernicious anaemia comes from the autoimmune disorder. The pathophysiology and pathogenesis of pernicious anaemia show that occurrence of autoimmune disorder affects secretion of the intrinsic factor leading to mal-absorption of vitamin B-12. Autoantibodies attach themselves to gastric H+/K+ââ¬âATPase that is a pump composed of a 100 a glycoprotein à ²-subunit (60-90 kDa) and a kDa catalytic à ±-subunit, and consequently prevent the proper functioning of parietal cells and intrinsic factor secretion (Toh et al., 1997). This report is an investigative analysis of the pathophysiology and pathogenesis of pernicious anaemia. It analyses the stomachââ¬â¢s morphology in relation to pernicious anaemia, diagnoses pernicious anaemia in sample stomachs through western blot technique, and seeks to detect antibodies in sample serums through immune-histochemistry staining. Aim The objective of this report is to detect antibodies of the proton pump of the stomach by applying immune-histochemistry and western blot techniques on serum samples. To set a good background for the study, the report first analyses the critical components of a healthy stomach.Advertising Looking for report on health medicine? Let's see if we can help you! Get your first paper with 15% OFF Learn More Material and Methods Procedure of Examining Morphology of the Stomach Stomach samples were placed in slides to prepare them for examination, and then stored in a safety cabinet. The slides were incubated in xylene for two minutes and further incubated in ethanol for another two minutes. Rinsing was done on the slides for 30 seconds after the slides were incubated in haematoxylin. Staining and washing was done, followed by soaking in 1% acid alcohol and rins ing. The stained slides were placed for 30 seconds in Scottââ¬â¢s tap to incubate, and thereafter, rinsed before staining them with eosin for a period of 4 minutes. The slides were subsequently placed for 30 seconds in 90% and 100% ethanol to fix them. After the fixation, the slides were put on ethanol for 2 minutes and dried in the air immediately. A drop of DPX medium was applied on fixed slide, preparing it for mounting, covered with cover slip, and a magnification of x400 used to view it. Diagrams focussing on the gastric gland and stomach wall were sketched using a pencil on an A4 paper. Procedure for Preparing SDS-PAGE and Western Blot For the stomach specimen, a protein sample of 48à µL was added into a microfuge with 12à µL of SDS-sample. Molecular weight markers for the assessment of the molecular weights were prepared in advance. To sediment the molecular markers and protein sample, the solutions were spun in a centrifuge for a few minutes. Next, lane 1 of the prepare d SDS-PAGE in electrophoresis apparatus was loaded with 10à µL of the molecular weight markers. Just like molecular weight markers, subsequent wells of the SDS-PAGE were loaded with 25à µL of the protein sample. Then the electrophoresis apparatus was started on a power of 200 volts and left for 45 minutes. After electrophoresis was over, the apparatus was switched off and dismantled, and a demonstrator was used to apply gel on a clean plate. Then the gel was placed on the iBlot device with the aim of assessing the separation of proteins according to their weights. 10uL of distilled water was used in the placement of one gel in iBlot device, which processed it for 7 minutes, and another gel was put in nitrocellulose film. The nitrocellulose membrane was stained using 50mL of 0.1% Ponceau solution and leaving it for 1 minute, which ensured that protein transformation had occurred. To detect sera of patients, the nitrocellulose membrane was labelled.Advertising We will write a c ustom report sample on Pernicious Anaemia: Causes and Curing Procedures specifically for you for only $16.05 $11/page Learn More Lanes were cut using a pair of scissors and subsequently rinsed using NaOH with an aim of removing stains. The membrane was washed in 50mL of TBS and stored for the subsequent practical, which would include 10mL of Tris-buffer at 40o C. Deionised water was used to clean membranes after they were retrieved from storage. The wash included the following three steps: blocking step involving 5mL of 5% skimmed milk blocking solution; incubation of primary antibody using 2.5mL of patient serum sample or the positive and the negative control; and secondary antibody incubation using 5mL of labelled antibody. The iBlot was put to use again in performing the aforementioned three steps. Consequently, membrane strips were removed from the device and washed with TBS 3 times for a period of five minutes. Several steps were subsequently performed before imaging using the Chemidoc. The results of the test were then uploaded to CloudDeakin. Procedure of Preparing Immuno-Peroxidase of Mouse Stomach As some steps of slides preparation was done beforehand, Incubation was done for 100à µL of serum for 20 minutes because some slide preparation steps had already been completed. Slides were washed using PBS horseradish peroxide, also known as anti-human Ig HRPO, was used to conjugate 50à µL of antibodies by incubating it for 45 minutes. Slides were then washed in PBS and water was used to rinse them before 100à µL of DAB was added, subsequently incubating them for 10 minutes. Before the slides were soaked in haematoxylin for 3 seconds, they were cleaned using PBS. After the cleaning, they were rinsed with water and incubated in ethanol for an approximate 2 minutes. DPX was applied on slides and they were viewed with a microscope magnification of Ãâ"400. Discussion Discussion: Morphology of the Stomach In order to understand the morphology of the stomach well, the mucosal and sub-mucosal layers of the mouseââ¬â¢s stomach are stained with eosin and haematoxylin as depicted in figure 1. Figure 2 shows gastric gland stained with the same, which together with the other parts are key determinants of the mouseââ¬â¢s stomach morphology.Advertising Looking for report on health medicine? Let's see if we can help you! Get your first paper with 15% OFF Learn More As Irvine (1965) notes, the major feature of pernicious anaemia is the atrophy of mucosal cells for example parietal cells. The latter are responsible for the secretion of gastric acid and intrinsic factor which aid in vitamin B-12 absorption in the end of the small intestine. ââ¬Å"Essentially, parietal cells have the H+/K+-ATPase pump, which consists of beta and alpha subunitsâ⬠(Rhoades Bell 2008). This is well illustrated in figure 2 whereby the parietal cells are pink stained while the other main cells (chief cells) are dark-pink stained. The chief cells play a significant role of pepsinogen secretion, which catabolizes proteins (Toh et al. 1997). In that case, the physiology of parietal cells and protein catabolism is dependent on the stomach morphology. Discussion: Western Blotting The experiment proved that a western blot technique is effective in determining whether patients are infected with pernicious anaemia. Results from the study were presented on CloudDeakin. I t is however important to mention that the western blot technique did not achieve the desired results in all experiments because its results were spurious. Some membranes in the study lacked bands, while some had patchy or uneven spots as shown in figure 3. Defects in antigens, buffers, or antibodies resulted in interference of definite bands in the membranes. Additionally, use of inappropriate antibodies, either primary or secondary, also resulted in vague bands or no bands at all. Likewise, the use of low concentrations of the antibody also results in invisible bands and lack of antigen or low concentrations of the same leads to an invisible signal. In this regard, it is necessary to use different antigens in order to determine whether invisibility is because of primary and/or secondary antibodies or antigens. The visibility of bands can also be affected by procedures like membrane washing. This is because prolonged washing is likely to diminish signal appearance. Contamination of buffers may also lead to invisible bands, hence the need to preserve their purity during the study (Lahner Annibale, 2009). The western blot procedure should be undertaken in such a way that buffers like TBST, ECL and PBS are not contaminated. The procedureââ¬â¢s transfer of protein component is perhaps the most sensitive because it may be interfered by air bubbles and partial transfer can occur (Andres Serraj, 2012). When such improper protein transfer occurs, then the blot comes out with uneven or patchy spots. In the event of trapped air bubbles within the membranes and gel, the output in film will be dark. Therefore, to prevent bubble formation, one should evenly distribute particles by incubating using a shaker. The results presented in figure 4 were prepared by the technical staff. The H+/K+-ATPase antigen is responsible for triggering an autoimmune disorder (Lahner Annibale, 2009), which interferes with the proper functioning of parietal cells and intrinsic-factor secr etion (Lahner Annibale, 2009). The parietal cellsââ¬â¢ autoantibodies act against H+/K+-ATPase, thereby affecting the function of parietal cells, and consequently the occurrence of pernicious anaemia (Andres Serraj, 2012). Structurally, the autoantigen measures an approximate 160 to 190 kDa, with the specific measurement being determined by its constituent protein subunits. The autoantigen is made up of a glycoprotein à ²-subunit (60-90 kDa) and a 100 kDa catalytic à ±-subunit (Toh et al., 1997). The subunits are the ones that determine the nature of the disorder affecting parietal cells and intrinsic factor secretion. The western blot analysis revealed a fragment of approximately 80 kDa. The fragment is a glycosylated à ²- subunit of the H+/K+-ATPase pump because the fragment detected corresponds a 80 kDa molecular marker. In the event of pernicious anaemia, H+/K+-ATPase is the autoantigen that is solely recognizable by parietal-cells antibodies. Therefore, it is vital in d iagnosing pernicious anaemia using western blot (Toh et al., 1997). Evidently, patient 1ââ¬â¢s stained strip in figure 4, indicates a positive diagnosis. The size of the autoantibody-bound subunit is approximately 80 kDa that is a glycosylated à ²-subunit of the H+/K+-ATPase pump. Therefore, the findings from the western blot procedure confirm that the patient is positive, and that the autoantigen subunit is glycosylated à ²- subunit of the H+/K+-ATPase pump. ELISA, which is an acronym for Enzyme linked immunosorbent assay, can also be utilized in detecting anti-proton pump antibodies that prove presence of pernicious anaemia in patients. H+/K+-ATPase is the main autoantigen, which is present in pernicious-anaemia patient sera because it induces autoimmune disorder that interferes with parietal cells and thereby inhibits secretion of the intrinsic factor (Lahner Annibale, 2009). When ELISA is used, the autoantigen on the micro titre plate is immobilized, and the serum sample t hat contains anti-proton pump antibodies is combined and incubated with the aforementioned antigen. Serum samples normally have antibodies that are specific to proton pump antigens. This implies that the antibodies bind to the proton pump à ± and à ² subunits. Sugiu et. al. explain that proteins and excess antibodies in serum are cleaned and the secondary antibody that is linked to the enzyme, and which is primary-antibody specific, is also added to the micro titre plate. A change in colouration is achieved by the use of the chromogenic substrate, indicating proton-pump antibodies presence. In order to be able to determine the amount of proton pump antibodies in sera by the use of ELISA, the assessment of electrical signal, intensity of colour and fluorescence is necessary. To measure colour intensity in absorbance of luminance, it is a spectrophotometer can be used (Sugiu et al., 2006). This accurately determines the level of proton pump antibodies in the sera. Therefore, determi nation of fluorescence or colour intensity is an important component of qualifying proton pump antibodies. The anti-human IG-HRP conjugate of the sheep is a secondary antibody, which links with the H+/K+ ATPase antigen indirectly because it uses the primary antibody to link to the antigen. Anti-human Ig-HRP is actually an antibody specific to the human Ig production of the antibody, and it results from the immunological response of a sheep to human Ig (Chevrier, Chateauneuf, Guerin Lemieux, 2004). The anti-human Ig is actually a secondary antibody and thus, immune-histochemistry and in western blot, horseradish is used to conjugate it. Horseradish is an enzyme, which catalyses chromogenic substrates. Discussion: Immuno-peroxidase staining of mouse stomach section In this section of the report, emphasis is put on the stomachââ¬â¢s immune-histology to determine whether the patient mouse tests positive or negative for pernicious anaemia. In case of positive results, the antibodies are seen attached on the proton pumps of the parietal cells (Irvine 1965). During this experiment, unknown patients were applied anti-human 1g to the human 1g, which were labelled P1 and P2. From the experiment results, it was depicted that there were no brown stains thus concluding that there was no antibody-attachment on the proton pumps of the parietal cell. The conclusion was that patient 2 had tested negative for pernicious anaemia. On the other hand, patient 1 tested positive for pernicious anaemia because there were brown stains, which indicated antibody-antigen attachment on the proton pumps of the parietal cells. Reference List Andres, E, Serraj, K 2012, ââ¬ËOptimal management of pernicious anaemiaââ¬â¢, Journal of Blood Medicine, vol. 3, no. 1, 97-103. Chevrier, C, Chateauneuf, I, Guerin, M, Lemieux, R 2004, ââ¬ËSensitive detection of human Ig in ELISA using a monoclonal anti-IgG-Peroxidase Conjugateââ¬â¢, Hybrid Hybridomics, vol. 23, no 6, 362-367. Irvine, J 1965, ââ¬ËImmunologic Aspects of Pernicious Anaemiaââ¬â¢, New England Journal of Medicine, vol. 273, no. 1, pp. 432-438. Lahner, E, Annibale, B 2009, ââ¬ËPernicious anaemia: New insights from a gastroenterological point of viewââ¬â¢, World Journal of Gastroenterology, vol. 15, no. 41, pp. 5121-5128. Rhoades, A, Bell, R 2008, Medical Physiology principles for clinical medicine. Lippincott Williams Wilkins, Philadelphia. Sugiu, K, Kamada, T, Ito, M, Kaya, S, Tanaka, A, Kusunoki, H, Haruma, K 2006, ââ¬ËEvaluation of an ELISA for detection of antiparietal cell antibody,ââ¬â¢ Hepatogastroenterology, vol. 53, no. 67, 11-14. Toh, H, Van Driel, R, Gleeson, A 1997, ââ¬ËPernicious Anaemiaââ¬â¢, New England Journal of Medicine, vol. 337, no. 20, pp. 1441-1448. This report on Pernicious Anaemia: Causes and Curing Procedures was written and submitted by user Frankl1nR1chards to help you with your own studies. You are free to use it for research and reference purposes in order to write your own paper; however, you must cite it accordingly. You can donate your paper here.
Wednesday, April 15, 2020
Using Samples of History Essay
Using Samples of History EssayIt is very common to come across students taking History classes where the assignments tend to require students to collect and make use of samples of history essay. This can be a valuable experience, and a great way to get a better feel for writing a paper.However, it is also important to understand that even the best of history papers will usually have some flaws. This can be frustrating, but knowing what these flaws are and how to correct them is vital. Using samples of history essay will help students realize just how important this is.The best place to look for samples of history essay is online. There are many places where one can find a sample essay that can be used as a guide. For example, if one searches for 'A Sample History Paper' on the top of Google, they will see a number of different samples of history essays.Some of these are available for free, while others can be purchased at a price. The websites that offer these samples are usually wel l-organized. Students should be able to quickly get their hands on a decent sample essay. Having one available to study from will help a student to get a feel for writing a decent essay.There are also a number of other sites that offer general guidelines. Some of these include the following: first paragraph should be more than one paragraph long, no more than 3 facts, be brief, and give a brief overview of what you want your paper to say. A student should be able to follow these basic guidelines when looking for samples of history essay.Many students do not realize that essay writing with research aids. The reason being is that they assume that they will write the best piece of history writing by following an outline. While there are templates for doing this, it is still best to learn what makes an outline and how to build a good one.Students can always take a look at examples that are freely available, but those that have their own ideas may not work out as well. This is especially true when looking for samples of history essay that have been written in the past. Past generations tend to write in ways that are different from the way we do.The best way to address this is to use examples that have a general outline. This way a student can follow along without too much trouble. Looking for samples of history essay may be a useful experience for students that are in need of one.
Tuesday, March 17, 2020
LDH Purification lab Report Essays
LDH Purification lab Report Essays LDH Purification lab Report Paper LDH Purification lab Report Paper OLD was purified from the ammonium sulfate precipitated protein mixture by affinity chromatography and its activity was studied by spectrophotometers determination of NADIA at 340 NM. From Pierce BCC assay of crude homogenate, initial protein concentration was shown to be 100 MGM/ml. The final protein concentration of the pooled affinity sample was shown to be 0. 2 MGM/ml. It was found that the total specific activity of OLD was 58. 5 mol/min/MGM, and yield of 0. 6%. Even though we were successful in purifying OLD enzyme, further steps can be taken to increase the yield. Materials and Methods Cell Lysine and Extraction of OLD: Approximately 40 g of minced chicken breast eat (40. 327 g) is blended with ml cold extraction buffer in four 30-seconds bursts for homogenate of the muscle tissue. The extraction buffer contained mm Tries-HCI (pH-7. 4), mm 2-Merchantable, mm Phenylmethylsulfonylflouride (AMPS), 1 mm Ethylene Dianne attracted acid (EDIT). The homogeneities procedure was carried out in the cold room to prevent the denomination of proteins. The homogenate was centrifuged at 15,000 RPM for 20 minutes at 40 C. The supernatant was filtered through two layers of cheesecloth to remove lipids from the supernatant. The total volume was noted and three 0. Ml aliquots (crude extract) were stored at -200 C. Ammonium sulfate precipitation: 60% ammonium sulfate concentration was used to precipitate proteins. 0. 39 g of ammonium sulfate per ml of the supernatant was added gradually to the supernatant for 15-20 min with continuous gentle stirring at 40 C. The mixture was centrifuged for 20 minutes at 1 5,000 RPM at 40 C. The supernatant was discarded and the pellet was stored at -200 c. Dialysis: Ammonium precipitation leads to high concentration of salts in protein mixture that can interfere with further purification steps. In order to remove excess salts, dialysis was performed. The pellet was suspended in Tries-AMPS buffer (10 rim Tries-HCI, pH 8. 6, 0. 5 mm 2-Merchantable, and mm ratio of EDIT) and mixed very gently until it dissolved at 40 C. Volume of ml protein mixture was added in the dialysis tubing and incubated twice overnight with two IL buffer changes (Same buffer as extraction buffer that was used for cell lysine). After two incubation, protein mixture was responded gently and centrifuged for 10 minutes at 15,RPM at ICC. Pellet was discarded, total volume of supernatant was noted and three 0. 1 ml aliquots were collected. Affinity Chromatography: Isobaric Blue column was used to separate OLD from the other proteins. Ml fractions were collected in thirteen test tubes. Column was first rinsed with Tries-AMPS buffer followed by addition of protein mixture. Then, ml AND Buffer (mm Tries-HCI pH-8. 6, 0. Mm 2-Merchantable, mm Lithium acetate and 1 mm AND+) was added followed by NADIA (mm Its- HCI PH 8. 6, mm NADIA and 0. Mm 2-Merchantable). Between each steps, column was washed with ml Tries-AMPS Buffer. Each fraction was subjected to absorbency reading of Mann. For absorbency above 1. NM, 1:10 dilutions were carried out. Activity Assay: We used OLD Enzyme assay to measure the amount of OLD activity in our protein mixture. OLD catalysts the conversion of lactate to private and AND+ to NADIA. The NADI A can be determined spectrophotometers at 340 NM. The OLD assay was performed in the crude homogenate, desalted fraction and six peak fractions from the Isobaric blue column. A cocktail solution was prepared by mixing lactate stock solution (120 rim lithium lactate, 10 mm Tries-HCI; pH 8. 6), AND+ stock solution (12 mm AND+, 10 mm Tries HCI; pH 8. 6) and bicarbonate stock solution (18 mm Enhance, 0. 5 M Nasal) in the ratio of in cavetti. 0 micrometers of the sample is then added and the assay absorption is measured at Mann. If absorbency was above 1. 5, samples were diluted. Protein Assay: The Pierce BCC Protein Assay (Thermo Scientific) is a detergent- compatible formulation based on bioscience acid (BCC) for the colorimetric detection and quantization of total protein concentration. A series of standard solution of Bovine Serum Albumin (BAS) ranging from 0-2000 pig/ml was prepared from a stock solution of 2 MGM/ml BAS. Lull of diluted crude (1:500, 1 :250), desalted (1:100, 1:50), and 6 peak fractions from isobaric blue column (1:10, 1:5) ere loaded in microscope along with lull of BCC working reagent. Microscope was incubated for mini at ICC and then the absorbency was measured at Mann. Results/Discussion The purpose of this experiment was to extract and purify OLD enzyme from chicken muscle tissue using a variety of techniques including homogeneities, ammonium sulfate precipitation, dialysis, and affinity chromatography. Activity and Protein assay were used to track the overall amount of OLD present in the samples. Crude Extraction: Chicken muscle tissue was homogeneity in a blender with cold extraction buffer in order to else cells, releasing OLD into slurry of tissue monuments. Centrifugation separated membranes, nuclei, and other large cellular components to a pellet leaving a supernatant of crude product. Controlling temperature was a major consideration after homogeneities since not only did this step releases proteins like OLD from the cell, but it also releases proteases that can now interact to degrade the OLD. Keeping samples on ice, pre-cooling the buffer, and avoiding excess kinetic energy through conservative blending were methods used to minimize activity of these proteases. After filtration through cheesecloth, our final volume of crude homogenate sample ml, much more volume than expected. Addition of more than ml of buffer volume could have increased the volume. Other possible explanation is that more solid components such as fats were present in the sample and hence, more than 20 minutes of centrifugation was required. Desalted Sample: 60% ammonium sulfate is added to the crude extract that precipitates OLD proteins. The resulting 40% pellet theoretically contains most of the original OLD, which is re-suspended in very less volume (ml) to create a more concentrated sample. This process leads to high concentration of salts in rotten mixture that can interfere with subsequent purification steps. Ml protein mixture underwent dialysis procedure that removes excess salts and our final volume after dialysis was ml. One possible explanation for increase in our volume could be that extraction buffer got mixed with protein mixture either due to tubing leaking or tubing clips not being properly tightened. Affinity Chromatography: Isobaric Blue column is an affinity column, which is specific to dehydrogenate type proteins, due to a compound structurally similar to NADIA being attached covalently attached to the column. 13 fractions were elected and absorbency was measured at Mann to check presence of OLD protein in the fractions. 1:10 dilution was performed if absorbency reading was above 1. NM since it spectrographically indicates saturation and less than 1% light reaching the detector. During the addition of protein mixture (fraction# 4), high absorbency reading of NM was obtained (Fig. 1). This could be due to lot of non-dehydrogenate-type proteins present in our sample that got eluted first during affinity chromatography. Second peak was seen after AND+ was added since AND solution results in the removal of the loosely bound protein. Third peak was seen after NADIA was added since NADIA solution results in release of maximum OLD proteins (Fig l) Enzyme Activity Assay: The OLD activity was measured spectrophotometers by measuring the absorbency of NADIA at 340 NM. Three peak fractions were selected for this assay based on their absorbency values obtained after adding AND+ (fraction# 6, 7, 8) and other three after adding NADIA in the affinity chromatography step (fraction# 10, 1 1 , 12). A huge activity of 141 mol/min/ml was seen at fraction# 7(PUFF ) which indicated that we had lot of proteins present in our sample. Second peak activity was seen t fraction indicating that more OLD proteins is present in this fraction than in fraction# 11 (PUFF) (fig. 1). Based on this information, we selected fraction #10 as for our protein assay. Desalted showed highest activity among all the samples (Tablet ) possible due to errors occurring during dialysis explained previously. Figure 1. Absorbency readings of eluted obtained from affinity chromatography with OLD activity for 6 peak fractions. The desalted fraction was loaded to the Isobaric blue column and proteins were eluted with Tries-AMPS, AND+ and NADIA wash subsequently. The absorbency at 280 NM of eluted were measured after ACH collected fractions. The OLD activity was calculated from the absorbency values obtained at Mann. Protein Assay: We used BCC Pierce Assay to determine protein concentrations in our protein mixture. BAS standard curve was created for series of dilutions ranging from 0-2000 pig/ml and linear graph equation was used to calculate protein concentrations for the samples (Table 1). Based on Table 1, with each subsequent purification step, protein concentration decreases as sample become more concentrated with only OLD protein. Specific activity should increase and total activity should decrease with very purification step as samples get less and less diluted.
Sunday, March 1, 2020
PODCAST Build a Marketing Strategy For 9,360% Bigger Results
PODCAST Build a Marketing Strategy For 9,360% Bigger Results Are you getting more out of your marketing than youââ¬â¢re putting into it? Nathan Ellering is the head of demand generation at , and he wants you to be able to say that youââ¬â¢re getting the results you want from your marketing, your content, and your social media. Nathan is also the author of The Ultimate Guide to Marketing Strategy, so heââ¬â¢s a bit of an expert on the topic! Weââ¬â¢re going to be talking to him today about how you can generate quality leads, get the traffic and traction that you want, and make your marketing strategy a success. What Nathan does at as the head of demand generation. The story behind the launch of the Ultimate Guide to Marketing Strategy: what it includes, why it was necessary to write it, how itââ¬â¢s different, and why you should check it out. Nathan also shares why he created it as a microsite as opposed to a blog. The importance of following the acronym SMART when youââ¬â¢re setting your goals. An explanation of what 10x content is and how Nathan makes it work for him. How Nathan recommends tracking marketing ROI to be sure that your content is doing what itââ¬â¢s supposed to do. How to prioritize within your marketing strategy. The concept of goals-driven budgeting. Nathanââ¬â¢s best advice for someone just starting a marketing strategy. Links: The Ultimate Guide to Marketing Strategy Google Analytics If you liked todayââ¬â¢s show, please subscribe on iTunes to The Actionable Content Marketing Podcast! The podcast is also available on SoundCloud, Stitcher, and Google Play. Quotes by Nathan: ââ¬Å"Marketers who document their strategy are 538 percent more likely to report success than those who donââ¬â¢t.â⬠ââ¬Å"The goals-driven budgeting methodology requires you to set your goals first, then plan your budget to make the goals a reality.â⬠ââ¬Å"As a marketer, your first blog post wonââ¬â¢t be good and your first ad is probably going to suck, but you need to do it. You need to ship something and you need to continuously improve.ââ¬
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